In a house that looks like any house in a residential area of the city of Piracicaba, State of São Paulo, about 100 miles from the capital, 60,000 larvae of the Diatraea saccharalis moth are put to death each day. One by one, they are exposed to their main natural enemy – the Cotesia flavipes wasp – by 12 employees of Bug Agentes Biológicos (Bug Biological Agents), a company specialized in producing insects to combat pests in cultures such as sugarcane, corn, tomato, etc. Known as sugarcane borer (broca-da-cana-de-açúcar in Portuguese), D. saccharalis larvae are the cause of red rot (in Portuguese podridão vermelha), a disease that affects sugarcane fields and causes great losses to sugarcane and alcohol producers. By exposing them to C. flavipes, Bug is able to get thousands of new wasps, which are then sold to sugarcane farmers.

In two other units – one in Piracicaba’s rural area, the other in the neighboring town of Saltinho, the company produces daily, through a similar method, 10 kilograms (22 lbs) of eggs of the Anagasta kuehniella clothes moth. Inside them are 360 million eggs of another wasp, Trichogramma spp, a natural enemy of pests that hit fields of various cultures, such as the sugarcane borer itself, the fall armyworm (Spodoptera frugiperda) (lagarta-do-cartucho, in Portuguese) and the corn earworm (Helicoverpa zea) (in Portuguese lagarta-da-espiga-do-milho). Thirty percent of that production is exported to seven European countries: Switzerland, Spain, France, Denmark, Britain, Italy and Belgium. With 68 employees, Bug’ sales in 2007 totaled some US$ 850,000, a figure that is expected to increase to about US$ 2.2 million in 2008.

The beginning

Bug was created in 1999 in Piracicaba, in the Escola Superior de Agricultura Luiz de Queiroz (Luiz de Queiroz College of Agriculture, Esalq), of the University of São Paulo (Universidade de São Paulo, USP). The initiative to create it came from agronomy engineers Danilo Scacalossi Pedrazzoli and Diogo Rodrigues Carvalho, at the time Master Degree’s candidates advised by professor José Roberto Postali Parra. The company was called CP2 (“C” from Carvalho, “P” from Pedrazzoli), which is Bug’s official name. “We chose the name Bug because of its meaning in English,” says Carvalho. “And also because it was a word known all over the world back then because of the infamous ‘millennium bug’, which was supposed to affect all the world’s computers. Another reason was to distinguish it from the ‘bio’ companies, such as Biocontrole, Biocontrol, Biocana, Bioagri, Bioagro, Bioresult, in Brazil, and Biocontrol, Biotop, Bio Bee abroad.”

The company was created in order to take advantage of a business opportunity generated by a joint research Esalq conducted in partnership with the Fundo de Defesa da Citricultura (Citriculture Defense Fund, Fundecitrus), an association of citrus planters and citrus processing industries, the Cooperativa dos Cafeicultores e Citricultores de São Paulo (Cooperative of Coffee and Citrus Planters of the State of São Paulo, Coopercitrus), and the University of California in Davis (UCDavis). One of the results was that the researchers were able to synthesize the pheromone – a chemical substance insects produce to communicate with each other or for females to attract males – of the citrus fruit borer (Ecdytolopha aurantiana) (bicho-furão-dos-citros in Portuguese), a plague whose worm attacks citrus fruits, such as oranges.

When a Japanese company industrialized the product as a pill the two agronomists saw the chance to make money and created Bug. The first objective of the company was to produce traps for the citrus fruit borer using the Japanese pills as bait. These traps are small boxes with open ends – like pipes, but triangular in shape – whose internal walls are smeared with glue, where the insects are trapped. Several of such traps are placed in the field, each with a pill inside. “But they’re not used for controlling the pest,” explains Carvalho. “Their function is simply to monitor the level of the citrus fruit borer’s infestation. If the average number of insects per trap reaches eight, the farmer knows it’s time to use insecticide. This method rationalizes the use of poison.”

PIPE

In 2001, the two partners decided to expand the company’s operations and requested a loan from the Programa Inovação Tecnológica em Pequenas Empresas (Technological Innovation in Small Businesses Program, PIPE) – the program of the State of São Paulo Research Foundation (Fundação de Amparo à Pesquisa do Estado de São Paulo, Fapesp) that provides support to innovating small companies. The project “Criação massal e comercialização de Trichogramma spp e Cotesia flavipes para o controle de pragas agrícolas” (mass breeding and commercialization of Trichogramma spp and Cotesia flavipes for agriculture pest control) was approved for the program’s Phase I and was carried out in the first semester of 2001. The approximately US$ 30,000 Fapesp granted were crucial for Bug. “With that money we bought permanent equipment, such as hot houses, climatized chambers, autoclaves, iceboxes, freezers and industrial blenders,” Carvalho tells. “In addition, we used it to buy products we’d need in the next six months, such as food for the worms and small flasks.”

