The virome of an endangered stingless bee suffering from annual mortality in southern Brazil.

Meliponiculture - the management of stingless bee colonies - is an expanding activity in Brazil with economic, social and environmental potential. However, unlike in apiculture, the pathogens that impact on meliponiculture remain largely unknown. In southern Brazil, every year at the end of the summer, managed colonies of the stingless bee Melipona quadrifasciata manifest a syndrome that eventually leads to collapse. Here we characterize the M. quadrifasciata virome using high-throughput sequencing, with the aim of identifying potentially pathogenic viruses, and test whether they are related to the syndrome outbreaks. Two paired viromes are explored, one from healthy bees and another from unhealthy ones. Each virome is built from metagenomes assembled from sequencing reads derived either from RNA or DNA. A total of 40 621 reads map to viral contigs of the unhealthy bees' metagenomes, whereas only 11 reads map to contigs identified as viruses of healthy bees. The viruses showing the largest copy numbers in the virome of unhealthy bees belong to the family Dicistroviridae - common pathogenic honeybee viruses - as well as Parvoviridae and Circoviridae, which have never been reported as being pathogenic in insects. Our analyses indicate that they represent seven novel viruses associated with stingless bees. PCR-based detection of these viruses in individual bees (healthy or unhealthy) from three different localities revealed a statistically significant association between viral infection and symptom manifestation in one meliponary. We conclude that although viral infections may contribute to colony collapses in the annual syndrome in some meliponaries, viruses spread opportunistically during the outbreak, perhaps due to colony weakness.

Physicochemical parameters and content of B-complex vitamins: an exploratory study of bee pollen from southern Brazilian states / Parámetros físico-químicos y contenido de vitaminas del complejo B: un estudio exploratorio del polen apícola del sur de Brasil

ABSTRACT This study aimed to determine the presence of B-complex vitamins and some physicochemical parameters in bee pollen samples from the southern Brazilian states of Paraná, Santa Catarina and Rio Grande do Sul and, then, to identify their correlations with the geographical and botanical origin of the samples using multivariate statistical techniques. B-complex vitamins were determined by HPLC and mean contents were 0.79 mg/100 g (vitamin B1), 0.88 mg/100 g (vitamin B2), 5.31 mg/100 g (sum of vitamin B3 vitamers) and 4.42 mg/100 g (sum of B6 vitamers). The physicochemical parameters of the samples were consistent with those reported in the literature. The results showed that bee pollen is an important source of B-complex vitamins and multivariate statistical exploratory techniques suggested its nutritional content should be evaluated locally.

RESUMEN El objetivo de este estudio fue determinar la presencia de vitaminas del complejo B y algunos parámetros físico-químicos en muestras de polen apícola oriundas de los Estados del Sur de Brasil (Paraná, Santa Catarina y Rio Grande do Sul); y correlacionar estos resultados con el origen botánico y geográfico, usando un análisis estadístico multivariado. Las vitaminas del complejo B fueron determinadas por HPLC y su contenido fue de 0,79 mg/100 g (vitamina B1); 0,88 mg/100 g (vitamina B2); 5,31 mg/100 g (suma de los compuestos de vitamina B3); y 4,42 mg/100 g (suma de los compuestos de vitamina B6). Los parámetros físico-químicos de las muestras fueron consistentes con los reportados en la literatura. Los resultados mostraron que el polen apícola fue una fuente importante de vitaminas del complejo B, y las técnicas multivariadas sugirieron que su contenido nutricional debe ser evaluado localmente.

Report on the microbiota of Melipona quadrifasciata affected by a recurrent disease.

Melipona quadrifasciata is an eusocial stingless bee traditionally used for honey production in Brazil. In the last decades, the species disappeared from the wild in Southern Brazil, being kept exclusively in managed colonies for commercial and recreational purposes. Stingless beekeepers from this region report annual losses of their colonies due to a syndrome of yet unknown causes. We investigate whether it is associated to pathogenic microorganisms already known to cause disease in bees. These results provide a starting point for future studies aimed at clarifying the relationship between the microbial community of stingless bees and their colony collapses.

