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glucosidasa cuya actividad fue casi la mitad de la observada en paja de
arroz shy I
La actividad endoglucanasa observada por Lowe 997 y las obtenidas en el
presente trabajo son bastante menores a las publicadas por Hebraud 1988
para Neocalimastix frontalis cultivado en fibras de sisal y menores a las
reportadas para Piromyces sp por Teunissen 1992 Sin embargo es
importante resaltar que en estos trabajos hay diferencias en relacioacuten con la
eMe utilizada por ejemplo el presente estudio al igual que el de Lowe
1997 se utilizoacute eMe comercial y de alta viscosidad mientras que Teunissen
1992 utilizoacute eMe de baja viscosidad y Hebraud no especificoacute las
caracteriacutesticas del compuesto utilizado Estas diferencias podriacutean estar
influenciando la accesibilidad del sustrato a los extractos enzimaacuteticos
De otro lado en el presente trabajo se utilizoacute celulosa cristalina Sigmacellreg y
no Avicel de la casa comercial Merck las cuales presentan caracteriacutesticas
fiacutesico-quiacutemicas distintas frente a la hidroacutelisis enzimaacutetica (Bemalier et al
1987) lo que debe tenerse en cuenta desde los valores de actividad
enzimaacutetica encontrados en este estudio
En concordancia con el bajo crecimiento en cascarilla de cafeacute las menores
actividades enzimaacuteticas se obtuvieron en este medio de cultivo auacuten cuando
la actividad exoglucanasa no es despreciable Sin embargo las actividades
especiacuteficas de las enzimas evaluadas en este medio de cultivo fueron mucho
mayores que las de paja y bagazo de cantildea Esto sugiere la necesidad de
realizar investigaciones que evaluacuteen pretratamientos para la eliminacioacuten de
dichos compuestos y posibiliten una mejor utilizacioacuten de este desecho por
los hongos
En todos los casos evaluados los valores de pH y temperatura obtenidos
para la maacutexima actividad celuloliacutetica concuerdan bien con las reportadas en
67
la literatura para hongos anaerobios ruminales (Teunissen MJ et al 1992
Lowe S E Et al 1987 Hebraud M amp Fevre M 1988 Tsai Cheng-Fang et al
2003) De igual manera la estabilidad del extracto enzimaacutetico de HARBR-1
cultivado en celulosa cristalina (Sigmacell 101) presentoacute un comportamiento
similar al reportado en la literatura (Lowe S E)987 Hebraud M 1988)
La termosensibilidad de los filtrados enzimaacuteticos se evidencioacute por la
disminucioacuten considerable de sus actividades a temperaturas de 40 y 50 oC
en un periacuteodo de 36 y 12 horas respectivamente y cuyos valores
concuerdan con los reportados en la literatura Lowe S E et al 1987
Hebraud M Fevre M 1988 Qiu Xiao et al 2000 Sin embargo esta
termosensibilidad no se observoacute en los cultivos del hongo en donde se
detectaron actividades enzimaacuteticas en filtrados colectados 4 diacuteas despueacutes de
haber alcanzado la fase estacionaria tiempo en el cual la actividad
enzimaacutetica del filtrado libre de ceacutelulas ha alcanzado valores cercanos a
cero Estos resultados sugieren que en los cultivos hay un recambio
continuo de las enzimas celuloliacuteticas lo cual es un factor importante para L
decidir si utilizar los productos enzimaacuteticos o los cultivos directamente
Bajo esta perspectiva el hongo HARBR-1 resulta promisorio para la
produccioacuten de celulasas Sin embargo se requieren estudios adicionales
para la optimizacioacuten de medios de cultivos que aumenten la produccioacuten
enzimaacutetica y permitan estimar su potencialidad en la industria biotecnoloacutegica
y otras aplicaciones industriales (Dijkerman Op den Camp HJM 1996)
El anaacutelisis conjunto de los datos y la habilidad para inducir la
) zoosporogeacutenesis de este hongo anaerobio ruminal indican que la paja de
-7) 1arroz y en menor proporcioacuten el bagazo de cantildea constituyen buenos
sustratos para el cultivo y produccioacuten de enzimas celuloliacuteticas
particularmente de exoglucanasas Este punto es importante si se tiene en
cuenta que ambos son residuos de cosecha en nuestro paiacutes y pueden ser
68
utilizados como un sustrato econoacutemico en lugar de la paja de trigo o de
cebada materiales de difiacutecil consecucioacuten en nuestro medio y uno de los
sustratos evaluados en otros paiacuteses por diversos autores (Dijkerman R Et
al 1997 Lowe S E et al 1987 Teunissen M J et al 1991 Teunissen M J
et al 1992 Rezaeian Mohammad et al 2005)
CONCLUSIONES Y CAMPOS POR EXPLORAR
Los resultados de estos trabajos aportan informacioacuten sobre la biologiacutea baacutesica
de microorganismos anaerobios ruminales del troacutepico y han permitido la
capacitacioacuten de recurso humano y la estandarizacioacuten de metodologiacuteas para
el estudio de bacterias y hongos celuloliacuteticos del rumen de ganado criollo
BON sobre los cuales existe informacioacuten muy incipiente hasta la fecha En
realidad constituyen parte de investigaciones iniciales que sirven de base
para estudios posteriores sobre estos micoorganismos en nuestro medio y
podriacutean resumirse en los siguientes puntos
bull Se aportoacute informacioacuten sobre la biologiacutea baacutesica cultivo y ciclo de vida
bull Se desarrolloacute una teacutecnica basada en PCR a partir de secuencias
diagnoacutesticas usadas como sondas que permiten la identificacioacuten raacutepida y
precisa de las tres principales bacterias celuloliacuteticas ruminales
bull Se disentildearon cebadores para la identificacioacuten molecular por PCR de
hongos anaerobios ruminales del geacutenero Orpinomyces
bull Se realizaron modimiddotficaciones metodoloacutegicas que mejoraron las teacutecnicas
de aislamiento cultivo mantenimiento y caracterizacioacuten morfoloacutegica de
los aislados que se veniacutean implementando en el laboratorio de BIORUM
bull Se estandarizaron teacutecnicas que permiten determinar la actividad
enzimaacutetica in vitro de los microorganismos
bull Se disentildeoacute un micropistilo metaacutelico adaptable a un taladro eleacutectrico para la
maceracioacuten de muestras bioloacutegicas
bull Se realizaron modificaciones en forma de ventosa a las pipetas Pasteur
para facilitar la purificacioacuten de la colonia fuacutengica a partir de tubos
Hungate
bull Se construyoacute en acriacutelico una caacutemara de cultivo de microorganismos
69
71
bull Produccioacuten de las enzimas celuloliacuteticas a una mayor escala
bull Utilizar las enzimas celuloliacuteticas o sus complejos como modelo
para realizar estudios de ingenieriacutea de proteiacutenas
bull Desarrollar estudios de diversidad geneacutetica microbiana yevolutivos
para esclarecer si los ganados criollos colombianos en efecto han ] Y-shy
seleccionado poblaciones microbianas con actividades maacutes
potentes que la de sus ancestros europeos
AGRADECIMIENTOS
Estos proyectos fueron financiados por la Direccioacuten de Investigaciones de la
Universidad Nacional de Colombia Sede Medelliacuten (DIME proyectos 030802530
030802530 030802603) El cultivo y mantenimiento de cepas nativas y de
referencia se realizoacute en el Laboratorio de Biotecnologiacutea Ruminal y las teacutecnicas
moleculares se realizaron en el Laboratorio de Estudios Moleculares y el Laboratorio
de Biologiacutea Molecular y Celular La autora agradece al Doctor Rafael Arango por sus
sugerencias en la estandarizacioacuten de algunos procedimientos experimentales a los
profesores Luis Alfonso Giraldo por su invitacioacuten a trabajar con los microorganismos
ruminales y su ayuda en el Laboratorio de Biotecnologiacutea Ruminal Oiga Ineacutes
Montoya Campuzano por su asesoriacutea en los meacutetodos de aislamiento y cultivo de
microorganismos y Orlando Simoacuten Ruiz Villadiego por su comentarios en la
evaluacioacuten de las actividades eeluloliacuteticas a los estudiantes que desarrollaron sus
trabajos de grado y tesis de maestriacutea Joseacute Carlos Meneo Gonzaacutelez Juan Aicardo
Segura Caro Diana Patricia Goacutemez Corteacutes Edgard de Jesuacutes Muacutenera Tuberquia al
Ingeniero Agroacutenomo Ricardo Mareel Landiacutenez por su apoyo experimental en la
estandarizacioacuten de la electroforesis de proteiacutenas en geles de poliacrilamida-SDS y
las bacterioacutelogas Juliana Botero y Yulime Monsalve quien realizaron su pasantiacutea en
el marco de uno de estos proyectos
bull Se estandarizaron teacutecnicas para el anaacutelisis molecular de DNA de
bacterias anaerobias rurninales y para analizar el nuacutemero y peso
molecular de proteiacutenas contenidas en el extracto de hongos anaerobios
Debido a que en nuestro paiacutes esta aacuterea de investigacioacuten es auacuten
incipiente los campos por explorar son bastante amplios de tal manera
que solo sentildealareacute algunos de los trabajos que pueden realizarse en el
futuro
bull Evaluacioacuten de los cebadores utilizados en este trabajo en un
nuacutemero mayor de microorganismos y en microbiota proveniente de
otras razas de ganado criollo
bull Evaluacioacuten de nuevos cebadores disentildeados en el marco de estos
proyectos que no alcanzaron a ser incorporados en el siguiente
informe
bull Evaluar nuevos protocolos de extraccioacuten de DNA de hongos
anaerobios ruminales que tengan en cuenta el caraacutecter refractario
de estos microorganismos
bull Identificacioacuten molecular de hongos anaerobios ruminales de varias
razas de ganado criollo otros ruminantes y no rumiantes En
particular seriacutea interesante explorar la microbiota anaerobia
celuloliacutetica en buacutefalos
bull Evaluacioacuten de la actividad celuloliacutetica en bacterias anaerobias
ruminales Caracterizacioacuten de enzimas o de los complejos
enzimaacuteticos celuloliacuteticos en bacterias y hongos anaerobios
ruminales
bull Optimizar las condiciones de cultivo para la produccioacuten de enzimas
celuloliticas de origen ruminal
70
72
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STEENBAKKERS PJ HARHANGI HR BOSSCHER MW VAN DER HOOFT MM KELTJENS JT VAN DER DRIFT C VOGELS GD OP DEN CAMP HJ (2003) ~shyGlucosidase in cellulosome of the anaerobic fungus Piromyces sp strain E2 is a family 3 glycoside hydrolase Biochem J 370 963 - 970
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67
la literatura para hongos anaerobios ruminales (Teunissen MJ et al 1992
Lowe S E Et al 1987 Hebraud M amp Fevre M 1988 Tsai Cheng-Fang et al
2003) De igual manera la estabilidad del extracto enzimaacutetico de HARBR-1
cultivado en celulosa cristalina (Sigmacell 101) presentoacute un comportamiento
similar al reportado en la literatura (Lowe S E)987 Hebraud M 1988)
La termosensibilidad de los filtrados enzimaacuteticos se evidencioacute por la
disminucioacuten considerable de sus actividades a temperaturas de 40 y 50 oC
en un periacuteodo de 36 y 12 horas respectivamente y cuyos valores
concuerdan con los reportados en la literatura Lowe S E et al 1987
Hebraud M Fevre M 1988 Qiu Xiao et al 2000 Sin embargo esta
termosensibilidad no se observoacute en los cultivos del hongo en donde se
detectaron actividades enzimaacuteticas en filtrados colectados 4 diacuteas despueacutes de
haber alcanzado la fase estacionaria tiempo en el cual la actividad
enzimaacutetica del filtrado libre de ceacutelulas ha alcanzado valores cercanos a
cero Estos resultados sugieren que en los cultivos hay un recambio
continuo de las enzimas celuloliacuteticas lo cual es un factor importante para L
decidir si utilizar los productos enzimaacuteticos o los cultivos directamente
Bajo esta perspectiva el hongo HARBR-1 resulta promisorio para la
produccioacuten de celulasas Sin embargo se requieren estudios adicionales
para la optimizacioacuten de medios de cultivos que aumenten la produccioacuten
enzimaacutetica y permitan estimar su potencialidad en la industria biotecnoloacutegica
y otras aplicaciones industriales (Dijkerman Op den Camp HJM 1996)
El anaacutelisis conjunto de los datos y la habilidad para inducir la
) zoosporogeacutenesis de este hongo anaerobio ruminal indican que la paja de
-7) 1arroz y en menor proporcioacuten el bagazo de cantildea constituyen buenos
sustratos para el cultivo y produccioacuten de enzimas celuloliacuteticas
particularmente de exoglucanasas Este punto es importante si se tiene en
cuenta que ambos son residuos de cosecha en nuestro paiacutes y pueden ser
68
utilizados como un sustrato econoacutemico en lugar de la paja de trigo o de
cebada materiales de difiacutecil consecucioacuten en nuestro medio y uno de los
sustratos evaluados en otros paiacuteses por diversos autores (Dijkerman R Et
al 1997 Lowe S E et al 1987 Teunissen M J et al 1991 Teunissen M J
et al 1992 Rezaeian Mohammad et al 2005)
CONCLUSIONES Y CAMPOS POR EXPLORAR
Los resultados de estos trabajos aportan informacioacuten sobre la biologiacutea baacutesica
de microorganismos anaerobios ruminales del troacutepico y han permitido la
capacitacioacuten de recurso humano y la estandarizacioacuten de metodologiacuteas para
el estudio de bacterias y hongos celuloliacuteticos del rumen de ganado criollo
BON sobre los cuales existe informacioacuten muy incipiente hasta la fecha En
realidad constituyen parte de investigaciones iniciales que sirven de base
para estudios posteriores sobre estos micoorganismos en nuestro medio y
podriacutean resumirse en los siguientes puntos
bull Se aportoacute informacioacuten sobre la biologiacutea baacutesica cultivo y ciclo de vida
bull Se desarrolloacute una teacutecnica basada en PCR a partir de secuencias
diagnoacutesticas usadas como sondas que permiten la identificacioacuten raacutepida y
precisa de las tres principales bacterias celuloliacuteticas ruminales
bull Se disentildearon cebadores para la identificacioacuten molecular por PCR de
hongos anaerobios ruminales del geacutenero Orpinomyces
bull Se realizaron modimiddotficaciones metodoloacutegicas que mejoraron las teacutecnicas
de aislamiento cultivo mantenimiento y caracterizacioacuten morfoloacutegica de
los aislados que se veniacutean implementando en el laboratorio de BIORUM
bull Se estandarizaron teacutecnicas que permiten determinar la actividad
enzimaacutetica in vitro de los microorganismos
bull Se disentildeoacute un micropistilo metaacutelico adaptable a un taladro eleacutectrico para la
maceracioacuten de muestras bioloacutegicas
bull Se realizaron modificaciones en forma de ventosa a las pipetas Pasteur
para facilitar la purificacioacuten de la colonia fuacutengica a partir de tubos
Hungate
bull Se construyoacute en acriacutelico una caacutemara de cultivo de microorganismos
69
71
bull Produccioacuten de las enzimas celuloliacuteticas a una mayor escala
bull Utilizar las enzimas celuloliacuteticas o sus complejos como modelo
para realizar estudios de ingenieriacutea de proteiacutenas
bull Desarrollar estudios de diversidad geneacutetica microbiana yevolutivos
para esclarecer si los ganados criollos colombianos en efecto han ] Y-shy
seleccionado poblaciones microbianas con actividades maacutes
potentes que la de sus ancestros europeos
AGRADECIMIENTOS
Estos proyectos fueron financiados por la Direccioacuten de Investigaciones de la
Universidad Nacional de Colombia Sede Medelliacuten (DIME proyectos 030802530
030802530 030802603) El cultivo y mantenimiento de cepas nativas y de
referencia se realizoacute en el Laboratorio de Biotecnologiacutea Ruminal y las teacutecnicas
moleculares se realizaron en el Laboratorio de Estudios Moleculares y el Laboratorio
de Biologiacutea Molecular y Celular La autora agradece al Doctor Rafael Arango por sus
sugerencias en la estandarizacioacuten de algunos procedimientos experimentales a los
profesores Luis Alfonso Giraldo por su invitacioacuten a trabajar con los microorganismos
ruminales y su ayuda en el Laboratorio de Biotecnologiacutea Ruminal Oiga Ineacutes
Montoya Campuzano por su asesoriacutea en los meacutetodos de aislamiento y cultivo de
microorganismos y Orlando Simoacuten Ruiz Villadiego por su comentarios en la
evaluacioacuten de las actividades eeluloliacuteticas a los estudiantes que desarrollaron sus
trabajos de grado y tesis de maestriacutea Joseacute Carlos Meneo Gonzaacutelez Juan Aicardo
Segura Caro Diana Patricia Goacutemez Corteacutes Edgard de Jesuacutes Muacutenera Tuberquia al
Ingeniero Agroacutenomo Ricardo Mareel Landiacutenez por su apoyo experimental en la
estandarizacioacuten de la electroforesis de proteiacutenas en geles de poliacrilamida-SDS y
las bacterioacutelogas Juliana Botero y Yulime Monsalve quien realizaron su pasantiacutea en
el marco de uno de estos proyectos
bull Se estandarizaron teacutecnicas para el anaacutelisis molecular de DNA de
bacterias anaerobias rurninales y para analizar el nuacutemero y peso
molecular de proteiacutenas contenidas en el extracto de hongos anaerobios
