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The Making of Senna part 9: The response in Brazil
7th June 2011, 9:19 2nd April 2015, 22:11 | Written by Keith Collantine
Ayrton Senna, Williams, Interlagos, 1994
One scene in Senna which is received with laughter by most audiences gets a very different reaction when shown in Brazil.
Why does one scene get two contrasting reactions?
The film's writer Manish Pandey explained some of the surprising responses to the film in Senna's home country.
"I should have kissed him for '94"
One of the more amusing moments in Senna sees him on stage in a Brazilian children's television show. He is smothered with kisses – and bright red lipstick – by the host Xuxa Meneghel.
But while this scene is viewed as comedy by many – including the filmmakers – in Brazil it carries more sombre undertones, as Pandey explains:
"Watching that in Sao Paulo, it was one of the darkest moments of the film.
"It just shows you, [what happens] when you're a bunch of foreigners and you make a film about somebody you never really knew.
"We turn up and Asif [Kapadia, director] and James [Gay-Rees, executive producer] and I watched the film at the premiere in a cinema full of Brazilians for the first time.
"Normally, when you watch it, you know where the funny moments are – like Alain Prost asking if they can be tied on points.
"When the Xuxa thing happens, everyone in England laughs: you've got this big world champion dancing badly with lipstick all over his face.
"What we didn't realise is that in Brazil there's a big myth about the number of times she kisses him. When she stops, she stops at 1993.
"It never occurred to me, it never occurred to Asif, it never occurred to James, it never occurred to anybody. We were laughing at it. She shook our hands at the end of the film and she was crying.
"And the next day in the Brazilian press she said 'I should have kissed him for '94′".
The Piquet rivalry
Ayrton Senna, Nelson Piquet, Jacarepagua, 1986
The critical reaction to the film in Brazil was also quite different: "They really wanted more of the Senna-[Nelson] Piquet rivalry, because our film is centred around Senna and Prost".
Pandey contrasts Senna's view of being Brazilian, given in an interview on a programme called Roda Viva, with Piquet's:
"He clearly felt that he was Brazilian, whereas Piquet wasn't. He clearly felt that he'd made this absolutely conscious decision to get on a plane after every race [to return home].
"Ron Dennis told us much the same thing that, at McLaren, even if they were travelling two days to be in Brazil for one, he just needed to do that.
"Whereas Piquet, as he says, lived in Italy, his friends are Italian and he's got very few Brazilians around him. Nelson Piquet's name was Piquet Souto Maior and he hid the Souto Maior part because he didn't want his mother to know he was motor racing.
"He's a great guy, a funny guy, but he was in some ways an extremely different human being to [Ayrton] Senna. I think the Brazilians loved that playboy image in the beginning but then they found something a bit more earnest. The thing about Senna is, if you're going to market the two, he's probably the easier one to, he's a bit more wholesome.
Ayrton Senna's F1 career
"But we couldn't get that into it, and it's a criticism in Brazil – they just feel, how can you talk about Senna without talking about Piquet. And that's absolutely fair".
But Pandey was less interested in another aspect of the story there was much appetite for in Brazil:
"I think the Brazilians also – because they've got quite an active tabloid press – they wanted to know why we hadn't gone into all his girlfriends. And our reaction to that was, well, how would you do it? Do you cut away from a race to a headline in a newspaper?
"Weirdly, I think we were desperately faithful. Because he does come across as a guy for whom, really, motor racing was it. Those girls were interchangeable and that's the truth of it.
"In the context of a guy who gave his all to become a motor racing driver, I think women, not matter how important they were to him in real life, could only ever have been incidental to him in the film".
This is the concluding part of "The Making of Senna". You can find all nine parts of the series here.
See the official website for more information on the film and the official Facebook page for a list of cinemas that are showing it.
Senna movie
The Making of Senna part 8: The Death of Ayrton Senna
The Making of Senna part 7: Imola 1994
The Making of Senna part 6: The perfect bad guy?
The Making of Senna part 5: The lost scenes
The Making of Senna part 4: 'You've never heard F1 sound like this'
The Making of Senna part 3: Inside the F1 archive
The Making of Senna part 2: Meeting the Sennas
The Making of Senna part 1: Life and death
"Senna" – the Ayrton Senna movie reviewed
Browse all articles on the Senna movie
Posted on 7th June 2011, 9:19 2nd April 2015, 22:11 Author Keith CollantineCategories Ayrton Senna, Interview, Senna movieTags Ayrton Senna, F1, f1 racing, formula 1, formula one, grand prix, motorsport, senna, senna movie
69 comments on "The Making of Senna part 9: The response in Brazil"
7th June 2011, 9:29
"He's a great guy, a funny guy, but he was in some ways an extremely different human being to [Ayrton] Senna."
I'll say. He's come a long way since then, what with his spiteful actions with his son in holding Renault hostage in 2009 instead of coming clean because it was the right ting to do. I wonder if, in seventeen years, we'll be seeing a documentary about the life of Nelson Piquet, looking at how he went from a World Champion to a bit player in one of the biggest controversies to mar the sport …
Idea for a guest article: Top Ten People/Events Who Need To Be The Subject Of A Documentary.
The less pleasant side of Piquet was evident while he was still active in F1. You're surely aware of the then infamous interview in which he suggests Senna doesn't like girls and that Nigel Mansell's wife is ugly?
So now Piquet Sr. is to be blamed by a cock up done by Briatori, Simmons and Junior in order to favor El Sonso (who nobody will ever convince me that he was not in the know)?
He played his part. He was, after all, involved in negotiations over his son's contract with Renault. And considering that Nelsinho chose to extort Briatore with his knowledge of Singapore 2008 rather than go straight to the FIA with his claims, I'd say that makes Piquet Snr. complicit in the scheme.
Give me a break. Piquet Sr. is no angel, but if anyone came out of the scandal with integrity left it was him.
zecks
Great report Keith, senna was such an enigma that no film, book or article could ever tell the entire story. For me the timeline jumps make the film feel very much as though a friend is recollecting shared memories. Inevitably it sparks a conversation becuase there are things that you remember as even more important to your own understanding of the man, but the friend missed. I for one really hope that a director's cut is released.
Thanks zecks!
I hate to rain on your parade but I do keep getting asked this question over and over again: all the footage they could get from FOM was used in the film, so there's not going to be a DVD with lots of extra bits of footage in unless they go and get more from FOM.
The DVD/Blu-Ray release includes all the original interviews they recorded cut into the film, which creates a feature with a longer running time, but there's no extra FOM footage in there.
More on that in parts three and five.
Keith, what about bits that were cut? (ie Senna/Tamburello at testing a month before his death?) won't they be on the film?
No, they're not in the film. Here's why. It mostly has to do with poor quality footage, footage that didn't convert well to the big screen, or a lack of footage that meant the film-makers couldn't portray both sides of the story.
I know they are not in the film, but surely they still have footage of it that could be an extra even if it was cut from the film?
Maybe. But I sincerely hope they are not included. That may sound like a strange thing to say, and even go against the grain of everything being a fan of the sport and Senna is supposed to represent, but there is a method to my madness.
It is a pet hate of mine when film-makers plan their entire project around the DVD extras they are going to include. JJ Abrams is particularly guilty of this (which is why I refuse to see Super 8). A narrative – whether a work of fiction or a biography like Senna – should speak for itself. The audience should not have to go through all the special features on a DVD in order to understand (or better understand) the story it is told. Any writer or director who feels the need to do this is very bad at their jobs. I have never watched any of the featurettes or extra footage for some of my all-time favourite films (notably Serenity and Inception, among others) simply because I believe that the films themselves are pretty much perfect. They don't need any extra content; they speak for themselves. And I honestly hope that Senna will be one of those films.
Keith: Surely they have much more footage already from FOM? They cut the movie down from 5 hours into 104 minutes. That means there is footage already from FOM.
Why do you think there will not be a directors cut?
The answer's in the question: Why buy five hours of footage and then leave two-thirds of it in the bin?
Answer: you wouldn't, you'd only request approval to use the material that's gone into the final cut.
Which, given Ecclestone's reputation for driving a hard bargain, makes sense.
Daniel Chico (@daniel-chico)
It does not make sense. FOM should release all the videos for anyone interested, increasing F1 popularity. More fans = more $… This logic is flawless…
Hi Keith, your point above regarding directors cuts and relying on additional material to better tell a story, thus make up for a bad job of portraying a story, for my mind is valid, previously I had not looked at it like that before. Sadly this movie fell way short of the mark in telling Ayrtons story. The extras do make up some ground, and assist the movie makers a little, but no where near enough coverage of Ayrton the man as opposed to Ayrton the racer. I was so disappointed leaving the cinema, but still purchased the bluray being the devoted Senna fan I am. I am not Brazilian but also did not laugh at the part of Xuxa kisses as immediately I realised the significance of her stopping at 93. Something lost on the film makers, and they claim perhaps because they are not Brazilian or understand the way Brazilians think. I rate the movie 5.5 / 10.
Interesting to think about Piquet there. He does seem to get lost a bit between Fittipaldi who was there earlier and then Senna who is more widely idolised.
Very nice piece to end the series with Keith. Thanks for that and thanks to makers of this movie to work with you on it.
Okay, so I've been thinking (always a dangerous statement from me). We've had all these attempts at making films about racing cars. Most of them don't turn out too well – you've got stuff like Talladega Nights and Driven (which originally supposed to be Stallone's Senna biopic), which never should have seen the light of day. Even the likes of Grand Prix and Le Mans aren't that great; lots of fantastic footage, but not much in the way of plot or characters. And fairly predictable. The biggest limitation of films is their running time – two hours is about the average.
