LUBILOSA
Encyclopedia
LUBILOSA was the name of a research
Research
Research can be defined as the scientific search for knowledge, or as any systematic investigation, to establish novel facts, solve new or existing problems, prove new ideas, or develop new theories, usually using a scientific method...

 programme
Program management
Program management or programme management is the process of managing several related projects, often with the intention of improving an organization's performance...

 that aimed at developing a biological
Biology
Biology is a natural science concerned with the study of life and living organisms, including their structure, function, growth, origin, evolution, distribution, and taxonomy. Biology is a vast subject containing many subdivisions, topics, and disciplines...

 alternative to the chemical control
Insecticide
An insecticide is a pesticide used against insects. They include ovicides and larvicides used against the eggs and larvae of insects respectively. Insecticides are used in agriculture, medicine, industry and the household. The use of insecticides is believed to be one of the major factors behind...

 of locust
Locust
Locusts are the swarming phase of short-horned grasshoppers of the family Acrididae. These are species that can breed rapidly under suitable conditions and subsequently become gregarious and migratory...

s. This name is an acronym of the French
French language
French is a Romance language spoken as a first language in France, the Romandy region in Switzerland, Wallonia and Brussels in Belgium, Monaco, the regions of Quebec and Acadia in Canada, and by various communities elsewhere. Second-language speakers of French are distributed throughout many parts...

 title of the programme: Lutte Biologique contre les Locustes et les Sauteriaux (biological control
Biological pest control
Biological control of pests in agriculture is a method of controlling pests that relies on predation, parasitism, herbivory, or other natural mechanisms...

 of locust
Locust
Locusts are the swarming phase of short-horned grasshoppers of the family Acrididae. These are species that can breed rapidly under suitable conditions and subsequently become gregarious and migratory...

s and grasshopper
Grasshopper
The grasshopper is an insect of the suborder Caelifera in the order Orthoptera. To distinguish it from bush crickets or katydids, it is sometimes referred to as the short-horned grasshopper...

s). During its 13 year life (November 1989 to December 2002), the programme identified an isolate
Isolate
Isolate may refer to:* Genetic isolate, a population of organisms that has little genetic mixing with other organisms of the same species* Isolate , 2007* Isolate , 1992...

 of Metarhizium
Metarhizium
Metarhizium is a genus of entomopathogenic fungi in the Clavicipitaceae family. With the advent of genetic profiling, it has now become possible to place these fungi in proper taxa. Most turn out to be the asexual forms of fungi in the phylum Ascomycota.- Species :Nine distinct species have now...

fungus
Fungus
A fungus is a member of a large group of eukaryotic organisms that includes microorganisms such as yeasts and molds , as well as the more familiar mushrooms. These organisms are classified as a kingdom, Fungi, which is separate from plants, animals, and bacteria...

 virulent
Virulence
Virulence is by MeSH definition the degree of pathogenicity within a group or species of parasites as indicated by case fatality rates and/or the ability of the organism to invade the tissues of the host. The pathogenicity of an organism - its ability to cause disease - is determined by its...

 to locusts and went through all the necessary steps to develop the commercial biopesticide product 'Green Muscle' based on its spore
Spore
In biology, a spore is a reproductive structure that is adapted for dispersal and surviving for extended periods of time in unfavorable conditions. Spores form part of the life cycles of many bacteria, plants, algae, fungi and some protozoa. According to scientist Dr...

s.

Collaborators

The programme was conceived by Chris Prior and David Greathead of the International Institute of Biological Control (IIBC), a former research institute of CAB International
CAB International
CAB International is a not-for-profit inter-governmental organisation based in the United Kingdom....

 that was based at Silwood Park
Silwood Park
Silwood Park is the rural campus of Imperial College London, England. It is situated near the village of Sunninghill, near Ascot in Berkshire. Since 1986, there have been major developments on the site with four new college buildings...

 in the UK
United Kingdom
The United Kingdom of Great Britain and Northern IrelandIn the United Kingdom and Dependencies, other languages have been officially recognised as legitimate autochthonous languages under the European Charter for Regional or Minority Languages...

 at that time. IIBC requested and obtained the collaboration of the former Biological Control Centre for Africa
Africa
Africa is the world's second largest and second most populous continent, after Asia. At about 30.2 million km² including adjacent islands, it covers 6% of the Earth's total surface area and 20.4% of the total land area...

 in Cotonou
Cotonou
-Demographics:*1979: 320,348 *1992: 536,827 *2002: 665,100 *2005: 690,584 The main languages spoken in Cotonou include the Fon language, Aja language, Yoruba language and French.-Transport:...

, Benin
Benin
Benin , officially the Republic of Benin, is a country in West Africa. It borders Togo to the west, Nigeria to the east and Burkina Faso and Niger to the north. Its small southern coastline on the Bight of Benin is where a majority of the population is located...

, that was owned and managed by the International Institute of Tropical Agriculture
International Institute of Tropical Agriculture
International Institute of Tropical Agriculture is one of the world's leading research partners in finding solutions to hunger, malnutrition, and poverty. IITA's award-winning research-for-development addresses the development needs of tropical countries...

