Micología Aplicada International ISSN: 1534-2581 [email protected] Colegio de Postgraduados México

Bermúdez, R. C.; García, N.; Gross, P.; Serrano, M. Cultivation of Pleurotus on agricultural substrates in Cuba Micología Aplicada International, vol. 13, núm. 1, january, 2001, pp. 25-29 Colegio de Postgraduados Puebla, México

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CULTIVATION OF PLEUROTUS ON AGRICULTURAL SUBSTRATES IN CUBA

R. C. BERMÚDEZ, N. GARCÍA, P. GROSS AND M. SERRANO

Centre of Studies for Industrial Biotechnology (CEBI), Faculty of Natural and Mathematical Sciences, University of Oriente, Patricio Lumumba s/n, CP 90500, Santiago de Cuba, Cuba. E-mail: [email protected]

Accepted for publication November 1, 2000

ABSTRACT

Pleurotus ostreatus f. sp. florida (P-184) was cultivated on several agricultural substrates, such as coffee pulp, cocoa shells, and coconut shells. These substrates were processed by solar drying, stored, pasteurized, and used for mushroom cultivation. The highest biological efficiencies were recorded on coffee pulp from Coffea arabica (168.5-179.4%), followed by coconut shells (90.0%), and cocoa shells (84.5%; particle size: > 4 mm).

Key words: Pleurotus ostreatus f. sp. florida, edible mushrooms, agricultural by- products, coffee pulp, cocoa shell, coconut shell, Cuba.

INTRODUCTION large amounts of organic by-products are generated, such as coffee pulp, shells, husks, Intensive research work has recently been among many others 4, 15. These lignocellulosic made for developing biological processes in materials have been shown to be suitable which agricultural and industrial by-products substrates for edible mushroom cultivation, can be used 10, 16. Agricultural activities which is an efficient biotechnological are economically important in Cuba. For process for their bioconversion and recycling example, the eastern mountainous region 7, 14. Furthermore, a sustainable model for produces about 80% of the high quality rural production and small-scale cultivation coffee in this country 2, while cocoa and technologies are available for promoting coconut are cultivated in the area of Baracoa, mushroom cultivation in developing McountyICOL .of A PLGuantánamo.. INT., 13(1), As 2001, a natural PP. 25-29result, countries 7, 13. 26 R. C. BERMÚDEZ ET AL. CULTIVATION OF PLEUROTUS IN CUBA 27

In Cuba, cultivation of the edible were also dried, and crushed in irregular mushroom Pleurotus started at ICIDCA fractions > 3 cm due to it highly fibrous (Cuban Research Institute for Sugar Cane texture. Rehydrated substrates (coffee pulp, Derivatives) in 1988, using sugar cane straw cocoa and coconut shells) were pasteurized and bagasse as growing substrates, and by immersion in hot water at 70o-80oC for 60 nowadays local technologies are available min. A solution of 0.02% benomyl (Bayer, 8. In this study, we have shown that several Germany) was added during pasteurization agricultural by-products available in the in order to reduce mould contamination. eastern region of Cuba can be used for the cultivation of Pleurotus ostreatus f.sp. Spawn preparation florida. It was performed according to Klibansky et al. 8. Wheat kernels, previously soaked during 15-17 h for reaching 40-50% MATERIALS AND METHODS moisture content, were sterilized in jars (200-400 g) at 121oC for 90 min. Sterilized Location kernels were inoculated with the mycelium Experiments were carried out in the of P. ostreatus f. sp. florida. mushroom research unit of the Centre of Studies for Industrial Biotechnology Substrate inoculation (CEBI), and in a rural mushroom farm of Spawn was homogeneously mixed with the Central Research Station of Coffee and pasteurized substrates, and placed in plastic Cacao, from the Santiago de Cuba county, bags of 2 kg (30x40 cm) and 4 kg (50x70 Cuba. cm). Plastic bags of 30x40 cm were used for the experiments with all substrates Microorganism studied, whereas those of 50x70 cm for A strain of Pleurotus ostreatus f. sp. florida coffee pulp experiments under different (P-184) was used in all experiments. This cultivation conditions. Every experiment strain is adapted to tropical conditions, had three replicates. Temperature and and is deposited at the CEBI’s culture relative humidity in the mushroom research collection. unit were managed according to Sánchez- Vázquez 19; while in the rural mushroom Substrate preparation farm they were managed according to the season of the year. Coefficients of variation All substrates studied were subjected to 17 solar drying before use. Coffee pulp was for both parameters were calculated . Biological efficiency (BE) was calculated obtained from a coffee-producing region 22 (Coffea arabica L., Coffea canephora according to Tschierpe and Hartmann . P.), III Frente in Santiago de Cuba, dried, Earliness was expressed as the number of and stored for a year. Dry coffee pulp was days from spawning to the appearance of rehydrated by immersion in water for 2- the first fruit-body primordia. 3 h. Cocoa shells (Theobroma cacao L.) gathered from a cacao orchard were dried, crushed, and screened. Two particle sizes RESULTS AND DISCUSSION were selected: 1) 1.25-4.00 mm, and 2) > 4.0 mm. Coconut shells (Cocos nucifera L.) Coffee pulp, as well as cocoa and coconut

