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Richard Stouthamer - One of the best experts on this subject based on the ideXlab platform.
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Genome-wide analyses of single nucleotide polymorphisms reveal the consequences of traditional mass-rearing on genetic variation in Aphytis melinus (Hymenoptera: Aphelinidae): the danger of putting all eggs in one basket.
Pest management science, 2019Co-Authors: Marco Gebiola, Joseph G. Morse, Jeffrey W. Streicher, Paul F. Rugman-jones, Richard StouthamerAbstract:Background Aphytis melinus DeBach (Hymenoptera: Aphelinidae) is a highly effective biocontrol agent of the California red scale Aonidiella aurantii (Maskell) (Hemiptera: Diaspididae). It is commercially reared and used for augmentative releases within integrated pest management programs. However, mass rearing of biocontrol agents can result in population bottlenecks and high levels of inbreeding and/or adaptation to the factitious rearing conditions. Although these factors can all negatively impact field performance of biocontrol agents, few empirical studies have examined the genetic consequences of mass rearing. We used double-digest RAD sequencing (ddRADseq) to investigate the effect of traditional mass rearing on genetic variation among insectary colonies of A. melinus relative to wild populations in native (Pakistan) and introduced (California) ranges. Results Analyses of up to 9700 single nucleotide polymorphisms (SNPs) revealed that insectary populations had less genomic variation than introduced populations. This was evidenced by fewer private alleles, reduced heterozygosity, and greater missing data in the insectary populations. Further, California Insectaries formed a distinct genomic cluster relative to the other samples, a surprising result given that the insectary colonies were putatively established at different times and from different source populations. These differences were evident across most data sets also after we filtered out contaminant DNA from the most common host species (Aspidiotus nerii Bouche and A. aurantii). Conclusion We hypothesize that this pattern would only result if: (i) directional selection for 'captive' phenotypes produces convergent patterns of genomic variation across Insectaries; or (ii) the California insectary colonies were all founded from a unifying source population and/or that the Insectaries regularly exchange 'genetic' stocks. We show that RADseq is an effective method to investigate the effects of mass rearing on genetics of biocontrol agents. © 2019 Society of Chemical Industry.
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quality assessment of trichogramma parasitoids trichogramma spp from six mexican Insectaries
Agrociencia, 2014Co-Authors: Jaime Gonzalezcabrera, Hugo C Arredondobernal, Richard StouthamerAbstract:La calidad de los agentes de control biologico producidos masivamente es vital. Existen reportes de que Trichogramma spp. no controla la plaga objetivo y su mala calidad puede explicar este fracaso. El primer objetivo de este estudio fue determinar que forma de reproduccion, ya sea sexual o asexual, se practico en las colonias de Trichogramma spp. de insectarios mexicanos, y el segundo objetivo fue determinar si esas colonias de Trichogramma spp. cumplen con los estandares de calidad sugeridos por la Organizacion Internacional para el Control Biologico/Comunidad Europea (IOBC/EC, por sus siglas en ingles). En marzo de 2010, se envio una carta a 27 productores de Trichogramma spp. Seis insectarios aceptaron participar y enviaron muestras muertas de sus colonias, un total de 10 colonias. Para el analisis estadistico estas colonias se agruparon por especies, Trichogramma pretiosum o fuentesi. Ninguna de las colonias de Trichogramma spp. estaba infectada de Wolbachia, que induce la partenogenesis, es decir, todas las colonias se reprodujeron sexualmente. Por ende, se usaron los estandares de calidad para especies sexuales: proporcion sexual (proporcion de hembras) ≥0.5, mortalidad embrionaria ≤20 % y fecundidad total ≥40 huevos. Entre las 10 colonias de Trichogramma spp. hubo gran uniformidad en la proporcion sexual, la mortalidad embrionaria y la fecundidad total, es decir, no hubo diferencias estadisticas entre las colonias por especie. En la proporcion sexual (0.552 ±0.038) y la mortalidad embrionaria (21± 4.48 %), nueve colonias de T. pretiosum apenas cumplieron con los estandares minimos sugeridos por la IOBC/EC. Con la longitud de la tibia posterior como proxy para fecundidad total, la fecundidad (37.85 huevos) de nueve colonias de T. pretiosum y dos colonias de T. fuentesi se ubico ligeramente debajo del estandar sugerido por la IOBC/EC. En Mexico se producen solo especies sexuales