But I've been reading the articles on the Senna film, and one of Manish Pandey's comments stands out: that they had five and a half extra hours' worth of content to include, but they had to cut it down to ninety minutes. So, what if, instead of making films about racing drivers … we started making television series? I'm a huge fan of "Buffy the Vampire Slayer" and "Angel", and both have year-long storylines (indeed, events in the first three seasons of "Angel" directly influence the fourth season). The standard quota of episodes for a television show is twenty-two to twenty-four (depending on the network). With twenty races in a season, it would be easy to fill that quota. The budget would be an issue, but a lot of the action could take place off the track. Instead of having all the melodrama that was all through Driven, the format of television episodes would allow the characters to be fully fleshed out because they have that time. After all, it has been said that where films are short stories, television shows are full novels. And just think: with all the controversy of recent years – Singapore, the McLaren-Ferrari affair, the Hamilton-Alonso rivalry, etc. – there's a lot of potential for storylines.
I actually did a university course in screenwriting a while ago, and we had to write a teleplay for a television show. One of my ideas was to be set in the paddock, but my lecturer didn't think there was a story there, so I went with something else instead because I didn't want to argue with him. But I reckon there is potential there. If a group of writers can crash a plane on an island and then spend six years writing about with no idea what's happening or where it's going, I see no reason why a television show set in the world of motorsport could be a hit.
Sorry, couldn't be a hit. I see no reason why a television show set in the world of motorsport couldn't be a hit.
You were right, dangerous to see you getting some thinking done here PM!
Actually I like the way this is going. Maybe it would be a start to get a nice storyline going first.
But it would be dangerous to include F1 politics, the scenarious are just so absolutely unbelievable (like that WMSC meeting and the interviews with Max, Bernie, Jean, … in the last day or two) :-D
Eh, there wouldn't be able Bernie and Jean could do to stop it. Especially if it was not set in the world of Formula 1, but the "Grand Prix World Championship".
As for a storyline, it wouldn't be ahrd to come up with something good. One the one hand, you'd have the established World Champion (based on Alain Prost) who races with his head, and the perfect rival, a young upstart who races with his heart (Ayrton Senna). But what could be sporting's greatest rivalry is undermined by a team trying to manipulate the rule book in their favour (Ferrari), by another team cheating (Renault), and by a third being embroiled in an espionage scandal (McLaren). But the big threat is organised crime – realising the potential for making money, a crime syndicate has managed to buy into a supplier of standardised parts, and has managed to incorporate powerful electromagnets into their parts, which fry the electronics of a car at the press of a button, allowing them to manipulate the outcome of the race.
We could have this hotheaded new kid as well, who keept thinking everyone is against him.
And we would need the playboy type stealing the others girls (those would boost the viewing figueres).
Don't forget about a team being cleverest to break the rules in spirit every time without ever failing a test.
And there would be loads of stories on backmarker teams with all sorts of bad joke troubles based on the greatest of all fails over time. Russian magnates, Italian Mafia, drugs smuggling in containers, Dodgy regime money (take your pick, Ghadaffi, Bahrain, Venezuela, … ).
And the odd rival championship thinking or media magnate going for PPV and control the sport himself.
By the way, I meant dangerous not for Bernie or Jean, but for its sheer amount of turns in plot :-)
Maybe, but it's a bit of a cliche. Especially since it was in Driven.
No, I'm thinking the main character could be a dignified older driver (like Fisichella) who never really got the shot he deserved (as Heidfeld's fans believe is the case). But rather than lower himself to the level of trundling around with a backmarker team (like Trulli), he decides to quit while he's still ahead (like Hakkinen). But after some time on the sidelines, he gets a second bite at the apple (like Glock) with a famous team going through tough times (like Williams). He gets partnered with an upstart young driver with a famous name (like Alonso with Piquet) who beat out other, more obvious choices for the seat (like Maldonado). But as the season goes on, he starts to realise that something is very, very wrong (like in 2007). By the halfway point of the season, he finds himself replaced (like Chandhok) and ends up fighting for his life (like … nobody, actually). At the end of the year, the team gets banned from the sport (like Andrea Moda), and its assets sold off (like Original Lotus). In the second season, the main character acquires the team and runs it as a team principal (like Prost). This naturally opens up a whole new set of challenges, and sets up a whole new set of storylines.
Maybe you would want some aspects of that storyline to go over 2 or even 3 seasons.
That way we can have the first season where the guy has his last season at a mid grid team before quitting (a halfway heroical drive for the second last race? and crashing out in the pracice session to his final race).
Then the next season we can show him on the sidelines and starting to get involved with getting back again.
And the third season can be about the new hope for the famous but trundling team.
Sounds pretty good. Maybe we should take it further on the forum?
Ah, but you run the risk of things going too slowly for audiences. You could reasonably have three or four episodes showing the final few races of his (planned) last season, another two on the sideline before he joins said famous team, and then finish the season with him buying up the team. The second season would be more political as another team keeps trying to buy the famous team out from under our hero as he tries to keep the team afloat (kind of like Arrows trying to cannibalise Prost through Phoenix Finance in 2002), and the governing body of the sport constantly changing the rules. However, while dealing with this, our hero is blind to the act that his drivers are squabbling amongst one another, threatening the entire team.
PaulK
You started ok, but the ending was terrible. I think a regular underdog story would be more appealing. Like his team surprisingly develops a good enough car so that he can show his talents and win races.
Then we could have some plots where someone crashes on purpose and some big shot is covering it but our hero finds out and try to tell everybody. Also his rivals would swap places whenever their teammates were ahead of them but the main guy wouldn't as it would not be honorous.
The season finale would be the last race of the championship where he needed to win the race to be the champion. You know the deal…
Yeah, but there is a TV show – every two weeks on BBC1!
And it's usually got more intrigue and twists than fiction ever could!
Yeah, but we don't see all the off-track stuff. We read about it, but we don't see it.
god it woul be like that awful show on sky about football http://en.wikipedia.org/wiki/Dream_Team_(TV_series)
419 shows and counting :-(
Not necessarily. "Friday Night Lights" is an American series based on football team, and has been a critical success.
well im sick of reality tv that follows around rich spoilt people who are famous for being famous, it would be great to have them follow a driver around, irvine did it in 99 when he almost won the title made a great doco, the shots where hes told he was diqualifed from malaysia and hakkain has the title then a week later when hes told to championship is still on where great. Or maybe even an AIRPORT style show that follow the fia and race stwards, marshals and track staff, before during and after a grand prix, id watch a 4 to 6 part show on preparing and staging the monaco grand prix.
In america they also do a series called "Hard Knocks" on HBO every year where they follow a team through training camp and pre season, its a fantastic series which really gives you an insight into how the sport works and who the players are as people. Im a big NFL fan and I have totally change my opinions of some players ive seen on that series compared to what i thought of them based on their game time and what we read about them.
Not to mention that this time, there could be someone who knows how to have the best shots end up on screen to tell the story of the race.
DavidS (@davids)
One thing I would like to see if a doco done like Truth in 24 that follows a team for the entire season, with a film crew in the garage getting great footage of drivers, crew and management with their guard down, and the viewer being able to get a great insight into what an F1 team is like.
Even if it focused on a small team, seeing what happens when drivers don't make the cut in qualifying, the disappointment when breaking down, the fight for survival. It could be possible to do one team per year before cycling back to a team which had been covered long enough ago for a significant amount to change to be interesting.
Dude, Driven was despicable but Talladega Nights was hilarious man, that movie is awesome.
That was one of the worst films I've ever seen. Looking back on the review I was far too kind!
debaser91
It would have been a weird if there was stronger focus on Piquet, when he won the title in '87 his main rival was Mansell, and in '86 Senna won races but faded near the end. After that Piquet didn't have a really competitive car.
Having watched the film I have to say I enjoyed it a lot, my only criticism being as you highlighted earlier in the series the fact that rivalry with Prost is too much in Senna's favour in my opinion.
For example if you didn't know about the history of the 1989 season you would infer that Senna had the title stripped from him, when in fact Prost would have won anyway as Senna crashed in Australia. Also not including the bits about Imola '89 and Estoril '88 makes it look like Prost simply couldn't handle a fast driver coming into the team, and doesn't explain why he refused to accommodate his behaviour any more.
The best bit about the film is without a doubt the sequence about Imola '94. I never saw it live as I was too young but have watched footage on youtube, but the horrendous impact of his crash is shocking, and so final. You can see him getting more and more upset over the course of the weekend at the terrible things happening, and the voiceovers from Sid Watkins really paid a fitting tribute to a great driver.
By the same token, Suzuka 1990 didn't really matter because Prost retired in the subsequent race in Australia.
yes, but the problem is that Senna put both their lives in danger.
Likewise, they didn't go into the Senna-Mansell rivalry as much as I would have liked either.
SirCoolbeans (@sircoolbeans)
When I watched the film in London a couple of weeks back I though "she should have kissed him for 94! It's all her fault!" as well. I thought it was a great scene for many reasons, including that.
Dan Selby
I thought exactly the same thing.
I was giggling to myself when she was asking what he wanted for Christmas, and he said he just couldn't say as it'd end up being censored! Then when the kisses began, I thought "wouldn't it be strange if she stopped before 1994…" – low and behold.
TommyB (@tommyb89)
I didn't think the film needed to show EVERYTHING that happened in F1. It doesn't even talk about Donington 1993 yet I didn't even notice.
The thing about the kisses I did funnily enough think about when she didn't say '94. But people can't seriously think that's why he was killed.
Adrian J (@adrian-j)
Saw the film last night. I'm lost for words to describe how good it was.