MICOL. APL. INT., 13(1), 2001, PP. 25-29 MICOL. APL. INT., 13(1), 2001, PP. 25-29 26 R. C. BERMÚDEZ ET AL. CULTIVATION OF PLEUROTUS IN CUBA 27

shells, were suitable substrates for mycelial Table 2. Average conditions of temperature and growth and fruiting of P. ostreatus f. sp. relative humidity within a rural mushroom farm. florida. Evident contamination was not observed on cocoa and coconut shells. Condition Period of Coefficient Temperature and relative humidity within the cultivation of variation mushroom research unit was relatively stable (Table 1), during substrate colonization (25.8o-26.1oC; 95%), and fruiting (19.8o- Temperature (oC): 21.1oC; 85.7-95.8%). However, these 7:00 a.m. 21.8 0.6 factors were more variable in the rural 12:00 m. 22.4 0.5 mushroom farm (Table 2), during the period 17:00 p.m. 22.7 0.4 of cultivation (temperature: 21.8o-22.7oC; Relative humidity (%): relative humidity: 92.0-100%). 7:00 a.m. 100 0.15 Fruit-body primordia began to appear 11 12:00 m. 92.0 2.73 days after spawning on coconut shells, 16 17:00 p.m. 96.7 3.02 days on cocoa shells, and 25 days on coffee pulp (Table 3). Mushroom yields were higher on coffee pulp (842.5 g), followed by cocoa shells (456.2 g), and coconut shells more vigorous and faster mycelial growth (270 g), which is in agreement with previous 1, 6, 11, 12, 18, 20, 21 rates were observed on cocoa and coconut research work . However, shells. Biological efficiencies (BE) on the coffee pulp varied in relation to cultivation conditions and the coffee shrub, which is in Table 1. Average conditions of temperature and accord with previous research from other relative humidity within the mushroom research authors 11, 12, 18, 19. Higher BEs were recorded unit during the experiments. on coffee pulp from Coffea arabica (Fig. 1), in either the mushroom research unit Condition Mushroom cultivation (168.5%) or the rural mushroom farm (179.4%) [Fig. 2]; by contrast, coffee pulp SC Cv Fruiting Cv from Coffea canephora showed lower BEs of 70.9% and 67.1%, respectively. Temperature (oC): This may be due to the different moisture 7:00 a.m. 26.1 0.06 21.1 0.07 content recorded for C. arabica (75-80%) 12:00 m. 25.8 0.06 19.8 0.07 and C. canephora (60-70%). Coconut shells 17:00 p.m. 25.9 0.06 19.8 0.06 showed a BE of 90.0%. In the case of cocoa Relative humidity (%): shells, the particle size had strong influence 7:00 a.m. 95 - 85.7 0.09 on the BE, being higher on larger particles 12:00 m. 95 - 95.8 0.04 (> 4 mm: 84.5%). 17:00 p.m. 95 - 93.1 0.05 The loss of organic matter is the simplest criterion adopted to evaluate the extent of a substrate biodegradation 5, 10. A decrease in SC= Substrate colonization. the content of organic matter is associated Cv= Coefficient of variation (%). with mushroom mycelial growth and fruiting

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Table 3. Cultivation of Pleurotus on agricultural by-products under different experimental conditions.

Substrate Earliness* Dry Yield Biological (days) weight (g) efficiency (kg) (%)

Coffee pulp: Experimental mushroom unit Coffea arabica 25 0.50 842.5 168.5 Coffea canephora 28 0.60 425.4 70.9 Rural mushroom farm Coffea arabica 25 0.50 897.0 179.4 Coffea canephora 30 0.60 402.6 67.1 Cocoa shells: Particle size: 1.2-4.0 mm 19 0.44 179.5 40.8 Particle size: > 4.0 mm 16 0.54 456.2 84.5 Coconut shells 11 0.30 270.0 90.0