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multiplex pcr assay to identify trichogramma parasitoids hymenoptera trichogrammatidae reared from mexican Insectaries
Agrociencia, 2014Co-Authors: Jaime Gonzalezcabrera, Hugo C Arredondobernal, Richard StouthamerAbstract:La identificacion precisa de insectos beneficos es el primer paso en el establecimiento de programas exitosos de control biologico. En Mexico, los parasitoides Trichogramma se identifican mediante caracteres morfologicos, lo cual es un procedimiento laborioso realizado solo por expertos; ademas, algunas estructuras morfologicas son susceptibles a cambios debido a las diferencias en la dieta o las condiciones ambientales, y por tanto existe el riesgo latente de una identificacion taxonomica erronea. Mas aun, la identificacion morfologica requiere la presencia de machos, dejando a las poblaciones conformadas por solo hembras (partenogeneticas) sin identificar. Como alternativa a la identificacion morfologica, el proposito de este estudio fue desarrollar un metodo molecular para la identificacion de parasitoides Trichogramma de insectarios mexicanos. Las especies de Trichogramma reproducidas en seis insectarios se identificaron por ADN y despues, con base en diferencias de las secuencias de ITS2, se diseno una prueba PCR-multiple para distinguir entre las especies reproducidas. Asi, se encontraron discrepancias entre la identidad reportada y la determinada por ADN: se suponia que las muestras de todos los insectarios contenian tres especies de Trichogramma (T. pretiosum, T. exiguum y T. platneri), pero solo hubo dos especies (T. pretiosum y T. fuentesi) presentes. Ademas, se encontro la presencia de especies de reemplazo. Afortunadamente, utilizando la prueba PCR ADN-multiple generada en este estudio puede hacerse una identificacion taxonomica exacta de las especies de Trichogramma. Ademas, esta metodologia puede identificar las especies nativas de Trichogramma, cuyo uso en proyectos de reproduccion masiva es rara
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QUALITY ASSESSMENT OF TRICHOGRAMMA PARASITOIDS (Trichogramma spp.) FROM SIX MEXICAN Insectaries EVALUACIÓN DE CALIDAD DEL PARASITOIDE TRICOGRAMA ( Trichogramma spp.) PROVENIENTE DE SEIS INSECTARIOS MEXICANOS
2014Co-Authors: Jaime Gonzalez-cabrera, Hugo César Arredondo-bernal, Richard StouthamerAbstract:Quality of mass-reared biological control agents is vital. There are reports that Trichogramma spp. failed to control the target pest, and poor quality may explain these failures. The first goal of this study was to determine which way of reproduction was practiced by Trichogramma spp. colonies from Mexican Insectaries, either sexual or asexual, and the second goal was to determine whether those Trichogramma spp. colonies meet the quality standards suggested by the International Organization for Biological Control/ European Community (IOBC/EC). In March 2010, a letter was sent to 27 Trichogramma spp. producers. Six Insectaries agreed to participate and sent dead samples of its colonies, a total of 10 colonies. For statistical analysis these colonies were grouped by species, either Trichogramma pretiosum or T. fuentesi. None of the Trichogramma spp. colonies were infected by parthenogenetic-inducing Wolbachia, i.e., all colonies reproduced sexually. Therefore, quality standards for sexual species were used: sex ratio (proportion females) 0.5, embryonic mortality 20 %, and total fecundity 40 eggs. Among the 10 Trichogramma spp. colonies there was great uniformity in sex ratio, embryonic mortality and total fecundity, i.e., per species there were not statistical differences among colonies. In sex ratio (0.552 0.038) and embryonic mortality (21 4.48 %) the nine colonies of T. pretiosum barely fulfilled the minimum standards suggested by the IOBC/EC. Using hind tibia length as a proxy of total fecundity, the fecundity (37.85 eggs) of nine colonies of T. pretiosum and two colonies of T. fuentesi fell slightly below the standard suggested by the IOBC/EC. In Mexico only sexual species of Trichogramma