Debido a que en nuestro paiacutes esta aacuterea de investigacioacuten es auacuten
incipiente los campos por explorar son bastante amplios de tal manera
que solo sentildealareacute algunos de los trabajos que pueden realizarse en el
futuro
bull Evaluacioacuten de los cebadores utilizados en este trabajo en un
nuacutemero mayor de microorganismos y en microbiota proveniente de
otras razas de ganado criollo
bull Evaluacioacuten de nuevos cebadores disentildeados en el marco de estos
proyectos que no alcanzaron a ser incorporados en el siguiente
informe
bull Evaluar nuevos protocolos de extraccioacuten de DNA de hongos
anaerobios ruminales que tengan en cuenta el caraacutecter refractario
de estos microorganismos
bull Identificacioacuten molecular de hongos anaerobios ruminales de varias
razas de ganado criollo otros ruminantes y no rumiantes En
particular seriacutea interesante explorar la microbiota anaerobia
celuloliacutetica en buacutefalos
bull Evaluacioacuten de la actividad celuloliacutetica en bacterias anaerobias
ruminales Caracterizacioacuten de enzimas o de los complejos
enzimaacuteticos celuloliacuteticos en bacterias y hongos anaerobios
ruminales
bull Optimizar las condiciones de cultivo para la produccioacuten de enzimas
celuloliticas de origen ruminal
70
72
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STEENBAKKERS J M et al (2003) I3-Glucosidase in ceflulosome of the anaerobic fungus Piromyces sp strain E2 is a family 3 glycoside hydrolase Biochem J 370963 - 970
STEENBAKKERS PJ HARHANGI HR BOSSCHER MW VAN DER HOOFT MM KELTJENS JT VAN DER DRIFT C VOGELS GD OP DEN CAMP HJ (2003) ~shyGlucosidase in cellulosome of the anaerobic fungus Piromyces sp strain E2 is a family 3 glycoside hydrolase Biochem J 370 963 - 970
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STEWART C S BRYANT M P 1988 The rumen bacteria In The rumen microbial ecosystem Edited By Hobson p Elsevier Applied Science New York 21-26
STEWART C S PANIAGUA C DINSDALE D CHENG K-J AND GARRAW S H 1981 Selective isolation and characterization of Bacteriodes succinogenes from the rumen of a cow Applied and Environmental Microbiology Vol 41 W 2 P 504-510
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TEUNISSEN M J V M DE KORT G OP DEN CAMP H J M HUIS INT VELD J H J (1992) Production of cellulolytic and xylanolytic enzymes during growth of the anaerobic fungus Piromyces sp on different substrates J of Gen Microb 138 1657-1664 l
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THOMPSON JD HIGGINS DG and GIBSON TJ CLUSTAL W Improving the sensitivity of progressive multiple sequence alignment through sequence weighting positions-specific gap penalties and weight matrix choice In Nucleic Acids Research Vol 22 (1994) p 225
TSAI CHENG-FANG et al (2003) A comparative analysis of two cDNA clones of the f cellulose gene family from anaerobic fungus Piromyces rhizinflata Anaerobe 9 131 - 140
68
utilizados como un sustrato econoacutemico en lugar de la paja de trigo o de
cebada materiales de difiacutecil consecucioacuten en nuestro medio y uno de los
sustratos evaluados en otros paiacuteses por diversos autores (Dijkerman R Et
al 1997 Lowe S E et al 1987 Teunissen M J et al 1991 Teunissen M J
et al 1992 Rezaeian Mohammad et al 2005)
CONCLUSIONES Y CAMPOS POR EXPLORAR
Los resultados de estos trabajos aportan informacioacuten sobre la biologiacutea baacutesica
de microorganismos anaerobios ruminales del troacutepico y han permitido la
capacitacioacuten de recurso humano y la estandarizacioacuten de metodologiacuteas para
el estudio de bacterias y hongos celuloliacuteticos del rumen de ganado criollo
BON sobre los cuales existe informacioacuten muy incipiente hasta la fecha En
realidad constituyen parte de investigaciones iniciales que sirven de base
para estudios posteriores sobre estos micoorganismos en nuestro medio y
podriacutean resumirse en los siguientes puntos
bull Se aportoacute informacioacuten sobre la biologiacutea baacutesica cultivo y ciclo de vida
bull Se desarrolloacute una teacutecnica basada en PCR a partir de secuencias
diagnoacutesticas usadas como sondas que permiten la identificacioacuten raacutepida y
precisa de las tres principales bacterias celuloliacuteticas ruminales
bull Se disentildearon cebadores para la identificacioacuten molecular por PCR de
hongos anaerobios ruminales del geacutenero Orpinomyces
bull Se realizaron modimiddotficaciones metodoloacutegicas que mejoraron las teacutecnicas
de aislamiento cultivo mantenimiento y caracterizacioacuten morfoloacutegica de
los aislados que se veniacutean implementando en el laboratorio de BIORUM
bull Se estandarizaron teacutecnicas que permiten determinar la actividad
enzimaacutetica in vitro de los microorganismos
bull Se disentildeoacute un micropistilo metaacutelico adaptable a un taladro eleacutectrico para la
maceracioacuten de muestras bioloacutegicas
bull Se realizaron modificaciones en forma de ventosa a las pipetas Pasteur
para facilitar la purificacioacuten de la colonia fuacutengica a partir de tubos
Hungate
bull Se construyoacute en acriacutelico una caacutemara de cultivo de microorganismos
69
71
bull Produccioacuten de las enzimas celuloliacuteticas a una mayor escala
bull Utilizar las enzimas celuloliacuteticas o sus complejos como modelo
para realizar estudios de ingenieriacutea de proteiacutenas
bull Desarrollar estudios de diversidad geneacutetica microbiana yevolutivos
para esclarecer si los ganados criollos colombianos en efecto han ] Y-shy
seleccionado poblaciones microbianas con actividades maacutes
potentes que la de sus ancestros europeos
AGRADECIMIENTOS
Estos proyectos fueron financiados por la Direccioacuten de Investigaciones de la
Universidad Nacional de Colombia Sede Medelliacuten (DIME proyectos 030802530
030802530 030802603) El cultivo y mantenimiento de cepas nativas y de
referencia se realizoacute en el Laboratorio de Biotecnologiacutea Ruminal y las teacutecnicas
moleculares se realizaron en el Laboratorio de Estudios Moleculares y el Laboratorio
de Biologiacutea Molecular y Celular La autora agradece al Doctor Rafael Arango por sus
sugerencias en la estandarizacioacuten de algunos procedimientos experimentales a los
profesores Luis Alfonso Giraldo por su invitacioacuten a trabajar con los microorganismos
ruminales y su ayuda en el Laboratorio de Biotecnologiacutea Ruminal Oiga Ineacutes
Montoya Campuzano por su asesoriacutea en los meacutetodos de aislamiento y cultivo de
microorganismos y Orlando Simoacuten Ruiz Villadiego por su comentarios en la
evaluacioacuten de las actividades eeluloliacuteticas a los estudiantes que desarrollaron sus
trabajos de grado y tesis de maestriacutea Joseacute Carlos Meneo Gonzaacutelez Juan Aicardo
Segura Caro Diana Patricia Goacutemez Corteacutes Edgard de Jesuacutes Muacutenera Tuberquia al
Ingeniero Agroacutenomo Ricardo Mareel Landiacutenez por su apoyo experimental en la
estandarizacioacuten de la electroforesis de proteiacutenas en geles de poliacrilamida-SDS y
las bacterioacutelogas Juliana Botero y Yulime Monsalve quien realizaron su pasantiacutea en
el marco de uno de estos proyectos
bull Se estandarizaron teacutecnicas para el anaacutelisis molecular de DNA de
bacterias anaerobias rurninales y para analizar el nuacutemero y peso
molecular de proteiacutenas contenidas en el extracto de hongos anaerobios
Debido a que en nuestro paiacutes esta aacuterea de investigacioacuten es auacuten
incipiente los campos por explorar son bastante amplios de tal manera
que solo sentildealareacute algunos de los trabajos que pueden realizarse en el
futuro
bull Evaluacioacuten de los cebadores utilizados en este trabajo en un
nuacutemero mayor de microorganismos y en microbiota proveniente de
otras razas de ganado criollo
bull Evaluacioacuten de nuevos cebadores disentildeados en el marco de estos
proyectos que no alcanzaron a ser incorporados en el siguiente
informe
bull Evaluar nuevos protocolos de extraccioacuten de DNA de hongos
anaerobios ruminales que tengan en cuenta el caraacutecter refractario
de estos microorganismos
bull Identificacioacuten molecular de hongos anaerobios ruminales de varias
razas de ganado criollo otros ruminantes y no rumiantes En
particular seriacutea interesante explorar la microbiota anaerobia
celuloliacutetica en buacutefalos
bull Evaluacioacuten de la actividad celuloliacutetica en bacterias anaerobias
ruminales Caracterizacioacuten de enzimas o de los complejos
enzimaacuteticos celuloliacuteticos en bacterias y hongos anaerobios
ruminales
bull Optimizar las condiciones de cultivo para la produccioacuten de enzimas
celuloliticas de origen ruminal
70
72
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THOMPSON JD HIGGINS DG and GIBSON TJ CLUSTAL W Improving the sensitivity of progressive multiple sequence alignment through sequence weighting positions-specific gap penalties and weight matrix choice In Nucleic Acids Research Vol 22 (1994) p 225
TSAI CHENG-FANG et al (2003) A comparative analysis of two cDNA clones of the f cellulose gene family from anaerobic fungus Piromyces rhizinflata Anaerobe 9 131 - 140
CONCLUSIONES Y CAMPOS POR EXPLORAR
Los resultados de estos trabajos aportan informacioacuten sobre la biologiacutea baacutesica
de microorganismos anaerobios ruminales del troacutepico y han permitido la
capacitacioacuten de recurso humano y la estandarizacioacuten de metodologiacuteas para
el estudio de bacterias y hongos celuloliacuteticos del rumen de ganado criollo
BON sobre los cuales existe informacioacuten muy incipiente hasta la fecha En
realidad constituyen parte de investigaciones iniciales que sirven de base
para estudios posteriores sobre estos micoorganismos en nuestro medio y
podriacutean resumirse en los siguientes puntos
bull Se aportoacute informacioacuten sobre la biologiacutea baacutesica cultivo y ciclo de vida
bull Se desarrolloacute una teacutecnica basada en PCR a partir de secuencias
diagnoacutesticas usadas como sondas que permiten la identificacioacuten raacutepida y
precisa de las tres principales bacterias celuloliacuteticas ruminales
bull Se disentildearon cebadores para la identificacioacuten molecular por PCR de
hongos anaerobios ruminales del geacutenero Orpinomyces
bull Se realizaron modimiddotficaciones metodoloacutegicas que mejoraron las teacutecnicas
de aislamiento cultivo mantenimiento y caracterizacioacuten morfoloacutegica de
los aislados que se veniacutean implementando en el laboratorio de BIORUM
bull Se estandarizaron teacutecnicas que permiten determinar la actividad
enzimaacutetica in vitro de los microorganismos
bull Se disentildeoacute un micropistilo metaacutelico adaptable a un taladro eleacutectrico para la
maceracioacuten de muestras bioloacutegicas
bull Se realizaron modificaciones en forma de ventosa a las pipetas Pasteur
para facilitar la purificacioacuten de la colonia fuacutengica a partir de tubos
Hungate
bull Se construyoacute en acriacutelico una caacutemara de cultivo de microorganismos
69
71
bull Produccioacuten de las enzimas celuloliacuteticas a una mayor escala
bull Utilizar las enzimas celuloliacuteticas o sus complejos como modelo
para realizar estudios de ingenieriacutea de proteiacutenas
bull Desarrollar estudios de diversidad geneacutetica microbiana yevolutivos
para esclarecer si los ganados criollos colombianos en efecto han ] Y-shy
seleccionado poblaciones microbianas con actividades maacutes
potentes que la de sus ancestros europeos
AGRADECIMIENTOS
Estos proyectos fueron financiados por la Direccioacuten de Investigaciones de la
Universidad Nacional de Colombia Sede Medelliacuten (DIME proyectos 030802530
030802530 030802603) El cultivo y mantenimiento de cepas nativas y de
referencia se realizoacute en el Laboratorio de Biotecnologiacutea Ruminal y las teacutecnicas
moleculares se realizaron en el Laboratorio de Estudios Moleculares y el Laboratorio
de Biologiacutea Molecular y Celular La autora agradece al Doctor Rafael Arango por sus
sugerencias en la estandarizacioacuten de algunos procedimientos experimentales a los
profesores Luis Alfonso Giraldo por su invitacioacuten a trabajar con los microorganismos
ruminales y su ayuda en el Laboratorio de Biotecnologiacutea Ruminal Oiga Ineacutes
Montoya Campuzano por su asesoriacutea en los meacutetodos de aislamiento y cultivo de
microorganismos y Orlando Simoacuten Ruiz Villadiego por su comentarios en la
evaluacioacuten de las actividades eeluloliacuteticas a los estudiantes que desarrollaron sus
trabajos de grado y tesis de maestriacutea Joseacute Carlos Meneo Gonzaacutelez Juan Aicardo
Segura Caro Diana Patricia Goacutemez Corteacutes Edgard de Jesuacutes Muacutenera Tuberquia al
Ingeniero Agroacutenomo Ricardo Mareel Landiacutenez por su apoyo experimental en la
estandarizacioacuten de la electroforesis de proteiacutenas en geles de poliacrilamida-SDS y
las bacterioacutelogas Juliana Botero y Yulime Monsalve quien realizaron su pasantiacutea en
el marco de uno de estos proyectos
bull Se estandarizaron teacutecnicas para el anaacutelisis molecular de DNA de
bacterias anaerobias rurninales y para analizar el nuacutemero y peso
molecular de proteiacutenas contenidas en el extracto de hongos anaerobios
Debido a que en nuestro paiacutes esta aacuterea de investigacioacuten es auacuten
incipiente los campos por explorar son bastante amplios de tal manera
que solo sentildealareacute algunos de los trabajos que pueden realizarse en el
futuro
bull Evaluacioacuten de los cebadores utilizados en este trabajo en un
nuacutemero mayor de microorganismos y en microbiota proveniente de
otras razas de ganado criollo
bull Evaluacioacuten de nuevos cebadores disentildeados en el marco de estos
proyectos que no alcanzaron a ser incorporados en el siguiente
informe
bull Evaluar nuevos protocolos de extraccioacuten de DNA de hongos
anaerobios ruminales que tengan en cuenta el caraacutecter refractario
de estos microorganismos
bull Identificacioacuten molecular de hongos anaerobios ruminales de varias
razas de ganado criollo otros ruminantes y no rumiantes En
particular seriacutea interesante explorar la microbiota anaerobia
celuloliacutetica en buacutefalos
bull Evaluacioacuten de la actividad celuloliacutetica en bacterias anaerobias
ruminales Caracterizacioacuten de enzimas o de los complejos
enzimaacuteticos celuloliacuteticos en bacterias y hongos anaerobios
ruminales
bull Optimizar las condiciones de cultivo para la produccioacuten de enzimas
celuloliticas de origen ruminal
70
72
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71
bull Produccioacuten de las enzimas celuloliacuteticas a una mayor escala
bull Utilizar las enzimas celuloliacuteticas o sus complejos como modelo
para realizar estudios de ingenieriacutea de proteiacutenas
bull Desarrollar estudios de diversidad geneacutetica microbiana yevolutivos
para esclarecer si los ganados criollos colombianos en efecto han ] Y-shy
seleccionado poblaciones microbianas con actividades maacutes
potentes que la de sus ancestros europeos
AGRADECIMIENTOS
Estos proyectos fueron financiados por la Direccioacuten de Investigaciones de la
Universidad Nacional de Colombia Sede Medelliacuten (DIME proyectos 030802530
030802530 030802603) El cultivo y mantenimiento de cepas nativas y de
referencia se realizoacute en el Laboratorio de Biotecnologiacutea Ruminal y las teacutecnicas
moleculares se realizaron en el Laboratorio de Estudios Moleculares y el Laboratorio
de Biologiacutea Molecular y Celular La autora agradece al Doctor Rafael Arango por sus
sugerencias en la estandarizacioacuten de algunos procedimientos experimentales a los
profesores Luis Alfonso Giraldo por su invitacioacuten a trabajar con los microorganismos
ruminales y su ayuda en el Laboratorio de Biotecnologiacutea Ruminal Oiga Ineacutes
Montoya Campuzano por su asesoriacutea en los meacutetodos de aislamiento y cultivo de
microorganismos y Orlando Simoacuten Ruiz Villadiego por su comentarios en la
evaluacioacuten de las actividades eeluloliacuteticas a los estudiantes que desarrollaron sus
trabajos de grado y tesis de maestriacutea Joseacute Carlos Meneo Gonzaacutelez Juan Aicardo
Segura Caro Diana Patricia Goacutemez Corteacutes Edgard de Jesuacutes Muacutenera Tuberquia al
Ingeniero Agroacutenomo Ricardo Mareel Landiacutenez por su apoyo experimental en la
estandarizacioacuten de la electroforesis de proteiacutenas en geles de poliacrilamida-SDS y
las bacterioacutelogas Juliana Botero y Yulime Monsalve quien realizaron su pasantiacutea en
el marco de uno de estos proyectos
bull Se estandarizaron teacutecnicas para el anaacutelisis molecular de DNA de
bacterias anaerobias rurninales y para analizar el nuacutemero y peso
molecular de proteiacutenas contenidas en el extracto de hongos anaerobios