The one thing that I hold as an iconic image of Senna that I thought would be included in the end-credits montage was that of Mansell giving him a lift back to the pits at Silverstone 1991.
Xuxa failing to kiss him for 94 is instantly perceived by brazilians… and it's certainly one of the darkest, hardest scenes…
I don't know if you have the precise notion about Xuxa's importance. Se is not just "a" host in a TV show for children, but THE most famous, loved and controversial – for her nude pictures and movies in her early days – TV show host in Brazil and, at the time, it was her appex. She had also dated Pele before… She still makes eventual TV shows, but nothing like it was before…
So every brazilian who sees Xuxa kissing Senna for happy new years until 93 has in mind two equally famous pop icons that dated each other during the peak of their careers…
Also, you can't dissociate that kissing scene from her devastated face in the funeral. They weren't dating anymore… he was with Adriane Galisteu at the time, than a beginner, know a famous model in Brazil, that was harshly accused by critics and humorists for building her career around her "widow image", but most people in Brazil believe Xuxa was the love of Senna's life…
Unosposklin
This is not true, Xuxa is who was criticized by posing false widow at the funeral of Senna. Because he was the boyfriend with Adriane over a year. And people here in Brazil do not think Xuxa was the great love of Ayrton, they dated in 89 and 90. Little seen, she working in Brazil and he with F1 in Europe.
What happens, unfortunately, is that Xuxa is always privileged, being famous and is hired TV GLOBO. The same TV GLOBO that transmits F1 races here in Brazil, since that time, until today.
Good news for those in the UK from Manish:
"Huge UK demand means that Universal are expanding Senna film into more cinemas this week. Details to follow. Thank you British fans!"
http://twitter.com/mpandey69/statuses/78060108671754240
Let's just hope it opens in regional Australia.
Cacarella
Cluffy_Wedge
They can probably thank Top Gear for that…
I always find it interesting when people talk to a couple of Brazilians and extend their opnions to be "what Brazilians think about" this or that (the 200 million of them). I am Brazilian and the last thing I would like to see in a movie about Senna are sections his girlfriends or any aspect of his personal life. I was a fan of the driver, I didn't know the man and besides that, I wouldn't watch a documentary talking about the private life of the best friends of mine, let alone about a sporting figure. Regarding his relationship with Piquet, they hated each others guts – in fact Piquet would not miss an opportunity to insinuate that Senna was gay (not a bad or good thing in itself, but Piquet meant to disparage him in doing so). Perhaps the whole subject came to fore because quite some people, mostly Piquet fans, would suggest that Xuxa was a sort of PR girlfriend… Anyway, I would not pay to see movie talking about things like these in a million years.
Yeah, and a deeper focus on Senna-Piquet relationship would certainly have to show these insinuations…
Anyway, the documentary is mostly focused on the driver, but I don't see any problem in talking about the man behind the driver…
Xuxa and Senna, for their fame and their public appearences, is not exactly private life, it is, at worst, a "PR relationship", so its mention makes sense…
But the most important thing in the film is, certainly, his career, his many success, his incredible stories…
So go see the film and don't be afraid to throw out your money… you won't regret.
I had read on this site (I think) about how many kisses Xuxa gives him, i'm not a suspicious guy but I see how it can be construed that way. That part of the film was quite weird, I just imagined seeing one of todays crew doing something similar!
I think it was a good idea to not include that much information about the women in his life. This film was about his motorsport career, well, F1 career. Anything else would have felt a bit shoe-horned in and could have cheapened the film.
Thanks for this series Keith, it's been a good read.
Truly a great film and a really good series of articles Keith. Thank you.
The film really did change my perception of Senna. I never fully appreciated the politics he had to fight against in his duels with Prost and I always saw him as the bad guy. I better understand the reasons for some of his actions now, however right/wrong the may have been.
If you haven't seen it, seriously, what are you waiting for? It's that good.
If you haven't seen it, seriously, what are you waiting for?
Uh, for it to be released?
Malibu_GP
Hmmm, as Someone that has worked in the entertainment business for nearly twenty years (including Universal music) I find the speculation fascinating. Perhaps You Guys could get Me a treatment that I could pitch. Seriously though, there are several motor-sports stories floating around/being considered. Unfortunately, they all wish to depict NASCAR. How fun…, NOT!
Your on!
Lets put this in. Seriously, I think it will be hard to do a really nice show (what studio has a range of pit boxes in their facilities?). But TV has been putting in some very high quality shows lately, a lot better than most hollywood movies for less budget.
During the television show scene in the movie, you could see she loved him very much. I thought it was powerful and the saddest scene in the movie for me, and I'm a hot dog eating American. I cant possibly begin to imagine the grief Xuxa felt when she saw that part of the movie, or even the Brazilians for that matter.
Nelson Piquet's name was Piquet Souto Maior
Reading it in the paragraph talking about Italy, I thought about "Sotomayor", an expression used as nonsense: http://www.youtube.com/watch?v=SofsI4j6i2g&feature=player_detailpage#t=334s
Probably no one will understand this!
These 9 articels really offer an excellent guide to the film. Thanks Keith!
You're welcome, glad you enjoyed it.
Well, as a brazilian I can say there's some misinformation here. First, I don't think anybody here paid much attention to the Xuxa scene, other than it being funny/awkward. Xuxa made some TV success in the 80's and then became quite a decadent personality. Most people here don't even know she dated Senna for a while, and i never heard the story about the 1993 kisses before. But in case anyone's interested, there's a Senna interview for Playboy mag in 91 or 92, where he talks a lot about women, and of how important some of they were, contrary to what Pandy suggests.
As for Piquet, it's really disapointing that the movie doesn't talk more about him. This may surprise some of you, but Piquet is by and large hated for most people here in Brazil, mainly because he was seen as a enemy of Senna, and jeaolous of his talent. They fought a lot on the press, and Piquet even said Senna was gay. By the way Pandy is incorrect when he says Piquet didn't use his last name because of his mother (that wouldnt make sense!). It was because of his father, a politician, that totally disapproved racing. SO he used his mother's maiden name.
I've put together a series of Piquet interviews with english subs here, if anyone's interested:
http://www.youtube.com/watch?v=p7Z1_sQIxbw
The 'Roda Viva' programs mentioned are also very interesting, there's one shot in 86 with Senna, when he talks about how Piquet was different from him and arrogant. And there's another one, from 94, with Piquet. He had already been confirmed in the show, and then, in a strange coincidence, Senna died the day before, so it's interesting to see Piquet having to talk about his disaffect the day after he died.
I am disappointed that the filmmakers didn't realize the '93 kisses connection – I thought that was the reason it was included in the film. The scene was absolutely chilling, to me, not even close to being a 'funny moment', so it isn't just those in Brasil who reacted in a different way, I think the filmakers have completely misunderstood or misappropriated that footage.
btw if you watch Senna, try and fine the long cut. The short cut is a very different film and not as good as the festival version
There's a lot of misinformation about the length of the film, largely spread by people who have obtained illegal copies of it.
For an explanation see this article:
And this comment:
http://www.f1fanatic.co.uk/2011/06/02/making-senna-part-5/#comment-704108
edmuiruf1 (@edmuiruf1)
How can we get hold of senna DVD in Africa (Nairobi).
I have seen the film twice, in the Duke of York cinema in Brighton, and I have to say neither time did anyone laugh at the scene where Senna was kissed good luck for every year up to 1993. There was just silence in the cinema, the majority of the people saw the "significance" immediately.
DennisKY
21st November 2012, 1:39
I've find it odd that all the key players in making the film were oblivious to the "prophetic" undertones of the Xuxa New Year's kisses incident. Sure, at the time it was probably funny to everyone, but in retrospect, it does seem a bit eerie that she stopped at "Happy 1993". I'm not even Brazilian and I thought that the first time I saw the film! How could those guys have missed it completely until they premiered the film in Brazil? Strange.
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BMC Biotechnology
Ethanol effect on metabolic activity of the ethalogenic fungus Fusarium oxysporum
Thomas Paschos1,
Charilaos Xiros2 &
Paul Christakopoulos3
BMC Biotechnology volume 15, Article number: 15 (2015) Cite this article
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Fusarium oxysporum is a filamentous fungus which has attracted a lot of scientific interest not only due to its ability to produce a variety of lignocellulolytic enzymes, but also because it is able to ferment both hexoses and pentoses to ethanol. Although this fungus has been studied a lot as a cell factory, regarding applications for the production of bioethanol and other high added value products, no systematic study has been performed concerning its ethanol tolerance levels.
In aerobic conditions it was shown that both the biomass production and the specific growth rate were affected by the presence of ethanol. The maximum allowable ethanol concentration, above which cells could not grow, was predicted to be 72 g/L. Under limited aeration conditions the ethanol-producing capability of the cells was completely inhibited at 50 g/L ethanol. The lignocellulolytic enzymatic activities were affected to a lesser extent by the presence of ethanol, while the ethanol inhibitory effect appears to be more severe at elevated temperatures. Moreover, when the produced ethanol was partially removed from the broth, it led to an increase in fermenting ability of the fungus up to 22.5%. The addition of F. oxysporum's system was shown to increase the fermentation of pretreated wheat straw by 11%, in co-fermentation with Saccharomyces cerevisiae.
The assessment of ethanol tolerance levels of F. oxysporum on aerobic growth, on lignocellulolytic activities and on fermentative performance confirmed its biotechnological potential for the production of bioethanol. The cellulolytic and xylanolytic enzymes of this fungus could be exploited within the biorefinery concept as their ethanol resistance is similar to that of the commercial enzymes broadly used in large scale fermentations and therefore, may substantially contribute to a rational design of a bioconversion process involving F. oxysporum. The SSCF experiments on liquefied wheat straw rich in hemicellulose indicated that the contribution of the metabolic system of F. oxysporum in a co-fermentation with S. cerevisiae may play a secondary role.