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MULTIPLEX PCR ASSAY TO IDENTIFY Trichogramma PARASITOIDS (HYMENOPTERA, TRICHOGRAMMATIDAE) REARED FROM MEXICAN Insectaries PCR MÚLTIPLE PARA IDENTIFICAR ESPECIES DE Trichogramma (HYMENOPTERA, TRICHOGRAMMATIDAE) PARASITOIDES CRIADAS EN INSECTARIOS MEXI
2014Co-Authors: Jaime Gonzalez-cabrera, Hugo César Arredondo-bernal, Richard StouthamerAbstract:Accurate identification of species as biological candidates is the first step in establishing successful biological control programs. In Mexico, Trichogramma parasitoids are identified using morphological characters, which is a laborious procedure made only by experts; in addition, some morphological structures are susceptible to changes due to differences in diet or environment, and consequently there is a latent risk of misidentification. Moreover, morphological identification requires the presence of males, leaving female-only populations unidentifiable. As an alternative to morphological identification, the purpose of this work was to develop a molecular method for the identification of Trichogramma parasitoids from Mexican Insectaries. Trichogramma species reared in six Insectaries were DNA-identified and subsequently, based on differences in ITS2 sequences, a DNA-multiplex PCR assay was designed to distinguish among those reared species. Thus, discrepancies were found between the reported and DNA-determined species identity, whereas the sample of all the Insectaries together was supposed to contain three species of Trichogramma (T. pretiosum, T. exiguum and T. platneri), only two species ( T. pretiosum and T. fuentesi) were present. In addition, it was found the presence of unnoticed species replacement. Fortunately, an accurate taxonomic identification of Trichogramma species can be made by using the DNA-multiplex PCR assay that was generated in this work. Additionally, this methodology may identify the native Trichogramma species, whose use in mass rearing projects is uncommon.
Francois Noireau - One of the best experts on this subject based on the ideXlab platform.
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Cryptic speciation in the Triatoma sordida subcomplex (Hemiptera, Reduviidae) revealed by chromosomal markers
Parasites & Vectors, 2015Co-Authors: Francisco Panzera, Sebastian Pita, Cleber Galvao, Tamara Chavez, Antonieta Rojas De Arias, Lourdes Cardozo Téllez, Yanina Panzera, Julieta Nattero, Francois NoireauAbstract:Background Chagas disease vectors (Hemiptera-Reduviidae) comprise more than 140 blood-sucking insect species of the Triatominae subfamily. The largest genus is Triatoma , subdivided in several complexes and subcomplexes according to morphology, ecology and genetic features. One of them is the sordida subcomplex, involving four species: Triatoma sordida, T. guasayana , T. garciabesi and T. patagonica . Given the great morphological similarity of these species, their taxonomic identification, evolutionary relationships and population differentiation have been controversial for many years and even today remain under discussion. Methods We simultaneously analyzed two chromosomal markers, C-heterochromatin distribution and 45S ribosomal genes chromosomal position, of 139 specimens from several sordida subcomplex populations from Argentina, Bolivia, Brazil and Paraguay, collected both in nature and from several established Insectaries. Our results were compared with COI sequences deposited in GenBank. Results We recognized five chromosomal taxa with putative hybrids, which each differ in at least one chromosome marker. Most of them present significant differences in their mtDNA sequences. Conclusion The chromosomal taxa here show a significant chromosome differentiation involving changes in the C-heterochromatin content and in the ribosomal clusters position. This paper identifies several erroneously classified populations by morphological methods, delimits the geographical distribution of each taxon and proposes the existence of a new cryptic species, widely distributed in Argentina. We also suggest that sordida sibling species involve closely related as well as evolutionary distant species. Taxonomic status of each chromosomal taxon is discussed considering phenotypic and genetic results previously published.
Francisco Panzera - One of the best experts on this subject based on the ideXlab platform.