Debido a que en nuestro paiacutes esta aacuterea de investigacioacuten es auacuten
incipiente los campos por explorar son bastante amplios de tal manera
que solo sentildealareacute algunos de los trabajos que pueden realizarse en el
futuro
bull Evaluacioacuten de los cebadores utilizados en este trabajo en un
nuacutemero mayor de microorganismos y en microbiota proveniente de
otras razas de ganado criollo
bull Evaluacioacuten de nuevos cebadores disentildeados en el marco de estos
proyectos que no alcanzaron a ser incorporados en el siguiente
informe
bull Evaluar nuevos protocolos de extraccioacuten de DNA de hongos
anaerobios ruminales que tengan en cuenta el caraacutecter refractario
de estos microorganismos
bull Identificacioacuten molecular de hongos anaerobios ruminales de varias
razas de ganado criollo otros ruminantes y no rumiantes En
particular seriacutea interesante explorar la microbiota anaerobia
celuloliacutetica en buacutefalos
bull Evaluacioacuten de la actividad celuloliacutetica en bacterias anaerobias
ruminales Caracterizacioacuten de enzimas o de los complejos
enzimaacuteticos celuloliacuteticos en bacterias y hongos anaerobios
ruminales
bull Optimizar las condiciones de cultivo para la produccioacuten de enzimas
celuloliticas de origen ruminal
70
72
BIBLlOGRAFIA
ALTSCHUL STEPHEN F THOMAS L MADDEN ALEJANDRO A SCHAacuteFFER JINGHUI lHANG lHENG lHANG WEBB MILLER AND DAVID J L1PMAN (1997) Gapped BLAST and PSI-BLAST a new generation of protein database search programs In Nucleic Acids Research Vol 25 (1997) p 3389-3402
AMANN R l KRUMHOLTl l STAHL D A 1990 Fluorescent oligonucleotide probing of whole cells for detenninative phylogenetic and environmental studies in microbiology Journal Bacteriology 172 762-770
AMANN R l ZARDA B STAHL D A SCHLEIFER K H 1992 Identification the individual procariotic cells with enzime laveled rRNA-tergeted oligonucleotide probes Applied and Environmental Microbiology 58 3007-3011
ATCC American Type Culture Collection [Organizacioacuten en liacutenea] Disponible en interne www ATCCorg
AVGERINOS G amp WANG D 1 C (1980) Annuals Reports of Fennentation Processes 4 165
BAUCHOP T 1979 Rumen anaerobic fungi colonizer of plant fiber Ann Rech Vet 10246
BAUCHOP T 1989 Colonization of plant fragments by protozoa and fungi IN The reole of protozoa and fungi in ruminant digestion Ed J Nolan R Leng D Demeyer Proceeding of an international seminar held at the University of New England Annidale Austria Penambul Books p 83-96
BAUCHOP T Rumen anaeobic fungi of cattle and sheep In Applied and Environmental Microbiology Vo138 (1979) p148-158
BAXEVANIS A D and OUELLETIE BF 2001 Bioinformatics A Practical Guide to the Analysis of Genes and Proteins Segunda Edicioacuten John Wiley amp Sons Inc Ed p 455
BERNALlER A FONTY G AND GOUET PH (1987) Fennentation properties of four strictly Anaerobic rumen Fungal Species H2 - Producing Micoorganisms Laboratoire de Microbiologie-Inra-CR de Clemonnont-Theix 63122 - ST Genes-Chamnpanelle-Frabce
BERNALlER A FONTY G BONNEMOY F GOWET P Degradation and fennentation of cellulose by the rumen anaerobic fungi in axenic cultures or in association with cellulolytic bacteria Current Microbiology Vol 25 1995 P 143-148
BHAT M K (2000) Cellulases and related enzymes in biotechnology Biotechnology Advances 18 355-383 Research review paper
BHAT M K AND BHAT S (1997) Cellulose degrading and their potential industrial applications Biotechnology Advances 15 3 Y4 583-620 Review
BIOEDIT A user-friendly biological sequence alignment editor and analysis program for Windows 9598NT Hall TA Nucl [CD-RUM) In Acids Symposer Vol 41 (1999) p 95shy98
BIOSYSTEM - Glucose Reagent amp Instruments Kit Glucose enzymatic - spectrphotometric glucose oxidaseperoxidase
73
BOHORQUEZ G Claudis M GAVIRIA M Gloria E Cambios cuantitativos de los microorganismos rum inales por la utilizacioacuten de flavophospholipol Tesis de grado 1991 Universidad Nacional de Colombia Sede Medelliacuten
BRIESACHER SL MAY T GRIGSBY KN KERLEY MS ANTHONY RV PATERSON JA (1992) Use of DNA probes to monitor nutritional effects on ruminal prokaryotes and Fibrobacter succinogenes S85 In Journal Animal Science Vol 70 p 289-295
BRIESACHER S L MAY T GRIGSBY K N KERLEY M S ANTHONY R V AND PATERSON J A 1992 Use of DNA probes to monitor nutritional efects on ruminal prokaryotes and fibrobacter succinogenes S85 Journal of Animal Science 70 289
BROWN T A Gen Cloning an introduction 1990 Great Britain Ed Edmundsbury Press Segunda Edicioacuten P 64-71
BRYANT M P Bacterial species of the rumen Bacteriologicla reviews 23 125 1959
BRYANT M P AND BURKEY L A 1953 Numbers and some predominant groups in the rumen of cows fed different ratios Journal of Animal Science Vol 36 p 218-224
CHENG J AND WEIMER P J 2001 Competition among three predominant ruminal cellulolytic bacteria in the absence or presence of non-cellulolytic bacteria Microbiology Vol 147 p 21-30
CHESSON A AND FORSBERG C W 1988 In The rumen Microbial Ecosystem Edites by P N Hobson Elsevier Science Publishers L TB Pp 251
CHURCH D C 1974 Fisiologia digestiva y nutricioacuten de los rumiantes Vol 1 Editorial Acribia Zaragoza Espantildea
COEN D M The polimerase chain reaction In Shorts Protocols in Molecular Biology P 15-4 a 15-5
COUGHLAN MICHAEL P (1990) Cellulose degradation by Fungi in Microbial Enzymes and Biotechnology Second edition Ed By William M Fogarty and Catherine 1 Kelly
COUTHINHO PEDRO M and Henrissat Bernard (1999) The Modular Structure of Cellulases and Other Carbohydrate-active Enzymes an Integrated Data Approach In Genettics Biochemistry and Ecology of Cellulose Degradation Ohmiya K et al eds Uni Publishers Co Tokyo pp 15 - 23
DANCOURT M BOLLET C CARLlOZ A MARTELlN R GAYRAL J-P AND RAOULT D 2000 16S ribosomal Dtla sequence analysis of a large collection of environmental and clinical unidentifiable bacterial isolates Journal of Clinical Microbiology Vol 38 W 10 p 3623-3630
DE LONG E F WICKHAM G S PACE N R 1989 Phylogenetic stains ribosomalshybased for the indentification of single cells Science Vol 243 1360-1363
DENMAN S XUE G-P PATEL B (1996) Characterization of a Neocallimastix patriciarum Cellulase cDNA (celA) Homologous to Trichoderma reesei Cellobiohydrolase 11 Appl And Env Microb 62(6) 1889-1896
DEVILLARD E NEWBOLD CJ SCOTT KP FORANO E WALLACE RJ JOUANY JP FLlNT HJ (1999) A xylanase produced by the rumen anaerobic protozoan Polyplastron multivesiculatum shows close sequence similarity to family 11 xylanases from Gram - positive bacteria In FEMS Microbiology Letters Vol 181 p 145-152
74
- ------
DIJKERMAN R OP DEN CAMP HJM (1996) Cultivation of anaerobic fungi in a 10-L fermenter system for the production of (hemi) cellulolytic enzymes Appl Microbiol Biotechnol 4685-91
EBERHARDT R Y GILBERT H J HAZLEWOOD G P (2000) Primary sequence and enzymes properties of two modular endoglucanases celA and Ce145A from the anaerobic fungus Piromyces equi Microbiology 146 1999 - 2008
FONTY G AND JOBLlN K N (1991) Rumen Anaerobic Fungi Their Role and Interactions with Other Rumen Microorganisms in Relation to Fiber Digestion in Physiological Aspects of Digestion and Metabolism in Rumiants Proceedings of the Seventh International Symposium on Ruminant Physiology
FORSBERG C W CRPSBY B and THOMAS D Y 1986 Potential for manipulation of the rumen fermentation through the use of recombinant DNA techniques Journal of Animal Science Vol 63 310-321
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GARRYTI George BOONE David R and CASTENHOLZ Richard W Editores 2001 Bergeys Manual of Systematic Bacteriology Pg744
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GILBERT HJ AND HASLEWOOD (1993) Bacterial cellulases and xylanases J of Gen Microb 139 187-194 Review
GILBERT HJ HAZLEWOOD GP LAURIE JI ORPIN CG XUE GP (1992) Homologous catalytic domains in a rumen fungal xylanase evidence for gene duplication and prokaryotic origino In Molecular Microbiology Vol 6 p 2065-2072
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HALL TA 1999 BioEdit a user-friendly biological sequence alignment editor and analysis program for Windows 9598NT Nucl Acids Symp Ser 41 95-98
HARHANGI HR FREELOVE AC UBHAYASEKERA W VAN DINTHER M STEENBAKKERS PJ AKHMANOVA A VAN DER DRIFT C JETTEN MS MOWBRAY SL GILBERT HJ OP DEN CAMP HJ (2003) CeI6A a major exoglucanasa from the cellulosome of the anaerobic fungi Piromyces sp E2 and Piromyces equi Biochimica et Biophysica Acta 1628 30 - 39
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HO W Y KHOO 1 Y S TAN S G ABDULLAH N JALALUDIN S KUDO H (1994) Isozyme analysis of anaerobic rumen fungi and their relationship to aerobic chytrids Microbiology 140 1495-1504
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HOLDEMAN LV EP CATO AND WEC MOORE (1977) Anaerobe Laboratory Manual 4 th Edition
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JANE A l BRYANT Marvin P and HASPELL R B 1982 Diurnal variations in bacterial numbers and fluid parameters in ruminal contents of animal fed low-or high-foraje diets Applied and Environmental Microbiolegy Vol 44 Ndeg 2 P 402-412
JEFFRIES T W 1987Production and applications of cellulase laboratory procedures USDA Forest products Laboratory Madison Wisconsin February
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JULLlAND V VAUX A MILLET L FONTY G 1999 Identificaction of Rumnococcus favefaciens as the predominant cellulolytic bacterial species of the equine cecum Applied and Environmental Microbiolegy Vol 65 N 8 P 3738-3741
KARLlN S and ALTSCHUL S F (1990) Methods for assessing the statistical significance of molecular sequence features by using general scoring schemes Proc Natl Acad Sci USA 87 2264-2268
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Domains for a Cellulase-Hemicellulase Complex Appli And Env Microb 63(12) 4721-4728 Dec
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UN Chuzao y STHAL David A 1995 Taxon-Specific Probes for the Cellulolitic Genus Fibrobacter Reveal Abundant and Novel EquineAssociated Populations En Applied and Environmental Microbiology Vol 61 Ndeg4 pg 1348-1351
-iquest LOWE S E THEODOROU M K AND TRINCI A P (1987) ) cellulases and Xylanases of an liexcl anaerobic rumen fungus grown on wheat straw wlleat---straw holocellulose cellulose and xylan Applied and environmental microbiology June p 1216 - 1223
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MURRAY AND THOMSON Q980 Citados por WILSON K Preparation of genomic DNA from bacteria In Short Protocols in Biology Molecular P 2-11 a 2-12
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ODENYO A A MACKIE R l STHAL D A WHITE B A 1994b The use of 16S rRNAshytargeted oligonucleotide probes for study competition between ruminal fibrolitic bacteria pure culture-studies with cellulose and alkaline peroxide-trated wheat straw Applied and Environmental Microbiolegy Vol 60 p 3697-3703
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OSSA F RIDRIGUEZ J ARCOS M DIAZ T 1999 Identificacioacuten molecular de las bacterias celuloliacuteticas Fibrobacter sucinogenes y Ruminococcus flavefaciens Revista
--- - - ---~ - ------====-------------------------------------------------------------------------shy
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Colombiana de Ciencias Pecuarias Memorias V Encuentro Nacional de Investigaciones de las Ciencias Pecuarias ENICIP Vol 12 P 120
PALMQVIST E AND HAN-HAGERDAL B (2000) Fermentation of lignocellulosic hydrolysates 1 inhibition and detoxification Bioresource Technology 74 17 - 24
PENtildeA Francisco 1982 El Rumen Aspectos de fisiologia y metabolismo microbioloacutegico Instituto colombiana Agropecuario ICA Divisioacuten de Ciencias Animales Bogotaacute P 78
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PRIMER 3 White head institutel MIl [Programa de computador] Disponible en internet www-genomewimiteducgi-binprimerprimer3-wwcgi
PROMEGA Manual tecnico Promega Kit wizard USA 1999
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RANADE DR and GRADE RV Microbiological aspects of anaerobic digestion laboratory manual Edited Maharashtra Association for the Cultivation of Science In Research Institute Pune 1988 102 p
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RODRIGUEZ V Fernando 1994 La microbiota ruminal y su importancia en la degradacioacuten de los forrajes tropicales En Seminario sobre Biotecnologia del Rumen Universidad Nacional de Colombia Sede Medelliacuten Nov 11-12
RODRIGUEZ DIAZ T (1996) Aislamiento patroacuten de fermentacioacuten de carbohidratos y caracterizacioacuten morfoloacutegica de bacterias celuloliacuteticas del rumen de bovinos alimentados con heno de Raigraacutes en Colombia En Revista CORPOICA Vol 1 p 123-28
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SRINIVASAN K MURAKAMI M NAKASHIMADA Y NISHIO N (2001) Efficient Production of Cellulolytic and Xylanolitic Enzymes by the Rumen Anaerobic Fungus Neocallimastix frontalis in a Repetead Batch Culture Journal of Bioscience and Bioengineering 91 2 153-158
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STEENBAKKERS J M et al (2003) I3-Glucosidase in ceflulosome of the anaerobic fungus Piromyces sp strain E2 is a family 3 glycoside hydrolase Biochem J 370963 - 970
STEENBAKKERS PJ HARHANGI HR BOSSCHER MW VAN DER HOOFT MM KELTJENS JT VAN DER DRIFT C VOGELS GD OP DEN CAMP HJ (2003) ~shyGlucosidase in cellulosome of the anaerobic fungus Piromyces sp strain E2 is a family 3 glycoside hydrolase Biochem J 370 963 - 970
STEPHEN A MADDEN T SCHAFFER A lHANG J lHANG l MILLER W AND LlPMAN D 1997 Gapped BLAST and PSI-BLAST a new generation of protein database search programs Nucleic Acids Res Vol 25 p 3389-3402
STEWART C S 1991 The Rumen Bacteria in Rumen Microbial Metabolism and Rumiant Digestion Editd by J p Jouany INRA Editions Paris pp 131
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STEWART C S PANIAGUA C DINSDALE D CHENG K-J AND GARRAW S H 1981 Selective isolation and characterization of Bacteriodes succinogenes from the rumen of a cow Applied and Environmental Microbiology Vol 41 W 2 P 504-510
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TEUNISSEN M J V M DE KORT G OP DEN CAMP H J M HUIS INT VELD J H J (1992) Production of cellulolytic and xylanolytic enzymes during growth of the anaerobic fungus Piromyces sp on different substrates J of Gen Microb 138 1657-1664 l
THEODOROU K M DAVIS R D NIELSEN B B LAWRENCE G l TRINCI P J (1990) Determination of growth of anaerobic fungi on soluble and cellulosic substrates using a pressure transducer Microbiology 141 671-678
THOMPSON JD HIGGINS DG and GIBSON TJ CLUSTAL W Improving the sensitivity of progressive multiple sequence alignment through sequence weighting positions-specific gap penalties and weight matrix choice In Nucleic Acids Research Vol 22 (1994) p 225
TSAI CHENG-FANG et al (2003) A comparative analysis of two cDNA clones of the f cellulose gene family from anaerobic fungus Piromyces rhizinflata Anaerobe 9 131 - 140
bull Se estandarizaron teacutecnicas para el anaacutelisis molecular de DNA de
bacterias anaerobias rurninales y para analizar el nuacutemero y peso
molecular de proteiacutenas contenidas en el extracto de hongos anaerobios
Debido a que en nuestro paiacutes esta aacuterea de investigacioacuten es auacuten
incipiente los campos por explorar son bastante amplios de tal manera
que solo sentildealareacute algunos de los trabajos que pueden realizarse en el
futuro
bull Evaluacioacuten de los cebadores utilizados en este trabajo en un
nuacutemero mayor de microorganismos y en microbiota proveniente de
otras razas de ganado criollo
bull Evaluacioacuten de nuevos cebadores disentildeados en el marco de estos
proyectos que no alcanzaron a ser incorporados en el siguiente
informe
bull Evaluar nuevos protocolos de extraccioacuten de DNA de hongos
anaerobios ruminales que tengan en cuenta el caraacutecter refractario
de estos microorganismos
bull Identificacioacuten molecular de hongos anaerobios ruminales de varias
razas de ganado criollo otros ruminantes y no rumiantes En
particular seriacutea interesante explorar la microbiota anaerobia
celuloliacutetica en buacutefalos
bull Evaluacioacuten de la actividad celuloliacutetica en bacterias anaerobias
ruminales Caracterizacioacuten de enzimas o de los complejos
enzimaacuteticos celuloliacuteticos en bacterias y hongos anaerobios
ruminales
bull Optimizar las condiciones de cultivo para la produccioacuten de enzimas
celuloliticas de origen ruminal
70
72
BIBLlOGRAFIA
ALTSCHUL STEPHEN F THOMAS L MADDEN ALEJANDRO A SCHAacuteFFER JINGHUI lHANG lHENG lHANG WEBB MILLER AND DAVID J L1PMAN (1997) Gapped BLAST and PSI-BLAST a new generation of protein database search programs In Nucleic Acids Research Vol 25 (1997) p 3389-3402