Fusarium oxysporum is a filamentous fungus, capable of producing ethanol not only from hexoses but also from pentoses. The presence of many cellulases and hemicellulases in its secretome is reflected to the ability to grow on many lignocellulosic substrates under submerged or solid state conditions [1-3]. The fungus is capable of degrading and fermenting a wide variety of different substrates under anaerobic or limited oxygen conditions in the presence of inhibitory compounds such as furan derivatives, phenolic compounds and weak acids [4]. The low ethanol production rate and in some cases the formation of significant amount of acetic acid as by-product, have been considered as obstacles for its industrial exploitation. However, it has been shown that the efficient lignocellulolytic secretome of this fungus combined with its ability to ferment xylose could significantly improve the ethanol production from lignocellulosic substrates in co-culture with Saccharomyces serevisiae [5]. The beneficial properties of F. oxysporum can be fully exploited in consolidated bioprocess (CBP) where a microbial system able to degrade lignocellulose and ferment the released sugars in a single reactor is used. CBP is decreasing not only the overall cost but also the environmental impact of the process by minimizing the addition of commercial lignocellulolytic enzymes. The economics of the process can be further improved by operating at high gravity (HG) conditions (operating at total solids content above 20% w/v). Although HG technology is associated with challenges such as the insufficient mixing of the lignocellulosic slurries, the absence of significant amounts of free water and the presence of high amounts of microbial inhibitors, it has gained much attention as a promising technology due to the possibility to achieve final ethanol concentrations above 4% w/v [6]. The higher the ethanol concentration is, the lower the distillation cost (per g of ethanol produced) is [7]. The tolerance to ethanol of microorganisms used in such processes is therefore crucial for their use in large scale.
Ethanol affects the cellular membranes and influences cell metabolism and macromolecular biosynthesis by inducing the production of heat shock-like proteins, lowering the rate of RNA and protein accumulation [8], enhancing the frequency of petite mutations, altering metabolism, denaturing intracellular proteins and glycolytic enzymes and reducing their activity [9]. These inhibitory effects are reflected to the decreased cell division rate, to the decreased cell volume and to low specific growth rates, while high ethanol concentration reduces cell vitality and increases cell death [10]. Robust fermenting microorganisms like S cerevisiae, have developed appropriate mechanisms to deal with several types of damages caused by increased ethanol concentration. The yeast stress response is a transient reprogramming of cellular activities to ensure survival in challenging conditions, protect essential cell components and enable cells to resume their normal metabolic conditions [11].
Although most of the published articles on the field deal with yeasts' ethanol stress and tolerance, there is some evidence concerning ethanol stress and cell response in fungi as well. The addition of ethanol to submerged cultures of Phanerochaete chrysosporium affected both the mycelial morphology and the fungal wall permeability and led to decreased pellet diameter and fungal biomass net weight [12]. The presence of ethanol (0.5–2%, v/v) hampered the secretion of cellulases by Trichoderma reesei [13]. The authors suggested that ethanol inhibition occurred at a pre-translational level by interfering with either the formation or the stability of cellulase mRNA. Asiimwe et al. reported indications that cytosolic aldehyde dehydrogenases play an important role in ethanol tolerance of the mycorrhizal fungus Tricholoma vaccinum [14]. The abundance of certain intracellular metabolites has also been connected to the fungal ethanol stress response: Evidence about the role of sterol glycosides and cerebrosides in the cell response to elevated ethanol levels was provided by the strong increase in the content of sterol glucoside that was observed after the treatment of fungal cells with increased ethanol concentrations [15].
Studying the effects of ethanol not only on the final yields but also on the biomass and ethanol rates is essential in order to evaluate a microorganism with regard to its implementation in biofuels production processes. Furthermore, the possibility of using lignocellulolytic activities in simultaneous saccharification and fermentation (SSF) processes is highly dependent on their stability during such a process. Ethanol is considered to be a non-competitive inhibitor for cellulases and this inhibition probably results from reversible enzyme denaturation [16]. Ethanol, like other organic molecules such as butanol or acetone, binds on the non-catalytic region of the enzyme, causing changes in the shape of the protein molecule, which in turn affects the catalytic activity. However, not all the enzymes are affected by ethanol in the same way. As shown by Chen and Jin the presence of ethanol had a positive effect on β-glucosidase from Penicillium decumbens, especially at temperatures above 40°C [17]. Therefore, the study of the effects of ethanol on the cellulolytic activities is of major importance for the evaluation of a cellulolytic system to be used in SSF processes for ethanol production.
Although many scientific reports have been published on both the hydrolytic and the fermentative performance of F. oxysporum on lignocellulosic substrates, little study has been published on the ethanol tolerance of this fungus. Early works on the fermentative performance of F. oxysporum refer to the effects of ethanol on this fungus claiming a relatively high ethanol tolerance but those statements were only based on the increase in ethanol concentration during the fermentation stage [18-20]. Recently, Hennessy et al., [3] showed that Agrobacterium tumefaciens-mediated transformation could be exploited as a tool to generate significant degrees of phenotypic diversity in F. oxysporum strain 11C in response to alcohol stress. In the same study, the importance of further investigation of the ethanol tolerance of this fungus is underlined.
The present work is studying for the first time the effects of ethanol on the cellulolytic and xylanolytic activities of F. oxysporum, on growth on glucose and xylose under aerobic conditions, and on ethanol production from glucose and xylose during micro-aerobic fermentation. This work complimented with the resent work of Hennessy et al., [3] gives to the scientific community a new prospect for the investigation of the lignocellulose degrading and C5, C6 fermenting fungus F. oxysporum as a potential CBP microorganism. Furthermore to overcome the ethanol limits, this study explored an alternative operating condition for fermentation with stepwise partial removal of ethanol. Finally, SSF under high dry matter conditions has been performed in order to explore the contribution of the metabolic system of F. oxysporum along with S. cerevisiae in ethanol production.
Effect of ethanol on growth (aerobic conditions) and fermentative performance (micro-aerobic conditions) of F. oxysporum
The ability of F. oxysporum to grow in the presence of ethanol under aerobic conditions was investigated. Various amounts of ethanol (0–60 g/L) were added to the medium prior to inoculation and the fungal growth was monitored over time. The effect of ethanol on growth on glucose was evident even 15 h after inoculation, while it was maximized after 44 h. As shown in Figure 1, the maximum biomass concentration decreased as the initial ethanol concentration increased. In the presence of 30 g/L of ethanol, the biomass produced corresponded to the 77% of the one in the absence of ethanol, while the decrease was dramatic when 40 g/L of ethanol were added to the medium. Almost no growth was observed when the experiments were performed in the presence of 60 g/L ethanol. The inhibitory effect of ethanol appears more severe when F. oxysporum was grown on xylose as carbon source. Figure 2 indicates that, the produced biomass was severely diminished when ethanol concentration exceeded 30 g/L and actually no growth was observed over 40 g/L initial ethanol in the broth.
Ethanol effect on F. oxysporum growth on glucose. Ethanol was initially added at concentrations up to 6% w/v in the cultivations prior to inoculation, using glucose as carbon source. All experiments were carried out in duplicate, vertical bars indicate the error levels. The symbols stand for: No ethanol added (●), 1% w/v initial ethanol (○), 2% w/v initial ethanol (▼), 3% w/v initial ethanol (Δ), 4% w/v initial ethanol (■),5% w/v initial ethanol (□), 6% w/v initial ethanol (♦).
Ethanol effect on F. oxysporum growth on xylose. Ethanol was initially added at concentrations up to 6% w/v in the cultivations prior to inoculation, using xylose as carbon source. All experiments were carried out in duplicate, vertical bars indicate the error levels. The symbols stand for: No ethanol added (●), 1% w/v initial ethanol (○), 2% w/v initial ethanol (▼), 3% w/v initial ethanol (Δ), 4% w/v initial ethanol (■),5% w/v initial ethanol (□), 6% w/v initial ethanol (♦).
Furthermore, the inhibitory effect of ethanol on fungal growth was clearly reflected not only on biomass yield but also on the growth rate. As shown in Table 1, the specific growth rate decreased dramatically as the ethanol concentration in the broth increased. In order to fully describe the kinetic pattern of ethanol inhibition on cell growth, the kinetic model proposed by Luong [21] was applied:
$$ \frac{\mu_i}{\mu_o}=1-{\left(\frac{P}{P_m}\right)}^a $$
Where, μ0 and μi are the maximum specific growth rate and maximum specific growth rate in the presence of ethanol respectively. Pm is the critical ethanol concentration above which cells cannot grow. The relationship between specific growth rate and critical ethanol concentration could be described by the ethanol tolerance index "α" proposed by the same model (a linear relationship for α = 1, a hyperbolic relationship when α > 1 and a parabolic relationship when α <1) [21]. The average specific growth rates were calculated and plotted against ethanol concentrations to estimate Pm and α of our experimental data, extracting values of 7.2% and 1.05 for Pm and α, respectively when glucose was used as substrate, and Pm = 3.8% and α = 2.5 when the fungus was grown on xylose.