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Cryptic speciation in the Triatoma sordida subcomplex (Hemiptera, Reduviidae) revealed by chromosomal markers
Parasites & Vectors, 2015Co-Authors: Francisco Panzera, Sebastian Pita, Cleber Galvao, Tamara Chavez, Antonieta Rojas De Arias, Lourdes Cardozo Téllez, Yanina Panzera, Julieta Nattero, Francois NoireauAbstract:Background Chagas disease vectors (Hemiptera-Reduviidae) comprise more than 140 blood-sucking insect species of the Triatominae subfamily. The largest genus is Triatoma , subdivided in several complexes and subcomplexes according to morphology, ecology and genetic features. One of them is the sordida subcomplex, involving four species: Triatoma sordida, T. guasayana , T. garciabesi and T. patagonica . Given the great morphological similarity of these species, their taxonomic identification, evolutionary relationships and population differentiation have been controversial for many years and even today remain under discussion. Methods We simultaneously analyzed two chromosomal markers, C-heterochromatin distribution and 45S ribosomal genes chromosomal position, of 139 specimens from several sordida subcomplex populations from Argentina, Bolivia, Brazil and Paraguay, collected both in nature and from several established Insectaries. Our results were compared with COI sequences deposited in GenBank. Results We recognized five chromosomal taxa with putative hybrids, which each differ in at least one chromosome marker. Most of them present significant differences in their mtDNA sequences. Conclusion The chromosomal taxa here show a significant chromosome differentiation involving changes in the C-heterochromatin content and in the ribosomal clusters position. This paper identifies several erroneously classified populations by morphological methods, delimits the geographical distribution of each taxon and proposes the existence of a new cryptic species, widely distributed in Argentina. We also suggest that sordida sibling species involve closely related as well as evolutionary distant species. Taxonomic status of each chromosomal taxon is discussed considering phenotypic and genetic results previously published.
Hugo César Arredondo-bernal - One of the best experts on this subject based on the ideXlab platform.
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Molecular characterization of Chrysoperla carnea (Neuroptera: Chrysopidae) from commercial Insectaries in Mexico.
Molecular biology reports, 2019Co-Authors: Martín Palomares-pérez, Cecilia Moreno-rodriguez, Yadira Contreras-bermúdez, Hugo César Arredondo-bernal, Adrien GallouAbstract:The larvae of the Chrysoperla carnea-group (Neuroptera: Chrysopidae) are recognized among the most effective larval predators of various phytophagous arthropods. Therefore, green lacewings are commonly grown by commercial Insectaries and released as biological control agents. Previous work has shown that commercial laboratories frequently supply indeterminate species of the large C. carnea cryptic species complex. In Mexico, at least 20 biological control companies have commercialized the species C. carnea, but none of the products reared by those companies have been analyzed scientifically. Thus, the goal of this work was to molecularly characterize nine C. carnea populations from Mexican Insectaries using the most efficient molecular markers available: the mitochondrial genes COI, COII, ND2, and ND5. Phylogenetic analysis demonstrated a unique mitochondrial haplotype in seven commercial Insectaries showing 100% similarity to the reference specimen C. plorabunda E100. In contrast, we observed two and four different mitochondrial haplotypes of the carnea-group in two commercial Insectaries. More precisely, three specimens possessed the mitochondrial haplotype of the species C. zastrowi, suggesting possible natural occurrence of this haplotype in Mexico. Consequently, this study demonstrated the need for an extensive survey of the different laboratories and Insectaries producing C. carnea in Mexico, including unambiguous species identification by song recordings to confirm the species identity of the observed mitochondrial haplotypes.
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QUALITY ASSESSMENT OF TRICHOGRAMMA PARASITOIDS (Trichogramma spp.) FROM SIX MEXICAN Insectaries EVALUACIÓN DE CALIDAD DEL PARASITOIDE TRICOGRAMA ( Trichogramma spp.) PROVENIENTE DE SEIS INSECTARIOS MEXICANOS