 (IITA). When IIBC asked for financial support from the Dutch
Netherlands
The Netherlands is a constituent country of the Kingdom of the Netherlands, located mainly in North-West Europe and with several islands in the Caribbean. Mainland Netherlands borders the North Sea to the north and west, Belgium to the south, and Germany to the east, and shares maritime borders...

 Directoraat Generaal voor Internationale Samenwerking (DGIS: Directorate General of International Cooperation), DGIS contributed by paying for a Dutch locust expert based at the Département de Formation en Protection des Végétaux (DFPV: Department of Crop Protection Training) of the AGRHYMET Regional Centre (ARC) in Niamey
Niamey
-Population:While Niamey's population has grown steadily since independence, the droughts of the early 1970s and 1980s, along with the economic crisis of the early 1980s, have propelled an exodus of rural inhabitants to Niger's largest city...

, Niger
Niger
Niger , officially named the Republic of Niger, is a landlocked country in Western Africa, named after the Niger River. It borders Nigeria and Benin to the south, Burkina Faso and Mali to the west, Algeria and Libya to the north and Chad to the east...

, which is part of the Comité permanent Inter-états de Lutte contre la Sécheresse dans le Sahel
Sahel
The Sahel is the ecoclimatic and biogeographic zone of transition between the Sahara desert in the North and the Sudanian Savannas in the south.It stretches across the North African continent between the Atlantic Ocean and the Red Sea....

 (CILSS: Permanent Interstate Committee for Drought Control in the Sahel). Since DFPV was given its own budget within LUBILOSA, it became a full partner within the programme. For a number of years, LUBILOSA collaborated with GTZ's locust programme. Permanent collaboration was established with the crop protection
Crop protection
Crop protection is the branch of horticulture concerned with protecting crops from pests, weeds, disease and theft.It encompasses:* Pesticide-based approaches such as herbicides, insecticides and fungicides...

 agencies of Niger
Niger
Niger , officially named the Republic of Niger, is a landlocked country in Western Africa, named after the Niger River. It borders Nigeria and Benin to the south, Burkina Faso and Mali to the west, Algeria and Libya to the north and Chad to the east...

, Benin
Benin
Benin , officially the Republic of Benin, is a country in West Africa. It borders Togo to the west, Nigeria to the east and Burkina Faso and Niger to the north. Its small southern coastline on the Bight of Benin is where a majority of the population is located...

, Burkina Faso
Burkina Faso
Burkina Faso – also known by its short-form name Burkina – is a landlocked country in west Africa. It is surrounded by six countries: Mali to the north, Niger to the east, Benin to the southeast, Togo and Ghana to the south, and Côte d'Ivoire to the southwest.Its size is with an estimated...

, Chad
Chad
Chad , officially known as the Republic of Chad, is a landlocked country in Central Africa. It is bordered by Libya to the north, Sudan to the east, the Central African Republic to the south, Cameroon and Nigeria to the southwest, and Niger to the west...

, Mali
Mali
Mali , officially the Republic of Mali , is a landlocked country in Western Africa. Mali borders Algeria on the north, Niger on the east, Burkina Faso and the Côte d'Ivoire on the south, Guinea on the south-west, and Senegal and Mauritania on the west. Its size is just over 1,240,000 km² with...

, Senegal
Senegal
Senegal , officially the Republic of Senegal , is a country in western Africa. It owes its name to the Sénégal River that borders it to the east and north...

 and The Gambia
The Gambia
The Republic of The Gambia, commonly referred to as The Gambia, or Gambia , is a country in West Africa. Gambia is the smallest country on mainland Africa, surrounded by Senegal except for a short coastline on the Atlantic Ocean in the west....

.

Donors

The LUBILOSA programme was supported financially by the following donors:
  • CIDA: Canadian International Development Agency
    Canadian International Development Agency
    The Canadian International Development Agency was formed in 1968 by the Canadian government. CIDA administers foreign aid programs in developing countries, and operates in partnership with other Canadian organizations in the public and private sectors as well as other international organizations...

  • DFID: Department for International Development
    Department for International Development
    The Department For International Development is a United Kingdom government department with a Cabinet Minister in charge. It was separated from the Foreign and Commonwealth Office in 1997. The goal of the department is "to promote sustainable development and eliminate world poverty". The current...

    , UK
  • DGIS: Dutch Directorate General of International Cooperation
  • SDC: Swiss Agency for Development and Cooperation
    Swiss Agency for Development and Cooperation
    The Swiss Agency for Development and Cooperation is an office-level agency in the federal administration of Switzerland, and a part of the Federal Department of Foreign Affairs...

  • USAID: United States Agency for International Development
    United States Agency for International Development
    The United States Agency for International Development is the United States federal government agency primarily responsible for administering civilian foreign aid. President John F. Kennedy created USAID in 1961 by executive order to implement development assistance programs in the areas...



Over the life of the programme, these donors contributed UK£10,200,000 (approximately US$17,000,000 at the time).