Table 1 Growth rates and lag phases of F.oxysporum' s aerobic growth in the presence and absence of ethanol
Fermentation stage
In order to investigate the ethanol effect on the ability of the fungus to convert sugars to ethanol, F. oxysporum submerged cultivations previously grown under aerobic conditions, were subsequently shifted to micro-aerobic conditions, while various ethanol concentrations (0–6% w/v) were added at the start-up of fermentation stage. Figures 3 and 4 show the net ethanol amounts produced by F. oxysporum cells at different initial ethanol concentrations with glucose or xylose as carbon sources respectively. The total ethanol concentration to which the cells are exposed during the fermentation course was the initial amount added to the broth plus the produced ethanol. The experimental results obtained on glucose fermentation showed that both ethanol production rate and final ethanol concentration were affected by the initial ethanol level added in the culture. When glucose used as carbon source, the addition of 2%, 3% and 4% w/v ethanol in the culture media resulted in reduction of 46%, 65% and 74% in the net ethanol production, respectively, while no fermenting activity was observed after the addition of 5% or 6% (w/v) of ethanol. Using xylose as carbon source, the reductions in ethanol yields were 26%, 50% and 68% when 2%, 3% and 4% w/v ethanol was added with no observed ethanol production over 5% (w/v) of initially added ethanol.
Ethanol effect on F. oxysporum fermetative performance on glucose. Ethanol was initially added at concentrations up to 6% w/v in the fermentation broth using glucose as carbon source. All experiments were carried out in duplicate, vertical bars indicate the error levels. The symbols stand for: No ethanol added (●), 1% w/v initial ethanol (○), 2% w/v initial ethanol (▼), 3% w/v initial ethanol (Δ), 4% w/v initial ethanol (■), 5% w/v initial ethanol (□), 6% w/v initial ethanol (♦).
Ethanol effect on F. oxysporum fermetative performance on xylose. Ethanol was initially added at concentrations up to 6% w/v in the fermentation broth using glucose as carbon source. All experiments were carried out in duplicate, vertical bars indicate the error levels. The symbols stand for: No ethanol added (●), 1% w/v initial ethanol (○), 2% w/v initial ethanol (▼), 3% w/v initial ethanol (Δ), 4% w/v initial ethanol (■), 5% w/v initial ethanol (□), 6% w/v initial ethanol (♦).
Effect of ethanol on the cellulolytic and xylanolytic systems of F. oxysporum
To evaluate the potential use of F. oxysporum cellulolytic system in consolidated or SSF bioprocesses, the effect of ethanol on the secreted hydrolytic activities was studied by measuring both the FPA and the xylanase activity in the presence of several ethanol levels. To evaluate the obtained data we performed similar studies using commercial enzymatic preparations. The results are shown in Figures 5 and 6. The assay reactions were carried out in two different temperatures, 30°C and 50°C. The first is the most likely temperature for a consolidated or SSF bioprocess, while the second is the standard temperature assay for the most lignocellulolytic enzymatic activities. The cellulolytic activity of F. oxysporum at 30°C was not significantly affected in ethanol concentrations up to 2% w/v (Figure 5). Moreover, despite the obvious reduction in higher ethanol concentrations, the residual activity at 6% and 8% (w/v) was measured at 72% and 63% of the initial, respectively. The ethanol inhibition was, as expected, more severe at 50°C where the enzymatic activity was reduced to 64% and 47% of the initial in the presence of 6% (w/v) and 8% (w/v) of ethanol, respectively. Finally, under the presence of 10% (w/v) ethanol the enzymes exhibited 50% and 70% of their initial activity at 50°C and 30°C, respectively. The decrease in cellulolytic activity showed a similar trend for both the commercial and the F. oxysporum enzyme mixtures (Figure 5).
Ethanol effect on cellulolytic enzymatic activities. The activity assays were carried out under the addition of ethanol in the reaction buffer. F. oxysporum enzymes compared to mixture of commercial enzymes Celluclast 1,5 L – Novozyme 188. All assays were carried out in duplicate, vertical bars indicate the error levels. (▼) indicates F. oxysporum enzyme complex at 50°C and (Δ) at 30°C. (●) indicates commercial enzymes at 50°C and (○) at 30°C.
Ethanol effect on hemicellulolytic enzymatic activities. The activity assays were carried out under the addition of ethanol in the reaction buffer. F. oxysporum enzymes compared to mixture of commercial enzymes Celluclast 1,5 L – Novozyme 188. All assays were carried out in duplicate, vertical bars indicate the error levels. (▼) indicates F. oxysporum enzyme complex at 50°C and (Δ) at 30°C. (●) indicates commercial enzymes at 50°C and (○) at 30°C.
The ethanol effect on xylanolytic activities is shown in Figure 6. Similarly with cellulases the hemicellulolytic activity of F. oxysporum was more intensely affected at 50°C. As shown, under most of the ethanol concentrations tested, the commercial mixture retained slightly higher activities (5% to 10%).
The stability of the lignocellulolytic enzymes in the presence of ethanol is another important parameter regarding their suitability for SSF or CBP processes where they have to withstand elevated ethanol concentrations for many hours. Therefore, ethanol stability tests were performed using F. oxysporum enzymes in comparison with commercial enzymes at 30°C in the presence of 25 g/L and 50 g/L ethanol. As shown (Figures 7 and 8), commercial enzymes retained 70% and 65% of their activity at 2.5% and 5% of ethanol over 48 hours of incubation while F. oxysporum enzymes retained 52% and 45% of their activity under the same conditions. It is worth to be noticed that the commercially available enzymatic preparations often contain additives in order to increase their stability.
Ethanol effect on lignocellulolytic enzymatic stabilities of F. oxysporum . F. oxysporum enzymes were incubated at 30°C in the presence of 0% w/v (●), 2,5%w/v (○) and 5% w/v (▼) ethanol.
Ethanol effect on lignocellulolytic enzymatic stabilities of commercial enzyme mixture. The commercial enzyme system Celluclast 1,5 L – Novozyme 188 were incubated at 30°C in the presence of 0% w/v (●), 2,5%w/v (○) and 5% w/v (▼) ethanol.
Removal of ethanol during fermentation process
As proved in the present study (Figure 3 and 4), the presence of ethanol had an inhibitory effect on F. oxysporum's fermenting ability. To examine whether the presence of ethanol has irreversible effects on the fungal metabolism, the ethanol was partially removed from the broth every 48 hours, as described in Methods section. Fermentation under vacuum has been performed in the past in order to overcome the inhibitory effects of ethanol [22,23]. Figure 9 shows that the ethanol production of F. oxysporum was increased by 22.5% when the ethanol was partially removed from the broth, showing that when the excessive ethanol was removed from the broth, F. oxysporum regained its metabolic activity confirming the reversible character of ethanol inhibition on F. oxysporum cells under the conditions of the experiment.
Effect of ethanol removal during the fermentation process. The produced ethanol during fermentation process was removed from the broth every 48 hours. All measurements were carried out in duplicate, vertical bars indicate the error levels. Ethanol present in broth (●), total ethanol produced (ethanol present in broth plus the removed ethanol) (♦), glucose consumption (▲), ethanol production in control experiment (∇) glucose consumption in control experiment (∆).
Evaluation of F. oxysporum potential during co-fermentation of liquefied wheat straw
Finally, in order to test the hypothesis that F. oxysporum can be efficiently used in fermentation processes contributing in xylose fermentation and minimization of the amount of externally added enzymes by exploiting the hydrolytic enzymes produced by the fungus, 50 mg/g DM of F. oxysporum cells and/or 5 FPU/g DM enzymes were added along with S. cerevisiae in the start-up of fermentation stage. As control experiments, fermentations of the material with (a) S. cerevisiae and commercial enzymes, (b) S. cerevisiae and no addition of enzymes, (c) S. cerevisiae and F. oxysporum biomass, (d) S. cerevisiae and F. oxysporum enzymes were conducted. Results are presented in Table 2. It can be pointed out that the addition of F. oxysporum enzyme system can adequately replace the commercial enzyme system reaching 94% of the ethanol yield. Furthermore, when there was no enzyme addition at the startup of the fermentation process, the combined effect of both microorganisms resulted in a slight increase in ethanol production of about 7%, compared to the fermentation only with S. cerevisiae. Finally, the addition of F. oxysporum's system in the SSF process increased the ethanol production by about 11%, compared to the fermentation conducted with S. cerevisiae and commercial enzymes.
Table 2 Fermentation of liquefied pretreated wheat straw, with and without the addition of F. oxysporum 's system
The results presented here showed that the response of F. oxysporum to ethanol can be compared to that of other xylose fermenting microorganisms used in bioethanol production processes. Studies on Pichia stipites showed that no growth was observed when the initial ethanol concentration was over 3.5% w/v during cultivation on glucose or xylose [24,25]. Additionally, there was no ethanol production measured on either source over the same ethanol concentration. Similarly, the growth of Kloeckera africana on Agave (tequila juice) at ethanol concentration above 2.5% w/v was associated with reduced biomass production [26]. Another study by Bajpai and Margaritis, made on Kluyveromyces maxianus, indicated that ethanol caused inhibitory effects on cell growth and fermentation ability [27]. As shown in that study, it was mainly the growth and ethanol production rates that were decreased, while the biomass and ethanol yields were almost not affected by the presence of ethanol. In accordance with the results presented here, a linear negative correlation of the specific growth rate with the initial ethanol concentration and a similar linear relationship between μmax and ethanol concentration was reported [27].
The effects of ethanol on growth of F. oxysporum (aerobic conditions) were more severe when xylose was the carbon source in contrast with the ethanol effects in the productive anaerobic stage where the inhibition was stronger on glucose. The difference in the effects on the fermentative performance could be explained by the higher net ethanol production in the case of glucose. Although the net amount produced did not exceed 9 g/L, we must take into account the total ethanol levels. Furthermore, it has been proposed that the toxic effect of the endogenously produced ethanol to the microorganisms is more severe than the effect of the externally added ethanol [28,29]. However, other studies explained similar observations by the cumulative toxic action of ethanol together with other toxic by-products of fermentation, such as organic acids [30,31]. Although organic acids' concentrations were not measured during the present study, it has been shown that acetic acid is a major by-product of the metabolism of F. oxysporum grown on xylose at similar conditions [32].
