First completely synthesized eukaryotic chromosome

Once again our world gets one step closer to the first extinct species to be cloned*. Remember when I reported the fully resolved genome of a British Mesolithicaurochs? I wrote that it is not yet technically possible to clone an aurochs because it is not yet figured out how to de novo synthesize a complete functional set of chromosomes which is needed to express the DNA after it was implanted into the blastocyst of a surrogate mother. All that we have of the aurochs’ genome yet is the reconstructed full sequence of this individual.

On April 4^th this year however, a team of scientists published the total synthesis of a functional eukaryotic chromosome (go here for the abstract). The same worked with a prokaryotic chromosome previously. It will probably take some more years to synthesize longer chromosomes and eventually a full chromosome set, but I see this as a step forwards to genetically reconstruct and revive extinct animals of which enough genetic material is resolved.

* One extinct animal was cloned already and lived for 7 minutes. It was a Pyrenean ibex, C. p. pyrenaica. Go here for the abstract. 

Interview with Henri Kerkdijk-Otten on the Uruz Project

Probably not unnoticed by followers of “aurochs rebreeding”, a new aurochs project exists since last year. Henri Kerkdijk-Otten, formerly manager of the Tauros Project, and his True Nature Foundation split of from the Tauros Project and initiated the Uruz Project. Finally, I and Henri had the time to do a little interview that provides you extra information on this most recent aurochs project. Enjoy!

> - First of all: Which cattle breeds is the Uruz Project going to use and in how far will the set of breeds be different from that of the Tauros Project?

We want to diminish the amount of breeds and combinations. There are multiple ways to Rome of course, but all the Pajuna, Limia, Tudanca and Scottish Highland cattle are the cause of too much variation and unwanted genes. At the same time in Portugal we will start with a selection program with Barrosã and Maronesa cattle. In Spain and Portugal hopefully with Toro de Lidia as well. That is a closed world unfortunately, but we do have some contacts.

  

> - I am very happy to hear about Toro de Lidia, their phenotype is simply too excellent (despite being small) to not be used in breeding-back. So the breeds the Uruz Project is going to use are Texas Longhorn, Barrosa, Chianina, Maronesa and Lidia? 

We will start with two breeding lines: Chianina x Watussi and Sayaguesa x Maremmana/Hungarian grey. In Romania we will probably start using Hungarian Grey influenced animals. For Romania we have a farmer with cows that we can use to implant embryos in, that we will produce in a lab. So that embryo will already be a F1 crossbred.

Barrosa, Maronesa and Lidia will be selected within their breed.

 

The Uruz Project's Chianina cow at Kloster Lorsch. Useful animals:  Tall and good body

> - And what about the Maltese Ox, do you still consider to use it? It seems like a "better Chianina" (as long as they are the same size) since it has a less diluted coat, more aurochs-shaped body and long and slender faces.

Yes, but difficult to get our hands on. They never experience frosts and they are stabled all day with little to no movement, so they are a little in the minus regarding those aspects.

Regarding the Watussi we use: they have gotten used to our climate. Farmers in the US and zoos in Europe have them out in the open in winter without problems. They are tough animals, can survive on poor forage and are overall resilient. And they bring in large horns to compensate for the Chianina and Sayaguesa influence...

 

Young Watussi bull Ubutaha at Kloster Lorsch, he will cover the Chianina cows in the first years. He has wild type E+ as base colour, plus that weird wine-red Watussi modifer. 

> - How many crossbred animals will be there in 2015?

No telling. We are busy building up contacts, relationships, live herds, embryo production and getting more funding.


> - How many breeding locations will the Uruz project have, and where?

Fluid thing. Several in the Netherlands (negotiating with national forestry, municipalities and forest group right now), 2 in Germany, 2 in the Czech Republic, 1 in Poland, 2 in the Ukraine, 1 to 3 in Romania, 3 or 4 in Portugal, several in Spain. In Germany our contact person is having talks next week with another municipality.


> - What are the selection criteria based on, and for how many years are you going to select the animals?

Same as Tauros project. The Aurochs hasn't changed, so criteria have stayed the same.
 We focus on phenotype a lot because we want to have a strong economic foundation underneath every rewilding project. For that we need to have a cattle breed that resembles the Aurochs as closely as possible. Safari-based tourism and other revenue streams is that which attracts investors to our foundation. Relying on Government funding is not a sustainable way of protecting nature and wilderness.

> - Does the Uruz Project have all the founding individuals of all the breeds to start crossbreeding already?