Four phases

The LUBILOSA programme has been funded in phases of three or four years each:
  • First phase 1990–1992
  • Second phase 1993–1995
  • Third phase 1996–1998
  • Fourth phase 1999–2002

First phase

The project was started late in 1989, with Chris Prior and David Greathead, obtaining funding and forming a team to develop a biological means of controlling locusts and grasshoppers. While examining the various options for biological control, it soon became apparent that oil formulations
Pesticide formulation
The biological activity of a pesticide, be it chemical or biological in nature, is determined by its active ingredient . Pesticide products very rarely consist of pure technical material. The AI is usually formulated with other materials and this is the product as sold, but it may be further...

 of the spores of certain fungi belonging to the form phylum
Phylum
In biology, a phylum The term was coined by Georges Cuvier from Greek φῦλον phylon, "race, stock," related to φυλή phyle, "tribe, clan." is a taxonomic rank below kingdom and above class. "Phylum" is equivalent to the botanical term division....

 Deuteromycota (Anamorphic fungi) offered the most promising option. Such fungi will grow on artificial substrates and so can be mass produced relatively quickly in large quantities. Their spores are lipophilic
Lipophilic
Lipophilicity, , refers to the ability of a chemical compound to dissolve in fats, oils, lipids, and non-polar solvents such as hexane or toluene. These non-polar solvents are themselves lipophilic — the axiom that like dissolves like generally holds true...

 and therefore suspend much more readily in oils than in water. As most locust control is carried out using oil formulations at ultra-low volume
Ultra-low volume
The term Ultra-Low Volume is used in the context of pesticide application.Ultra-low volume application of pesticides has been defined as spraying at a of les than 5 L/ha for field crops or les than 50 L/ha for tree/bush crops...

 (ULV) rates of application, developing an oil formulation of fungal spores enabled operators to use the same equipment for applying
Pesticide application
Pesticide application refers to the practical way in which pesticides, are delivered to their biological targets...

 a chemical pesticide and maximise efficacy.

Usually, fungi are most active under moist conditions and in early attempts to use them as control agents, they were therefore applied in water formulations. However, the results in practice were often disappointing and the approach was largely abandoned during most of the 20th century. When interest in microbial control was revived, water formulations of fungi started to be used successfully in greenhouse
Greenhouse
A greenhouse is a building in which plants are grown. These structures range in size from small sheds to very large buildings...

s, but the results in open fields continued to disappoint. In the 1980s, Prior had previously discovered that some anamorphic fungi are more effective when applied in oil.

During the first phase of LUBILOSA, the technical feasibility of using such oil formulations against locusts was demonstrated in the laboratory
Laboratory
A laboratory is a facility that provides controlled conditions in which scientific research, experiments, and measurement may be performed. The title of laboratory is also used for certain other facilities where the processes or equipment used are similar to those in scientific laboratories...

, then field cage and "arena" trials. An extensive survey was launched in West Africa
West Africa
West Africa or Western Africa is the westernmost region of the African continent. Geopolitically, the UN definition of Western Africa includes the following 16 countries and an area of approximately 5 million square km:-Flags of West Africa:...

 and the Arabian Peninsula
Arabian Peninsula
The Arabian Peninsula is a land mass situated north-east of Africa. Also known as Arabia or the Arabian subcontinent, it is the world's largest peninsula and covers 3,237,500 km2...

 to look for isolates
Primary isolate
Primary isolate is a pure microbial or viral sample that has been obtained from an infected individual, rather than grown in a laboratory. In chemistry and bacteriology, the verb isolate means to obtain a pure chemical, bacteriological or viral sample...

 virulent to locusts and grasshoppers, because few such isolates were available in public collections. The survey made use of a network of collaborators and came up with some 180 isolates, many of which belonged to Metarhizium acridum
Metarhizium acridum
Metarhizium acridum is the new name given to a group of fungal isolates that are known to be virulent and specific to Acrididea. Previously, this species has had variety status in Metarhizium anisopliae and before that, reference was made to M. flavoviride.- Biology :M...

, some to other Metarhizium species and to Beauveria bassiana
Beauveria bassiana
Beauveria bassiana is a fungus that grows naturally in soils throughout the world and acts as a parasite on various arthropod species, causing white muscardine disease; it thus belongs to the entomopathogenic fungi. It is being used as a biological insecticide to control a number of pests such as...

with a few records of Syngliocladium acridiorum (syn. Sorosporella). Laboratory screening of these isolates showed that M. acridum was easily the most virulent species under warm conditions (30°C), and confirmed the selection of isolate IMI 330189 for further development. Subsequent trials in cages and arenas confirmed that oil formulations were infective even under very dry conditions. IMI 330189 and similar isolates were assigned to Metarhizium flavoviride
Metarhizium flavoviride
Metarhizium flavoviride is the name given to a group of fungal isolates that are known to be virulent against Hemiptera and some Coleoptera...

in early papers, then Metarhizium anisopliae
Metarhizium anisopliae
Metarhizium anisopliae, formerly known as Entomophthora anisopliae , is a fungus that grows naturally in soils throughout the world and causes disease in various insects by acting as a parasite. Ilya I. Mechnikov named it after the insect species it was originally isolated from: the beetle...

var. acridum, but are now described as Metarhizium acridum
Metarhizium acridum
Metarhizium acridum is the new name given to a group of fungal isolates that are known to be virulent and specific to Acrididea. Previously, this species has had variety status in Metarhizium anisopliae and before that, reference was made to M. flavoviride.- Biology :M...