Once the project was proven economically viable – the purpose of PIPE’s Phase I –, the company requested funding for Phase II. Again the request was approved. This time Bug received approximately US$ 100,000. This stage lasted two years, from the second semester of 2001 to the first semester of 2003, and the purpose was to launch the product in the market. “Actually we were already selling Cotesias and Trichogrammas, but wanted to expand the business,” explains Carvalho. Later the company got yet another grant from PIPE, this time for Phase III, amounting to some US$ 200,000. This stage lasted two years as well, from 2004 and 2005. The money was used to increase production and expand the company.

In August of 2006 Bug requested funding for a second project, entitled “Criação massal e comercialização dos parasitóides de ovos Trissolcus basalis e Telenomus podisi para o controle de percevejos da soja” (mass breeding and commercialization of egg parasitoids Trissolcus basalis and Telenomus podisi for the control of the soybean stink bug). Approved in the beginning of 2007 directly to the program’s Phase II, the project is expected to continue to be developed through 2008. Fapesp granted around US$ 200,000, money that is being used in research and in the increase of the production scale of those natural enemies of the soybean stink bug. “We’re not producing commercially those parasitoids,” says Pedrazzoli. “We’re finishing up the adaptation of the technology we’ve developed in laboratory scale to an industrial process.”

A successful strategy

Thus Bug repeats the successful strategy that turned it one of Brazil’s top producers of insects for biological control of pests. To get there the company didn’t have to develop new technologies – it just used age old knowledge. For even though only now it’s gaining importance in Brazil, biological control of pests is much older than many people might think: back in the third century BC the Chinese already used ants to control pests in citrus.

In modern times, the use of natural enemies has a landmark in 1888, when Americans adopted the vedalia lady bug beetle (Rodolia cardinalis) to combat the cottony cushion scale (Icerya purchasi). “It was the first successful example of biological control, and has become a classic in the literature on the subject,” says agronomy engineer Wilson Carlos Pazini, of the School of Agrarian Sciences (Faculdade de Ciências Agrárias), of the Jaboticabal campus of the São Paulo State University (Universidade Estadual Paulista, Unesp). “Two years after the lady bug beetle was releases the cottony cushion scale had been controlled.” In the case of Bug, what the company did was to improve breeding techniques and equipment, as well as the insects’ diet. “In general terms we generate only productive processes, which are used internally and don’t need to be patented because they won’t be commercialized,” explains Pedrazzoli.

Cotesia flavipes is originally from Trinidad and Tobago and was introduced in Brazil in the 1970s. Since then it has been successfully combating in Brazilian fields its natural enemy sugarcane borer. Until the 1980s the damages this worm caused to sugar and alcohol producers reached US$ 100 million a year in the State of São Paulo alone. “With the introduction of the Cotesia flavipes and its release in the State the infestation intensity of the sugarcane borer, which was between 8 and 10 percent, dropped to 2 percent,” says Unesp’s Pazini. “This resulted in savings of around US$ 80 million a year, with a reduction of annual losses from US$ 100 million to US$ 20 million.”

Most of that loss was caused by red rot. The worm itself doesn’t cause the disease, but by digging a hole in the sugarcane stem in order to complete its life cycle, that is, to become a moth, it opens the way for fungi, which are the true cause of red rot. The disease causes chemical changes in the plant that reduce sugar and alcohol output. “For a productivity of 80 tons of sugarcane per hectare, losses from each 1 percent of sugarcane borer infestation amounts to 616 kilograms (1,356 lbs) of sugarcane, 28 kilograms (62 lbs) of sugar and 16 liters (4.2 gallons) of alcohol,” informs Mauro Sampaio Benedini, regional manager of Products of the Sugar & Alcohol Technology Center (Centro de Tecnologia Canavieira, CTC), a private non-profit organization dedicated to the technological development of the sugarcane, sugar, alcohol and bio-energy industries.

For that reason, says Bendini, the industry is already conscious that the rational control of pests through insects helps reduce production costs. While the cost of combating pests with insecticides is around US$ 26 per hectare, control through the Cotesias flavipes costs less than US$ 9, and through Trichogrammas close to US$ 21. Considering that in Brazil 5 million hectares are planted with sugarcane, with a total output of 387 million tons, the gains are significant.

But the benefits of biological control are not just in lower costs. Chemical products in general reduce pest infection quickly only in the beginning. It’s an emergency control indicated for situations in which the level of infection is alarming or if what’s wanted is the pest’s quick extermination. The problem is that insecticides cause environmental imbalances, because they kill both the pest and its natural enemy. After a while the pest is back, this time without its enemy. Biological control, in contrast, maintains the pest in check without causing damages.

Production lines

Bug’s three units operate like true insect factories, with production lines and everything else. The one in Piracicaba produces both Cotesia flavipes, which are the “good guys”, and the pest, sugarcane borers, the “bad guys”. The other two units produce Trichogramma wasps. In addition, thanks to PIPE the company maintains a partnership with Esalq, through which it’s allowed to conduct research and control of quality of insect production in the school’s laboratories.