No, we are building up and expanding. We are able to acquire all founder animal (breeds) we need.

> - Something that I really miss in the Tauros Program is a useful web presence that tells you how many crossbred animals and in which combinations there are and photos of how they look like. The data interested people are able to access is very scanty. Is the Uruz Project going to provide us more information than that?

We will keep people up to date via our facebook page. Tauros is lacking in that, but they do not have the time to do that, so I find it understandable. Although they recognize that some people want to see the results, they put all their resources in the project itself, next to all the daily jobs everyone has. We do have a closed-off Uruz project page that is for project members only. We are not going to put a herd book only. No project does that as far as I know. Takes too much time. So Facebook will be the communication tool we are going to use. Maybe we will set up a separate Uruz facebook page.


> - I guess the Uruz Project will try to release the cattle into the wild as far as possible when they are ready for. How do you plan to overcome the legal problems of releasing domestic animals into the wild and which reserves do you have in mind?

Cattle have the legal status of kept animals in the European Union. We do have contacts with Brussels to talk about this. There is an investigation going on if and when some cattle (herds) can get the legal status of a wild animal. That will take time (politics). In the meantime we will have to make do with what we have; keep cattle in a semi-wild state and try to have as little government control. In the Oostvaardersplassen animal carcasses have to be carried away, but the area is too big to monitor all that.

Regarding areas; in eastern Europe there are more possibilities. Problem with national parks in Europe is that they mostly do not want areas to be fenced. If you do not fence off areas, you will have to keep animal populations at an unnatural low. See for instance the maximum of 25 Wisents on an area of about 5000 hectares in Germany. This is going nowhere. They animals are not able to shape landscape and the population is too small to be viable. So therefore we opt to set up private game reserves. So far, that option is working pretty well, mostly in Spain. In Romania some national parks are more eager to fence off large grazing areas.


>
- What is the current relationship with the Tauros Project?

None.

> - When will you use genome editing?

These things take time. Long Now Foundation has contacts for this. They want to do it, but have a lot on their mind. Then we need to set up a procedure, identify what to do and how to do it. When the genome is altered, it has to be put in stem-cells that are able to divide to eggs or semen. Etcetera. For instance, it is feasible to knock the dilutor genes off the Chianina genome, so the wildtype coloration surfaces again. And that will save generations of recessive genes. A problem that plagues the Heck cattle population. Lot to be done. In the meantime, we will start with good old fashioned breeding.

> - Since the genome of one aurochs is fully sequenced, why not doing genome editing by directly inserting aurochs alleles instead of cattle alleles on loci that are known to be different? 

The genome sequencing was done by sequencing pieces 7 times. For a perfect genome or parts of a genome you need about 30 times. So yes, we want to use Aurochs genome parts to be transplanted onto a cattle genome, but we need better sequencing results. Professor Hofreiter, who just set up that new ancient DNA lab at Potsdam University, want to do that, but costs could go towards 500.000 euro. The Neanderthal genome took about 2 or 3 million. This is difficult to fund, because private investors do not fund this, because there is not a clear revenue model underneath it. We are talking to the son of Ted Turner, the wealthy FOX Media owner, who has millions of acres of land in the US with lots of buffalo and other wildlife. But this is really future talk. We might scale this down to start with. Hofreiter's aim is to deliver Aurochs DNA that we can use for genome editing or even for cloning an Aurochs. But that will take lots of more money => something for the future. Still, it is worthwhile to start with this path. 

> - As a last question and moving to horses, I think the looks of those Exmoor x Konik crossbreeds looked very interesting. Their bay-dun short-maned exterieur probably is representative for many European wild horses, and the fact that one in three of them had standing manes is really stunning. Why did they stop breeding them?

I think the Konik x Exmoor crossbreds of the Taurus Foundation* looked cool in a sense. I don't know why the stopped crossbreeding Exmoors and Koniks.
I wouldn't mind starting a crossbreeding program of some sort. I don't want to use Przewalski through, since it has been proven they broke off from the caballus lineage some time ago.

* Stichting Taurus, a Dutch foundation running grazing projects with cattle and horses and partner of the Tauros Project, no connection to Taurus cattle; just to avoid confusions.

Heck cattle outside Germany Pt. III: Austria

There is only one Austrian grazing project for conservational purposes like in Germany, where Hungarian Grey cattle is used (more on that in a future post). All Heck cattle in Austria are kept in zoos, game parks or private farms. There are three zoos/game parks that keep Heck cattle, two of which I already visited several times and I am going to present a number of photos I took there here. 