(Driver & Milner) J.F. Bisch., Rehner & Humber.

Second phase

During phase 2, the oil formulation was tested in the field and shown to be effective. Field trials proved to be difficult to carry out on such highly mobile insects as grasshoppers and locusts. With locusts, fixed plots, unless several square kilometres in size, cannot be used, so it is necessary to follow hopper bands. Although there were many locust infestations during this phase, of both desert locust
Desert locust
Plagues of the desert locust have threatened agricultural production in Africa, the Middle East, and Asia for centuries. The livelihood of at least one-tenth of the world’s human population can be affected by this voracious insect...

s (Schistocerca gregaria) and brown locust
Brown locust
The brown locust, Locustana pardalina, is a medium-sized small locust species found in Southern Africa and shows classic gregarious behaviour with phase polymorphism) on crowding.-Biology:...

s (Locustana pardalina), the chemical treatment teams were also very active and little headway was therefore made in developing the techniques necessary to measure the effect of Metarhizium on unrestrained locusts. However, it was possible to scale up treatments against the variegated grasshopper (Zonocerus variegatus) in the forest zone and against Sahelian grasshoppers, especially the Senegalese grasshopper
Senegalese grasshopper
Oedaleus senegalensis is a medium-sized grasshopper species found in the Sahel region of Africa, the Canary Islands, Cape Verde Islands and West Asia. Although not called a locust in English, this species shows gregarious behaviour and some morphological change on crowding...

 (Oedaleus senegalensis).

Contacts were made with several internationally renowned biopesticide
Biopesticide
Biopesticides include "naturally occurring substances that control pests , microorganisms that control pests , and pesticidal substances produced by plants containing added genetic material or PIPs."Biopesticides are biochemical pesticides that are naturally occurring substances that control pests...

 producers to ascertain whether they would be able and willing to reliably meet LUBILOSA's spore requirements during phase 3, but none were able to make that commitment. Accordingly, part of the IITA station in Cotonou was converted into a spore production unit and the human and technical capacity enhanced to meet the expected spore requirements. The facility proved an excellent research unit and made it possible to refine realistic technical specifications and to test production, contamination control, spore separation, drying and packaging.

Field trials

The LUBILOSA team continued to look for opportunities to test Metarhizium against locusts and grasshoppers. Field trials were conducted against Senegalese grasshoppers, African rice grasshoppers (Hieroglyphus daganensis
Hieroglyphus daganensis
The African rice grasshopper, Hieroglyphus daganensis is a medium-sized grasshopper species found in the Sahel region. Although not called a locust in English, this species shows gregarious behaviour and some morphological change on crowding and may become a moderately important pest species for...

), variegated grasshoppers, Sahelian tree locusts (Anacridium melanorhodon), brown locusts and desert locusts. These trials demonstrated that Metarhizium at a dosage of 50 g/ha could reduce grasshopper population
Population
A population is all the organisms that both belong to the same group or species and live in the same geographical area. The area that is used to define a sexual population is such that inter-breeding is possible between any pair within the area and more probable than cross-breeding with individuals...

s and those of tree locusts by 80-90% within two to three weeks. Population level control was more difficult to achieve with other locusts, but a significant effect was demonstrated on hopper bands notwithstanding the great difficulties in tracking individual bands. Some trials against Senegalese grasshoppers included the chemical insecticide
Insecticide
An insecticide is a pesticide used against insects. They include ovicides and larvicides used against the eggs and larvae of insects respectively. Insecticides are used in agriculture, medicine, industry and the household. The use of insecticides is believed to be one of the major factors behind...

 fenitrothion
Fenitrothion
Fenitrothion is a phosphorothioate insecticide....

 for comparison. Because of its rather short persistence, this chemical turned out to be much less effective than commonly believed. It was highly effective in producing an immediate sharp reduction in population densities, but continued hatch
Hatch
Hatch may refer to:* Hatching, also called "cross-hatching", an artistic technique used to create tonal or shading effects using closely spaced parallel lines* Hatching, the emergence of a young animal from an egg...

ing and remigration into the treated plots caused these densities to start increasing again after less than a week, until they reached or exceeded the original levels after about two to three weeks. Metarhizium, on the other hand, reduced population levels at a slower rate, but maintained low levels for at least one and a half months. The half-life
Half-life
Half-life, abbreviated t½, is the period of time it takes for the amount of a substance undergoing decay to decrease by half. The name was originally used to describe a characteristic of unstable atoms , but it may apply to any quantity which follows a set-rate decay.The original term, dating to...

 of its spores on the vegetation was estimated at more than 7 days under rainy season conditions in the Sahel and, of course, some conidia must survive between seasons.