AMANN R l KRUMHOLTl l STAHL D A 1990 Fluorescent oligonucleotide probing of whole cells for detenninative phylogenetic and environmental studies in microbiology Journal Bacteriology 172 762-770
AMANN R l ZARDA B STAHL D A SCHLEIFER K H 1992 Identification the individual procariotic cells with enzime laveled rRNA-tergeted oligonucleotide probes Applied and Environmental Microbiology 58 3007-3011
ATCC American Type Culture Collection [Organizacioacuten en liacutenea] Disponible en interne www ATCCorg
AVGERINOS G amp WANG D 1 C (1980) Annuals Reports of Fennentation Processes 4 165
BAUCHOP T 1979 Rumen anaerobic fungi colonizer of plant fiber Ann Rech Vet 10246
BAUCHOP T 1989 Colonization of plant fragments by protozoa and fungi IN The reole of protozoa and fungi in ruminant digestion Ed J Nolan R Leng D Demeyer Proceeding of an international seminar held at the University of New England Annidale Austria Penambul Books p 83-96
BAUCHOP T Rumen anaeobic fungi of cattle and sheep In Applied and Environmental Microbiology Vo138 (1979) p148-158
BAXEVANIS A D and OUELLETIE BF 2001 Bioinformatics A Practical Guide to the Analysis of Genes and Proteins Segunda Edicioacuten John Wiley amp Sons Inc Ed p 455
BERNALlER A FONTY G AND GOUET PH (1987) Fennentation properties of four strictly Anaerobic rumen Fungal Species H2 - Producing Micoorganisms Laboratoire de Microbiologie-Inra-CR de Clemonnont-Theix 63122 - ST Genes-Chamnpanelle-Frabce
BERNALlER A FONTY G BONNEMOY F GOWET P Degradation and fennentation of cellulose by the rumen anaerobic fungi in axenic cultures or in association with cellulolytic bacteria Current Microbiology Vol 25 1995 P 143-148
BHAT M K (2000) Cellulases and related enzymes in biotechnology Biotechnology Advances 18 355-383 Research review paper
BHAT M K AND BHAT S (1997) Cellulose degrading and their potential industrial applications Biotechnology Advances 15 3 Y4 583-620 Review
BIOEDIT A user-friendly biological sequence alignment editor and analysis program for Windows 9598NT Hall TA Nucl [CD-RUM) In Acids Symposer Vol 41 (1999) p 95shy98
BIOSYSTEM - Glucose Reagent amp Instruments Kit Glucose enzymatic - spectrphotometric glucose oxidaseperoxidase
73
BOHORQUEZ G Claudis M GAVIRIA M Gloria E Cambios cuantitativos de los microorganismos rum inales por la utilizacioacuten de flavophospholipol Tesis de grado 1991 Universidad Nacional de Colombia Sede Medelliacuten
BRIESACHER SL MAY T GRIGSBY KN KERLEY MS ANTHONY RV PATERSON JA (1992) Use of DNA probes to monitor nutritional effects on ruminal prokaryotes and Fibrobacter succinogenes S85 In Journal Animal Science Vol 70 p 289-295
BRIESACHER S L MAY T GRIGSBY K N KERLEY M S ANTHONY R V AND PATERSON J A 1992 Use of DNA probes to monitor nutritional efects on ruminal prokaryotes and fibrobacter succinogenes S85 Journal of Animal Science 70 289
BROWN T A Gen Cloning an introduction 1990 Great Britain Ed Edmundsbury Press Segunda Edicioacuten P 64-71
BRYANT M P Bacterial species of the rumen Bacteriologicla reviews 23 125 1959
BRYANT M P AND BURKEY L A 1953 Numbers and some predominant groups in the rumen of cows fed different ratios Journal of Animal Science Vol 36 p 218-224
CHENG J AND WEIMER P J 2001 Competition among three predominant ruminal cellulolytic bacteria in the absence or presence of non-cellulolytic bacteria Microbiology Vol 147 p 21-30
CHESSON A AND FORSBERG C W 1988 In The rumen Microbial Ecosystem Edites by P N Hobson Elsevier Science Publishers L TB Pp 251
CHURCH D C 1974 Fisiologia digestiva y nutricioacuten de los rumiantes Vol 1 Editorial Acribia Zaragoza Espantildea
COEN D M The polimerase chain reaction In Shorts Protocols in Molecular Biology P 15-4 a 15-5
COUGHLAN MICHAEL P (1990) Cellulose degradation by Fungi in Microbial Enzymes and Biotechnology Second edition Ed By William M Fogarty and Catherine 1 Kelly
COUTHINHO PEDRO M and Henrissat Bernard (1999) The Modular Structure of Cellulases and Other Carbohydrate-active Enzymes an Integrated Data Approach In Genettics Biochemistry and Ecology of Cellulose Degradation Ohmiya K et al eds Uni Publishers Co Tokyo pp 15 - 23
DANCOURT M BOLLET C CARLlOZ A MARTELlN R GAYRAL J-P AND RAOULT D 2000 16S ribosomal Dtla sequence analysis of a large collection of environmental and clinical unidentifiable bacterial isolates Journal of Clinical Microbiology Vol 38 W 10 p 3623-3630
DE LONG E F WICKHAM G S PACE N R 1989 Phylogenetic stains ribosomalshybased for the indentification of single cells Science Vol 243 1360-1363
DENMAN S XUE G-P PATEL B (1996) Characterization of a Neocallimastix patriciarum Cellulase cDNA (celA) Homologous to Trichoderma reesei Cellobiohydrolase 11 Appl And Env Microb 62(6) 1889-1896
DEVILLARD E NEWBOLD CJ SCOTT KP FORANO E WALLACE RJ JOUANY JP FLlNT HJ (1999) A xylanase produced by the rumen anaerobic protozoan Polyplastron multivesiculatum shows close sequence similarity to family 11 xylanases from Gram - positive bacteria In FEMS Microbiology Letters Vol 181 p 145-152
74
- ------
DIJKERMAN R OP DEN CAMP HJM (1996) Cultivation of anaerobic fungi in a 10-L fermenter system for the production of (hemi) cellulolytic enzymes Appl Microbiol Biotechnol 4685-91
EBERHARDT R Y GILBERT H J HAZLEWOOD G P (2000) Primary sequence and enzymes properties of two modular endoglucanases celA and Ce145A from the anaerobic fungus Piromyces equi Microbiology 146 1999 - 2008
FONTY G AND JOBLlN K N (1991) Rumen Anaerobic Fungi Their Role and Interactions with Other Rumen Microorganisms in Relation to Fiber Digestion in Physiological Aspects of Digestion and Metabolism in Rumiants Proceedings of the Seventh International Symposium on Ruminant Physiology
FORSBERG C W CRPSBY B and THOMAS D Y 1986 Potential for manipulation of the rumen fermentation through the use of recombinant DNA techniques Journal of Animal Science Vol 63 310-321
_ FUJII SIMONE ET AL (2004) Atividade fungistaacutetica in Vitro de cafeina em fungos asociados com graos de cafeacute Maringaacute vol 26 no 3 279 - 285
GACESA P HUBBLE (1990) Tecnologia de las Enzimas Ed Acribia- Espantildea Paacuteginas 1 shy25
GARCIA-VALLVEacute S ROMEU A AND PALAU J 2000 Horizontal gene transfer of glycosyl hydrolases of the rumen fungi Molecular Biology and Evolution Vol 17 W 3 P 352shy361
GARRYTI George BOONE David R and CASTENHOLZ Richard W Editores 2001 Bergeys Manual of Systematic Bacteriology Pg744
GHOSE T K (1987) Measurements of cellulase activities IUPAC Applied chemistry division commission on biotechnology Pure amp Appl Chem Vol 59 N 2 p 257 - 268
GILBERT HJ AND HASLEWOOD (1993) Bacterial cellulases and xylanases J of Gen Microb 139 187-194 Review
GILBERT HJ HAZLEWOOD GP LAURIE JI ORPIN CG XUE GP (1992) Homologous catalytic domains in a rumen fungal xylanase evidence for gene duplication and prokaryotic origino In Molecular Microbiology Vol 6 p 2065-2072
GIOVANNONI S J DeLONG E F OLSEN G J AND PACE N R 1988 Phylogenetic group specific oligodeoxinucleotide probe for identification of single microbial cells Journal of Bacteriology Vol 170 W 2 p720-726
GUETTLER MV RUMLER D and JAIN MK 1976 Actinobacillus succinogenes sp nov a novel succinic-acid-producing strain from the bovine rumen Applied and Environmental Microbiolegy Vol 32 Ndeg 4 P 623-632
HALL TA 1999 BioEdit a user-friendly biological sequence alignment editor and analysis program for Windows 9598NT Nucl Acids Symp Ser 41 95-98
HARHANGI HR FREELOVE AC UBHAYASEKERA W VAN DINTHER M STEENBAKKERS PJ AKHMANOVA A VAN DER DRIFT C JETTEN MS MOWBRAY SL GILBERT HJ OP DEN CAMP HJ (2003) CeI6A a major exoglucanasa from the cellulosome of the anaerobic fungi Piromyces sp E2 and Piromyces equi Biochimica et Biophysica Acta 1628 30 - 39
75
HEAD 1 M SAUNDERS J R AND PICKUP R W 1998 Microbial evolution diversity and ecology a decade of ribosomal RNA aalysis of uncultivate microorganisms Microbial Ecology Vol 35 P 1-21
HEBRAUD M FEVRE M (1988) Characterization of Glycoside Hydrolases Secreted by the Rumen Anaerobic Fungi Neocallimastix frontalis Sphaeromonas communis and Piromonas communis J of Gen Microb 134 1123-1129
HO W Y AND BARR Q D J S (1995) Classification of anaerobic gut fungi from herbivores whit emphasis on rumen fungi from Malaysia Mycologia 87(5) p 655 - 677
HO W Y KHOO 1 Y S TAN S G ABDULLAH N JALALUDIN S KUDO H (1994) Isozyme analysis of anaerobic rumen fungi and their relationship to aerobic chytrids Microbiology 140 1495-1504
HOBSON P N (1988) The rumen microbial ecosystem London and New York Elsevier Applied science 1989 p 530
HOLDEMAN LV EP CATO AND WEC MOORE (1977) Anaerobe Laboratory Manual 4 th Edition
HUNGATE R E (1966) The rumen and its microbes Ed Academic press New York and London 533 p
HUNGATE R E 1944 Studies on cellulose fermentation 1 The culture and physiology of an anaerobic cellulose-digesting bacterium Journal of Bacteriology Vol 48 p 499-513
INNIS M A AND GELFAND D H Optimization of PCRs In PCR protocols a guide to methods and applications 1990 Academic Press P 3-11
JANE A l BRYANT Marvin P and HASPELL R B 1982 Diurnal variations in bacterial numbers and fluid parameters in ruminal contents of animal fed low-or high-foraje diets Applied and Environmental Microbiolegy Vol 44 Ndeg 2 P 402-412
JEFFRIES T W 1987Production and applications of cellulase laboratory procedures USDA Forest products Laboratory Madison Wisconsin February
JOBLlN K N (1981) Isolation enumeration and maintenance of rumen anaerobic fungi in roll tube Applied and environmental microbiology p 119 - 1122
JULLlAND V VAUX A MILLET L FONTY G 1999 Identificaction of Rumnococcus favefaciens as the predominant cellulolytic bacterial species of the equine cecum Applied and Environmental Microbiolegy Vol 65 N 8 P 3738-3741
KARLlN S and ALTSCHUL S F (1990) Methods for assessing the statistical significance of molecular sequence features by using general scoring schemes Proc Natl Acad Sci USA 87 2264-2268
KOIKE S PAN J KOBAYASHI Y and TANAKA K 2003 Kinetics of in sacco fibershyattachment of representative ruminal cellulolytic bacteria monitored by competitive PCR Journal of Aniaml Science Vol 68 p 1429-1435
KOIKE Satoshi y KOBAYASHI Yasuo 2001 Development and use of competitive PCR assay for the rumen cellulolytic bacteria Fibrobacter succinogenes Ruminococcus albus and Ruminococcus favefaciens FEMS Microbiology Letter Vol 204 p361-366
LI X-L CHEN H LJUNGDAHL L G (1997 a) Two Cellulases Cel A and CelC from the polycentric Anaerobic Fungus Orpinomyces Strain PC-2 Contain N-Terminal Docking
76
Domains for a Cellulase-Hemicellulase Complex Appli And Env Microb 63(12) 4721-4728 Dec
LI X-L CHEN H LJUNGDAHL L G (1997 b) Monocentric and Polycentric Anaerobic Fungi Produce Structurally Related Cellulases and Xylanases Appl And Env Microb 63(2) 628shy635
UN Chuzao y STHAL David A 1995 Taxon-Specific Probes for the Cellulolitic Genus Fibrobacter Reveal Abundant and Novel EquineAssociated Populations En Applied and Environmental Microbiology Vol 61 Ndeg4 pg 1348-1351
-iquest LOWE S E THEODOROU M K AND TRINCI A P (1987) ) cellulases and Xylanases of an liexcl anaerobic rumen fungus grown on wheat straw wlleat---straw holocellulose cellulose and xylan Applied and environmental microbiology June p 1216 - 1223
LUDWIG W DORN S SPRINGER N KIRCHHOF G AND SCHLEIFER K H 1994 PCR Based preparation of 23S rRNA-targeted group-specific polynucleotide probes Applied and Environmental Microbiology Vol 61 3236-3244
MACKIE R 1 1996 The use of oligonucleotide probes to study the ecology of ruminal microbial populations Ann Zootech 45 Suppl 281-285 Elsevier INRA
MANUAL DE PROCEDIMIENTOS(2003) Laboratorio de Biotecnologiacutea Ruminal (BIORUM) Universidad Nacional de Colombia Sede Medelliacuten
MARTIN S A Nutrient transport by ruminal bacteria a review Journal of animal science 1994 74 3019-3031
MILLER G L (1959) Use of Dinitrosalisylic Acid Reagent for Determination of Reducing Sugar Anal Chem 426-428
MILLET L FONTY G GOUET P 1995 Study of the composition of the ruminal cellulolytic flora of bison and deer with rRNA-targetes oligonucleotide probes Ann Zootech 44 Supp 163 Elsevier INRA
MORITZ C AND HILLlS D M 1996 Molecular systematics contex and controversies In Molecular Systematic Edited by Hillis D M Moritz C and Mable B K Sunderland U S Sinauer Associates Segunda Edicioacuten p 1-13
MORRISON M 1996 Do ruminal bacteria exchange genetic material Journal of Dairy Science Vol 79 p 1476-1486
MURRAY AND THOMSON Q980 Citados por WILSON K Preparation of genomic DNA from bacteria In Short Protocols in Biology Molecular P 2-11 a 2-12
ODENYO A A MACKIE R l STHAL D A WHITE B A 1994a The use of 16S rRNAshytargeted oligonucleotide probes for Ruminococcus species and evidence for bacteriocin production Applied and Environmental Microbiolegy Vol 60 p 3688-3696
ODENYO A A MACKIE R l STHAL D A WHITE B A 1994b The use of 16S rRNAshytargeted oligonucleotide probes for study competition between ruminal fibrolitic bacteria pure culture-studies with cellulose and alkaline peroxide-trated wheat straw Applied and Environmental Microbiolegy Vol 60 p 3697-3703
OLSEN G J AND WOESE C R 1993 Ribosomal RNA a key to phlogeny FESAB Journal Vol 7 p 113-123
OSSA F RIDRIGUEZ J ARCOS M DIAZ T 1999 Identificacioacuten molecular de las bacterias celuloliacuteticas Fibrobacter sucinogenes y Ruminococcus flavefaciens Revista
--- - - ---~ - ------====-------------------------------------------------------------------------shy
77
Colombiana de Ciencias Pecuarias Memorias V Encuentro Nacional de Investigaciones de las Ciencias Pecuarias ENICIP Vol 12 P 120
PALMQVIST E AND HAN-HAGERDAL B (2000) Fermentation of lignocellulosic hydrolysates 1 inhibition and detoxification Bioresource Technology 74 17 - 24
PENtildeA Francisco 1982 El Rumen Aspectos de fisiologia y metabolismo microbioloacutegico Instituto colombiana Agropecuario ICA Divisioacuten de Ciencias Animales Bogotaacute P 78
PERCIVAL ZHANG YI-HENG AND LYND LEE R (2004) Toward an Agggregated Understanding of Enzymatic Hydrolysis of Cellulose Noncomplexed Cellulase Systems Biotechnology and Bioengineering vol 88 No 7 798 - 824
PRIMER 3 White head institutel MIl [Programa de computador] Disponible en internet www-genomewimiteducgi-binprimerprimer3-wwcgi
PROMEGA Manual tecnico Promega Kit wizard USA 1999
QIU XIAO (2000) Isolation and anaacutelisis of two cellulas cDNAs from Orpinomices joyonii Gene 245 119-126
RANADE D R AND GADRE R V 1988 Microbiological Aspects of Anaerobic Digestion Laboratory Manual Maharachtra Association for the Cultivation of Science Research Institute Pune India 107p
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EBERHARDT R Y GILBERT H J HAZLEWOOD G P (2000) Primary sequence and enzymes properties of two modular endoglucanases celA and Ce145A from the anaerobic fungus Piromyces equi Microbiology 146 1999 - 2008
FONTY G AND JOBLlN K N (1991) Rumen Anaerobic Fungi Their Role and Interactions with Other Rumen Microorganisms in Relation to Fiber Digestion in Physiological Aspects of Digestion and Metabolism in Rumiants Proceedings of the Seventh International Symposium on Ruminant Physiology
FORSBERG C W CRPSBY B and THOMAS D Y 1986 Potential for manipulation of the rumen fermentation through the use of recombinant DNA techniques Journal of Animal Science Vol 63 310-321
_ FUJII SIMONE ET AL (2004) Atividade fungistaacutetica in Vitro de cafeina em fungos asociados com graos de cafeacute Maringaacute vol 26 no 3 279 - 285
GACESA P HUBBLE (1990) Tecnologia de las Enzimas Ed Acribia- Espantildea Paacuteginas 1 shy25
GARCIA-VALLVEacute S ROMEU A AND PALAU J 2000 Horizontal gene transfer of glycosyl hydrolases of the rumen fungi Molecular Biology and Evolution Vol 17 W 3 P 352shy361
GARRYTI George BOONE David R and CASTENHOLZ Richard W Editores 2001 Bergeys Manual of Systematic Bacteriology Pg744
GHOSE T K (1987) Measurements of cellulase activities IUPAC Applied chemistry division commission on biotechnology Pure amp Appl Chem Vol 59 N 2 p 257 - 268
GILBERT HJ AND HASLEWOOD (1993) Bacterial cellulases and xylanases J of Gen Microb 139 187-194 Review
GILBERT HJ HAZLEWOOD GP LAURIE JI ORPIN CG XUE GP (1992) Homologous catalytic domains in a rumen fungal xylanase evidence for gene duplication and prokaryotic origino In Molecular Microbiology Vol 6 p 2065-2072
GIOVANNONI S J DeLONG E F OLSEN G J AND PACE N R 1988 Phylogenetic group specific oligodeoxinucleotide probe for identification of single microbial cells Journal of Bacteriology Vol 170 W 2 p720-726
GUETTLER MV RUMLER D and JAIN MK 1976 Actinobacillus succinogenes sp nov a novel succinic-acid-producing strain from the bovine rumen Applied and Environmental Microbiolegy Vol 32 Ndeg 4 P 623-632