During the consolidated and SSF processes the need of an active enzyme system is necessary to maintain high saccharification rates and yields. F. oxysporum secrets a multi-enzyme system, rich in cellulases and hemicellulases, which is very efficient in biomass saccharification [33]. F. oxysporum's enzymatic system showed an adequate tolerance to ethanol. Especially at 30°C, the most common temperature for SSF processes, (where F. oxysporum's enzymatic system is more likely to be exploited) the effect of ethanol on the cellulolytic and xylanolytic activities of the enzymatic system was significantly less severe than at 50°C. Different reasons could explain this result. Various researchers have previously shown that ethanol reduces the enzymatic activities of cellulases. Its inhibition effect is temperature depended and increases as the reaction temperature increases [34,35]. Although the inhibitory effects of ethanol on cellulases are in many cases demonstrated studies have reported also destabilizing effects [17,36,37]. It seems that at elevated temperatures, the destabilizing effect of ethanol may accelerate denaturation of the enzyme even if the temperature itself could not cause protein denaturation for such a short period (as in our case, 1 h) [38]. Skovgaard and Jorgensen working on a set of thermo tolerant enzyme mixture found that the effect of ethanol on the activity reduction as the reaction temperature increases is not related to inhibition but rather to destabilization of the enzymes [39].
As shown, the ethanol effect on F. oxysporum enzymes is temperature dependent. When the assay is performed at 30°C, F. oxysporum's cellulolytic and xylanolytic activity remains in high levels, in the presence of considerable amounts of ethanol. Even in the presence of 50 g/L of ethanol, F. oxysporum's enzymes retain 45% of their activity over 48 hours. This could be beneficial in a consolidated bioprocess where the amount of commercial enzymes added in the hydrolysis step could be reduced by the ability of F. oxysporum to produce ethanol tolerant enzymes, able to continue the saccharification during the fermentation stage. Another study indicated that, F. oxysporum in mixed culture with S. cerevisiae was successfully used to ferment carbohydrates of wet exploded pre-treated wheat straw to ethanol under an SSF mode [5].
Additionally, when the produced ethanol was partially removed from the fermentation system of F. oxysporum in a stepwise manner the final ethanol production level was increased by around 23% reaching 38.4 g/L. The increase in ethanol production achieved here is similar to that reported in earlier studies dealing with the in situ removal of ethanol from the fermentation broth, where an increase up to 30% in the overall ethanol production could be achieved in most cases [22,40-42]. As shown in Figure 9, the beneficial effect of ethanol removal is clear only when the ethanol present in the medium is above 3% w/v. When ethanol concentration in the broth tends to reach 3%, the fermentation seems to stop. This is consistent with our findings regarding the ethanol tolerance levels of F. oxysporum. In cultures where the ethanol was partially removed it is obvious that this removal allowed the total ethanol production to exceed the 3% barrier. These results, suggest that as the ethanol concentration diminishes sufficiently, any metabolic inhibitory effects are reversed. As a result, the fermenting capacity is recovered when ethanol is removed. The ethanol concentration achieved here is very close to the crucial ethanol concentration of 4% (w/v) in the broth which is considered as a minimum prerequisite for a feasible large-scale distillation technology [7].
The straw used at the present work is rich in hemicellulose, and thus suitable to evaluate the ability of F. oxysporum not only to hydrolyze it but also to ferment the released xylose. The addition of F. oxysporum cells in the fermentation process resulted in 11% increase of the ethanol production (Table 2). Specifically, if ethanol yields are based on the dry material, the mixed microbial culture fermentation led to 138 g ethanol per kg pretreated wheat straw, while fermentation only with S. cerevisiae resulted in 128 g ethanol per kg pretreated wheat straw. The results shown in Table 2 indicate that F. oxysporum's system is worthy of further investigation regarding its potential to enhance ethanol production during an SSF process along with S. cerevisiae when pentoses are present in the fermentation medium. These results are in accordance with Panagiotou et al. who reported in an earlier study that F. oxysporum multienzyme and microbial system had positive effect in ethanol production [5]. Moreno et al. [43] reported ethanol concentration 25 g/L at 20% solid content, which corresponds to 125 g ethanol per kg of pretreated material. Whereas, Jorgensen et al. [35] reported ethanol concentration up to 130 g/kg, applying a similar liquefaction process to that of the present study, in a high hemicellulose content material. At the present work, the liquefaction step took place for 6 hours, and 5 FPU/g DM were used at the liquefaction step supplemented with 5 FPU/g DM at the SSF process. Comparatively the enzyme loadings of Moreno et al., [43] and Jorgensen et al. [35] were 15 FPU/g DM and 7 FPU/g DM respectively. The saccharification times also vary, at the present study 6 hours of liquefaction were followed by 48 h of Simultaneous Saccharification and Co-Fermentation (SSCF), while Moreno et al. [43] conducted 144 h of SSCF and Jorgensen et al. [35] conducted 24 h of liquefaction and 48 h of SSF.
The complexity of SSCF process when dealing with lignocellulosic materials justifies the differences observed by different researchers. Many factors are affecting ethanol yields when operating at high solids fermentations, such as the nutrients deficiency, the presence of other inhibitors, the viscosity, and the inhomogeneity [6]. Moreover, differences between different raw materials and different pretreatments (The SF value of the pretreated wheat straw used in the present study was calculated at 3.58 (same as Jorgensen et al.), while Moreno et al. and presented a process of SF 3.34) make these processes difficult to control and to compare in many cases. Undoubtedly, many factors not being the objectives of this study, are affecting F. oxysporum ethanol yields and productivities and no clear conclusions regarding the contribution of F. oxysporum metabolic system to xylose fermentation during SSCF can be made.
Considering the above, the results of the present study show that F. oxysporum's addition led to a significant ethanol increase, exploiting the hydrolytic enzymes produced by the fungus and thus reducing the amount of commercial enzymes needed. On the other hand, the ethanol yield, of about 31% of the maximum theoretical based on total carbohydrates content, achieved in the present work indicates that further investigation of the process is required.
The results presented here assess three different aspects of F. oxysporum ethanol tolerance (effects on growth, enzymes, fermenting ability) for the use of a microorganism in a bioethanol production process and show that F. oxysporum can be used in a biorefinery both as a pentose fermenting and enzyme producing microorganism. Hemicellulose rich streams, produced after pretreatment of the raw material could be used for the production of necessary enzymes for the degradation of lignocellulosic biomass. Although the ethanol tolerance levels of F.oxysporum indicated that the fungus could be useful at the early stages of the fermentation, this was not confirmed by the SSCF and SSF results, in those cases F. oxysporum biomass did not demonstrate the expected contribution to the ethanol production, probably due to other stress factors not studied during this work (effect of high solids, presence of other inhibitory compounds). Nevertheless, the effect of F. oxysporum metabolic system could not be neglected as (even not as high as expected) the addition of F. oxyporum biomass increased the produced ethanol in an SSCF experiment along with F. oxysporum enzymes and S. cerevisiae compared to the one that only enzymes and yeast were added. The satisfactory ethanol tolerance levels of F. oxysporum enzymatic system allow the use of these enzymes also in high gravity bioconversion processes. This could lead to the reduction of the overall financial cost of the process, not only by achieving a high final ethanol concentration but also by reducing the amount of commercial enzymes added. To conclude with, this study as well as other studies made on F. oxysporum tolerance and adaptability [3,4] indicated that F. oxysporum is capable of dealing with ethanol, which makes it a very interesting microorganism for further study on CBP and SSCF processes.
Fusarium oxysporum F3 isolated from cumin [1], the fungus was grown on potato-dextrose-agar (PDA) slants for 5 days at 30°C. The slants were maintained as a stock culture at 4°C. Commercial dry baker's yeast (Yiotis, Athens, Greece) was used at the pretreated wheat straw fermentations.
Carbon sources – chemicals
Pretreated wheat straw (Triticum aestivum L) was used as raw material for ethanol production. The pretreatment of the straw (PWS) was performed in the Inbicon pilot plan at Skærbæk, Denmark. The residence time setpoint in the reactor was 12 min and the reactor temperature was maintained at 185°C by injection of steam. The severity effect (SF) was determined according to Garrote et al. [44] by the following equation:
$$ SF= \log (R)= log\left(t\cdot e\frac{T-100}{14.75}\right) $$
Pretreated wheat straw lignocellulose content was (%) cellulose 39.0 ± 1.5 and hemicellulose 21.0 ± 1.2. Structural carbohydrate content (cellulose and hemicellulose) was determined by a method adopted by the NREL protocol [45].
Brewers Grain (BG) and Corn Cobs (CC) were supplied by the Athens Brewery S.A. and the Agricultural University of Athens, respectively. BG composition was analyzed by Xiros et al. [46] and Corn Cobs was analyzed by Katapodis and Christakopoulos [47].
All chemicals and reagents were provided by Sigma-Aldrich (USA).
Commercial enzymes Celluclast 1.5 L and Novozyme 188 were provided by Novozymes (Denmark). The commercial enzymes were used in a 5:1 v/v ratio.