Grasshopper behaviour

In the course of these field trials, it became clear that locusts and grasshoppers are able to detect that they have become infected and alter their behaviour accordingly. In particular, they spend more time basking in the sun, even in the middle of the day. Uninfected grasshoppers also bask in the sun when their body temperature is below the preferred 38-40°C. However, detailed investigations showed that infected grasshoppers increase their body temperature to up to 4° higher. This effect has been dubbed “behavioural fever” and is thought to slow down the infection just like a fever in warm-blooded
Warm-blooded
The term warm-blooded is a colloquial term to describe animal species which have a relatively higher blood temperature, and maintain thermal homeostasis primarily through internal metabolic processes...

 animal
Animal
Animals are a major group of multicellular, eukaryotic organisms of the kingdom Animalia or Metazoa. Their body plan eventually becomes fixed as they develop, although some undergo a process of metamorphosis later on in their life. Most animals are motile, meaning they can move spontaneously and...

s does. This basking behaviour poses a potential problem for the fungus. The latter's preferred temperature is 28-30°C and it stops growing above 35°. Measurements of body temperatures in the field have shown that the basking behaviour reduces the time available for growth by many hours per day. Under certain conditions, this enables the insects to lay at least one egg
Egg (biology)
An egg is an organic vessel in which an embryo first begins to develop. In most birds, reptiles, insects, molluscs, fish, and monotremes, an egg is the zygote, resulting from fertilization of the ovum, which is expelled from the body and permitted to develop outside the body until the developing...

 pod before dying. Interestingly, healthy locusts and grasshoppers, especially females, take some time after fledging
Ecdysis
Ecdysis is the moulting of the cuticula in many invertebrates. This process of moulting is the defining feature of the clade Ecdysozoa, comprising the arthropods, nematodes, velvet worms, horsehair worms, rotifers, tardigrades and Cephalorhyncha...

 to reach maturity and lay eggs, during which time they build up sufficient fat to produce up to three egg pods. However, infected ones usually dispense with this fattening period and go on to produce eggs immediately after fledging. This generally wears them out so much that they die after laying one pod. So even if Metarhizium does not kill before or shortly after fledging, it usually allows the formation of only a single egg pod and thereby reduces the fecundity of infected females.

(Eco)toxicology

During phase 3, the programme started investigating the environmental
Natural environment
The natural environment encompasses all living and non-living things occurring naturally on Earth or some region thereof. It is an environment that encompasses the interaction of all living species....

 impact of the chosen Metarhizium isolate (IMI 330189). Standard toxicological
Toxicology
Toxicology is a branch of biology, chemistry, and medicine concerned with the study of the adverse effects of chemicals on living organisms...

 tests carried out by certified laboratories showed that the strain had practically no effect on mammal
Mammal
Mammals are members of a class of air-breathing vertebrate animals characterised by the possession of endothermy, hair, three middle ear bones, and mammary glands functional in mothers with young...

s, bird
Bird
Birds are feathered, winged, bipedal, endothermic , egg-laying, vertebrate animals. Around 10,000 living species and 188 families makes them the most speciose class of tetrapod vertebrates. They inhabit ecosystems across the globe, from the Arctic to the Antarctic. Extant birds range in size from...

s and fish
Fish
Fish are a paraphyletic group of organisms that consist of all gill-bearing aquatic vertebrate animals that lack limbs with digits. Included in this definition are the living hagfish, lampreys, and cartilaginous and bony fish, as well as various extinct related groups...

 (e.g. for rat
Rat
Rats are various medium-sized, long-tailed rodents of the superfamily Muroidea. "True rats" are members of the genus Rattus, the most important of which to humans are the black rat, Rattus rattus, and the brown rat, Rattus norvegicus...

s >2000 mg active ingredient per kg of body weight). Other toxicological and ecotoxicological
Ecotoxicology
Ecotoxicology is the study of the effects of toxic chemicals on biological organisms, especially at the population, community, ecosystem level. Ecotoxicology is a multidisciplinary field, which integrates toxicology and ecology....

 tests were performed by the LUBILOSA team. An important goal was to show that its isolate only infected locusts and grasshoppers. Infection tests were therefore performed on a large number of invertebrate
Invertebrate
An invertebrate is an animal without a backbone. The group includes 97% of all animal species – all animals except those in the chordate subphylum Vertebrata .Invertebrates form a paraphyletic group...

s. The great majority could not be infected even by high doses. However, some species, like termite
Termite
Termites are a group of eusocial insects that, until recently, were classified at the taxonomic rank of order Isoptera , but are now accepted as the epifamily Termitoidae, of the cockroach order Blattodea...

s, honey bee
Honey bee
Honey bees are a subset of bees in the genus Apis, primarily distinguished by the production and storage of honey and the construction of perennial, colonial nests out of wax. Honey bees are the only extant members of the tribe Apini, all in the genus Apis...

s, certain parasitoid wasps and silk worms, did get infected under laboratory conditions. Further experiments were carried out under semi-field and full field conditions, which showed that only silk worms suffered significant infection under those conditions. The conclusion from all these tests was that the LUBILOSA strain of Metarhizium was incapable of infecting vertebrates and infected only a small number of insect species under artificial conditions. The available studies indicate that under natural conditions, the isolate only infects species belonging to the Acrididea and the domesticated silk worm Bombyx mori.

