Fauna de abelhas nativa em plantações de tomate : uma comparação de métodos de amostragem ativa e de armadilha

O tomate é amplamente cultivado em todo o mundo e requer polinização por abelhas nativas ou manejadas para realizar o pleno potencial de produção dos frutos. Para investigar a riqueza e abundância de espécies de abelhas nativas em plantações de tomate do Centro-Oeste do Brasil, dois métodos de amostragem (armadilhas pan-trap e amostragem ativa) foram utilizados em nove propriedades de junho a setembro de 2011. Um total de 465 indivíduos de 44 espécies foi coletado. A composição das espécies de abelhas amostradas diferiu dependendo do método utilizado. Vinte e duas espécies foram capturadas exclusivamente em armadilhas, 13 outras por meio de amostragem ativa e nove por ambos os métodos. A maioria das espécies de abelhas capturadas neste estudo pode ser considerada polinizadores eficazes do tomate, porque elas podem executar a polinização por vibração. Vibrando seus músculos torácicos, essas abelhas podem liberar o pólen das anteras para seus próprios corpos e para os estigmas da mesma flor, uma vez que eles estão dentro do cone de anteras da variedade do tomate estudado. Ambos os métodos amostraram espécies exclusivas de abelhas vibradoras. No entanto, as armadilhas capturaram abelhas vibradoras e não vibradoras indiscriminadamente e o método ativo amostrou principalmente a abelha vibradora. As coletas utilizando armadilhas e amostragens ativas foram complementares. O uso de apenas um método de amostragem não fornece um entendimento completo da riqueza de espécies de polinizadores de tomate no campo.The tomato is widely cultivated throughout the world and requires pollination by wild or managed bees to realize its full-potential fruit production. Two different sampling methods (pan trapping and active sampling) were employed in nine different properties from June to September of 2011 to investigate the richness and abundance of native bee species present in tomato crops of Center-West Brazil. A total of 465 individuals of 44 species were collected, with the composition of sampled bee species differing between the methods used. Twenty-two species were exclusively captured in pan traps, 13 others through active sampling and nine by both methods. Most of the sampled bee species can be considered effective pollinators of the tomato because they can perform buzz-pollination. By vibration, these bees can liberate pollen from anthers into the air or onto their own bodies and the stigmas of the same flower because the stigmas of the studied tomato variety are within the anther cone. Both methods exclusively sampled some species of buzz-pollinating bees, however, pan-trapping captured buzzing and non-buzzing visitors indiscriminately while active sampling captured more buzzing bees. Pan-trapping and active sampling appear to complement each other, and so the use of only one or the other would not provide a full understanding of the species richness of tomato pollinators in the field

Interacción entre fungicidas biológicos y químicos con polinizadores de tomate

1 recurso en línea (páginas 425-435).El uso inapropiado de agroquímicos es perjudicial para las abejas que visitan los cultivos agrícolas, lo que reduce la producción por la afectación de la polinización y son pocos los estudios sobre este tema. El objetivo de este estudio fue verificar la incidencia de diferentes fungicidas sobre la visita de abejas en cultivos de tomate y sus efectos sobre la deposición de granos de polen en el estigma, número de semillas, masa y tamaño del fruto. Los experimentos consistieron en 10 tratamientos que fueron: (T1) tratamiento control sin agroquímicos; (T2 y T3) Bacillus subtilis en diferentes frecuencias de aplicación; (T4) hidróxido de cobre; (T5) B. subtilis e hidróxido de cobre; (T6) acibenzolar-S-metilo; (T7) trifloxistrobina+tebuconazol y B. subtilis; (T8) hidróxido de cobre + Mancozeb; (T9) propineb+(-trifloxistrobina+tebuconazol); (T10) trifloxistrobina+tebuconazol)+B. subtilis+hidróxido de cobre. Se determinó la presencia de la marca de polinización en la flor, la carga de polen en los estigmas, el número de semillas por fruto, y el tamaño y masa de los frutos en cada tratamiento. Posteriormente, se estimó la tasa de mortalidad de Melipona quadrifasciata expuesta a cuatro fungicidas (trifloxistrobina+tebuconazol, manganeso y zinc, hidróxido de cobre, Bacillus subtilis). La tasa de mortalidad de M. quadrifasciata en 24 horas de evaluación fue mayor en los tratamientos con hidróxido de cobre y trifloxistrobina+tebuconazol (75 y 50%, respectivamente). La tasa de mortalidad fue menor en los tratamientos con manganeso y zinc, Bacillus subtilis y el tratamiento de control. Los tratamientos con trifloxistrobina y tebuconazol redujeron la presencia de marcas de mordida y granos de polen en el estigma de las flores. Los frutos de los tratamientos control y con B. subtilis e hidróxido de cobre fueron más grandes y tuvieron mayor masa. Por lo tanto, un mayor número de aplicaciones de pesticidas en las plantas de tomate reducen las tasas de visitas de abejas en las flores y en consecuencia, la cantidad de granos de polen depositados en los estigmas afectando también la producción de los frutos.The use of agrochemicals is harmful to bees visiting agricultural crops, reducing production gains from pollination, but the effect of fungicides on these bees is not known. The objective of this study was to verify the effect of bee visitation influenced by different fungicides on the tomato crop and on the deposition of pollen grains on the stigma, number of seeds, mass and fruit size. The experiment was conducted with 10 treatments: (T1) control treatment, without application of agrochemicals; (T2 and T3) Bacillus subtilis in different application frequencies; (T4) copper hydroxide; (T5) B. subtilis and copper hydroxide; (T6) acibenzolar-S-methyl; (T7) (trifloxystrobin+tebuconazole) and B. subtilis; (T8) copper hydroxide+Mancozeb; (T9) propineb+(trifloxystrobin+ tebuconazole); (T10) (trifloxystrobin+tebuconazole)+B. subtilis+copper hydroxide. The presence of the pollination mark on the flower, the pollen load of the stigmas, the number of seeds per fruit, and the size and mass of the fruits were determined in each treatment. Subsequently, the mortality rate of Melipona quadrifasciata (Hymenoptera, Apidae) exposed to four fungicides (trifloxystrobin+tebuconazole; manganese and zinc; copper hydroxide; Bacillus subtilis) was estimated. The mortality rate of M. quadrifasciata over 24 h of evaluation was higher in the treatments with copper hydroxide and trifloxystrobin+tebuconazole (75 and 50%, respectively). The mortality rate was lower in the treatments with manganese and zinc and Bacillus subtilis and in the control treatment. The treatments with trifloxystrobin+tebuconazole reduced the presence of bite marks on the flowers and of pollen grains on the flower stigma. The fruits of the control treatments and treatments with B. subtilis and copper hydroxide were larger and had greater mass, as compared to other agrochemicals. Thus, a higher number of pesticide applications on the tomatoes reduced bee visitation rates to the flowers and, consequently, reduced the amount of pollen grains deposited on the stigmas, also reducing the fruit production.Bibliografía: páginas 434-43