2014Co-Authors: Jaime Gonzalez-cabrera, Hugo César Arredondo-bernal, Richard StouthamerAbstract:Quality of mass-reared biological control agents is vital. There are reports that Trichogramma spp. failed to control the target pest, and poor quality may explain these failures. The first goal of this study was to determine which way of reproduction was practiced by Trichogramma spp. colonies from Mexican Insectaries, either sexual or asexual, and the second goal was to determine whether those Trichogramma spp. colonies meet the quality standards suggested by the International Organization for Biological Control/ European Community (IOBC/EC). In March 2010, a letter was sent to 27 Trichogramma spp. producers. Six Insectaries agreed to participate and sent dead samples of its colonies, a total of 10 colonies. For statistical analysis these colonies were grouped by species, either Trichogramma pretiosum or T. fuentesi. None of the Trichogramma spp. colonies were infected by parthenogenetic-inducing Wolbachia, i.e., all colonies reproduced sexually. Therefore, quality standards for sexual species were used: sex ratio (proportion females) 0.5, embryonic mortality 20 %, and total fecundity 40 eggs. Among the 10 Trichogramma spp. colonies there was great uniformity in sex ratio, embryonic mortality and total fecundity, i.e., per species there were not statistical differences among colonies. In sex ratio (0.552 0.038) and embryonic mortality (21 4.48 %) the nine colonies of T. pretiosum barely fulfilled the minimum standards suggested by the IOBC/EC. Using hind tibia length as a proxy of total fecundity, the fecundity (37.85 eggs) of nine colonies of T. pretiosum and two colonies of T. fuentesi fell slightly below the standard suggested by the IOBC/EC. In Mexico only sexual species of Trichogramma
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MULTIPLEX PCR ASSAY TO IDENTIFY Trichogramma PARASITOIDS (HYMENOPTERA, TRICHOGRAMMATIDAE) REARED FROM MEXICAN Insectaries PCR MÚLTIPLE PARA IDENTIFICAR ESPECIES DE Trichogramma (HYMENOPTERA, TRICHOGRAMMATIDAE) PARASITOIDES CRIADAS EN INSECTARIOS MEXI
2014Co-Authors: Jaime Gonzalez-cabrera, Hugo César Arredondo-bernal, Richard StouthamerAbstract:Accurate identification of species as biological candidates is the first step in establishing successful biological control programs. In Mexico, Trichogramma parasitoids are identified using morphological characters, which is a laborious procedure made only by experts; in addition, some morphological structures are susceptible to changes due to differences in diet or environment, and consequently there is a latent risk of misidentification. Moreover, morphological identification requires the presence of males, leaving female-only populations unidentifiable. As an alternative to morphological identification, the purpose of this work was to develop a molecular method for the identification of Trichogramma parasitoids from Mexican Insectaries. Trichogramma species reared in six Insectaries were DNA-identified and subsequently, based on differences in ITS2 sequences, a DNA-multiplex PCR assay was designed to distinguish among those reared species. Thus, discrepancies were found between the reported and DNA-determined species identity, whereas the sample of all the Insectaries together was supposed to contain three species of Trichogramma (T. pretiosum, T. exiguum and T. platneri), only two species ( T. pretiosum and T. fuentesi) were present. In addition, it was found the presence of unnoticed species replacement. Fortunately, an accurate taxonomic identification of Trichogramma species can be made by using the DNA-multiplex PCR assay that was generated in this work. Additionally, this methodology may identify the native Trichogramma species, whose use in mass rearing projects is uncommon.
Anna-bella Failloux - One of the best experts on this subject based on the ideXlab platform.
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Persistent Wolbachia and Cultivable Bacteria Infection in the Reproductive and Somatic Tissues of the Mosquito Vector Aedes albopictus
2013Co-Authors: Karima Zouache, Denis Voronin, Van Tran-van, Laurence Mousson, Anna-bella FaillouxAbstract:Background: Commensal and symbiotic microbes have a considerable impact on the behavior of many arthropod hosts, including hematophagous species that transmit pathogens causing infectious diseases to human and animals. Little is known about the bacteria associated with mosquitoes other than the vectorized pathogens. This study investigated Wolbachia and cultivable bacteria that persist through generations in Ae. albopictus organs known to host transmitted arboviruses, such as dengue and chikungunya. Methodology/Principal Findings: We used culturing, diagnostic and quantitative PCR, as well as in situ hybridization, to detect and locate bacteria in whole individual mosquitoes and in dissected tissues. Wolbachia, cultivable bacteria of the genera Acinetobacter, Comamonas, Delftia and Pseudomonas co-occurred and persisted in the bodies of both males and females of Ae. albopictus initially collected in La Réunion during the chikungunya outbreak, and maintained as colonies in Insectaries. In dissected tissues, Wolbachia and the cultivable Acinetobacter can be detected in the salivary glands. The other bacteria are commonly found in the gut. Quantitative PCR estimates suggest that Wolbachia densities are highest in ovaries, lower than those of Acinetobacter in the gut, and approximately equal to those of Acinetobacter in the salivary glands. Hybridization using specific fluorescent probes successfully localized Wolbachia in all germ cells, including the oocytes, and in the salivary glands, whereas the Acinetobacter hybridizing signal was mostly located in the foregut and in the anterio
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Disseminated infection rate (A), Transmission rate (B) and mean titer of infectious viral particles present in saliva (C) of Culex pipiens challenged with WNV.