Zoo Haag 

I enjoyed the herd at Haag because of their aurochs-like colour, with the exception of some bull-coloured cows and a very lightly coloured individual that was born one and a half year ago. The bull of the herd is really heavy, but interestingly it has an S-curved back with a little shoulder hump. There is a cow that looks exactly like some Corsicana (uppermost). Like the herd at Lainz, this one is mediocre, but there are worse Heck cattle herds. I had the opportunity to measure them: the bull is 140 cm tall at the shoulders, and one of the cows 130 cm - normal Heck cattle size. They were very tame and allowed me to stroke them all over their back, neck and face and even to touch their horns. 

Lainzer Tiergarten

The Lainzer Tiergarten in Vienna is a large game park that also contains a protected forest. They have a herd of Heck cattle that has a pretty large paddock with bushes, trees and grassland. The founding population for Oostvaardersplassen also had Heck cattle from Austria, and I suspect that these were from this population. The herds breed freely and there are several bulls within the herd, what means that there must be some competition for the females. I don't know if the managers of the herd select individuals out. Three years ago, I discovered a piebald bull calf within the herd, I am going to check if this bull is still within the herd. Interestingly, one of the bulls has horns that face more forwards than usual in Heck cattle. At first, I thought this might be the result of the cattle breeding freely over the three decades they have been living in that park, but then I discovered that some un-crossed Heck cattle at game parks do have forwards-facing horns (like this bull at Rheingönheim), which is rather surprising. 
Apart from that, the herd is pretty mediocre. Although many individuals have long legs, their bodies are relatively heavy and, with some exceptions, not slender. The horns of all cows are upright and thin and they have juvenile skulls. The colour is good in most individuals. One young bull has these interesting curly hair in their winter coat, probably a legacy of Highland cattle. 

Tyrol 

There is a Heck cattle herd in commercial use in Tyrol in an outdoor museum at Umhausen. Perhaps they are bred for meat because one of the bulls is really really heavy (see this video here, the heaviest Heck bull I have ever seen, even heavier than many Highland bulls). The other one of their bulls is from Hellabrunn, which is a good thing because this herd has good colour and acceptable horns. Remember that Heck cattle is a normal breed, the breeders are free to do what they want and Heck cattle per se is not a breeding back project (anymore). Umhausen is the Heck breeding location at highest altitude, and they graze an alpine pastures like their relatives of the Braunvieh breed do, and they fit surprisingly well into this beautiful typically Austrian landscape (I did not take the photo down below myself). 

Social and reproductive behaviour of cattle and aurochs

All members of the Bovini, including domestic cattle, have a more or less similar social behaviour, and so the aurochs probably showed the same behavioural patterns that all its relatives and descendants do. Only few aspects of the aurochs’ behaviour have been documented, mostly in Schneeberger’s report to Conrad Gesner from 1602. But most of what is described in this document is congruent with those of domestic cattle, for example the flehmen gesture, the way cows look for their calves or impressing behaviour by the bulls. All cattle have the same and comparably complex social and reproductive behaviour that also resembles the other bovine species, indicating that it is not greatly altered by domestication.  Therefore I assume that the behaviour of the aurochs was very similar if not identical to that of domestic cattle. The following passages are based on observations of living domestic cattle (Camargue, Chillingham cattle, Heck cattle, Highland cattle, Galloway) and wild bovines as much as on Schneeberger’s report. I am going to use photos of more or less aurochs-like cattle on a relatively natural-looking background here to show the behaviour patterns described in the text.

Herd structure

Cattle form herds all year round, even tough Schneeberger reports that aurochs roam separately during summer [1]. Probably he spotted single bulls since these usually separate themselves from the herds that are formed by cows and juveniles. These herds have a matriarchal structure [2] and wander around looking for feeding grounds. The usual herd size for cattle is between 20 and 30 animals [3]. If the number of the animals within the herd grow larger, the herd splits up [2]. The cows form sub groups of two or three animals that graze and rest together. The distance between the individuals depends on the social rank and lies between 0,5 and 3 meters [4]. Friendly behaviour includes licking and stroking with the horns. Calves form “kindergarden groups” when their mothers are occupied with grazing and gestating, which are guarded by non-suckling cows or young bulls. The latter leave the herd at the age of 1,5 years when becoming sexually mature.