HALL TA 1999 BioEdit a user-friendly biological sequence alignment editor and analysis program for Windows 9598NT Nucl Acids Symp Ser 41 95-98
HARHANGI HR FREELOVE AC UBHAYASEKERA W VAN DINTHER M STEENBAKKERS PJ AKHMANOVA A VAN DER DRIFT C JETTEN MS MOWBRAY SL GILBERT HJ OP DEN CAMP HJ (2003) CeI6A a major exoglucanasa from the cellulosome of the anaerobic fungi Piromyces sp E2 and Piromyces equi Biochimica et Biophysica Acta 1628 30 - 39
75
HEAD 1 M SAUNDERS J R AND PICKUP R W 1998 Microbial evolution diversity and ecology a decade of ribosomal RNA aalysis of uncultivate microorganisms Microbial Ecology Vol 35 P 1-21
HEBRAUD M FEVRE M (1988) Characterization of Glycoside Hydrolases Secreted by the Rumen Anaerobic Fungi Neocallimastix frontalis Sphaeromonas communis and Piromonas communis J of Gen Microb 134 1123-1129
HO W Y AND BARR Q D J S (1995) Classification of anaerobic gut fungi from herbivores whit emphasis on rumen fungi from Malaysia Mycologia 87(5) p 655 - 677
HO W Y KHOO 1 Y S TAN S G ABDULLAH N JALALUDIN S KUDO H (1994) Isozyme analysis of anaerobic rumen fungi and their relationship to aerobic chytrids Microbiology 140 1495-1504
HOBSON P N (1988) The rumen microbial ecosystem London and New York Elsevier Applied science 1989 p 530
HOLDEMAN LV EP CATO AND WEC MOORE (1977) Anaerobe Laboratory Manual 4 th Edition
HUNGATE R E (1966) The rumen and its microbes Ed Academic press New York and London 533 p
HUNGATE R E 1944 Studies on cellulose fermentation 1 The culture and physiology of an anaerobic cellulose-digesting bacterium Journal of Bacteriology Vol 48 p 499-513
INNIS M A AND GELFAND D H Optimization of PCRs In PCR protocols a guide to methods and applications 1990 Academic Press P 3-11
JANE A l BRYANT Marvin P and HASPELL R B 1982 Diurnal variations in bacterial numbers and fluid parameters in ruminal contents of animal fed low-or high-foraje diets Applied and Environmental Microbiolegy Vol 44 Ndeg 2 P 402-412
JEFFRIES T W 1987Production and applications of cellulase laboratory procedures USDA Forest products Laboratory Madison Wisconsin February
JOBLlN K N (1981) Isolation enumeration and maintenance of rumen anaerobic fungi in roll tube Applied and environmental microbiology p 119 - 1122
JULLlAND V VAUX A MILLET L FONTY G 1999 Identificaction of Rumnococcus favefaciens as the predominant cellulolytic bacterial species of the equine cecum Applied and Environmental Microbiolegy Vol 65 N 8 P 3738-3741
KARLlN S and ALTSCHUL S F (1990) Methods for assessing the statistical significance of molecular sequence features by using general scoring schemes Proc Natl Acad Sci USA 87 2264-2268
KOIKE S PAN J KOBAYASHI Y and TANAKA K 2003 Kinetics of in sacco fibershyattachment of representative ruminal cellulolytic bacteria monitored by competitive PCR Journal of Aniaml Science Vol 68 p 1429-1435
KOIKE Satoshi y KOBAYASHI Yasuo 2001 Development and use of competitive PCR assay for the rumen cellulolytic bacteria Fibrobacter succinogenes Ruminococcus albus and Ruminococcus favefaciens FEMS Microbiology Letter Vol 204 p361-366
LI X-L CHEN H LJUNGDAHL L G (1997 a) Two Cellulases Cel A and CelC from the polycentric Anaerobic Fungus Orpinomyces Strain PC-2 Contain N-Terminal Docking
76
Domains for a Cellulase-Hemicellulase Complex Appli And Env Microb 63(12) 4721-4728 Dec
LI X-L CHEN H LJUNGDAHL L G (1997 b) Monocentric and Polycentric Anaerobic Fungi Produce Structurally Related Cellulases and Xylanases Appl And Env Microb 63(2) 628shy635
UN Chuzao y STHAL David A 1995 Taxon-Specific Probes for the Cellulolitic Genus Fibrobacter Reveal Abundant and Novel EquineAssociated Populations En Applied and Environmental Microbiology Vol 61 Ndeg4 pg 1348-1351
-iquest LOWE S E THEODOROU M K AND TRINCI A P (1987) ) cellulases and Xylanases of an liexcl anaerobic rumen fungus grown on wheat straw wlleat---straw holocellulose cellulose and xylan Applied and environmental microbiology June p 1216 - 1223
LUDWIG W DORN S SPRINGER N KIRCHHOF G AND SCHLEIFER K H 1994 PCR Based preparation of 23S rRNA-targeted group-specific polynucleotide probes Applied and Environmental Microbiology Vol 61 3236-3244
MACKIE R 1 1996 The use of oligonucleotide probes to study the ecology of ruminal microbial populations Ann Zootech 45 Suppl 281-285 Elsevier INRA
MANUAL DE PROCEDIMIENTOS(2003) Laboratorio de Biotecnologiacutea Ruminal (BIORUM) Universidad Nacional de Colombia Sede Medelliacuten
MARTIN S A Nutrient transport by ruminal bacteria a review Journal of animal science 1994 74 3019-3031
MILLER G L (1959) Use of Dinitrosalisylic Acid Reagent for Determination of Reducing Sugar Anal Chem 426-428
MILLET L FONTY G GOUET P 1995 Study of the composition of the ruminal cellulolytic flora of bison and deer with rRNA-targetes oligonucleotide probes Ann Zootech 44 Supp 163 Elsevier INRA
MORITZ C AND HILLlS D M 1996 Molecular systematics contex and controversies In Molecular Systematic Edited by Hillis D M Moritz C and Mable B K Sunderland U S Sinauer Associates Segunda Edicioacuten p 1-13
MORRISON M 1996 Do ruminal bacteria exchange genetic material Journal of Dairy Science Vol 79 p 1476-1486
MURRAY AND THOMSON Q980 Citados por WILSON K Preparation of genomic DNA from bacteria In Short Protocols in Biology Molecular P 2-11 a 2-12
ODENYO A A MACKIE R l STHAL D A WHITE B A 1994a The use of 16S rRNAshytargeted oligonucleotide probes for Ruminococcus species and evidence for bacteriocin production Applied and Environmental Microbiolegy Vol 60 p 3688-3696
ODENYO A A MACKIE R l STHAL D A WHITE B A 1994b The use of 16S rRNAshytargeted oligonucleotide probes for study competition between ruminal fibrolitic bacteria pure culture-studies with cellulose and alkaline peroxide-trated wheat straw Applied and Environmental Microbiolegy Vol 60 p 3697-3703
OLSEN G J AND WOESE C R 1993 Ribosomal RNA a key to phlogeny FESAB Journal Vol 7 p 113-123
OSSA F RIDRIGUEZ J ARCOS M DIAZ T 1999 Identificacioacuten molecular de las bacterias celuloliacuteticas Fibrobacter sucinogenes y Ruminococcus flavefaciens Revista
--- - - ---~ - ------====-------------------------------------------------------------------------shy
77
Colombiana de Ciencias Pecuarias Memorias V Encuentro Nacional de Investigaciones de las Ciencias Pecuarias ENICIP Vol 12 P 120
PALMQVIST E AND HAN-HAGERDAL B (2000) Fermentation of lignocellulosic hydrolysates 1 inhibition and detoxification Bioresource Technology 74 17 - 24
PENtildeA Francisco 1982 El Rumen Aspectos de fisiologia y metabolismo microbioloacutegico Instituto colombiana Agropecuario ICA Divisioacuten de Ciencias Animales Bogotaacute P 78
PERCIVAL ZHANG YI-HENG AND LYND LEE R (2004) Toward an Agggregated Understanding of Enzymatic Hydrolysis of Cellulose Noncomplexed Cellulase Systems Biotechnology and Bioengineering vol 88 No 7 798 - 824
PRIMER 3 White head institutel MIl [Programa de computador] Disponible en internet www-genomewimiteducgi-binprimerprimer3-wwcgi
PROMEGA Manual tecnico Promega Kit wizard USA 1999
QIU XIAO (2000) Isolation and anaacutelisis of two cellulas cDNAs from Orpinomices joyonii Gene 245 119-126
RANADE D R AND GADRE R V 1988 Microbiological Aspects of Anaerobic Digestion Laboratory Manual Maharachtra Association for the Cultivation of Science Research Institute Pune India 107p
RANADE DR and GRADE RV Microbiological aspects of anaerobic digestion laboratory manual Edited Maharashtra Association for the Cultivation of Science In Research Institute Pune 1988 102 p
RAY S 1986 Nitrogen and energy nutrition of rumiant Departament of animal Science University of Florida Academic Press Inc
REZAEIAN MOHAMMAD et al (2005) Relative fibrolytic activities of anaerobic rumen fungi on untreated and sodium hydroxide treated barley straw in in vitro culture Anaerobe 11 163 -175
RODRIGUEZ V Fernando 1994 La microbiota ruminal y su importancia en la degradacioacuten de los forrajes tropicales En Seminario sobre Biotecnologia del Rumen Universidad Nacional de Colombia Sede Medelliacuten Nov 11-12
RODRIGUEZ DIAZ T (1996) Aislamiento patroacuten de fermentacioacuten de carbohidratos y caracterizacioacuten morfoloacutegica de bacterias celuloliacuteticas del rumen de bovinos alimentados con heno de Raigraacutes en Colombia En Revista CORPOICA Vol 1 p 123-28
ROLFS A SCHULLER l FINCKH O AND WEBER-ROLFS 1 PCR Clinical diagnostic and reserch Alemania 1992 P 271 Ed Springer-Verlag
SAELEY P G AND SOUTHERN E M Gel electrophoresis the DNA In Acids a Practical Approach Por Rickwoo and Hames IRL Press 1982 Sin mas datos
SAIKI DH GELFAND AND S STOFFEL 1988 Citado por KRAMER M E AND COEN D M Enzimatic amplification of DNA by PCR standard procedures and optimization In Short Protocols in Biology Molecularp 15-3 a 15-4
SHIMADA M TAKAHASHI M (1990) Biodegradation of Cellulosic Materials in Wood and Cellulosic Chemistry edited by David N -S Hon and Nobuo Shiraishi Marcel Dekker NY
78
SHOFIELD P PELL A N AND KRAUSE D O 1997 Molecular Beacons Trial of a fluorescence-based solution hybridization technique for ecological studies with ruminal bacteria Applied and Environmental Microbiology Vol 63 W 3 p1143-1147
SLEAT R AND MAH R A (1984) Quantitative method for colorimetric determination of formate in fermentation media Applied and environmental microbiology p 884 - 885
SRINIVASAN K MURAKAMI M NAKASHIMADA Y NISHIO N (2001) Efficient Production of Cellulolytic and Xylanolitic Enzymes by the Rumen Anaerobic Fungus Neocallimastix frontalis in a Repetead Batch Culture Journal of Bioscience and Bioengineering 91 2 153-158
STAHL D A FLESHER B MANSFIELD H R AND MONTGOMERY L 1988 Use of phylogeneticafly based hybridization probes for studies of ruminal microbial ecology Applied and Environmental Microbiology Vol 54(5) 1079-1084
STEENBAKKERS J M et al (2003) I3-Glucosidase in ceflulosome of the anaerobic fungus Piromyces sp strain E2 is a family 3 glycoside hydrolase Biochem J 370963 - 970
STEENBAKKERS PJ HARHANGI HR BOSSCHER MW VAN DER HOOFT MM KELTJENS JT VAN DER DRIFT C VOGELS GD OP DEN CAMP HJ (2003) ~shyGlucosidase in cellulosome of the anaerobic fungus Piromyces sp strain E2 is a family 3 glycoside hydrolase Biochem J 370 963 - 970
STEPHEN A MADDEN T SCHAFFER A lHANG J lHANG l MILLER W AND LlPMAN D 1997 Gapped BLAST and PSI-BLAST a new generation of protein database search programs Nucleic Acids Res Vol 25 p 3389-3402
STEWART C S 1991 The Rumen Bacteria in Rumen Microbial Metabolism and Rumiant Digestion Editd by J p Jouany INRA Editions Paris pp 131
STEWART C S BRYANT M P 1988 The rumen bacteria In The rumen microbial ecosystem Edited By Hobson p Elsevier Applied Science New York 21-26
STEWART C S PANIAGUA C DINSDALE D CHENG K-J AND GARRAW S H 1981 Selective isolation and characterization of Bacteriodes succinogenes from the rumen of a cow Applied and Environmental Microbiology Vol 41 W 2 P 504-510
TEUNISSEN M J OP DEN CAMP H J ORPIN CG HUIS JH VOGELS GD (1991) Comparison of growth characteristic of anaerobic fungi isolated from ruminant and nonshyruminant hervivores during cultivation in a defined medium J of Gen Microb 137 1401 shy1408
TEUNISSEN M J V M DE KORT G OP DEN CAMP H J M HUIS INT VELD J H J (1992) Production of cellulolytic and xylanolytic enzymes during growth of the anaerobic fungus Piromyces sp on different substrates J of Gen Microb 138 1657-1664 l
THEODOROU K M DAVIS R D NIELSEN B B LAWRENCE G l TRINCI P J (1990) Determination of growth of anaerobic fungi on soluble and cellulosic substrates using a pressure transducer Microbiology 141 671-678
THOMPSON JD HIGGINS DG and GIBSON TJ CLUSTAL W Improving the sensitivity of progressive multiple sequence alignment through sequence weighting positions-specific gap penalties and weight matrix choice In Nucleic Acids Research Vol 22 (1994) p 225
TSAI CHENG-FANG et al (2003) A comparative analysis of two cDNA clones of the f cellulose gene family from anaerobic fungus Piromyces rhizinflata Anaerobe 9 131 - 140
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BOHORQUEZ G Claudis M GAVIRIA M Gloria E Cambios cuantitativos de los microorganismos rum inales por la utilizacioacuten de flavophospholipol Tesis de grado 1991 Universidad Nacional de Colombia Sede Medelliacuten
BRIESACHER SL MAY T GRIGSBY KN KERLEY MS ANTHONY RV PATERSON JA (1992) Use of DNA probes to monitor nutritional effects on ruminal prokaryotes and Fibrobacter succinogenes S85 In Journal Animal Science Vol 70 p 289-295
BRIESACHER S L MAY T GRIGSBY K N KERLEY M S ANTHONY R V AND PATERSON J A 1992 Use of DNA probes to monitor nutritional efects on ruminal prokaryotes and fibrobacter succinogenes S85 Journal of Animal Science 70 289
BROWN T A Gen Cloning an introduction 1990 Great Britain Ed Edmundsbury Press Segunda Edicioacuten P 64-71
BRYANT M P Bacterial species of the rumen Bacteriologicla reviews 23 125 1959
BRYANT M P AND BURKEY L A 1953 Numbers and some predominant groups in the rumen of cows fed different ratios Journal of Animal Science Vol 36 p 218-224
CHENG J AND WEIMER P J 2001 Competition among three predominant ruminal cellulolytic bacteria in the absence or presence of non-cellulolytic bacteria Microbiology Vol 147 p 21-30
CHESSON A AND FORSBERG C W 1988 In The rumen Microbial Ecosystem Edites by P N Hobson Elsevier Science Publishers L TB Pp 251
CHURCH D C 1974 Fisiologia digestiva y nutricioacuten de los rumiantes Vol 1 Editorial Acribia Zaragoza Espantildea
COEN D M The polimerase chain reaction In Shorts Protocols in Molecular Biology P 15-4 a 15-5
COUGHLAN MICHAEL P (1990) Cellulose degradation by Fungi in Microbial Enzymes and Biotechnology Second edition Ed By William M Fogarty and Catherine 1 Kelly
COUTHINHO PEDRO M and Henrissat Bernard (1999) The Modular Structure of Cellulases and Other Carbohydrate-active Enzymes an Integrated Data Approach In Genettics Biochemistry and Ecology of Cellulose Degradation Ohmiya K et al eds Uni Publishers Co Tokyo pp 15 - 23
DANCOURT M BOLLET C CARLlOZ A MARTELlN R GAYRAL J-P AND RAOULT D 2000 16S ribosomal Dtla sequence analysis of a large collection of environmental and clinical unidentifiable bacterial isolates Journal of Clinical Microbiology Vol 38 W 10 p 3623-3630
DE LONG E F WICKHAM G S PACE N R 1989 Phylogenetic stains ribosomalshybased for the indentification of single cells Science Vol 243 1360-1363
DENMAN S XUE G-P PATEL B (1996) Characterization of a Neocallimastix patriciarum Cellulase cDNA (celA) Homologous to Trichoderma reesei Cellobiohydrolase 11 Appl And Env Microb 62(6) 1889-1896
DEVILLARD E NEWBOLD CJ SCOTT KP FORANO E WALLACE RJ JOUANY JP FLlNT HJ (1999) A xylanase produced by the rumen anaerobic protozoan Polyplastron multivesiculatum shows close sequence similarity to family 11 xylanases from Gram - positive bacteria In FEMS Microbiology Letters Vol 181 p 145-152
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DIJKERMAN R OP DEN CAMP HJM (1996) Cultivation of anaerobic fungi in a 10-L fermenter system for the production of (hemi) cellulolytic enzymes Appl Microbiol Biotechnol 4685-91
EBERHARDT R Y GILBERT H J HAZLEWOOD G P (2000) Primary sequence and enzymes properties of two modular endoglucanases celA and Ce145A from the anaerobic fungus Piromyces equi Microbiology 146 1999 - 2008
FONTY G AND JOBLlN K N (1991) Rumen Anaerobic Fungi Their Role and Interactions with Other Rumen Microorganisms in Relation to Fiber Digestion in Physiological Aspects of Digestion and Metabolism in Rumiants Proceedings of the Seventh International Symposium on Ruminant Physiology
FORSBERG C W CRPSBY B and THOMAS D Y 1986 Potential for manipulation of the rumen fermentation through the use of recombinant DNA techniques Journal of Animal Science Vol 63 310-321
_ FUJII SIMONE ET AL (2004) Atividade fungistaacutetica in Vitro de cafeina em fungos asociados com graos de cafeacute Maringaacute vol 26 no 3 279 - 285
GACESA P HUBBLE (1990) Tecnologia de las Enzimas Ed Acribia- Espantildea Paacuteginas 1 shy25
GARCIA-VALLVEacute S ROMEU A AND PALAU J 2000 Horizontal gene transfer of glycosyl hydrolases of the rumen fungi Molecular Biology and Evolution Vol 17 W 3 P 352shy361
GARRYTI George BOONE David R and CASTENHOLZ Richard W Editores 2001 Bergeys Manual of Systematic Bacteriology Pg744
GHOSE T K (1987) Measurements of cellulase activities IUPAC Applied chemistry division commission on biotechnology Pure amp Appl Chem Vol 59 N 2 p 257 - 268
GILBERT HJ AND HASLEWOOD (1993) Bacterial cellulases and xylanases J of Gen Microb 139 187-194 Review
GILBERT HJ HAZLEWOOD GP LAURIE JI ORPIN CG XUE GP (1992) Homologous catalytic domains in a rumen fungal xylanase evidence for gene duplication and prokaryotic origino In Molecular Microbiology Vol 6 p 2065-2072
GIOVANNONI S J DeLONG E F OLSEN G J AND PACE N R 1988 Phylogenetic group specific oligodeoxinucleotide probe for identification of single microbial cells Journal of Bacteriology Vol 170 W 2 p720-726
