Sunday 3 May 2015

Mushroom Farm

Laurent Demuynck, CEO of Kigali Farms
The scientific part of the mission is over.  The time has come to deliver our samples to Kigali farms.  The Petri dishes contain material of wild varieties of edible mushrooms.  With this material Kigali farms can start to carry out tests that could eventually lead to the commercial exploitation of certain species.

We arrive in Kigali where we meet up with Laurent Demuynck, the founder and CEO of the company.  He told us about the aims of his enterprise.

Kigali farms is a social profit making enterprise that was set up in 2010.  Coming from an idea to reduce the level of malnutrition, they have concentrated on mushrooms because they have a high nutritional value.  The wild edible mushrooms of Rwanda interested us from the beginning although they had never previously been researched.  Our ambition was to discover the hidden treasures of the Rwandan forests which are eaten by the local population

Our goal was to make Rwanda an example of excellence when it comes to the cultivation of mushrooms.

We then make our way to the production facility of Byumba where we meet Adriane Mukeshimana (production and quality control manager) who will show us the various stages of production, starting with the wild varieties collected by our mycologists.  So let's follow the guide…


Step 1
Purification of the mycelium

A piece of mycelium is removed from the collected mother material and transferred to a new Petri dish with agar nutrient medium.  This step takes place under sterile conditions following a standard protocol.  The mycelium can now start to grow.


Step 2
Preparation of the inoculum

An agar plug with mycelium is taken from the Petri dish and transferred to a growth medium based on milled grain.  The mycelium continues to grow into what we call an inoculum.


Step 3
Seeding of the inoculum into grow bags

The inoculum is seeded into pasteurised cotton bags containing cotton fluff, wheat husk and hydrated lime.  This specially formulated growth medium is not only favourable for the growth of mycelium but also for the fruiting bodies (sporophores) of mushrooms.


Step 4
The grow bags go to the incubator
The grow bags are stored in the dark in the incubation room.  These conditions prevent the premature formation of fruiting bodies.


Step 5
The grow bags are buried

The grow bags are finally transferred to trays containing soil.  The air humidity is kept at a constant 95% with a temperature of 18-22°C.  Under these conditions we can shortly expect fruiting bodies to develop on the surface of the soil.  In this case Oyster mushrooms that can then be harvested.


Hopefully the mushrooms which we collected on our expedition will also grow here… but it’s far too soon to say… Fingers crossed.


Saturday 2 May 2015

Camp-base


The collection of edible mushrooms in the mountain forests is only the first stage in the work of our mycologists.  Back at base camp their working day has still many hours to go.

A sample consists of several mushrooms originating from the same mycelium.  The sample is given a number, a description and photos taken in the field.  Afterwards, each sample undergoes various procedures with three end goals:


1. Spore print and inoculation

Spore print 
The cap (pileus) of the mushroom is placed on a holder to collect the spores.  In mycological terminology, this is called taking a spore print.  The following day the spores are harvested and seeded onto a Petri dish containing growth medium (agar).  After a few days incubation the mycelium in the Petri dish are sufficiently grown to be passed onto to Kigali farms to grow on.


2. Living stems and DNA samples

Inoculation

A fragment of stem (stipe) 2mm is placed in a test tube containing CTAB, a substance that blocks the degradation of DNA.  Once in Belgium, these samples are for the collections of the Botanic Garden, Meise, whilst the living stems go to the mushroom collection of the UCL (Catholic University of Louvain la Neuve, Belgium).  The latter receive fungal samples from all over the world.  These samples are mainly used for taxonomical research.



3. Collections of the Botanic Garden

DNA samples
The rest of the sample is dried and placed in a plastic bag and deposited by RDB.  A duplicate goes to the herbarium collection in the Botanic Garden, together with a description, the field photos and a sample of the spore print.  A microscopic examination is necessary to confirm the identity of a species.


A mission day starts at dawn but seldom ends before 7p.m.


Collections for the Botanic Garden









Thursday 30 April 2015

Surprises in between the bamboos

Sabyinyo volcano
The deciduous forests at the foot of the Bisoke volcano had a diversity of mushrooms.  But how is it in the bamboo forests? To answer the question our team will go on an exploratory mission to three of the volcanoes (Sabyinyo, Gahinga and Karisimbi).