Mass production

The spore production unit in Cotonou made it possible to optimise the mass production of spores of the LUBILOSA strain at an appropriate level of technology. The production process chosen was one with a liquid phase and a solid phase. During the liquid phase, the fungus biomass
Biomass
Biomass, as a renewable energy source, is biological material from living, or recently living organisms. As an energy source, biomass can either be used directly, or converted into other energy products such as biofuel....

 is bulked up in a solution of sugar
Sugar
Sugar is a class of edible crystalline carbohydrates, mainly sucrose, lactose, and fructose, characterized by a sweet flavor.Sucrose in its refined form primarily comes from sugar cane and sugar beet...

 and yeast
Yeast
Yeasts are eukaryotic micro-organisms classified in the kingdom Fungi, with 1,500 species currently described estimated to be only 1% of all fungal species. Most reproduce asexually by mitosis, and many do so by an asymmetric division process called budding...

 and may produce submerged conidia . Subsequently, the resulting broth is used to inoculate partially boiled and sterilised rice
Rice
Rice is the seed of the monocot plants Oryza sativa or Oryza glaberrima . As a cereal grain, it is the most important staple food for a large part of the world's human population, especially in East Asia, Southeast Asia, South Asia, the Middle East, and the West Indies...

. The fungal mycelium
Mycelium
thumb|right|Fungal myceliaMycelium is the vegetative part of a fungus, consisting of a mass of branching, thread-like hyphae. The mass of hyphae is sometimes called shiro, especially within the fairy ring fungi. Fungal colonies composed of mycelia are found in soil and on or within many other...

 invades the rice and then starts to sporulate. At this point, rice and fungus are slowly dried before separating the spores from the rice. A novel machine was invented to achieve this in an efficient way, which was subsequently further developed into the 'MycoHarvester'. The study and optimisation of the mass production process led to the adoption of quality standards for the produced spores, which could be imposed on those who wanted to produce the spores commercially under licence. Various aspects of quality control
Quality control
Quality control, or QC for short, is a process by which entities review the quality of all factors involved in production. This approach places an emphasis on three aspects:...

 are important, including: levels of contamination (especially the absence of human pathogens), virulence to target pests, particle size spectrum and, not least, viable spore count. Extensive research was carried out on optimising storage of spores, which should be dry (<5% moisture content) and ideally be maintained under cool conditions .

Commercialisation

Two commercial companies agreed to look into the feasibility and economic viability of producing Metarhizium for the locust control market. These were Biological Control Products (BCP) near Durban
Durban
Durban is the largest city in the South African province of KwaZulu-Natal and the third largest city in South Africa. It forms part of the eThekwini metropolitan municipality. Durban is famous for being the busiest port in South Africa. It is also seen as one of the major centres of tourism...

, South Africa
South Africa
The Republic of South Africa is a country in southern Africa. Located at the southern tip of Africa, it is divided into nine provinces, with of coastline on the Atlantic and Indian oceans...

, and Natural Plant Protection (NPP) of Noguères
Noguères
Noguères is a commune in the Pyrénées-Atlantiques department in south-western France.-References:*...

, France
France
The French Republic , The French Republic , The French Republic , (commonly known as France , is a unitary semi-presidential republic in Western Europe with several overseas territories and islands located on other continents and in the Indian, Pacific, and Atlantic oceans. Metropolitan France...

 (part of Calliope S.A.). The former registered the product under the name Green Muscle (deposited by CAB International), first in South Africa and subsequently in other southern and eastern African countries. BCP is the leading producer of biopesticides in Africa. Its main market is South Africa but its products are increasingly being sold in other countries. The company continues its production and promotion of Green Muscle. NPP, however, only obtained a temporary sales permit for the CILSS countries in West Africa, but was unable to produce Green Muscle using its own production process.

Fourth phase

The last phase of LUBILOSA was characterised by promotion activities and stewardship of the commercial producer. Launching the product on the market and informing potential clients was not sufficient. The market for locust control products was (and still is) peculiar and difficult to penetrate. Most users are government or intergovernmental agencies involved in locust and/or grasshopper control. Especially in the beginning, it proved difficult to convince the professionals in these agencies of the need for an alternative product and to convince them that the product was as effective as those they had been using so far. Moreover, the large pesticide companies offered all kinds of incentives to those in charge of procurement in order that they buy their products, a practice that was difficult to follow for a small company like BCP. Until the end of the programme and even beyond, there was a constant need to do more field trials or demonstrations. Towards the end, the Locust Group of the Food and Agriculture Organization of the United Nations (FAO) finally became interested and involved in some of the trials: especially against desert locust
Desert locust
Plagues of the desert locust have threatened agricultural production in Africa, the Middle East, and Asia for centuries. The livelihood of at least one-tenth of the world’s human population can be affected by this voracious insect...

s.