The factory in which Cotesias are produced is located in a 400-square meter (4,300 sq ft) two-story house. The main production line is on the top floor, which is divided into four rooms with controlled temperature and humidity. The process starts in the Adults Room (Sala de Adultos), with temperature of 22° Celsius (72º F), where pupas of the Diatraea saccharalis moth about to become adults are put inside cages. Four to seven days later, the adult insects – yellowish moths with nocturnal habits – are removed from the cages, paired up in couples and placed inside PVC pipes 20 centimeters (7.9 in) high by 10 centimeters (3.9 in) in diameter lined with carton paper. Each pipe holds between 30 and 50 couples, depending on the time of the year. In four days each female is able to lay up to 450 eggs.

After those four days the carton paper is removed and cut into pieces with some 250 eggs each – more or less the equivalent of a female’s egg production. Each piece goes into a glass pipe or flask that the company has developed (the pipes carry only 25 eggs), where there’s a special menu for the worms that will hatch, based on soybean protein, wheat germ, sugar and vitamins. Those flasks and pipes – hundreds of them – are stored, in shelves that resemble a wine cellar, in the Development Room (Sala de Desenvolvimento), where the temperature is kept at 30º C (143º F) and humidity is kept around 80 percent. Fifteen days after they hatch, 5 percent of the worms are removed from the glass pipes or flasks. They are the lucky ones that are going to complete their life cycles, become matrix and continue the breeding cycle. The remaining worms are kept in the flasks or pipes for 30 days; when they’re about 1 centimeter (.4 in) long, they are removed to be parasited by the Cotesia wasp.

The able hands of the well-trained employees take the worms out of the pipes and flasks and press them against a minute hole in the center of small transparent flasks. Inside each of these flasks some 50 small wasps fly frenetically. When they have contact with the worms through the hole, the wasps lay their eggs inside the worm. It sounds like a slow process. However, a Cotesia needs just a fraction of a second to get to the hole and introduce its ovipositor in the worm, leaving inside it about 50 eggs. It’s so fast that each employee is able to expose between 3,000 and 5,000 worms a day.

Next, the sugarcane borers are taken to the Mass Room (Sala de Massa), where the temperature is kept at 25º C (77º F). There they are kept on acrylic plates inside small flat, transparent flasks. On the plates there’s a special diet for the worms so that they feed while the wasps’ eggs develop inside them until they hatch. When that takes place the small larvae eat their host from within. Ten or eleven days after they had parasited, the larvae leave the dead worm and get together in masses of pupas, forming small cocoons that together look like white foam.

The cocoons are one of Bug’s products. The company sells them in small plastic cups. Each cup holds 30 cocoons with about 50 pupas of Cotesia flavipes each – in other words, it gives origin to 1,500 wasps. The customer gets the cups and waits until the pupas turn into insects. Then he/she goes to the field, opens the cup and walks with it 25 meters (27 yards) so that the wasps fly out. Then the wasps themselves are going to do the same thing that is done at the factory: lay their eggs inside sugarcane borers. And the process starts all over. “Six thousands wasps are needed to control the pest in 1 hectare,” explains Pedrazzoli. “Today we produce 250 million Cotesia a month, which is enough to treat 40,000 hectares of sugarcane.”

Clothes moth eggs

The other product in Bug’s portfolio is a set of eggs of the Anagasta kuehniella clothes moth already parasited by the Trichogramma wasp. This wasp is more versatile than the Cotesia and controls a larger number of pests, such as the sugarcane borer itself, the fall armyworm and the corn earworm, as well as pests in vegetables such as tomatoes. It is also easy to be produced because it may be bred in alternative hosts. “It’s different from what takes place with the Cotesia, which can only be multiplied in its natural host, that is, the very pest it combats, the sugarcane borer,” Carvalho explains. “In the case of the Trichogramma, we use the Anagasta, which lives in stored grain and is happy with a simpler diet, dry, comprised of just integral wheat flour and yeast.”

The production process of both wasps, however, is similar. First, adult couples of the Anagasta clothes moth are placed in cages to mate. There the females lay eggs, which are removed. Part of them is mixed with the diet based on flour and yeast that will feed the colony. The rest – 95 percent – is sterilized with ultraviolet rays and then exposed in trays to Trichogramma wasps, which in turn lay their eggs inside the clothes moths’.

In another room the parasited eggs are placed in containers with holes, which make possible for the parasitoids to leave for the fields. The containers are just biodegradable blister packs comprised of three layers of cardboard. But there’s a trick: the middle layer has small “tunnels” 2 millimeters (.04 in) in diameter. When the other layers are superposed on top of it these “tunnels” form small recipients or capsules – each with capacity to store 2,000 eggs. Developed by Bug, this innovative packaging is already protected by a patent – the only one the company has.

The company came to it after several unsuccessful experiments. Previous versions were vulnerable to ants, which ate the eggs, thus preventing the wasps from hatching. Besides protecting the eggs from these predators, the capsule provides more safety regarding temperature changes and rain. The blister packages are sold to the producers, who put them in the plant itself or place them in braces. When the wasps hatch, they leave through the holes and go parasite the pests’ eggs in the field.

Customer satisfaction can be measured by the expansion plans Bug has. “We’re increasing production and have hired many people in the past few days,” says Pedrazzoli. “We’re soon going to move to a building ten times bigger than the one we have today. We hope to be in the short run one of the largest biological control companies in the world. Our target markets are Brazil, Argentina and Uruguay, as well as Europe.”