A new local guide accompanied us.  His name is Damascene and he knows a lot about the forest.  No wonder as he lived there for 18 years before leaving, now some twenty years ago. 

First to the Sabyinyo volcano.  Many paths wind their way through the bamboo.  They are covered with the footprints of ungulates. ‘Buffalo’ said Raymond.  Judging by the amount of manure there must be a lot of them.

The edible mushrooms are ready for inspection.  Damascene does not know their names but firmly claims that he has eaten them – also species that mycologists don’t regard as being edible or very tasty, but probably he did not have the luxury to be that selective.








What is striking said Jerome are the mushrooms that grow on bamboos, but their diversity is not so great; maximum a few Marasmius and Collybia species.  In the straw layer of fallen bamboo leaves almost no mushrooms are to be found there.

Although the diversity of species is not so great, the bamboos have a few special surprises for us.  First surprise: two species of honey fungus.  The true honey fungus (Armillaria mellea) which we know as a pathogen in European forests and its relation Armillaria heimii.


Collybia aurea
Still more surprises were to come: in the bamboo forests of the Karisimbi volcano our mycologists find the golden collybia (Collybia aurea), a warm yellow-coloured species that is considered as a delicacy in the neighbouring country of Burundi.  A little further on we find Termitomyces robustus, the famous mushroom that lives in symbiosis with termites.  The discovery of this edible species in the bamboo forests comes as a big surprise.  It’s one for the list of exciting discoveries, we were told by a clearly satisfied mycologist



Tuesday 28 April 2015

Mycologists in the mist

Pleurotus cystidiosus
To see mushrooms you need to get up early in the morning.  Its 7 a.m.  Up we go direction Kiningi, the entrance to the Volcanoes National Park.  We have an appointment with Raymond, a ranger who will accompany us during our mycological expedition on the slopes of the volcanoes.  We are not alone – a visitors centre with a fleet of 4x4s and tourists all hoping to catch a glimpse of the mountain gorillas. 

Between the Lobelias

Our first stop: the Bisoke or Vasuki volcano (3711 m) shrouded in the mist.  A few soldiers were waiting for us.  To protect us during our ascent from the buffaloes they told us.
Our journey was painfully slow between the giant Lobelias because the underground was wet and spongy.  

The magnificent trees are completely covered with mosses and lichens… including a nice variety of edible mushrooms.  We also found a species of oyster mushroom that we had not so far found on our mission Pleurotus cystidiosus; certainly one to try at Kigali farms.

Raymond was very interested in our mission and in wild mushrooms.  He knew some of the species which are edible or not, but regretted that he knew no more than that.  No problem said Jérôme, because we will make an illustrated guide of edible mushrooms.  This will also be useful for people such as the rangers of the park.


The harvest is good.  In the basket of the mycologist are 20 plus different species including a light green coloured Agaricus mushroom.  Maybe a new species for science but still be confirmed on our return says our mycologist with hope but also with caution.

Camp Dian Fossey

After a few hours plodding through the mud we reach the remains of the camp Dian Fossey who was murdered here in 1985.  Here rests ‘Nyiranachabelli’ the ‘lonely spirit of the wood’ it reads on a memorial gravestone, next to the remains of ‘Digit’ one of her favourite gorillas.

The day went well and it was time to start our descent to the base camp where our scientists had some evening work to do.  On our way back we see a fresh footprint of a gorilla … one can dream…if only we could see one.

Sunday 26 April 2015

To the Volcanoes!


After visiting Bweyeye and Gishwali our third trip, the Volcanoes National Park.  A national Park along the northern border of Rwanda with the Democratic Republic of Congo and Uganda which was set up in 1925 (originally Albert park: named after the Belgian King) to protect the last of the mountain gorillas.
 Karisimbi (4507 m)

Since it was set up, the boundaries of this park have been continuously eroded by agriculture, cattle farmers but also on more than one occasion since 1979 by the government as a solution to the rapidly increasing population but also to allow for the cultivation of pyrethrum (a bio insecticide).  In 1960 it was 34.000 hectares, but now only 16.000 ha remain which now come under Rwandan State protection.

The park is known for its five extinct volcanoes that belong to the Virunga chain of volcanoes (a total of 8 African volcanoes);

For the record
  • The Karisimbi (4507 m)
  • The Bisoke (3711 m) 
  • The Sabyinyo (3634 m)
  • The Gahingha (3474 m)
  • The Muhabura (4127 m)


From the foot of volcanoes to the summit four distinct vegetation zones can be differentiated.