Apart from the Sahelian zone, grasshoppers are generally not considered serious pests in Africa and are therefore rarely controlled with pesticides. For that reason, it was only in the Sahel that serious efforts were undertaken to promote Green Muscle for grasshopper control. In a number of countries, and especially Niger, cereal
Cereal
Cereals are grasses cultivated for the edible components of their grain , composed of the endosperm, germ, and bran...

 growers are organised in village brigades that are charged with controlling pests that surpass the capacity of individual growers. Green Muscle was given to some of these after training in its correct application. After the initial disappointment with its slow action, the villagers usually became satisfied with the final results. However, the price proved to be a serious obstacle. At that time, the cost of producing one kg of Green Muscle was about $200. At a dosage of 50 g/ha, that translated into a price of $10 per hectare
Hectare
The hectare is a metric unit of area defined as 10,000 square metres , and primarily used in the measurement of land. In 1795, when the metric system was introduced, the are was defined as being 100 square metres and the hectare was thus 100 ares or 1/100 km2...

. The price of conventional products was then approximately $5/ha. Though growers were prepared to pay a premium of up to $2 for the fact that Green Muscle was harmless to their health and that of their livestock
Livestock
Livestock refers to one or more domesticated animals raised in an agricultural setting to produce commodities such as food, fiber and labor. The term "livestock" as used in this article does not include poultry or farmed fish; however the inclusion of these, especially poultry, within the meaning...

, they were clearly not going to pay $10. Another problem was that no producer had yet been identified for the West African market and the availability of the product could therefore not be guaranteed.

At the end of the programme in December 2002, the LUBILOSA collaborators were faced with a situation where commercial uptake of Green Muscle was still non-existent, all quantities purchased so far exclusively for trial purposes. BCP was left alone to promote the product with little support from CABI and IITA, who had run out of funds for serious activities. Fortunately, the FAO had become interested, which made it possible to carry out a few more field trials.

Post-LUBILOSA

After the termination of the LUBILOSA programme, activities to promote the use of Green Muscle continued, including field trials against red locusts and desert locusts with support from the FAO. A new project in West Africa, PRéLISS (Regional Project for Integrated Grasshopper Control in the Sahel), aimed at developing an integrated pest management
Integrated Pest Management
Integrated pest management is an ecological approach to agricultural pest control that integrates pesticides/herbicides into a management system incorporating a range of practices for economic control of a pest...

 (IPM) strategy that included the use of Green Muscle. This project conducted field trials with reduced dosages of Green Muscle and proved that 25 g/ha was as effective as the registered dosage of 50 g/ha. It also tested mixtures with lambda-cyhalothrin (a pyrethroid
Pyrethroid
A pyrethroid is an organic compound similar to the natural pyrethrins produced by the flowers of pyrethrums . Pyrethroids now constitute a major commercial household insecticides...

 insecticide) to try to solve the problem of the slow action of Green Muscle. It was demonstrated that application of both products together, each at a quarter of its registered dose, caused mortality almost as quick as lambda-cyhalothrin alone and maintained low population levels for weeks, and all this at a much lower cost.

After it had become clear that NPP could not produce Green Muscle, the search for a second producer continued. A number of candidate producers were approached, but none was ready to take up the challenge. This situation only changed when desert locusts started invading West African countries again in late 2003. The president of Senegal, Abdoulaye Wade
Abdoulaye Wade
Abdoulaye Wade is the third and current President of Senegal, in office since 2000. He is also the Secretary-General of the Senegalese Democratic Party and has led the party since it was founded in 1974...

, and his wife Viviane
Viviane Wade
Viviane Wade is the current First Lady of Senegal. Born at Besançon, she was raised in France. In 1963 she married Abdoulaye Wade, a Senegalese man who later became active as an opposition leader in Senegal and was eventually elected as President of Senegal in 2000. They have two children, son...

 were very concerned about the prospect of huge quantities of chemical insecticides being sprayed to contain the upsurge. The first lady instructed her technical advisor, Sébastien Couasnet, to find out whether an alternative existed. Apparently, the information about Green Muscle had remained stuck at the professional level of the crop protection
Crop protection
Crop protection is the branch of horticulture concerned with protecting crops from pests, weeds, disease and theft.It encompasses:* Pesticide-based approaches such as herbicides, insecticides and fungicides...

 agency of Senegal (and most likely other countries as well), notwithstanding all the efforts of LUBILOSA and PRéLISS to make the existence of the new product widely known.

Fortunately, Couasnet stumbled upon the LUBILOSA website, which was still hosted by IITA and maintained by the then project leader of PRéLISS, Christiaan Kooyman. The two met in Cotonou and immediately set about to design a production plant for Green Muscle. Couasnet was able to convince the first lady to organise the financing and construction of the plant started in 2005. It took a while to start up the production and get rid of all sources of contamination, but this was eventually achieved with the backstopping of IITA, CAB International and Roy Bateman of IPARC, who installed the latest version of the MycoHarvester v.3. The production licence was granted in late 2007. For the time being, production and marketing are taken care of by the Fondation Agir pour l'Education et la Santé, of which Mme. Wade is president.