2013Co-Authors: Fadila Amraoui, Ghazi Krida, Ali Bouattour, Adel Rhim, Jabeur Daaboub, Zoubir Harrat, Said-chawki Boubidi, Mhamed Tijane, Mhammed Sarih, Anna-bella FaillouxAbstract:F1 mosquitoes (autogenous AU and anautogenous AN) were orally challenged with WNV at a titer of 107.8 PFU/mL using an artificial feeding system. After completion of the blood-meal, mosquitoes were maintained in BSL-3 Insectaries at 28°C. At day 14 pi, saliva was collected from surviving females using the forced salivation technique. The number of infectious viral particles present in saliva was estimated by plaque assay on Vero cells. After salivation, females were tested for the presence of WNV on head squashes by IFA. p
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Disseminated infection rate, Transmission rate and mean titer of infectious viral particles present in saliva of Culex pipiens at day 14 (A,B and C) and 21 (D,E and F) post-infection with RVFV.
2013Co-Authors: Fadila Amraoui, Ghazi Krida, Ali Bouattour, Adel Rhim, Jabeur Daaboub, Zoubir Harrat, Said-chawki Boubidi, Mhamed Tijane, Mhammed Sarih, Anna-bella FaillouxAbstract:F1 mosquitoes (autogenous AU and anautogenous AN) were orally challenged with RVFV at a titer of 108.5 PFU/mL using an artificial feeding system. After completion of the blood-meal, mosquitoes were maintained in BSL-3 Insectaries at 28°C. At day 14 pi and day 21 pi, saliva was collected from surviving females using the forced salivation technique. The number of infectious viral particles present in saliva was estimated by plaque assay on Vero cells. After salivation, females were tested for the presence of RVFV on head squashes by IFA. In brackets, the number of mosquitoes tested. Error bars show the confidence interval (95%) for DIR and TR, and the standard deviation for Log10 pfu/saliva.
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Persistent Wolbachia and Cultivable Bacteria Infection in the Reproductive and Somatic Tissues of the Mosquito Vector Aedes albopictus
PLoS ONE, 2009Co-Authors: Karima Zouache, Anna-bella Failloux, Denis Voronin, Van Tran-van, Laurence Mousson, Patrick MavinguiAbstract:BACKGROUND: Commensal and symbiotic microbes have a considerable impact on the behavior of many arthropod hosts, including hematophagous species that transmit pathogens causing infectious diseases to human and animals. Little is known about the bacteria associated with mosquitoes other than the vectorized pathogens. This study investigated Wolbachia and cultivable bacteria that persist through generations in Ae. albopictus organs known to host transmitted arboviruses, such as dengue and chikungunya. METHODOLOGY/PRINCIPAL FINDINGS: We used culturing, diagnostic and quantitative PCR, as well as in situ hybridization, to detect and locate bacteria in whole individual mosquitoes and in dissected tissues. Wolbachia, cultivable bacteria of the genera Acinetobacter, Comamonas, Delftia and Pseudomonas co-occurred and persisted in the bodies of both males and females of Ae. albopictus initially collected in La Réunion during the chikungunya outbreak, and maintained as colonies in Insectaries. In dissected tissues, Wolbachia and the cultivable Acinetobacter can be detected in the salivary glands. The other bacteria are commonly found in the gut. Quantitative PCR estimates suggest that Wolbachia densities are highest in ovaries, lower than those of Acinetobacter in the gut, and approximately equal to those of Acinetobacter in the salivary glands. Hybridization using specific fluorescent probes successfully localized Wolbachia in all germ cells, including the oocytes, and in the salivary glands, whereas the Acinetobacter hybridizing signal was mostly located in the foregut and in the anterior midgut. CONCLUSIONS/SIGNIFICANCE: Our results show that Proteobacteria are distributed in the somatic and reproductive tissues of mosquito where transmissible pathogens reside and replicate. This location may portend the coexistence of symbionts and pathogens, and thus the possibility that competition or cooperation phenomena may occur in the mosquito vector Ae. albopictus. Improved understanding of the vectorial system, including the role of bacteria in the vector's biology and competence, could have major implications for understanding viral emergences and for disease control.