  1. The bamboo forest zone (Yushania alpina) between 2500 and 3200 m above sea level.
  2. The Hagenia – Hypericum zone (3100-3500 m)
  3. The Senecio- Lobelia zone (3500-4200 m) – this is also the tree limit zone
  4. The alpine zone where only a few grass species, mosses and lichens can be found.


The next few days we stayed in the slopes of the volcanoes looking for edible mushrooms from the rainforest.  Looking out for the local gorillas!

Saturday 25 April 2015

Local mycology in Gishwati



After a long journey we arrived in Nyundo, almost one and a half hour bumpy roads until we finally arrive in the forest of Gushwati.  During the journey you cannot miss the tea plantations, the pride of the region and if you believe the bill boards, the best tea in whole Rwanda!  

This morning we meet up with Charles and Martin, two Rwandans.  They are our guides for the next 2 days.  They know the Gishwali forest like the back of their hands, as Charles told us, who lived there until 1985 with her family under a green leaf roof.  Since then she has been forced to live in villages because their native home areas in the forest have been out of bounds because they have been given a sacred status.  

We were curious.  Does the knowledge of mushrooms make a part of their collective cultural memories? Do they still eat these mushrooms? Do they have their own local names for these mushrooms?  It’s up to us to see what we can find out during our mission.


Into the forest via one of the small paths, we are greeted with the ear bashing chorus of crickets and a concert of some exotic birds and in the distance the chatter of chimps.  After a few minutes into the forest we make our first find – two species of Judas ears (Auricularia cornea and A. delicata )

Auricularia cornea
According to our guides they are named ‘ Ikinyagutwi’ in Kinyarwanda. Both of them know these species very well, but, surprisingly they don’t eat them so much which is striking since they are considered elsewhere in Africa as a delicacy.  We quickly make some photos, take some samples and continue on our way.

Using machetes our guides cut a path through the thick undergrowth.  A little further on another find the mushroom Agaricus cf. bingensis.  In Uganda this relative of the 'champignon de Paris' is highly appreciated as a culinary delicacy.  Not here though.  Local tradition has it that you will become deaf if you eat it!

The local name leaves no doubt over.  ‘Ikizibamatwi’ means ‘mouth’ on the ear!
Continuing on our journey we find around 20 other edible species that are collectively known as ‘Ubuzuruzuru’.  Possibly a generic name that indicates that they are edible.
Amongst our finds an oyster mushroom (Pleurotus djamor), an interesting find with potential for Kigali farms.

Although Charles and Martin have a considerable knowledge of native forest mushrooms, the same cannot be said for the younger generations who have never know life in the forest.
Unless we do something now, all of this local knowledge will be lost forever.  It’s for this reason that local mycology is such an important part of our mission here in Rwanda and it underlines the commitment of the Botanic Garden, Meise to contribute, in a positive way, to keep this knowledge from being lost forever. If we succeed, the future prospects for the local people will be improved giving them a sustainable means to improve their nutrition and their lives.  The potential for eco-tourism should also not be underestimated as an additional source of income.

Thursday 23 April 2015

The Gishwati Forest

Gishwati Forest
Gudula, a young Rwandan scientist joins the team.  She will replace Assoumpta for the rest of our stay.  After the Amanita saga of Bweyeye, it’s the second stage of our mission:  the study of mushrooms in the Gishwati Forest, of course, the edible species.  

What remains of the forest is only a small garden in comparison to the original surface area of 28.000 hectares in 1970.  Today, the forest is little more than 1.500 hectares of its former glory… an enormous loss.  The causes are well known, agriculture, free grazing of cattle, and… relocation of refugees after the genocide and the introduction of non-native tree species.  Also, well know are the very negative consequences of such a huge loss of virgin montane forest – soil erosion, reduction in soil fertility and perhaps most significantly the loss of biodiversity.  We will never know what species have been lost since they never been studied by scientists. The few remaining chimpanzees and golden apes must feel very crowded in such a small living space.

Nevertheless, there is still hope!  All may not be lost forever for these isolated animals.  There are current plans to create a ‘green corridor’ to connect this isolated forest with the Nyungwe National Park.  If these plans ever become reality, this would offer a real lifeline for the continued existence of the Gishwati Forest and its endangered inhabitants.