Programme staff

  • Yvonne Abraham (IIBC/CABI): 1990-1996, entomologist
  • Serge Attignon (BCCA/IITA): 1996–1998, socioeconomist
  • Roy Bateman (IIBC/CABI): 1989–2002, spray application
    Pesticide application
    Pesticide application refers to the practical way in which pesticides, are delivered to their biological targets...

     and formulation
    Pesticide formulation
    The biological activity of a pesticide, be it chemical or biological in nature, is determined by its active ingredient . Pesticide products very rarely consist of pure technical material. The AI is usually formulated with other materials and this is the product as sold, but it may be further...

     specialist
  • Dave Batt (IIBC/CABI): 1990-2000, laboratory technician
  • Michelle Carey (IIBC/CABI): 1990-1993, laboratory technician
  • Andy Cherry (BCCA/IITA and NRI): 1996–2002, entomopathologist
    Entomopathogenic fungus
    An entomopathogenic fungus is a fungus that can act as a parasite of insects and kills or seriously disables them.-Typical life cycle:These fungi usually attach to the external body surface of insects in the form of microscopic spores...

     and mass production coordinator
  • Hugo de Groote (BCCA/IITA): 1996–1998, socioeconomist
  • David Dent (IIBC/CABI): 1996–2000, programme manager
  • Orou-Kobi Douro-Kpindou (BCCA/IITA): 1990–2002, entomopathologist
    Entomopathogenic fungus
    An entomopathogenic fungus is a fungus that can act as a parasite of insects and kills or seriously disables them.-Typical life cycle:These fungi usually attach to the external body surface of insects in the form of microscopic spores...

  • Comlan Gbongboui (BCCA/IITA): 1993–2002, field technician
  • Ignace Godonou (BCCA/IITA): 1990–1998, entomopathologist
  • David Greathead (IIBC/CABI): 1989–1992 Director
  • Jane Gunn (IIBC/CABI): 1996–2000, laboratory technician
  • Nina Jenkins (IIBC/CABI): 1996–2002, microbiologist
    Microbiology
    Microbiology is the study of microorganisms, which are defined as any microscopic organism that comprises either a single cell , cell clusters or no cell at all . This includes eukaryotes, such as fungi and protists, and prokaryotes...

     and mass production specialist
  • Christiaan Kooyman (DFPV/AGRHYMET and IIBC/CABI): 1990–2002, acridologist
    Acrididae
    The Acrididae are the predominant family of grasshoppers, comprising some 10,000 of the 11,000 species of the entire suborder Caelifera. The Acrididae are best known because all locusts are of the Acrididae. The subfamily Oedipodinae is sometimes classified as a distinct family Oedipodidae in the...

     and entomopathologist, project coordinator West Africa (1990–1995) and East Africa (1996–2002)
  • Jürgen Langewald (BCCA/IITA): 1993–2002, entomologist
    Entomology
    Entomology is the scientific study of insects, a branch of arthropodology...

     and programme leader 1999–2002
  • Chris Lomer (IIBC/CABI and BCCA/IITA): 1990–1999, entomopathologist
    Entomopathogenic fungus
    An entomopathogenic fungus is a fungus that can act as a parasite of insects and kills or seriously disables them.-Typical life cycle:These fungi usually attach to the external body surface of insects in the form of microscopic spores...

     and programme leader 1993–1999
  • Belinda Luke (IIBC/CABI): 1996–2002, research assistant
  • Dave Moore (IIBC/CABI): 1990–2002, entomologist
  • Peter Neuenschwander: 1989–2002 Director, IITA Biological Control Centre for Africa
  • Zakaria Ouambama (DFPV/AGRHYMET): 1990–2002, laboratory assistant and field trial organiser
  • Armand Paraïso (BCCA/IITA): 1990–1995, NARS network coordinator
  • Ralf Peveling (University of Basel): 1993–1998, ecotoxicologist
    Ecotoxicology
    Ecotoxicology is the study of the effects of toxic chemicals on biological organisms, especially at the population, community, ecosystem level. Ecotoxicology is a multidisciplinary field, which integrates toxicology and ecology....

  • Chris Prior (IIBC/CABI): 1989–1995, phytopathologist and programme leader 1989–1993
  • Paresh Shah (IIBC/CABI and IITA): 1989–1993, grasshopper field studies 1989–1993
  • Bill Steele (IIBC/CABI): 1990–1996, programme manager
  • Matthew Thomas (IIBC/CABI and Imperial College): 1996–2002, ecologist
    Ecology
    Ecology is the scientific study of the relations that living organisms have with respect to each other and their natural environment. Variables of interest to ecologists include the composition, distribution, amount , number, and changing states of organisms within and among ecosystems...

  • Jeff Waage (IIBC/CABI): 1992–2000 Director
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