Amelioration of Streptozotocin-Induced Diabetes in Mice with Cells Derived from Human Marrow Stromal Cells

Pluri-potent bone marrow stromal cells (MSCs) provide an attractive opportunity to generate unlimited glucose-responsive insulin-producing cells for the treatment of diabetes. We explored the potential for human MSCs (hMSCs) to be differentiated into glucose-responsive cells through a non-viral genetic reprogramming approach.Two HMSC lines were transfected with three genes: PDX-1, NeuroD1 and Ngn3 without subsequent selection, followed by differentiation induction in vitro and transplantation into diabetic mice. Human MSCs expressed mRNAs of the archetypal stem cell markers: Sox2, Oct4, Nanog and CD34, and the endocrine cell markers: PDX-1, NeuroD1, Ngn3, and Nkx6.1. Following gene transfection and differentiation induction, hMSCs expressed insulin in vitro, but were not glucose regulated. After transplantation, hMSCs differentiated further and approximately 12.5% of the grafted cells expressed insulin. The graft bearing kidneys contained mRNA of insulin and other key genes required for the functions of beta cells. Mice transplanted with manipulated hMSCs showed reduced blood glucose levels (from 18.9+/-0.75 to 7.63+/-1.63 mM). 13 of the 16 mice became normoglycaemic (6.9+/-0.64 mM), despite the failure to detect the expression of SUR1, a K(+)-ATP channel component required for regulation of insulin secretion.Our data confirm that hMSCs can be induced to express insulin sufficient to reduce blood glucose in a diabetic mouse model. Our triple gene approach has created cells that seem less glucose responsive in vitro but which become more efficient after transplantation. The maturation process requires further study, particularly the in vivo factors influencing the differentiation, in order to scale up for clinical purposes

Bone Marrow Mesenchymal Stem Cells for Improving Hematopoietic Function: An In Vitro and In Vivo Model. Part 2: Effect on Bone Marrow Microenvironment

The aim of the present study was to determine how mesenchymal stem cells (MSC) could improve bone marrow (BM) stroma function after damage, both in vitro and in vivo. Human MSC from 20 healthy donors were isolated and expanded. Mobilized selected CD34+ progenitor cells were obtained from 20 HSCT donors. For in vitro study, long-term bone marrow cultures (LTBMC) were performed using a etoposide damaged stromal model to test MSC effect in stromal confluence, capability of MSC to lodge in stromal layer as well as some molecules (SDF1, osteopontin,) involved in hematopoietic niche maintenance were analyzed. For the in vivo model, 64 NOD/SCID recipients were transplanted with CD34+ cells administered either by intravenous (IV) or intrabone (IB) route, with or without BM derived MSC. MSC lodgement within the BM niche was assessed by FISH analysis and the expression of SDF1 and osteopontin by immunohistochemistry. In vivo study showed that when the stromal damage was severe, TP-MSC could lodge in the etoposide-treated BM stroma, as shown by FISH analysis. Osteopontin and SDF1 were differently expressed in damaged stroma and their expression restored after TP-MSC addition. Human in vivo MSC lodgement was observed within BM niche by FISH, but MSC only were detected and not in the contralateral femurs. Human MSC were located around blood vessels in the subendoestal region of femurs and expressed SDF1 and osteopontin. In summary, our data show that MSC can restore BM stromal function and also engraft when a higher stromal damage was done. Interestingly, MSC were detected locally where they were administered but not in the contralateral femur