Inoculum
For inoculum production, spores were extracted from the stock slants using 5 ml of sterile distilled water and then cultivated in 250 mL Erlenmeyer flasks containing 100 mL of the following mineral medium:(in g L-1) 1.00 KH2PO4, 0.30 CaCl2•2H2O, 0.30 MgSO4•7H2O, 10.00 (NH4)2HPO4, 6.94 NaH2PO4° 2H2O, 9.52 Na2HPO4° 2H2O [2], supplemented with either 40 g L-1 BG-CC (2/1) or 20 g L-1 glucose or xylose, depending on the experimental procedure. The pH was adjusted to 6.0. The flasks were incubated at 30°C for 2 days in an orbital shaker at 200 rpm (Zhicheng ZHWY-211C) for mycelium production.
Aerobic submerged cultivation
Aerobic submerged cultures were carried out in 250 mL Erlenmeyer flasks containing 2 g of glucose or xylose and 100 mL of the above described mineral medium (pH 6.0). Medium and sugars were sterilized separately at 109°C for 40 min. The culture medium was inoculated with 10 ml of 48 h-old inoculum (prepared as described above). The flasks were incubated at 30°C for 7 days at 200 rpm. The flasks were capped with cotton that allowed aerobic conditions and assured sterile conditions. At different time intervals aliquots were aseptically withdrawn and used for biomass and sugar consumption estimation.
For analyzing the potential impact, of the presence of ethanol, on the growth of F. oxysporum the following experiment was carried out. In submerged cultures as described above ethanol concentrations up to 6% w/v were exogenously added in the broth. At different time intervals aliquots were aseptically withdrawn and used for biomass and sugar consumption estimation.
Biomass estimation
Samples of the aerobic cultures were filtered using 0.2 μm pure size filter paper, Millipore (USA). The biomass content was measured by weighing the dried samples.
Enzyme production
Crude enzyme extract production was carried out in 3 L Erlenmeyer flasks containing 40 g of carbon sources (BG-CC 2/1) and 1 L of mineral medium. Prior to sterilization, the initial pH of the medium was adjusted to 6.0. The medium was sterilized at 121°C for 20 min and was inoculated with 100 mL of 72 h old inoculum (prepared as described above). The flasks were incubated at 30°C for 5 days at 200 rpm. At the end of the enzyme production stage, the culture was centrifuged (14,000 rpm, 4°C, 40 min) and the clarified supernatant was concentrated using ultra filtration membranes (10,000 kDa) Millipore (USA), there was no buffer addition during ultrafiltration process.
One unit (U) of enzyme activity was defined as the amount of enzyme required to liberate 1 μmol of product per minute, at assay temperature.
Filter paper activity (FPA) was determined as described by Wood and Bhat [48]. In a capped 2 ml Eppendorf tube, 1 mL of buffer solution, 0.5 mL of enzyme (with the appropriate dilution so to release less than 1 mg product per mL) and a filter paper (Watman #1 1x6 cm, approximately 50 mg) were added. The mixture was incubated at 50°C for 1 hour at 1000 rpm. The released reducing sugars were measured by the 3,5-dinitrosalicylic acid (DNS) method [49].
The β-1,4-d-endoxylanase activity was determined by incubating the enzyme for 10 minutes at 50°C with 1% birchwood xylan [50]. The released reducing sugars were measured by the 3,5-dinitrosalicylic acid (DNS) method [49].
Ethanol effect on enzymatic activity
For the investigation of the effect of ethanol concentration on the cellulolytic and xylanolytic activity of the enzymes used in this study, assays were carried out in the presence of initial ethanol concentrations up to 10% w/v. The ethanol was added in the reaction buffer solution prior to the enzyme addition. The assay reactions were carried out as described above at either 30°C or 50°C.
Ethanol stability
The ethanol stability of the enzymes was performed at 30°C the SSF temperature at which the enzymes withstand the presence of ethanol for many hours. The enzymes were incubated at 30°C with the addition of two ethanol concentrations, 25 g/L and 50 g/L. At different time intervals aliquots were taken and the cellulolytic activity was measured by the Filter Paper method.
Reducing sugars concentration was determined according to dinitro-3.5-salicilic acid (DNS) method [47]. The effect of ethanol on the sugar measuring method was determined by control measurements, containing all the ethanol concentrations used and a standard sugar concentration. Ethanol appeared to have no effect on the reducing sugar determining method. Glucose was measured according to commercial enzyme solution of GOD/PAP (glucose oxidase/ peroxidase assay) (Biosis, Greece). Xylose was measured according to commercial D-xylose assay kit (Megazyme, Ireland) Ethanol was analyzed using an HPLC system (Szimadju) equipped with an Aminex HPX-87H column (Bio-Rad, 300 x 7.8 mm, particle size 9 μm) using a Refractive Index (RI) detector. Mobile phase was 5 mM H2SO4 in HPLC grade water at 0.6 mL/min flow rate, column temperature was 40°C, injection volume was 50 μl and total runtime was 30 min. All samples were filtered (0,2 μm, Macherey-Nagel) prior to the analysis.
Ethanol production
Ethanol production experiments under micro aerobic conditions were carried out in 250 mL Erlenmeyer flasks containing 2 g of glucose or xylose, sterilized at 109°C for 40 min. 100 mL of culture previously grown on glucose as carbon source for about 4 days (until substrate was fully consumed) at 30°C, were aseptically transferred in the Erlenmeyer flasks provided with needle-pierced rubber stoppers, which ensured micro-aerobic conditions and allowed the release of produced carbon dioxide [51] and the cultures were incubated at 30°C for 7 days and 80 rpm (Zhicheng ZHWY-211C). At different time intervals, aliquots were aseptically withdrawn and used for ethanol production and substrate consumption estimation.
For analyzing the potential impact, of the presence of ethanol, on the fermenting ability of F. oxysporum, micro aerobic cultivations of F. oxysporum were carried out, as mention before, in the presence of extracellular, exogenously added ethanol. The initial ethanol concentrations were up to 6% w/v. At different time intervals aliquots were aseptically withdrawn and used for ethanol and substrate consumption estimation.
All ethanol production experiments were carried out as described by Xiros et al. [4], Dogaris et al. [51] and Matsakas and Christakopoulos [52]. After each sampling, nitrogen (0.1 vvm) was flushed in the cultures for 10 minutes to assure the anaerobic conditions.
Partial ethanol removal
Ethanol production experiments under micro aerobic conditions were carried out in 250 mL Erlenmeyer flasks containing 9.5 g of glucose sterilized at 109°C for 40 min. 100 mL of culture previously grown with glucose as carbon source for about 4 days (until glucose was fully consumed) at 30°C, were aseptically transferred in the Erlenmeyer flasks and the cultures were incubated at 30°C for 7 days at 80 rpm (Zhicheng ZHWY-211C). Every two days the broth was aseptically centrifuged and transferred to a rotary evaporator (Rotavapor RE 11, Buchi (Switzerland)) where the contained ethanol was partially removed. The vacuum process was carried out at 75°C for approximately 10 minutes. Consequently, the broth was returned in the Erlenmeyer flasks along with the centrifuged fungal cells. At this stage, nitrogen was flushed in the cultures to assure the anaerobic conditions. For avoiding broth condensation, the loss of water during the evaporation process was estimated by weighting. Equal amount of sterile water (less than 3 mL each time) was added in the culture.
Liquefaction of hydrothermaly treated wheat straw
For the liquefaction and saccharification of the pretreated wheat straw a reactor was manufactured in house. The reactor consists of a cylinder drum 25 cm wide and 60 cm diameter, and a rotating shaft for mixing the material. A 0,55 kW motor was used for the rotation of the shaft. The rotation speed could be controlled from 0 up to 20 rpm, and the motor can be programmed to shift its rotation clockwise and anti-clockwise. An external jacket filled with oil was used for temperature control up to 90°C. The experiments were executed at 50°C, using 30% DM of straw supplemented with buffer (phosphate-citrate, 50 mM, pH 5,5) and an enzyme mixture of 5 FPU per g DM, consisting of Celluclast 1.5 L and Novozyme 188 (both from Novozymes A/S, Bagsværd, Denmark) in ratio of 5:1 v/v. The activity of this mixture was measured at 81 FPU/ml by the filter paper assay. The mixing speed was 7 rpm shifting clockwise and anti-clockwise twice a minute. The duration of the liquefaction was 6 h.
Fermentation of liquefied wheat straw
The liquefied at 30% DM material was used as carbon source for ethanol production. S. cerevisiae (Dry Baker's Yeast) and F. oxysporum (previously grown on glucose for 4 days under submerged cultivation) were used as the fermentative microorganisms. 5 mg/g DM dry yeast and 50 mg/g DM F. oxysporum centrifuged cells were added. Moreover 5 FPU/g DM of either commercial enzymes (Celluclast 1.5 L and Novozym 188) or F. oxysporum's enzymes were added to enhance the hydrolysis. The final DM in the mixed fermentation was 25% w/w due to the addition of the enzymes and the fungal biomass. All experiments were carried out, in duplicate, under submerged cultivation at pH 5.5 and T = 30°C in 250 mL Erlenmeyer flasks.
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The research leading to these results has received funding from the European Community's 7th Framework Program (FP7/2007-2013) under grant agreement 213139 – the HYPE project.
Thomas Paschos would like to thank the Hellenic Scholarship Foundation for a grand.
Paul Christakopoulos would like to thank Bio4Energy, a strategic research environment appointed by the Swedish government, for supporting this work.
Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens, 9 Iroon Polytechniou Str, Zografou Campus, Athens, 5780, Greece
Thomas Paschos
Industrial Biotechnology, Department of Chemical and Biological Engineering, Chalmers University of Technology, Kemivägen 10, Gothenburg, 41296, Sweden
Charilaos Xiros
Biochemical Process Engineering, Division of Chemical Engineering, Department of Civil, Environmental and Natural Resources Engineering, Luleå University of Technology, Luleå, SE–971 87, Sweden
Paul Christakopoulos
Correspondence to Paul Christakopoulos.