* Number of days from spawning to the appearance of the first fruit-body primordia. on lignocellulosic by-products, in which 2. Bermúdez, R. C., J. R. Cárdenas, M. Serrat, N. García, CO and H O are liberated. According to P. Gross and T. Orberá. 1999. Caracterización 2 2 técnica-socio-económica y ambiental de las this, there was a good degradation of organic despulpadoras de la provincia Santiago de Cuba. materials studied, as the proportion of spent Proyecto Nacional Valorización de los Residuales substrate was 57.8% for the coffee pulp, 27.9 del Café. Ministerio CITMA, Cuba. 75 pp. 3. Bermúdez, R. C., N. García, I. Ramos, C. Martínez, Y. % for cocoa shells, and 70.3% for coconut Pérez and M. Serrano. 1999. Utilización de la shells. Similar results have previously been pleurotina como fuente de bioabono. In: Simposio reported for the coffee pulp 12, 18. Internacional de Café y Cacao (Abstracts). The implementation of this technology CubaCafé’99. Santiago de Cuba. 4. Braham, J. E. and R. Bressani. 1979. Pulpa de café, on a large or small scale in agricultural composición, tecnología y utilización. CIID, communities of Cuba would generate a Bogotá, Colombia. 152 pp. human food (mushrooms) for regional 5. Brock, T., M. Madigan, J. Martinko and J. Parker. 1994. consumption or tourism, as well as spent Biology of microorganisms. Seventh edition. Prentice-Hall International, Inc., Englewood substrates useful as organic fertilizers or Cliffs, New Jersey. 909 pp. animal feed, and local jobs 3, 7, 9, 13. 6. Calvo, L. and J. Sánchez-Vázquez. 1993. Producción de hongos comestibles en condiciones rústicas bajo un cacaotal y utilizando cáscara de coco como sustrato. Proceed. 11th Int. Cocoa Res. Conf. July LITERATURE CITED 18-24. Cocoa Producers Alliance, Yamoussoukro, Ivory Coast. 1. Bermúdez, R. C., J. A. Traba, M. Verdecía and P. Gross. 1994. 7. Chang, S. T. and P. G. Miles. 1989. Edible mushrooms and Producción de Pleurotus sp. cfr. florida sobre los their cultivation. CRC Press, Boca Raton. 345 residuales de la agroindustria cafetalera en Cuba. pp. Micol. Neotrop. Apl. 7: 47-50.

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Fig. 2. Pleurotus cultivation in a rural mushroom farm, using coffee pulp as a growing substrate.

13. Martínez-Carrera, D., A. Aguilar, W. Martínez, P. Morales, M. Sobal, M. Bonilla and A. Larqué-Saavedra. 1998. A sustainable model for rural production of edible mushrooms in Mexico. Micol. Neotrop. Apl. 11: 77-96. 14. Martínez-Carrera, D., P. Morales, R. Leben, M. Sobal and A. Larqué-Saavedra. 1991. Historia del cultivo comercial de hongos comestibles en México. Ciencia y Desarrollo 65: 41-48. Fig. 1. Fruit bodies of Pleurotus ostreatus f. sp. 15. MINAGRI. 1993. Subproductos del cacao: posibilidades florida cultivated on coffee pulp from Coffea y perspectivas de su uso. Ministerio de la arabica. Agricultura, La Habana. 10 pp. 16. Olguín, E. 1994. El potencial de la biotecnología ambiental dentro de un contexto de desarrollo sustentable. Instituto de Ecología, Xalapa, Mexico. 110 pp. 17. Ostle, B. 1980. Estadística aplicada. Ed. Revolucionarias, 8. Klibansky, M., M. Mansur and I. Gutiérrez. 1993. Productions La Habana. 629 pp. of Pleurotus ostreatus mushrooms on sugar cane 18. Rodríguez-Valencia, N. and V. Zuluaga. 1994. Cultivo agrowastes. Acta Biotechnol. 13(1): 71-78. de Pleurotus pulmonarius en pulpa de café. 9. Labarére, J. E. and U. G. Menini. 1998. Global policy CENICAFE 45(3): 81-92. profile related to the collection, characterization, 19. Sánchez-Vázquez, J. E. 1994. Producción de hongos conservation and utilization of mushroom genetic comestibles. Centro de Investigaciones Ecológicas resources for food and agriculture. F.A.O. Global del Sureste, Chiapas, México. 80 pp. Network on Mushrooms, Newsletter No. 2, 20. Senyah, J. K. 1988. Mushrooms from waste materials. In: Bordeaux. Developments in food microbiology –4 (edited 10. Martin, A. 1994. Biodegradation and bioremediation. by R. K. Robinson). Pp. 1-22. Elsevier Applied Academic Press, New York. 247 pp. Science Publihers Ltd. Barking, Essex, UK. 11. Martínez-Carrera, D. 1987. Design of a mushroom farm for 21. Soto, C., G. Guzmán, D. Salmones and G. Mata. 1993. growing Pleurotus on coffee pulp. Mush. J. Tropics El cultivo de hongos comestibles. Instituto 7: 13-23. Politécnico Nacional, Mexico, D. F. 245 pp. 12. Martínez-Carrera, D. 1989. Simple technology to cultivate 22. Tschierpe, H. J. and K. Hartmann. 1977. A comparison of Pleurotus on coffee pulp in the tropics. Mushroom different growing methods. Mushroom Journal Science 12 (II): 169–178. 60: 404-416.

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