Stingless Bees as Alternative Pollinators of Canola.

Alternative pollinators can ensure pollination services if the availability of the managed or most common pollinator is compromised. In this study, the behavior and pollination efficiency of Apis mellifera L. and two species of stingless bees, Plebeia emerina Friese and Tetragonisca fiebrigi Schwarz, were evaluated and compared in flowers of Brassica napus L. 'Hyola 61'. A. mellifera was an efficient pollinator when collecting nectar because it effectively touched the reproductive organs of the flower. In contrast, stingless bees were efficient pollinators only when collecting pollen. The number of pollen grains deposited on the stigma after a single visit by worker bees of the three species was greater than the number of grains resulting from pollination without the bee visits. On average, the three species deposited enough pollen grains to fertilize all of the flower ovules. A. mellifera and P. emerina had similar pollination efficiency because no significant differences were observed in the characteristics of the siliques produced. Although T. fiebrigi is also an effective pollinator, the seed mass produced by their pollination was lower. Native bees promoted similar rates of fruit set compared with A. mellifera. Thus, P. emerina has potential to be used for pollination in canola crops.

Nosema ceranae has been present in Brazil for more than three decades infecting Africanized honey bees.

Until the mid-1990s, the only microsporidium known to infect bees of the genus Apis was Nosema apis. A second species, Nosema ceranae, was first identified in 1996 from Asian honey bees; it is postulated that this parasite was transmitted from the Asian honey bee, Apis cerana, to the European honey bee, Apis mellifera. Currently, N. ceranae is found on all continents and has often been associated with honey bee colony collapse and other reports of high bee losses. Samples of Africanized drones collected in 1979, preserved in alcohol, were analyzed by light microscopy to count spores and were subjected to DNA extraction, after which duplex PCR was conducted. All molecular analyses (triplicate) indicated that the drones were infected with both N. ceranae and N. apis. PCR products were sequenced and matched to sequences reported in the GenBank (Acc. Nos. JQ639316.1 and JQ639301.1). The venation pattern of the wings of these males was compared to those of the current population living in the same area and with the pattern of drones collected in 1968 from Ribeirão Preto, SP, Brazil, from a location close to where African swarms first escaped in 1956. The morphometric results indicated that the population collected in 1979 was significantly different from the current living population, confirming its antiquity. Considering that the use of molecular tools for identifying Nosema species is relatively recent, it is possible that previous reports of infections (which used only light microscopy, without ultrastructural analysis) wrongly identified N. ceranae as N. apis. Although we can conclude that N. ceranae has been affecting Africanized honeybees in Brazil for at least 34 years, the impact of this pathogen remains unclear.

Genetic parameters for five traits in Africanized honeybees using Bayesian inference.

Heritability and genetic correlations for honey (HP) and propolis production (PP), hygienic behavior (HB), syrup-collection rate (SCR) and percentage of mites on adult bees (PMAB) of a population of Africanized honeybees were estimated. Data from 110 queen bees over three generations were evaluated. Single and multi-trait models were analyzed by Bayesian Inference using MTGSAM. The localization of the hive was significant for SCR and HB and highly significant for PP. Season-year was highly significant only for SCR. The number of frames with bees was significant for HP and PP, including SCR. The heritability estimates were 0.16 for HP, 0.23 for SCR, 0.52 for HB, 0.66 for PP, and 0.13 for PMAB. The genetic correlations were positive among productive traits (PP, HP and SCR) and negative between productive traits and HB, except between PP and HB. Genetic correlations between PMAB and other traits, in general, were negative, except with PP. The study permitted to identify honeybees for improved propolis and honey production. Hygienic behavior may be improved as a consequence of selecting for improved propolis production. The rate of syrup consumption and propolis production may be included in a selection index to enhance honeybee traits.