Reproductive success of Cabralea canjerana (Meliaceae) in Atlantic forest fragments, Brazil

In Brazil, the Atlantic forest remnants have high biological diversity and a high level of endemism, but very little is known about the reproductive success of native species. Cabralea canjerana is a common tree in the Montane Atlantic forest, and its reproduction is highly dependent on pollinators. In order to contribute with the particular knowledge on this species, we collected data in three fragmented and three continuous forest sites, where the effects of fragmentation on both mutualistic (pollination) and antagonistic (seed predation) interactions were analysed. We determined fruit production and weight of 25 trees per site. The number of seeds and the percentage of predated and aborted seeds were also accessed for seven fruits of 10 trees per site. Pollinator visitation frequencies to flowers were recorded in two forest fragments and in two sites of the continuous forest. Our data showed that plants of C. canjerana produced more fruits (z-value=-8.24; p<0.0001) and seeds per fruit (z-value=-6.58; p=0.002) in the continuous than in the fragmented sites. This was likely due to differences in pollination, because the number of pollinator visits was higher in the continuous forest than in the fragments. Seed abortion (z-value=4.08, p<0.001) and predation (z-value=3.72, p=0.0002), on the other hand, were higher in the fragmented than in the continuous sites. Then, mutualistic and antagonistic interactions were affected by fragmentation, decreasing the reproductive success of the study tree. This study was the first to show a decrease in the reproductive output in forest fragments in an Atlantic forest tree species. This decrease may threaten the population structure and viability of C. canjerana in forest fragments.In Brazil, the Atlantic forest remnants have high biological diversity and a high level of endemism, but very little is known about the reproductive success of native species. Cabralea canjerana is a common tree in the Montane Atlantic forest, and its reproduction is highly dependent on pollinators. In order to contribute with the particular knowledge on this species, we collected data in three fragmented and three continuous forest sites, where the effects of fragmentation on both mutualistic (pollination) and antagonistic (seed predation) interactions were analysed. We determined fruit production and weight of 25 trees per site. The number of seeds and the percentage of predated and aborted seeds were also accessed for seven fruits of 10 trees per site. Pollinator visitation frequencies to flowers were recorded in two forest fragments and in two sites of the continuous forest. Our data showed that plants of C. canjerana produced more fruits (z-value=-8.24; p<0.0001) and seeds per fruit (z-value=-6.58; p=0.002) in the continuous than in the fragmented sites. This was likely due to differences in pollination, because the number of pollinator visits was higher in the continuous forest than in the fragments. Seed abortion (z-value=4.08, p<0.001) and predation (z-value=3.72, p=0.0002), on the other hand, were higher in the fragmented than in the continuous sites. Then, mutualistic and antagonistic interactions were affected by fragmentation, decreasing the reproductive success of the study tree. This study was the first to show a decrease in the reproductive output in forest fragments in an Atlantic forest tree species. This decrease may threaten the population structure and viability of C. canjerana in forest fragments. 

Optimisation of mesenchymal stromal cells karyotyping analysis: implications for clinical use

[Purpose]: The aim of this study was to optimise the yield of metaphases in mesenchymal stromal cells (MSC) in vitro cultures and to study the karyotype of MSC expanded in good manufacturing practice (GMP) conditions for clinical use. [Background]: MSC are being increasingly used in clinical trials for a number of diseases. Biosafety demonstration in all cases is mandatory. Unfortunately, current standard karyotyping methods fail to obtain enough number of evaluable metaphases. [Methods and materials]: In the present work, to optimise the yield of metaphases in MSC expanded in vitro, we have tested several conditions by modifying colcemid concentration (we have tested 0·01, 0·05 and 0·1 μg mL -1) and exposure time (during 5, 15 and 24 h). We further applied these optimised conditions to 61 MSC expansions in GMP conditions for clinical use. [Results]: Our results show that the highest number of metaphases was obtained when MSC were incubated with 0·05 μg mL -1 of colcemid overnight (15 h), compared to the remaining experimental conditions. In most cases (59/61 cases) enough number of metaphases was obtained. And what is more relevant, only in one case a karyotypic abnormality was found (trisomy of chromosome 10), and cells were subsequently discarded for clinical use. [Conclusion]: We describe here an optimal method to obtain enough number of metaphases for karyotype analysis of in vitro expanded MSCs, what is essential for their clinical use in cell therapy programmes. © 2012 The Authors. Transfusion Medicine © 2012 British Blood Transfusion Society.This project had been partially supported by a grant from Ministerio de Ciencia e Innovación (reference code PLE2009-0094).Peer Reviewe