In the next days the team will have to work in an already difficult context. Will they find here a diversity of mushrooms? And will it be as rich as in the mountains of Nyungwe and on the Volcanoes?

Only the future will tell…


Sunset on Lake Kivu


Tuesday 21 April 2015

On route to Gishwati




A transition day...

We travel by jeep to the Gishwati Forest Reserve, which rises majestically above the Lake Kivu, half a kilometer from the town of Gisenyi.  In this virgin montane forest of 1500 hectares we make our way through difficult terrain looking for edible mushrooms that could be grown by our project partner Kigali farms.  

On route our 4X4 broke down.  But then, lifting our spirits, Jérôme cried out ‘Look Termitomyces robustus!'


On the side of the road a young girl was selling two nice bunches of this mushroom (see photo).  This edible mushroom, known through the whole of central Africa, is not included in our mission.  This is because there is virtually no chance that it could ever be commercially grown.  Why not?  

The reason is simple.  Termitomyces robustus grow in termite hills forming a symbiosis (partnership) with termites.  This partnership is a win-win situation.  By farming the mycelium (or fungal hyphae) inside the termite hill, the ants are helped to breakdown woody material by enzymes secreted by the fungus.  This means that if we want to cultivate this particular mushroom we will have to find a way of ‘domesticating’ the termites because they are essential for the development of the fungal mycelium.  This is perhaps future music but, for the meantime, we are a long way from achieving this goal.  It is for this reason that our mission is primarily focused on the search for edible saprotrophic mushrooms (like Pleurotes) that are far easier to cultivate commercially.  More on this later…

Friday 17 April 2015

Back to Bweyeye

This morning we repeated our trip of yesterday to Bweyeye.  Two hours of bone shaking driving by jeep in the forest of Nyungwe, until we meet up again with our local guide of yesterday evening, Damascene.  We had given him the task to find for us a nice group of Amanites, so that we could carry out a toxicological investigation.  Considering that we already found 4 examples yesterday we were confident that our search of today would be successful.

Damascene returned to the village after his fungal foray with a trog full of mushrooms.  A quick look by one of our mycologists (mushroom expert) confirmed that they were indeed Amanitas and enough of them for us to investigate.

A satisfied Jérôme decided to continue the search for other species by criss-crossing the hills surrounding the village.  Whether the Amanitas are edible or not we leave in the capable hands of Assoumpta who will carry out a survey amongst the local population.  He is essential to analyse the various claims made.

A young girl, Louise says she will let us see how she prepares these mushrooms.  We return to the village followed by a group of laughing children who quickly take their place in the limited space in her home.

Louise confirms our first witness: the cuticle of the mushroom cap is removed.  Once all of the fibres of the mushroom cap have been peeled, the flesh of the cap is cut into pieces and mixed with other vegetables (tomato, onion…) and cooked in water.
Assoumpta asked Louise why she peeled the mushroom.  Was it to avoid the poisonous chemicals that could be present in the cuticle or for another reason.  Louise answered that it was because the cuticle was sticky and not because it was poisonous.  Other witnesses were not in agreement with Louise and confirmed that it was poisonous, a family of 4 were presumably killed after the consumption of unpeeled mushrooms.  But there is no evidence that mushrooms were the cause of these fatalities.  The only thing for sure is that they remove the cuticle before cooking the mushrooms.

At this stage there are three possible hypotheses about non-toxic Amanitas which belong to the group of ‘phalloides/marmorata’ which are known or suspected of containing fatal fungal toxins.
  • Theory 1 – the local inhabitants are immune to the fungal toxins and their bodies can break down the toxins of the mushrooms
  • Theory 2 The fungal toxins are only present in the cuticle and the rest of the mushroom is edible
  • Theory 3 – all parts of the Amanite are edible



The mystery of the Amanita is still not solved, but witnesses have already given an indication to the possible answer to our investigation.  The Amanitas are separted into 2 parts: those which are completely intact with a complete cap, the other a cap without cuticle.  There will be two samples cut, separately dried for a toxicological analysis which is planned for our return.  The Amanitas of Bweyeye has not yet given up all of its secrets... to be continued !







Thursday 16 April 2015

The mystery of the Amanite of Bweyeye!