PC conceived the study. PC and CX designed the experiments. TP carried on the experimental work. PC, TP and CX drafted the manuscript. PC and CX revised the manuscript. All authors read and approved the final manuscript.
Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made.
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To view a copy of this licence, visit https://creativecommons.org/licenses/by/4.0/.
The Creative Commons Public Domain Dedication waiver (https://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
Paschos, T., Xiros, C. & Christakopoulos, P. Ethanol effect on metabolic activity of the ethalogenic fungus Fusarium oxysporum . BMC Biotechnol 15, 15 (2015). https://doi.org/10.1186/s12896-015-0130-3
Ethanol inhibition
Ethanol tolerance
Ethanol removal
Submission enquiries: bmcbiotechnology@biomedcentral.com


This interview is with Hilary Berg, the editor of the Oregon Wine Press. In this interview, she talks about the start of OWP, learning about the Oregon wine industry and how she finds the right stories for publication. Hilary finishes the talk with a discussion of the future of the Oregon wine industry and the OWP.
This interview was conducted by Rich Schmidt on May 11, 2017 at Linfield College in Nicholson Library.


also featuring Billy Strings, The California Honeydrops, Trout Steak Revival, Daniel Rodriguez, Tierro Band with Bridget Law, Bonfire Dub and more.
The tradition of Campout for the Cause has always been rooted in connection, conversation, passion, sport, music and feeling more inspired upon exit than arrival. The goal of the festival is to focus on the purpose, the gathering and the evolution of human connection. Come for the music, conversation, yoga, bonfire connections, rafting, family time or just to take your shoes off and run in the grass!
The Cause: As has been Campout's cause for two years, this year profits are once again dedicated to local grassroots fund, Emmy's Friends. Created in 2017, Emmy's Friends is a fund benefiting Emery Welle, a 2-year-old from Red Cliff, CO who was diagnosed with a rare childhood cancer calledembryonal rhabdomyosarcoma. She and her family have been fighting this disease the majority of her life and Campout is proud to continue to aid her in that fight.


If this is your first foray to the World's biggest X-wing event, this may be a helpful little guide to survival. Even if a veteran of such, there might be a useful nugget below.
Due to population density and relative ease of travel, (ignoring the terrible road systems and poor public transport), the UK system open often attracts a fair old number of participants. One of the reasons it has sold out quicker and with more players than Uk Nationals and Euros over the years is that for all but the first event it has not been hosted at a gaming Expo and with the additional fees and accommodation issues that entails. Added to that, the system open events have the best participation prizes and a decent tournament structure for players of all abilities.
You get a great bang for your buck in terms of games and goodies.
You will see this in most tournament survival guides, but as always I recommend making sure you have plenty of water with you and sensible energy-boosting snacks. A chocolate bar whilst providing energy will be a quick hit followed by a slump. I would also recommend being aware that there may be gaps of 30 to 45 minutes between rounds due to the large numbers. If you are going to eat food on site, tactically decide the best moment to gather it, especially if you finish a round early.
Last year was impressive with 500 players getting match ups pretty smoothly and on time but expect to finish the sixth round at about 9pm on Saturday. With the likelihood of a pre-cut elimination round following for those on 5-1 records.
Last year the Sunday caused issues in terms of pairing times due to players dropping out of the side event, but I imagine the TO team have looked at their systems again for that this year with increased numbers taking part. This could still be a mildly late finish.
Templates, check, dice, check, dials, cards, space ships check as well as excuses like muh dice, bad matchup, road works on Chertsey road put me off, check! Anything else? Ticket, hotel accommodation, travel plans, printed off list, spare lists, options for Hyperspace, pen & paper, a bottle of Campari, cash or maybe rare 1.0 Alt art Omnicron group pilot cards for bartering in case a no deal Brexit happens on April 12th.
On a serious note, it might be worth to keep an eye on the news, especially if travelling from abroad. Its unlikely anything odd will happen, like the millennium bug a while back, whenever that was supposed to happen.
This depends on what one's own objectives for having a good time or fun is. A recent blog by Paul Braggins looks at this and is worth a read.
We at Zombie Squadron cover a broad church of players from the winner of the most prestigious invitational event so far, the Corescant invitational 2018, to jank players and those just happy to dodge the rocks and roll dice making pew pew noises inside their heads. It's worth putting your personal aspirations into some form of perspective before the event to gain maximum fun.
For example one of our guys who made the cut last year going 5-1 is having trouble 'feeling it' at the moment with a list that they are happy with flying. So in this regard, he has altered his expectations to not even to think about making the cut again but hopefully, a respectable 3-3 or 4-2 might be attainable.
More so just to enjoy six games on the Saturday and six on the Sunday with a list that might be fun to fly. Personally, I'd be happy with 4-2, and if it were better than that, it would be a real bonus.
Hyperspace list or Extended list?
If you feel that the Hyperspace event on Sunday might be more your jam then maybe playing the same list on both days could be a good option. Certainly, a variety of Hyperspace lists look competitive in extended. Some Zombies are looking to do this as they are looking to play either CIS, Galactic Republic, Resistance or First Order.
There has been little local data outside of Hyperspace to go on in regards to Extended in the UK of late. We know from recent System opens that quad Tie Phantom is good, Drea Renthal and friends are good, as well as Imperial Aces. We have also seen some breakthrough into cuts of First Order and Resistance. I would imagine we may see more Hyperspace styled lists in the extended event.
If we look at what has gone 5-1 at the recent Hyperspace events in the UK, we get an idea of what is probably good in this format going forward. The following lists at most lost only 1 game. All bar one set of results comprised of 6 round events, the other was a 5 round. This excludes the recent Dark Sphere event as I was already past writing it (and should have published this last week).
Interestingly Ollie Pocknell's two wins with Fenn and 3 protectorates comes from a 7th and 11th place standing in the Swiss on 4-2. This may mean that the list in the right hands is a bit like double IG in 1.0. If you can navigate through the Swiss, you may be able to feast on the elimination bracket. Fang Fighter variance is something you may have to live with if taking it.
Perhaps the interesting thing when looking at this small data snapshot is that both the winning lists and top of Swiss lists all look capable of doing the job in Extended.
Resistance 5s won the Dark sphere event which to me says that in the right hands they are quite potent, topping two in Swiss and taking the win in another.
I'm slightly torn at the moment. I put Poe, Nein and Lulo down on the table to have a play and it turned out to be really good and fun. I have put a fair amount of time into the Imperial ace and mini swarm, and I'm still undecided on whether I take Duchess or Whisper as the accompanying ace in extended. Duchess is better with Fifth Brother, but you get to the points region of Whisper pretty quickly who has in-built shield upgrade comparatively. Soontir could be an option, also now hyperspace legal.
Last year in a room of 500 players, over 6 rounds, I managed to play Ed from Firestorm Squadron from down the road, in round 2 and a Zombie off versus Dave in round 6. After getting knocked out of the cut by Jesper 1.0 from the 186th, I had a further Zombie off in round 3 of the Hyperspace on Sunday. Twelve of us made the trip last year and we had four inter-squad games in the main competitions. I expect to face a familiar face at some point again.
This in many ways is why we go to these big events. To catch up in person with some of our sparring partners from across the country or further afield. Most of the time, especially when your weekends are more limited to local events for local people, these are the people you only get to see in person a couple of times a year.
But it did the trick. Settled some nerves, confirmed their own expectations and he went and had a good time and won one game, which he was delighted with.
At the end of the day, there can be only one tournament winner out of 630 odd players. Although a win for anyone is playing 12 games of one's favourite game over the course of the weekend in the company of friends.
I have been really enjoying the Sith takers podcasts. They are intentionally on the short side, but as the old saying goes, less is more. Really worth a listen.
April 5, 2019 in Article.
Points Rebalance January 2019: What's going to happen?
Oh, dear God. I went to the Mynock Open (unofficial, of course) last November and there were just over 100 people there. It was run super smooth.
Went to the Atlanta regional last spring, about 160 players. It went relatively smooth.
I can't even imagine over 600 players. I'm going to two official SOs this year (in the states, of course), and I'm expecting 200 to be on the high end.
The guys who have run it each year, starting with 412 players in 2016 have always done a good job and have looked to improve year on year with their methods. I was impressed last year at their handling if 509 players.


AM- Woke up pretty early, ate a light breakfast, and drove up to the starting line of the 5k. I was worried that I wasn't going to get to the start of the race, because of all the road blockage for the other races, but ended up not being a problem at all. Did a mile warmup around the neighborhoods and several drills and strides afterward. First two miles are downhill, 5:31 and 5:34 for those. Once it leveled out, my third mile was slower with a 5:47. The last stretch of this race doesn't have a significant amount of uphill, but after having the legs pounded on the downhill, I could definitely feel it. I need to work on being able to finish stronger in the last mile, because during most of my 5k races, I have been able to hold the pace that I would like to maintain for the about the first mile or two, and then the third mile is a huge struggle. Now that my summer 5k races are done, I think that I am going to take about two weeks of down time from intense training, and do some other activities besides running. After that I will begin to focus on my fall half marathon and marathon training, and try to get a good PR in one or both of those distances. Overall a really fun race, and I am considering doing the half next year, but I will just see what is going on during that time next year. After the race, watched the parade, went to the Days of 47 Rodeo, and toured the University of Utah campus. Very tired now.