GUETTLER MV RUMLER D and JAIN MK 1976 Actinobacillus succinogenes sp nov a novel succinic-acid-producing strain from the bovine rumen Applied and Environmental Microbiolegy Vol 32 Ndeg 4 P 623-632
HALL TA 1999 BioEdit a user-friendly biological sequence alignment editor and analysis program for Windows 9598NT Nucl Acids Symp Ser 41 95-98
HARHANGI HR FREELOVE AC UBHAYASEKERA W VAN DINTHER M STEENBAKKERS PJ AKHMANOVA A VAN DER DRIFT C JETTEN MS MOWBRAY SL GILBERT HJ OP DEN CAMP HJ (2003) CeI6A a major exoglucanasa from the cellulosome of the anaerobic fungi Piromyces sp E2 and Piromyces equi Biochimica et Biophysica Acta 1628 30 - 39
75
HEAD 1 M SAUNDERS J R AND PICKUP R W 1998 Microbial evolution diversity and ecology a decade of ribosomal RNA aalysis of uncultivate microorganisms Microbial Ecology Vol 35 P 1-21
HEBRAUD M FEVRE M (1988) Characterization of Glycoside Hydrolases Secreted by the Rumen Anaerobic Fungi Neocallimastix frontalis Sphaeromonas communis and Piromonas communis J of Gen Microb 134 1123-1129
HO W Y AND BARR Q D J S (1995) Classification of anaerobic gut fungi from herbivores whit emphasis on rumen fungi from Malaysia Mycologia 87(5) p 655 - 677
HO W Y KHOO 1 Y S TAN S G ABDULLAH N JALALUDIN S KUDO H (1994) Isozyme analysis of anaerobic rumen fungi and their relationship to aerobic chytrids Microbiology 140 1495-1504
HOBSON P N (1988) The rumen microbial ecosystem London and New York Elsevier Applied science 1989 p 530
HOLDEMAN LV EP CATO AND WEC MOORE (1977) Anaerobe Laboratory Manual 4 th Edition
HUNGATE R E (1966) The rumen and its microbes Ed Academic press New York and London 533 p
HUNGATE R E 1944 Studies on cellulose fermentation 1 The culture and physiology of an anaerobic cellulose-digesting bacterium Journal of Bacteriology Vol 48 p 499-513
INNIS M A AND GELFAND D H Optimization of PCRs In PCR protocols a guide to methods and applications 1990 Academic Press P 3-11
JANE A l BRYANT Marvin P and HASPELL R B 1982 Diurnal variations in bacterial numbers and fluid parameters in ruminal contents of animal fed low-or high-foraje diets Applied and Environmental Microbiolegy Vol 44 Ndeg 2 P 402-412
JEFFRIES T W 1987Production and applications of cellulase laboratory procedures USDA Forest products Laboratory Madison Wisconsin February
JOBLlN K N (1981) Isolation enumeration and maintenance of rumen anaerobic fungi in roll tube Applied and environmental microbiology p 119 - 1122
JULLlAND V VAUX A MILLET L FONTY G 1999 Identificaction of Rumnococcus favefaciens as the predominant cellulolytic bacterial species of the equine cecum Applied and Environmental Microbiolegy Vol 65 N 8 P 3738-3741
KARLlN S and ALTSCHUL S F (1990) Methods for assessing the statistical significance of molecular sequence features by using general scoring schemes Proc Natl Acad Sci USA 87 2264-2268
KOIKE S PAN J KOBAYASHI Y and TANAKA K 2003 Kinetics of in sacco fibershyattachment of representative ruminal cellulolytic bacteria monitored by competitive PCR Journal of Aniaml Science Vol 68 p 1429-1435
KOIKE Satoshi y KOBAYASHI Yasuo 2001 Development and use of competitive PCR assay for the rumen cellulolytic bacteria Fibrobacter succinogenes Ruminococcus albus and Ruminococcus favefaciens FEMS Microbiology Letter Vol 204 p361-366
LI X-L CHEN H LJUNGDAHL L G (1997 a) Two Cellulases Cel A and CelC from the polycentric Anaerobic Fungus Orpinomyces Strain PC-2 Contain N-Terminal Docking
76
Domains for a Cellulase-Hemicellulase Complex Appli And Env Microb 63(12) 4721-4728 Dec
LI X-L CHEN H LJUNGDAHL L G (1997 b) Monocentric and Polycentric Anaerobic Fungi Produce Structurally Related Cellulases and Xylanases Appl And Env Microb 63(2) 628shy635
UN Chuzao y STHAL David A 1995 Taxon-Specific Probes for the Cellulolitic Genus Fibrobacter Reveal Abundant and Novel EquineAssociated Populations En Applied and Environmental Microbiology Vol 61 Ndeg4 pg 1348-1351
-iquest LOWE S E THEODOROU M K AND TRINCI A P (1987) ) cellulases and Xylanases of an liexcl anaerobic rumen fungus grown on wheat straw wlleat---straw holocellulose cellulose and xylan Applied and environmental microbiology June p 1216 - 1223
LUDWIG W DORN S SPRINGER N KIRCHHOF G AND SCHLEIFER K H 1994 PCR Based preparation of 23S rRNA-targeted group-specific polynucleotide probes Applied and Environmental Microbiology Vol 61 3236-3244
MACKIE R 1 1996 The use of oligonucleotide probes to study the ecology of ruminal microbial populations Ann Zootech 45 Suppl 281-285 Elsevier INRA
MANUAL DE PROCEDIMIENTOS(2003) Laboratorio de Biotecnologiacutea Ruminal (BIORUM) Universidad Nacional de Colombia Sede Medelliacuten
MARTIN S A Nutrient transport by ruminal bacteria a review Journal of animal science 1994 74 3019-3031
MILLER G L (1959) Use of Dinitrosalisylic Acid Reagent for Determination of Reducing Sugar Anal Chem 426-428
MILLET L FONTY G GOUET P 1995 Study of the composition of the ruminal cellulolytic flora of bison and deer with rRNA-targetes oligonucleotide probes Ann Zootech 44 Supp 163 Elsevier INRA
MORITZ C AND HILLlS D M 1996 Molecular systematics contex and controversies In Molecular Systematic Edited by Hillis D M Moritz C and Mable B K Sunderland U S Sinauer Associates Segunda Edicioacuten p 1-13
MORRISON M 1996 Do ruminal bacteria exchange genetic material Journal of Dairy Science Vol 79 p 1476-1486
MURRAY AND THOMSON Q980 Citados por WILSON K Preparation of genomic DNA from bacteria In Short Protocols in Biology Molecular P 2-11 a 2-12
ODENYO A A MACKIE R l STHAL D A WHITE B A 1994a The use of 16S rRNAshytargeted oligonucleotide probes for Ruminococcus species and evidence for bacteriocin production Applied and Environmental Microbiolegy Vol 60 p 3688-3696
ODENYO A A MACKIE R l STHAL D A WHITE B A 1994b The use of 16S rRNAshytargeted oligonucleotide probes for study competition between ruminal fibrolitic bacteria pure culture-studies with cellulose and alkaline peroxide-trated wheat straw Applied and Environmental Microbiolegy Vol 60 p 3697-3703
OLSEN G J AND WOESE C R 1993 Ribosomal RNA a key to phlogeny FESAB Journal Vol 7 p 113-123
OSSA F RIDRIGUEZ J ARCOS M DIAZ T 1999 Identificacioacuten molecular de las bacterias celuloliacuteticas Fibrobacter sucinogenes y Ruminococcus flavefaciens Revista
--- - - ---~ - ------====-------------------------------------------------------------------------shy
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Colombiana de Ciencias Pecuarias Memorias V Encuentro Nacional de Investigaciones de las Ciencias Pecuarias ENICIP Vol 12 P 120
PALMQVIST E AND HAN-HAGERDAL B (2000) Fermentation of lignocellulosic hydrolysates 1 inhibition and detoxification Bioresource Technology 74 17 - 24
PENtildeA Francisco 1982 El Rumen Aspectos de fisiologia y metabolismo microbioloacutegico Instituto colombiana Agropecuario ICA Divisioacuten de Ciencias Animales Bogotaacute P 78
PERCIVAL ZHANG YI-HENG AND LYND LEE R (2004) Toward an Agggregated Understanding of Enzymatic Hydrolysis of Cellulose Noncomplexed Cellulase Systems Biotechnology and Bioengineering vol 88 No 7 798 - 824
PRIMER 3 White head institutel MIl [Programa de computador] Disponible en internet www-genomewimiteducgi-binprimerprimer3-wwcgi
PROMEGA Manual tecnico Promega Kit wizard USA 1999
QIU XIAO (2000) Isolation and anaacutelisis of two cellulas cDNAs from Orpinomices joyonii Gene 245 119-126
RANADE D R AND GADRE R V 1988 Microbiological Aspects of Anaerobic Digestion Laboratory Manual Maharachtra Association for the Cultivation of Science Research Institute Pune India 107p
RANADE DR and GRADE RV Microbiological aspects of anaerobic digestion laboratory manual Edited Maharashtra Association for the Cultivation of Science In Research Institute Pune 1988 102 p
RAY S 1986 Nitrogen and energy nutrition of rumiant Departament of animal Science University of Florida Academic Press Inc
REZAEIAN MOHAMMAD et al (2005) Relative fibrolytic activities of anaerobic rumen fungi on untreated and sodium hydroxide treated barley straw in in vitro culture Anaerobe 11 163 -175
RODRIGUEZ V Fernando 1994 La microbiota ruminal y su importancia en la degradacioacuten de los forrajes tropicales En Seminario sobre Biotecnologia del Rumen Universidad Nacional de Colombia Sede Medelliacuten Nov 11-12
RODRIGUEZ DIAZ T (1996) Aislamiento patroacuten de fermentacioacuten de carbohidratos y caracterizacioacuten morfoloacutegica de bacterias celuloliacuteticas del rumen de bovinos alimentados con heno de Raigraacutes en Colombia En Revista CORPOICA Vol 1 p 123-28
ROLFS A SCHULLER l FINCKH O AND WEBER-ROLFS 1 PCR Clinical diagnostic and reserch Alemania 1992 P 271 Ed Springer-Verlag
SAELEY P G AND SOUTHERN E M Gel electrophoresis the DNA In Acids a Practical Approach Por Rickwoo and Hames IRL Press 1982 Sin mas datos
SAIKI DH GELFAND AND S STOFFEL 1988 Citado por KRAMER M E AND COEN D M Enzimatic amplification of DNA by PCR standard procedures and optimization In Short Protocols in Biology Molecularp 15-3 a 15-4
SHIMADA M TAKAHASHI M (1990) Biodegradation of Cellulosic Materials in Wood and Cellulosic Chemistry edited by David N -S Hon and Nobuo Shiraishi Marcel Dekker NY
78
SHOFIELD P PELL A N AND KRAUSE D O 1997 Molecular Beacons Trial of a fluorescence-based solution hybridization technique for ecological studies with ruminal bacteria Applied and Environmental Microbiology Vol 63 W 3 p1143-1147
SLEAT R AND MAH R A (1984) Quantitative method for colorimetric determination of formate in fermentation media Applied and environmental microbiology p 884 - 885
SRINIVASAN K MURAKAMI M NAKASHIMADA Y NISHIO N (2001) Efficient Production of Cellulolytic and Xylanolitic Enzymes by the Rumen Anaerobic Fungus Neocallimastix frontalis in a Repetead Batch Culture Journal of Bioscience and Bioengineering 91 2 153-158
STAHL D A FLESHER B MANSFIELD H R AND MONTGOMERY L 1988 Use of phylogeneticafly based hybridization probes for studies of ruminal microbial ecology Applied and Environmental Microbiology Vol 54(5) 1079-1084
STEENBAKKERS J M et al (2003) I3-Glucosidase in ceflulosome of the anaerobic fungus Piromyces sp strain E2 is a family 3 glycoside hydrolase Biochem J 370963 - 970
STEENBAKKERS PJ HARHANGI HR BOSSCHER MW VAN DER HOOFT MM KELTJENS JT VAN DER DRIFT C VOGELS GD OP DEN CAMP HJ (2003) ~shyGlucosidase in cellulosome of the anaerobic fungus Piromyces sp strain E2 is a family 3 glycoside hydrolase Biochem J 370 963 - 970
STEPHEN A MADDEN T SCHAFFER A lHANG J lHANG l MILLER W AND LlPMAN D 1997 Gapped BLAST and PSI-BLAST a new generation of protein database search programs Nucleic Acids Res Vol 25 p 3389-3402
STEWART C S 1991 The Rumen Bacteria in Rumen Microbial Metabolism and Rumiant Digestion Editd by J p Jouany INRA Editions Paris pp 131
STEWART C S BRYANT M P 1988 The rumen bacteria In The rumen microbial ecosystem Edited By Hobson p Elsevier Applied Science New York 21-26
STEWART C S PANIAGUA C DINSDALE D CHENG K-J AND GARRAW S H 1981 Selective isolation and characterization of Bacteriodes succinogenes from the rumen of a cow Applied and Environmental Microbiology Vol 41 W 2 P 504-510
TEUNISSEN M J OP DEN CAMP H J ORPIN CG HUIS JH VOGELS GD (1991) Comparison of growth characteristic of anaerobic fungi isolated from ruminant and nonshyruminant hervivores during cultivation in a defined medium J of Gen Microb 137 1401 shy1408
TEUNISSEN M J V M DE KORT G OP DEN CAMP H J M HUIS INT VELD J H J (1992) Production of cellulolytic and xylanolytic enzymes during growth of the anaerobic fungus Piromyces sp on different substrates J of Gen Microb 138 1657-1664 l
THEODOROU K M DAVIS R D NIELSEN B B LAWRENCE G l TRINCI P J (1990) Determination of growth of anaerobic fungi on soluble and cellulosic substrates using a pressure transducer Microbiology 141 671-678
THOMPSON JD HIGGINS DG and GIBSON TJ CLUSTAL W Improving the sensitivity of progressive multiple sequence alignment through sequence weighting positions-specific gap penalties and weight matrix choice In Nucleic Acids Research Vol 22 (1994) p 225
TSAI CHENG-FANG et al (2003) A comparative analysis of two cDNA clones of the f cellulose gene family from anaerobic fungus Piromyces rhizinflata Anaerobe 9 131 - 140
74
- ------
DIJKERMAN R OP DEN CAMP HJM (1996) Cultivation of anaerobic fungi in a 10-L fermenter system for the production of (hemi) cellulolytic enzymes Appl Microbiol Biotechnol 4685-91
EBERHARDT R Y GILBERT H J HAZLEWOOD G P (2000) Primary sequence and enzymes properties of two modular endoglucanases celA and Ce145A from the anaerobic fungus Piromyces equi Microbiology 146 1999 - 2008
FONTY G AND JOBLlN K N (1991) Rumen Anaerobic Fungi Their Role and Interactions with Other Rumen Microorganisms in Relation to Fiber Digestion in Physiological Aspects of Digestion and Metabolism in Rumiants Proceedings of the Seventh International Symposium on Ruminant Physiology
FORSBERG C W CRPSBY B and THOMAS D Y 1986 Potential for manipulation of the rumen fermentation through the use of recombinant DNA techniques Journal of Animal Science Vol 63 310-321
_ FUJII SIMONE ET AL (2004) Atividade fungistaacutetica in Vitro de cafeina em fungos asociados com graos de cafeacute Maringaacute vol 26 no 3 279 - 285
GACESA P HUBBLE (1990) Tecnologia de las Enzimas Ed Acribia- Espantildea Paacuteginas 1 shy25
GARCIA-VALLVEacute S ROMEU A AND PALAU J 2000 Horizontal gene transfer of glycosyl hydrolases of the rumen fungi Molecular Biology and Evolution Vol 17 W 3 P 352shy361
GARRYTI George BOONE David R and CASTENHOLZ Richard W Editores 2001 Bergeys Manual of Systematic Bacteriology Pg744
GHOSE T K (1987) Measurements of cellulase activities IUPAC Applied chemistry division commission on biotechnology Pure amp Appl Chem Vol 59 N 2 p 257 - 268
GILBERT HJ AND HASLEWOOD (1993) Bacterial cellulases and xylanases J of Gen Microb 139 187-194 Review
GILBERT HJ HAZLEWOOD GP LAURIE JI ORPIN CG XUE GP (1992) Homologous catalytic domains in a rumen fungal xylanase evidence for gene duplication and prokaryotic origino In Molecular Microbiology Vol 6 p 2065-2072
GIOVANNONI S J DeLONG E F OLSEN G J AND PACE N R 1988 Phylogenetic group specific oligodeoxinucleotide probe for identification of single microbial cells Journal of Bacteriology Vol 170 W 2 p720-726
GUETTLER MV RUMLER D and JAIN MK 1976 Actinobacillus succinogenes sp nov a novel succinic-acid-producing strain from the bovine rumen Applied and Environmental Microbiolegy Vol 32 Ndeg 4 P 623-632
HALL TA 1999 BioEdit a user-friendly biological sequence alignment editor and analysis program for Windows 9598NT Nucl Acids Symp Ser 41 95-98
HARHANGI HR FREELOVE AC UBHAYASEKERA W VAN DINTHER M STEENBAKKERS PJ AKHMANOVA A VAN DER DRIFT C JETTEN MS MOWBRAY SL GILBERT HJ OP DEN CAMP HJ (2003) CeI6A a major exoglucanasa from the cellulosome of the anaerobic fungi Piromyces sp E2 and Piromyces equi Biochimica et Biophysica Acta 1628 30 - 39
75
HEAD 1 M SAUNDERS J R AND PICKUP R W 1998 Microbial evolution diversity and ecology a decade of ribosomal RNA aalysis of uncultivate microorganisms Microbial Ecology Vol 35 P 1-21
HEBRAUD M FEVRE M (1988) Characterization of Glycoside Hydrolases Secreted by the Rumen Anaerobic Fungi Neocallimastix frontalis Sphaeromonas communis and Piromonas communis J of Gen Microb 134 1123-1129
HO W Y AND BARR Q D J S (1995) Classification of anaerobic gut fungi from herbivores whit emphasis on rumen fungi from Malaysia Mycologia 87(5) p 655 - 677
HO W Y KHOO 1 Y S TAN S G ABDULLAH N JALALUDIN S KUDO H (1994) Isozyme analysis of anaerobic rumen fungi and their relationship to aerobic chytrids Microbiology 140 1495-1504
HOBSON P N (1988) The rumen microbial ecosystem London and New York Elsevier Applied science 1989 p 530
HOLDEMAN LV EP CATO AND WEC MOORE (1977) Anaerobe Laboratory Manual 4 th Edition
HUNGATE R E (1966) The rumen and its microbes Ed Academic press New York and London 533 p
HUNGATE R E 1944 Studies on cellulose fermentation 1 The culture and physiology of an anaerobic cellulose-digesting bacterium Journal of Bacteriology Vol 48 p 499-513
INNIS M A AND GELFAND D H Optimization of PCRs In PCR protocols a guide to methods and applications 1990 Academic Press P 3-11
JANE A l BRYANT Marvin P and HASPELL R B 1982 Diurnal variations in bacterial numbers and fluid parameters in ruminal contents of animal fed low-or high-foraje diets Applied and Environmental Microbiolegy Vol 44 Ndeg 2 P 402-412
JEFFRIES T W 1987Production and applications of cellulase laboratory procedures USDA Forest products Laboratory Madison Wisconsin February
JOBLlN K N (1981) Isolation enumeration and maintenance of rumen anaerobic fungi in roll tube Applied and environmental microbiology p 119 - 1122