Cercopithecus l'hoesti
Today, we have a meeting in Bweyeye, a village on the edge of the Nyungwe National Park (970 km²), located in the south west of Rwanda on the border with Burundi, not far from the Lake Kivu.  The mountain forest of Nyungwe is perhaps one of the best preserved in Central Africa.  It has an exceptionally rich fauna and flora: around 13 species of primates, 32 species of amphibians and 38 species of reptiles and not to forget a thousand species of plants.  The mushroom flora is not so well known, just like in the surrounding countries and regions.  This is the motivation for our scientific mission which has as a goal of gaining a better knowledge of edible mushrooms.


Rendez-vous in Batwa-land

After two hours hiking through a maze of paths in the montane forest of Nyungwe, we arrive in Bweyeye.  Here the local population are hunter- gatherers.  Jean Marie, a forest ranger we met from an earlier scientific mission in October of last year, is waiting for us.
The first exploratory mission in October already indicated the presence of strange Amanites – from the group ‘phalloides’ or ‘marmorata’.  This remains to be confirmed but it has almost certainly been introduced with Eucalyptus trees originating from Australia.  This is not so exceptional.  Many mushrooms have been spread across the entire world in the way.  The hyphae or mycelium of the fungus were hidden on the roots of the Eucalyptus or Fir trees.  But most surprising is that this Amanite is considered as edible by the local population.  To science species of this group are unanimously considered as deadly poisonous !

Is the Amanite edible?

Jean-Marie found a local inhabitant that would act as a guide in our search for the Amanite.  The aim: to collect large quantities of Amanite to carry out genetic and toxicological analyses.  In this way we can determine the species and check the presence or not of deadly poisons.  In the meantime, Assoumpta, remained in the village to find out how local people prepared these mushrooms for eating.

Without some effort you will not reach your goal said Jérôme Degreef, the scientific coordinator of the expedition.  Ahead of us a steep climb to the top of the hill where these famous mushrooms can be found.  We are looking for mushrooms with a volva, which is a sort of sack that remains on the base of the stem (stipe), a characteristic morphological feature of Amanites.  After three hours of trekking we find some perfect specimens of other mushrooms – but no Amanites.  They nevertheless grow in large numbers in October.  A look of despair could be seen on Jérôme’s face until finally our local guide leads us to a plot of ground where he believes we could find what we were looking for.  ‘Yes that’s it’ shouted Jérôme, and as quick as lightning the mushrooms were photographed, listed and harvested for our collection.


After some ten minutes we find another three examples in various stages of growth (see photo).  
Sadly, the amount of material is far too limited to be able to carry out a toxicological investigation.  
We take the opportunity to ask our guide if he eats these mushrooms and how he prepares them.  His answer is yes, but he removes the cuticle of the cap before cooking them.  This confirms what Assoumpta was told by the village women.


Tomorrow we will return to continue our harvest but to be sure of success we ask our guide and the local village children to help.  What will the result be? That will see on Friday.





On route to Nyungwe

On route to Nyungwe and Bweyeye

During the first day in Kigali, we finalised our preparations for the mission.  It took no less than 12 hours hard driving on bad roads (made even worse by the rainy season) – with the inevitable unexpected surprises en-route- before we reached south-west Rwanda.


Our first trip was to the buffer zone in the  Nyungwe National Park.  For the scientists Assoumpta and Jérôme: they were hoping to solve the mystery surrounding the Amanita of Bweyeye… this saga to be continued


Friday 10 April 2015

Mission Rwanda Fungi 2015

From 13 April until 4 May , the Botanic Garden Meise (Belgium) undertook a scientific expedition to Rwanda to make an inventory of edible mushrooms from the mountain forests and national parks: Volcanoes National Park, Gishwati Forest Reserve, Nyungwe Forest…During an exploratory expedition in November 2014, a whole range of mushrooms were identified on-site.

Within the group of edible mushrooms are a number of species that have a definite economic value for local communities.  These species will be collected and then propagated by Kigali Farms, a for enterprise with a social purpose. This unique project combines fundamental and applied research.  It also seeks to have immediate but sustainable development by generating income for local communities and commercial concerns.

For the mission, the project will be communicated from this blog.  We will publish as many reports as possible about the various phases of the project.  This media will also serve as a diary of the mission.