Adolescent bulls form bull groups, which is common to most bovines (therefore I was a little bit upset when an individual on deviantArt called this behaviour “speculative” commenting this illustration). These bull groups circle around the cow herds, and each group mates with cows of different herds each respective year, what can be interpreted as a mechanism against inbreeding [2]. Bulls only join the herd during mating season. The common presence of bulls within cow/calf herds in some free-ranging domestic cattle populations might be explained by the loss of seasonality in the reproductive circle of domestic cattle [3]. If the place is confined, cattle form a herd of all age classes and both sexes [3], so the herding behaviour apparently is plastic.

Adolescent bull group

Mixed-sex herd

Older bulls from an age of 10 to 12 separate from the bull group to avoid the constant competition with the other bulls. These old bulls are solitary, territorial and do not partake in reproduction anymore. The maximum life expectancy of cattle is about 20 years [5]. In wild aurochs it probably was a little shorter, and also Schneeberger reported that aurochs lived for only 15 years [1].

When cattle are alarmed or start to flee, they do not stray up but stay close together. Like many other large ungulates, they form a defensive circle around their calves [1].

Agonistic behaviour

Cattle are hierarchic animals and there is a dynamic social order within a herd. This order is not linear but polygonal, f.e. C < A < B < C. The higher the rank of an animal the higher the quality of food and resting places, and the lower the level of stress because a lower rank in the hierarchy means that is constantly dominated by other herd members. Dominating behaviour includes display like standing in front of the opponent in an angle of 90° and elevating the head to show off its full size, or hits with the horns and pushing the individual aside. If the conflict does not end here, a combat fight follows, which is carried out head-to-head by pushing and pulling the opponent with the horns until it retreats. The winner might chase the loser for a few seconds. Harsh fights take place by bulls as much as cows [2,3,4,5]. Free ranging cattle, especially bulls, often show the results of such fights: scars along the neck and shoulders, broken-off horn tips, ear cuffs ripped apart, and sometimes even a stabbed eye. Superiority does not necessarily depend on mass and strength only – the size and shape of the horns plays a role as well, as much as the psyche of the animal [3,4].

Reproductive behaviour

Bulls also show off their strength by throwing dust and plant material into the air with their horns and hooves (which is also reported by Schneeberger), omitting a growling sound. Bulls in a rut also make repetitive, trumpeting sounds which I guess are used to attract cows and intimidate other bulls. For videos showing the behaviours I just described, see here, here, here and here.

Cattle form harems during the mating season, which under natural circumstances usually takes place during late august* [1]. The traditional image that one bull conquers a harem and is the only male who covers the cows is not correct. In fact there is a number of other bulls that seize their chances when the leader of the harem is busy at the moment, but it is the cow’s choice which bull mates with her. Bulls in heat show the so-called flehmen gesture by curling back their upper lip and stretching out the tip of the tongue in order to detect pheromones and other scents with their vomeronasal organ. After three days of testing the cow’s scent and urine, the bull mates with the cow. Friendly behaviour between bull and cow like licking and stroking with the horns usually are restricted to the mating season.

* Domestic cattle in man’s custody mate all year round, but if exposed to natural factors and natural selection in particular, their reproductive circle becomes seasonal again like in all large wild mammals. Aurochs apparently had the same rhythm.

After mating, a nine month-long pregnancy starts. The behaviour of the cow immediately before birth probably depends on habitat [3]. If forest edges and bushes are available as shelter, the cow separates from the herd a few days or hours before birth [3,5]. On open grassland, the cows might tend to calf near to the herd [4]. The new-born calf immediately tries to make its first step while the mother is licking its wet fur [5]. The placenta arrives a few hours after birth and is consumed by the mother. As soon as the calf is able to walk, the cow takes it away from the birth site to not attract predators. The first milk the calf consumes is called colostrum and is very alimentary. The mother-calf bond is formed within three days. Mother and calf recognize each other by scent, voice and looks. While the mother grazes in the herd, the calf is hidden between bushes or at a forest edge, where it is exposed to predators [1,5].

Literature

[1] van Vuure, Cis: Retracing the Aurochs - History, Morphology and Ecology of an extinct wild Ox. 2005

[2] Meissner & Limpens: Dedomestikation – wilde Herden zwischen den Menschen. 2001

[3] Julia Poettinger, 2011: Vergleichende Studie zur Haltung und zum Verhalten des Wisents und des Heckrinds.

[4] Annette Perrey: Die Sozialstruktur einer Herde Auerochsen im Wildgehege Neandertal. 1999

[5] Frisch, Walter: Der Auerochs – das europäische Rind. 2010.