JULLlAND V VAUX A MILLET L FONTY G 1999 Identificaction of Rumnococcus favefaciens as the predominant cellulolytic bacterial species of the equine cecum Applied and Environmental Microbiolegy Vol 65 N 8 P 3738-3741
KARLlN S and ALTSCHUL S F (1990) Methods for assessing the statistical significance of molecular sequence features by using general scoring schemes Proc Natl Acad Sci USA 87 2264-2268
KOIKE S PAN J KOBAYASHI Y and TANAKA K 2003 Kinetics of in sacco fibershyattachment of representative ruminal cellulolytic bacteria monitored by competitive PCR Journal of Aniaml Science Vol 68 p 1429-1435
KOIKE Satoshi y KOBAYASHI Yasuo 2001 Development and use of competitive PCR assay for the rumen cellulolytic bacteria Fibrobacter succinogenes Ruminococcus albus and Ruminococcus favefaciens FEMS Microbiology Letter Vol 204 p361-366
LI X-L CHEN H LJUNGDAHL L G (1997 a) Two Cellulases Cel A and CelC from the polycentric Anaerobic Fungus Orpinomyces Strain PC-2 Contain N-Terminal Docking
76
Domains for a Cellulase-Hemicellulase Complex Appli And Env Microb 63(12) 4721-4728 Dec
LI X-L CHEN H LJUNGDAHL L G (1997 b) Monocentric and Polycentric Anaerobic Fungi Produce Structurally Related Cellulases and Xylanases Appl And Env Microb 63(2) 628shy635
UN Chuzao y STHAL David A 1995 Taxon-Specific Probes for the Cellulolitic Genus Fibrobacter Reveal Abundant and Novel EquineAssociated Populations En Applied and Environmental Microbiology Vol 61 Ndeg4 pg 1348-1351
-iquest LOWE S E THEODOROU M K AND TRINCI A P (1987) ) cellulases and Xylanases of an liexcl anaerobic rumen fungus grown on wheat straw wlleat---straw holocellulose cellulose and xylan Applied and environmental microbiology June p 1216 - 1223
LUDWIG W DORN S SPRINGER N KIRCHHOF G AND SCHLEIFER K H 1994 PCR Based preparation of 23S rRNA-targeted group-specific polynucleotide probes Applied and Environmental Microbiology Vol 61 3236-3244
MACKIE R 1 1996 The use of oligonucleotide probes to study the ecology of ruminal microbial populations Ann Zootech 45 Suppl 281-285 Elsevier INRA
MANUAL DE PROCEDIMIENTOS(2003) Laboratorio de Biotecnologiacutea Ruminal (BIORUM) Universidad Nacional de Colombia Sede Medelliacuten
MARTIN S A Nutrient transport by ruminal bacteria a review Journal of animal science 1994 74 3019-3031
MILLER G L (1959) Use of Dinitrosalisylic Acid Reagent for Determination of Reducing Sugar Anal Chem 426-428
MILLET L FONTY G GOUET P 1995 Study of the composition of the ruminal cellulolytic flora of bison and deer with rRNA-targetes oligonucleotide probes Ann Zootech 44 Supp 163 Elsevier INRA
MORITZ C AND HILLlS D M 1996 Molecular systematics contex and controversies In Molecular Systematic Edited by Hillis D M Moritz C and Mable B K Sunderland U S Sinauer Associates Segunda Edicioacuten p 1-13
MORRISON M 1996 Do ruminal bacteria exchange genetic material Journal of Dairy Science Vol 79 p 1476-1486
MURRAY AND THOMSON Q980 Citados por WILSON K Preparation of genomic DNA from bacteria In Short Protocols in Biology Molecular P 2-11 a 2-12
ODENYO A A MACKIE R l STHAL D A WHITE B A 1994a The use of 16S rRNAshytargeted oligonucleotide probes for Ruminococcus species and evidence for bacteriocin production Applied and Environmental Microbiolegy Vol 60 p 3688-3696
ODENYO A A MACKIE R l STHAL D A WHITE B A 1994b The use of 16S rRNAshytargeted oligonucleotide probes for study competition between ruminal fibrolitic bacteria pure culture-studies with cellulose and alkaline peroxide-trated wheat straw Applied and Environmental Microbiolegy Vol 60 p 3697-3703
OLSEN G J AND WOESE C R 1993 Ribosomal RNA a key to phlogeny FESAB Journal Vol 7 p 113-123
OSSA F RIDRIGUEZ J ARCOS M DIAZ T 1999 Identificacioacuten molecular de las bacterias celuloliacuteticas Fibrobacter sucinogenes y Ruminococcus flavefaciens Revista
--- - - ---~ - ------====-------------------------------------------------------------------------shy
77
Colombiana de Ciencias Pecuarias Memorias V Encuentro Nacional de Investigaciones de las Ciencias Pecuarias ENICIP Vol 12 P 120
PALMQVIST E AND HAN-HAGERDAL B (2000) Fermentation of lignocellulosic hydrolysates 1 inhibition and detoxification Bioresource Technology 74 17 - 24
PENtildeA Francisco 1982 El Rumen Aspectos de fisiologia y metabolismo microbioloacutegico Instituto colombiana Agropecuario ICA Divisioacuten de Ciencias Animales Bogotaacute P 78
PERCIVAL ZHANG YI-HENG AND LYND LEE R (2004) Toward an Agggregated Understanding of Enzymatic Hydrolysis of Cellulose Noncomplexed Cellulase Systems Biotechnology and Bioengineering vol 88 No 7 798 - 824
PRIMER 3 White head institutel MIl [Programa de computador] Disponible en internet www-genomewimiteducgi-binprimerprimer3-wwcgi
PROMEGA Manual tecnico Promega Kit wizard USA 1999
QIU XIAO (2000) Isolation and anaacutelisis of two cellulas cDNAs from Orpinomices joyonii Gene 245 119-126
RANADE D R AND GADRE R V 1988 Microbiological Aspects of Anaerobic Digestion Laboratory Manual Maharachtra Association for the Cultivation of Science Research Institute Pune India 107p
RANADE DR and GRADE RV Microbiological aspects of anaerobic digestion laboratory manual Edited Maharashtra Association for the Cultivation of Science In Research Institute Pune 1988 102 p
RAY S 1986 Nitrogen and energy nutrition of rumiant Departament of animal Science University of Florida Academic Press Inc
REZAEIAN MOHAMMAD et al (2005) Relative fibrolytic activities of anaerobic rumen fungi on untreated and sodium hydroxide treated barley straw in in vitro culture Anaerobe 11 163 -175
RODRIGUEZ V Fernando 1994 La microbiota ruminal y su importancia en la degradacioacuten de los forrajes tropicales En Seminario sobre Biotecnologia del Rumen Universidad Nacional de Colombia Sede Medelliacuten Nov 11-12
RODRIGUEZ DIAZ T (1996) Aislamiento patroacuten de fermentacioacuten de carbohidratos y caracterizacioacuten morfoloacutegica de bacterias celuloliacuteticas del rumen de bovinos alimentados con heno de Raigraacutes en Colombia En Revista CORPOICA Vol 1 p 123-28
ROLFS A SCHULLER l FINCKH O AND WEBER-ROLFS 1 PCR Clinical diagnostic and reserch Alemania 1992 P 271 Ed Springer-Verlag
SAELEY P G AND SOUTHERN E M Gel electrophoresis the DNA In Acids a Practical Approach Por Rickwoo and Hames IRL Press 1982 Sin mas datos
SAIKI DH GELFAND AND S STOFFEL 1988 Citado por KRAMER M E AND COEN D M Enzimatic amplification of DNA by PCR standard procedures and optimization In Short Protocols in Biology Molecularp 15-3 a 15-4
SHIMADA M TAKAHASHI M (1990) Biodegradation of Cellulosic Materials in Wood and Cellulosic Chemistry edited by David N -S Hon and Nobuo Shiraishi Marcel Dekker NY
78
SHOFIELD P PELL A N AND KRAUSE D O 1997 Molecular Beacons Trial of a fluorescence-based solution hybridization technique for ecological studies with ruminal bacteria Applied and Environmental Microbiology Vol 63 W 3 p1143-1147
SLEAT R AND MAH R A (1984) Quantitative method for colorimetric determination of formate in fermentation media Applied and environmental microbiology p 884 - 885
SRINIVASAN K MURAKAMI M NAKASHIMADA Y NISHIO N (2001) Efficient Production of Cellulolytic and Xylanolitic Enzymes by the Rumen Anaerobic Fungus Neocallimastix frontalis in a Repetead Batch Culture Journal of Bioscience and Bioengineering 91 2 153-158
STAHL D A FLESHER B MANSFIELD H R AND MONTGOMERY L 1988 Use of phylogeneticafly based hybridization probes for studies of ruminal microbial ecology Applied and Environmental Microbiology Vol 54(5) 1079-1084
STEENBAKKERS J M et al (2003) I3-Glucosidase in ceflulosome of the anaerobic fungus Piromyces sp strain E2 is a family 3 glycoside hydrolase Biochem J 370963 - 970
STEENBAKKERS PJ HARHANGI HR BOSSCHER MW VAN DER HOOFT MM KELTJENS JT VAN DER DRIFT C VOGELS GD OP DEN CAMP HJ (2003) ~shyGlucosidase in cellulosome of the anaerobic fungus Piromyces sp strain E2 is a family 3 glycoside hydrolase Biochem J 370 963 - 970
STEPHEN A MADDEN T SCHAFFER A lHANG J lHANG l MILLER W AND LlPMAN D 1997 Gapped BLAST and PSI-BLAST a new generation of protein database search programs Nucleic Acids Res Vol 25 p 3389-3402
STEWART C S 1991 The Rumen Bacteria in Rumen Microbial Metabolism and Rumiant Digestion Editd by J p Jouany INRA Editions Paris pp 131
STEWART C S BRYANT M P 1988 The rumen bacteria In The rumen microbial ecosystem Edited By Hobson p Elsevier Applied Science New York 21-26
STEWART C S PANIAGUA C DINSDALE D CHENG K-J AND GARRAW S H 1981 Selective isolation and characterization of Bacteriodes succinogenes from the rumen of a cow Applied and Environmental Microbiology Vol 41 W 2 P 504-510
TEUNISSEN M J OP DEN CAMP H J ORPIN CG HUIS JH VOGELS GD (1991) Comparison of growth characteristic of anaerobic fungi isolated from ruminant and nonshyruminant hervivores during cultivation in a defined medium J of Gen Microb 137 1401 shy1408
TEUNISSEN M J V M DE KORT G OP DEN CAMP H J M HUIS INT VELD J H J (1992) Production of cellulolytic and xylanolytic enzymes during growth of the anaerobic fungus Piromyces sp on different substrates J of Gen Microb 138 1657-1664 l
THEODOROU K M DAVIS R D NIELSEN B B LAWRENCE G l TRINCI P J (1990) Determination of growth of anaerobic fungi on soluble and cellulosic substrates using a pressure transducer Microbiology 141 671-678
THOMPSON JD HIGGINS DG and GIBSON TJ CLUSTAL W Improving the sensitivity of progressive multiple sequence alignment through sequence weighting positions-specific gap penalties and weight matrix choice In Nucleic Acids Research Vol 22 (1994) p 225
TSAI CHENG-FANG et al (2003) A comparative analysis of two cDNA clones of the f cellulose gene family from anaerobic fungus Piromyces rhizinflata Anaerobe 9 131 - 140
75
HEAD 1 M SAUNDERS J R AND PICKUP R W 1998 Microbial evolution diversity and ecology a decade of ribosomal RNA aalysis of uncultivate microorganisms Microbial Ecology Vol 35 P 1-21
HEBRAUD M FEVRE M (1988) Characterization of Glycoside Hydrolases Secreted by the Rumen Anaerobic Fungi Neocallimastix frontalis Sphaeromonas communis and Piromonas communis J of Gen Microb 134 1123-1129
HO W Y AND BARR Q D J S (1995) Classification of anaerobic gut fungi from herbivores whit emphasis on rumen fungi from Malaysia Mycologia 87(5) p 655 - 677
HO W Y KHOO 1 Y S TAN S G ABDULLAH N JALALUDIN S KUDO H (1994) Isozyme analysis of anaerobic rumen fungi and their relationship to aerobic chytrids Microbiology 140 1495-1504
HOBSON P N (1988) The rumen microbial ecosystem London and New York Elsevier Applied science 1989 p 530
HOLDEMAN LV EP CATO AND WEC MOORE (1977) Anaerobe Laboratory Manual 4 th Edition
HUNGATE R E (1966) The rumen and its microbes Ed Academic press New York and London 533 p
HUNGATE R E 1944 Studies on cellulose fermentation 1 The culture and physiology of an anaerobic cellulose-digesting bacterium Journal of Bacteriology Vol 48 p 499-513
INNIS M A AND GELFAND D H Optimization of PCRs In PCR protocols a guide to methods and applications 1990 Academic Press P 3-11
JANE A l BRYANT Marvin P and HASPELL R B 1982 Diurnal variations in bacterial numbers and fluid parameters in ruminal contents of animal fed low-or high-foraje diets Applied and Environmental Microbiolegy Vol 44 Ndeg 2 P 402-412
JEFFRIES T W 1987Production and applications of cellulase laboratory procedures USDA Forest products Laboratory Madison Wisconsin February
JOBLlN K N (1981) Isolation enumeration and maintenance of rumen anaerobic fungi in roll tube Applied and environmental microbiology p 119 - 1122
JULLlAND V VAUX A MILLET L FONTY G 1999 Identificaction of Rumnococcus favefaciens as the predominant cellulolytic bacterial species of the equine cecum Applied and Environmental Microbiolegy Vol 65 N 8 P 3738-3741
KARLlN S and ALTSCHUL S F (1990) Methods for assessing the statistical significance of molecular sequence features by using general scoring schemes Proc Natl Acad Sci USA 87 2264-2268
KOIKE S PAN J KOBAYASHI Y and TANAKA K 2003 Kinetics of in sacco fibershyattachment of representative ruminal cellulolytic bacteria monitored by competitive PCR Journal of Aniaml Science Vol 68 p 1429-1435
KOIKE Satoshi y KOBAYASHI Yasuo 2001 Development and use of competitive PCR assay for the rumen cellulolytic bacteria Fibrobacter succinogenes Ruminococcus albus and Ruminococcus favefaciens FEMS Microbiology Letter Vol 204 p361-366
LI X-L CHEN H LJUNGDAHL L G (1997 a) Two Cellulases Cel A and CelC from the polycentric Anaerobic Fungus Orpinomyces Strain PC-2 Contain N-Terminal Docking
76
Domains for a Cellulase-Hemicellulase Complex Appli And Env Microb 63(12) 4721-4728 Dec
LI X-L CHEN H LJUNGDAHL L G (1997 b) Monocentric and Polycentric Anaerobic Fungi Produce Structurally Related Cellulases and Xylanases Appl And Env Microb 63(2) 628shy635
UN Chuzao y STHAL David A 1995 Taxon-Specific Probes for the Cellulolitic Genus Fibrobacter Reveal Abundant and Novel EquineAssociated Populations En Applied and Environmental Microbiology Vol 61 Ndeg4 pg 1348-1351
-iquest LOWE S E THEODOROU M K AND TRINCI A P (1987) ) cellulases and Xylanases of an liexcl anaerobic rumen fungus grown on wheat straw wlleat---straw holocellulose cellulose and xylan Applied and environmental microbiology June p 1216 - 1223
LUDWIG W DORN S SPRINGER N KIRCHHOF G AND SCHLEIFER K H 1994 PCR Based preparation of 23S rRNA-targeted group-specific polynucleotide probes Applied and Environmental Microbiology Vol 61 3236-3244
MACKIE R 1 1996 The use of oligonucleotide probes to study the ecology of ruminal microbial populations Ann Zootech 45 Suppl 281-285 Elsevier INRA
MANUAL DE PROCEDIMIENTOS(2003) Laboratorio de Biotecnologiacutea Ruminal (BIORUM) Universidad Nacional de Colombia Sede Medelliacuten
MARTIN S A Nutrient transport by ruminal bacteria a review Journal of animal science 1994 74 3019-3031
MILLER G L (1959) Use of Dinitrosalisylic Acid Reagent for Determination of Reducing Sugar Anal Chem 426-428
MILLET L FONTY G GOUET P 1995 Study of the composition of the ruminal cellulolytic flora of bison and deer with rRNA-targetes oligonucleotide probes Ann Zootech 44 Supp 163 Elsevier INRA
MORITZ C AND HILLlS D M 1996 Molecular systematics contex and controversies In Molecular Systematic Edited by Hillis D M Moritz C and Mable B K Sunderland U S Sinauer Associates Segunda Edicioacuten p 1-13
MORRISON M 1996 Do ruminal bacteria exchange genetic material Journal of Dairy Science Vol 79 p 1476-1486
MURRAY AND THOMSON Q980 Citados por WILSON K Preparation of genomic DNA from bacteria In Short Protocols in Biology Molecular P 2-11 a 2-12
ODENYO A A MACKIE R l STHAL D A WHITE B A 1994a The use of 16S rRNAshytargeted oligonucleotide probes for Ruminococcus species and evidence for bacteriocin production Applied and Environmental Microbiolegy Vol 60 p 3688-3696
ODENYO A A MACKIE R l STHAL D A WHITE B A 1994b The use of 16S rRNAshytargeted oligonucleotide probes for study competition between ruminal fibrolitic bacteria pure culture-studies with cellulose and alkaline peroxide-trated wheat straw Applied and Environmental Microbiolegy Vol 60 p 3697-3703
OLSEN G J AND WOESE C R 1993 Ribosomal RNA a key to phlogeny FESAB Journal Vol 7 p 113-123
OSSA F RIDRIGUEZ J ARCOS M DIAZ T 1999 Identificacioacuten molecular de las bacterias celuloliacuteticas Fibrobacter sucinogenes y Ruminococcus flavefaciens Revista
--- - - ---~ - ------====-------------------------------------------------------------------------shy
77
Colombiana de Ciencias Pecuarias Memorias V Encuentro Nacional de Investigaciones de las Ciencias Pecuarias ENICIP Vol 12 P 120
PALMQVIST E AND HAN-HAGERDAL B (2000) Fermentation of lignocellulosic hydrolysates 1 inhibition and detoxification Bioresource Technology 74 17 - 24
PENtildeA Francisco 1982 El Rumen Aspectos de fisiologia y metabolismo microbioloacutegico Instituto colombiana Agropecuario ICA Divisioacuten de Ciencias Animales Bogotaacute P 78
PERCIVAL ZHANG YI-HENG AND LYND LEE R (2004) Toward an Agggregated Understanding of Enzymatic Hydrolysis of Cellulose Noncomplexed Cellulase Systems Biotechnology and Bioengineering vol 88 No 7 798 - 824
PRIMER 3 White head institutel MIl [Programa de computador] Disponible en internet www-genomewimiteducgi-binprimerprimer3-wwcgi
PROMEGA Manual tecnico Promega Kit wizard USA 1999
QIU XIAO (2000) Isolation and anaacutelisis of two cellulas cDNAs from Orpinomices joyonii Gene 245 119-126
RANADE D R AND GADRE R V 1988 Microbiological Aspects of Anaerobic Digestion Laboratory Manual Maharachtra Association for the Cultivation of Science Research Institute Pune India 107p
RANADE DR and GRADE RV Microbiological aspects of anaerobic digestion laboratory manual Edited Maharashtra Association for the Cultivation of Science In Research Institute Pune 1988 102 p
RAY S 1986 Nitrogen and energy nutrition of rumiant Departament of animal Science University of Florida Academic Press Inc
REZAEIAN MOHAMMAD et al (2005) Relative fibrolytic activities of anaerobic rumen fungi on untreated and sodium hydroxide treated barley straw in in vitro culture Anaerobe 11 163 -175
RODRIGUEZ V Fernando 1994 La microbiota ruminal y su importancia en la degradacioacuten de los forrajes tropicales En Seminario sobre Biotecnologia del Rumen Universidad Nacional de Colombia Sede Medelliacuten Nov 11-12
RODRIGUEZ DIAZ T (1996) Aislamiento patroacuten de fermentacioacuten de carbohidratos y caracterizacioacuten morfoloacutegica de bacterias celuloliacuteticas del rumen de bovinos alimentados con heno de Raigraacutes en Colombia En Revista CORPOICA Vol 1 p 123-28
ROLFS A SCHULLER l FINCKH O AND WEBER-ROLFS 1 PCR Clinical diagnostic and reserch Alemania 1992 P 271 Ed Springer-Verlag
SAELEY P G AND SOUTHERN E M Gel electrophoresis the DNA In Acids a Practical Approach Por Rickwoo and Hames IRL Press 1982 Sin mas datos
SAIKI DH GELFAND AND S STOFFEL 1988 Citado por KRAMER M E AND COEN D M Enzimatic amplification of DNA by PCR standard procedures and optimization In Short Protocols in Biology Molecularp 15-3 a 15-4
SHIMADA M TAKAHASHI M (1990) Biodegradation of Cellulosic Materials in Wood and Cellulosic Chemistry edited by David N -S Hon and Nobuo Shiraishi Marcel Dekker NY
78
SHOFIELD P PELL A N AND KRAUSE D O 1997 Molecular Beacons Trial of a fluorescence-based solution hybridization technique for ecological studies with ruminal bacteria Applied and Environmental Microbiology Vol 63 W 3 p1143-1147
SLEAT R AND MAH R A (1984) Quantitative method for colorimetric determination of formate in fermentation media Applied and environmental microbiology p 884 - 885
SRINIVASAN K MURAKAMI M NAKASHIMADA Y NISHIO N (2001) Efficient Production of Cellulolytic and Xylanolitic Enzymes by the Rumen Anaerobic Fungus Neocallimastix frontalis in a Repetead Batch Culture Journal of Bioscience and Bioengineering 91 2 153-158
STAHL D A FLESHER B MANSFIELD H R AND MONTGOMERY L 1988 Use of phylogeneticafly based hybridization probes for studies of ruminal microbial ecology Applied and Environmental Microbiology Vol 54(5) 1079-1084
STEENBAKKERS J M et al (2003) I3-Glucosidase in ceflulosome of the anaerobic fungus Piromyces sp strain E2 is a family 3 glycoside hydrolase Biochem J 370963 - 970
STEENBAKKERS PJ HARHANGI HR BOSSCHER MW VAN DER HOOFT MM KELTJENS JT VAN DER DRIFT C VOGELS GD OP DEN CAMP HJ (2003) ~shyGlucosidase in cellulosome of the anaerobic fungus Piromyces sp strain E2 is a family 3 glycoside hydrolase Biochem J 370 963 - 970
STEPHEN A MADDEN T SCHAFFER A lHANG J lHANG l MILLER W AND LlPMAN D 1997 Gapped BLAST and PSI-BLAST a new generation of protein database search programs Nucleic Acids Res Vol 25 p 3389-3402
STEWART C S 1991 The Rumen Bacteria in Rumen Microbial Metabolism and Rumiant Digestion Editd by J p Jouany INRA Editions Paris pp 131
STEWART C S BRYANT M P 1988 The rumen bacteria In The rumen microbial ecosystem Edited By Hobson p Elsevier Applied Science New York 21-26
STEWART C S PANIAGUA C DINSDALE D CHENG K-J AND GARRAW S H 1981 Selective isolation and characterization of Bacteriodes succinogenes from the rumen of a cow Applied and Environmental Microbiology Vol 41 W 2 P 504-510
TEUNISSEN M J OP DEN CAMP H J ORPIN CG HUIS JH VOGELS GD (1991) Comparison of growth characteristic of anaerobic fungi isolated from ruminant and nonshyruminant hervivores during cultivation in a defined medium J of Gen Microb 137 1401 shy1408
TEUNISSEN M J V M DE KORT G OP DEN CAMP H J M HUIS INT VELD J H J (1992) Production of cellulolytic and xylanolytic enzymes during growth of the anaerobic fungus Piromyces sp on different substrates J of Gen Microb 138 1657-1664 l
THEODOROU K M DAVIS R D NIELSEN B B LAWRENCE G l TRINCI P J (1990) Determination of growth of anaerobic fungi on soluble and cellulosic substrates using a pressure transducer Microbiology 141 671-678
THOMPSON JD HIGGINS DG and GIBSON TJ CLUSTAL W Improving the sensitivity of progressive multiple sequence alignment through sequence weighting positions-specific gap penalties and weight matrix choice In Nucleic Acids Research Vol 22 (1994) p 225
TSAI CHENG-FANG et al (2003) A comparative analysis of two cDNA clones of the f cellulose gene family from anaerobic fungus Piromyces rhizinflata Anaerobe 9 131 - 140
76
Domains for a Cellulase-Hemicellulase Complex Appli And Env Microb 63(12) 4721-4728 Dec
LI X-L CHEN H LJUNGDAHL L G (1997 b) Monocentric and Polycentric Anaerobic Fungi Produce Structurally Related Cellulases and Xylanases Appl And Env Microb 63(2) 628shy635
UN Chuzao y STHAL David A 1995 Taxon-Specific Probes for the Cellulolitic Genus Fibrobacter Reveal Abundant and Novel EquineAssociated Populations En Applied and Environmental Microbiology Vol 61 Ndeg4 pg 1348-1351
-iquest LOWE S E THEODOROU M K AND TRINCI A P (1987) ) cellulases and Xylanases of an liexcl anaerobic rumen fungus grown on wheat straw wlleat---straw holocellulose cellulose and xylan Applied and environmental microbiology June p 1216 - 1223
LUDWIG W DORN S SPRINGER N KIRCHHOF G AND SCHLEIFER K H 1994 PCR Based preparation of 23S rRNA-targeted group-specific polynucleotide probes Applied and Environmental Microbiology Vol 61 3236-3244
MACKIE R 1 1996 The use of oligonucleotide probes to study the ecology of ruminal microbial populations Ann Zootech 45 Suppl 281-285 Elsevier INRA
MANUAL DE PROCEDIMIENTOS(2003) Laboratorio de Biotecnologiacutea Ruminal (BIORUM) Universidad Nacional de Colombia Sede Medelliacuten
MARTIN S A Nutrient transport by ruminal bacteria a review Journal of animal science 1994 74 3019-3031
MILLER G L (1959) Use of Dinitrosalisylic Acid Reagent for Determination of Reducing Sugar Anal Chem 426-428
MILLET L FONTY G GOUET P 1995 Study of the composition of the ruminal cellulolytic flora of bison and deer with rRNA-targetes oligonucleotide probes Ann Zootech 44 Supp 163 Elsevier INRA
MORITZ C AND HILLlS D M 1996 Molecular systematics contex and controversies In Molecular Systematic Edited by Hillis D M Moritz C and Mable B K Sunderland U S Sinauer Associates Segunda Edicioacuten p 1-13
MORRISON M 1996 Do ruminal bacteria exchange genetic material Journal of Dairy Science Vol 79 p 1476-1486
MURRAY AND THOMSON Q980 Citados por WILSON K Preparation of genomic DNA from bacteria In Short Protocols in Biology Molecular P 2-11 a 2-12
ODENYO A A MACKIE R l STHAL D A WHITE B A 1994a The use of 16S rRNAshytargeted oligonucleotide probes for Ruminococcus species and evidence for bacteriocin production Applied and Environmental Microbiolegy Vol 60 p 3688-3696
ODENYO A A MACKIE R l STHAL D A WHITE B A 1994b The use of 16S rRNAshytargeted oligonucleotide probes for study competition between ruminal fibrolitic bacteria pure culture-studies with cellulose and alkaline peroxide-trated wheat straw Applied and Environmental Microbiolegy Vol 60 p 3697-3703
OLSEN G J AND WOESE C R 1993 Ribosomal RNA a key to phlogeny FESAB Journal Vol 7 p 113-123
OSSA F RIDRIGUEZ J ARCOS M DIAZ T 1999 Identificacioacuten molecular de las bacterias celuloliacuteticas Fibrobacter sucinogenes y Ruminococcus flavefaciens Revista
--- - - ---~ - ------====-------------------------------------------------------------------------shy
77
Colombiana de Ciencias Pecuarias Memorias V Encuentro Nacional de Investigaciones de las Ciencias Pecuarias ENICIP Vol 12 P 120
PALMQVIST E AND HAN-HAGERDAL B (2000) Fermentation of lignocellulosic hydrolysates 1 inhibition and detoxification Bioresource Technology 74 17 - 24
PENtildeA Francisco 1982 El Rumen Aspectos de fisiologia y metabolismo microbioloacutegico Instituto colombiana Agropecuario ICA Divisioacuten de Ciencias Animales Bogotaacute P 78
PERCIVAL ZHANG YI-HENG AND LYND LEE R (2004) Toward an Agggregated Understanding of Enzymatic Hydrolysis of Cellulose Noncomplexed Cellulase Systems Biotechnology and Bioengineering vol 88 No 7 798 - 824
PRIMER 3 White head institutel MIl [Programa de computador] Disponible en internet www-genomewimiteducgi-binprimerprimer3-wwcgi
PROMEGA Manual tecnico Promega Kit wizard USA 1999
QIU XIAO (2000) Isolation and anaacutelisis of two cellulas cDNAs from Orpinomices joyonii Gene 245 119-126
RANADE D R AND GADRE R V 1988 Microbiological Aspects of Anaerobic Digestion Laboratory Manual Maharachtra Association for the Cultivation of Science Research Institute Pune India 107p
RANADE DR and GRADE RV Microbiological aspects of anaerobic digestion laboratory manual Edited Maharashtra Association for the Cultivation of Science In Research Institute Pune 1988 102 p
RAY S 1986 Nitrogen and energy nutrition of rumiant Departament of animal Science University of Florida Academic Press Inc
REZAEIAN MOHAMMAD et al (2005) Relative fibrolytic activities of anaerobic rumen fungi on untreated and sodium hydroxide treated barley straw in in vitro culture Anaerobe 11 163 -175
RODRIGUEZ V Fernando 1994 La microbiota ruminal y su importancia en la degradacioacuten de los forrajes tropicales En Seminario sobre Biotecnologia del Rumen Universidad Nacional de Colombia Sede Medelliacuten Nov 11-12
RODRIGUEZ DIAZ T (1996) Aislamiento patroacuten de fermentacioacuten de carbohidratos y caracterizacioacuten morfoloacutegica de bacterias celuloliacuteticas del rumen de bovinos alimentados con heno de Raigraacutes en Colombia En Revista CORPOICA Vol 1 p 123-28
ROLFS A SCHULLER l FINCKH O AND WEBER-ROLFS 1 PCR Clinical diagnostic and reserch Alemania 1992 P 271 Ed Springer-Verlag
SAELEY P G AND SOUTHERN E M Gel electrophoresis the DNA In Acids a Practical Approach Por Rickwoo and Hames IRL Press 1982 Sin mas datos
SAIKI DH GELFAND AND S STOFFEL 1988 Citado por KRAMER M E AND COEN D M Enzimatic amplification of DNA by PCR standard procedures and optimization In Short Protocols in Biology Molecularp 15-3 a 15-4
SHIMADA M TAKAHASHI M (1990) Biodegradation of Cellulosic Materials in Wood and Cellulosic Chemistry edited by David N -S Hon and Nobuo Shiraishi Marcel Dekker NY
78
SHOFIELD P PELL A N AND KRAUSE D O 1997 Molecular Beacons Trial of a fluorescence-based solution hybridization technique for ecological studies with ruminal bacteria Applied and Environmental Microbiology Vol 63 W 3 p1143-1147
SLEAT R AND MAH R A (1984) Quantitative method for colorimetric determination of formate in fermentation media Applied and environmental microbiology p 884 - 885
SRINIVASAN K MURAKAMI M NAKASHIMADA Y NISHIO N (2001) Efficient Production of Cellulolytic and Xylanolitic Enzymes by the Rumen Anaerobic Fungus Neocallimastix frontalis in a Repetead Batch Culture Journal of Bioscience and Bioengineering 91 2 153-158
STAHL D A FLESHER B MANSFIELD H R AND MONTGOMERY L 1988 Use of phylogeneticafly based hybridization probes for studies of ruminal microbial ecology Applied and Environmental Microbiology Vol 54(5) 1079-1084
STEENBAKKERS J M et al (2003) I3-Glucosidase in ceflulosome of the anaerobic fungus Piromyces sp strain E2 is a family 3 glycoside hydrolase Biochem J 370963 - 970
STEENBAKKERS PJ HARHANGI HR BOSSCHER MW VAN DER HOOFT MM KELTJENS JT VAN DER DRIFT C VOGELS GD OP DEN CAMP HJ (2003) ~shyGlucosidase in cellulosome of the anaerobic fungus Piromyces sp strain E2 is a family 3 glycoside hydrolase Biochem J 370 963 - 970
STEPHEN A MADDEN T SCHAFFER A lHANG J lHANG l MILLER W AND LlPMAN D 1997 Gapped BLAST and PSI-BLAST a new generation of protein database search programs Nucleic Acids Res Vol 25 p 3389-3402
STEWART C S 1991 The Rumen Bacteria in Rumen Microbial Metabolism and Rumiant Digestion Editd by J p Jouany INRA Editions Paris pp 131
STEWART C S BRYANT M P 1988 The rumen bacteria In The rumen microbial ecosystem Edited By Hobson p Elsevier Applied Science New York 21-26
STEWART C S PANIAGUA C DINSDALE D CHENG K-J AND GARRAW S H 1981 Selective isolation and characterization of Bacteriodes succinogenes from the rumen of a cow Applied and Environmental Microbiology Vol 41 W 2 P 504-510
TEUNISSEN M J OP DEN CAMP H J ORPIN CG HUIS JH VOGELS GD (1991) Comparison of growth characteristic of anaerobic fungi isolated from ruminant and nonshyruminant hervivores during cultivation in a defined medium J of Gen Microb 137 1401 shy1408
TEUNISSEN M J V M DE KORT G OP DEN CAMP H J M HUIS INT VELD J H J (1992) Production of cellulolytic and xylanolytic enzymes during growth of the anaerobic fungus Piromyces sp on different substrates J of Gen Microb 138 1657-1664 l
THEODOROU K M DAVIS R D NIELSEN B B LAWRENCE G l TRINCI P J (1990) Determination of growth of anaerobic fungi on soluble and cellulosic substrates using a pressure transducer Microbiology 141 671-678
THOMPSON JD HIGGINS DG and GIBSON TJ CLUSTAL W Improving the sensitivity of progressive multiple sequence alignment through sequence weighting positions-specific gap penalties and weight matrix choice In Nucleic Acids Research Vol 22 (1994) p 225
TSAI CHENG-FANG et al (2003) A comparative analysis of two cDNA clones of the f cellulose gene family from anaerobic fungus Piromyces rhizinflata Anaerobe 9 131 - 140
--- - - ---~ - ------====-------------------------------------------------------------------------shy
77
Colombiana de Ciencias Pecuarias Memorias V Encuentro Nacional de Investigaciones de las Ciencias Pecuarias ENICIP Vol 12 P 120
PALMQVIST E AND HAN-HAGERDAL B (2000) Fermentation of lignocellulosic hydrolysates 1 inhibition and detoxification Bioresource Technology 74 17 - 24
PENtildeA Francisco 1982 El Rumen Aspectos de fisiologia y metabolismo microbioloacutegico Instituto colombiana Agropecuario ICA Divisioacuten de Ciencias Animales Bogotaacute P 78
PERCIVAL ZHANG YI-HENG AND LYND LEE R (2004) Toward an Agggregated Understanding of Enzymatic Hydrolysis of Cellulose Noncomplexed Cellulase Systems Biotechnology and Bioengineering vol 88 No 7 798 - 824
PRIMER 3 White head institutel MIl [Programa de computador] Disponible en internet www-genomewimiteducgi-binprimerprimer3-wwcgi
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SRINIVASAN K MURAKAMI M NAKASHIMADA Y NISHIO N (2001) Efficient Production of Cellulolytic and Xylanolitic Enzymes by the Rumen Anaerobic Fungus Neocallimastix frontalis in a Repetead Batch Culture Journal of Bioscience and Bioengineering 91 2 153-158
STAHL D A FLESHER B MANSFIELD H R AND MONTGOMERY L 1988 Use of phylogeneticafly based hybridization probes for studies of ruminal microbial ecology Applied and Environmental Microbiology Vol 54(5) 1079-1084
STEENBAKKERS J M et al (2003) I3-Glucosidase in ceflulosome of the anaerobic fungus Piromyces sp strain E2 is a family 3 glycoside hydrolase Biochem J 370963 - 970
STEENBAKKERS PJ HARHANGI HR BOSSCHER MW VAN DER HOOFT MM KELTJENS JT VAN DER DRIFT C VOGELS GD OP DEN CAMP HJ (2003) ~shyGlucosidase in cellulosome of the anaerobic fungus Piromyces sp strain E2 is a family 3 glycoside hydrolase Biochem J 370 963 - 970
STEPHEN A MADDEN T SCHAFFER A lHANG J lHANG l MILLER W AND LlPMAN D 1997 Gapped BLAST and PSI-BLAST a new generation of protein database search programs Nucleic Acids Res Vol 25 p 3389-3402
STEWART C S 1991 The Rumen Bacteria in Rumen Microbial Metabolism and Rumiant Digestion Editd by J p Jouany INRA Editions Paris pp 131
STEWART C S BRYANT M P 1988 The rumen bacteria In The rumen microbial ecosystem Edited By Hobson p Elsevier Applied Science New York 21-26
STEWART C S PANIAGUA C DINSDALE D CHENG K-J AND GARRAW S H 1981 Selective isolation and characterization of Bacteriodes succinogenes from the rumen of a cow Applied and Environmental Microbiology Vol 41 W 2 P 504-510
TEUNISSEN M J OP DEN CAMP H J ORPIN CG HUIS JH VOGELS GD (1991) Comparison of growth characteristic of anaerobic fungi isolated from ruminant and nonshyruminant hervivores during cultivation in a defined medium J of Gen Microb 137 1401 shy1408
TEUNISSEN M J V M DE KORT G OP DEN CAMP H J M HUIS INT VELD J H J (1992) Production of cellulolytic and xylanolytic enzymes during growth of the anaerobic fungus Piromyces sp on different substrates J of Gen Microb 138 1657-1664 l
THEODOROU K M DAVIS R D NIELSEN B B LAWRENCE G l TRINCI P J (1990) Determination of growth of anaerobic fungi on soluble and cellulosic substrates using a pressure transducer Microbiology 141 671-678
THOMPSON JD HIGGINS DG and GIBSON TJ CLUSTAL W Improving the sensitivity of progressive multiple sequence alignment through sequence weighting positions-specific gap penalties and weight matrix choice In Nucleic Acids Research Vol 22 (1994) p 225
TSAI CHENG-FANG et al (2003) A comparative analysis of two cDNA clones of the f cellulose gene family from anaerobic fungus Piromyces rhizinflata Anaerobe 9 131 - 140
