Variant A of the Deformed Wings Virus Alters the Olfactory Sensitivity and the Expression of Odorant Binding Proteins on Antennas of Apis mellifera.

Insects have a highly sensitive sense of smell, allowing them to perform complex behaviors, such as foraging and peer recognition. Their sense of smell is based on the recognition of ligands and is mainly coordinated by odorant-binding proteins (OBPs). In Apis mellifera, behavior can be affected by different pathogens, including deformed wing virus (DWV) and its variants. In particular, it has been shown that variant A of DWV (DWV-A) is capable of altering the ultra-cellular structure associated with olfactory activity. In this study was evaluated olfactory sensitivity and the expression of OBP genes in honey bees inoculated with DWV-A. Electroantennographic analyses (EAG) were carried out to determine the olfactory sensitivity to the essential oils Eucalyptus globulus and Mentha piperita. The expression of nine antenna-specific OBP genes and DWV-A load in inoculated bees was also quantified by qPCR. We observed an inverse relationship between viral load and olfactory sensitivity and the expression of some OBP proteins. Thus, high viral loads reduced olfactory sensitivity to essential oils and the gene expression of the OBP2, OBP5, OBP11, and OBP12 proteins on the antennas of middle- and forager-age bees. These results suggest that DWV-A could have negative effects on the processes of aroma perception by worker bees, affecting their performance in tasks carried out in and outside the colony.

Impact of Mixed Infections of Gut Parasites Lotmaria passim and Nosema ceranae on the Lifespan and Immune-related Biomarkers in Apis mellifera.

Lotmaria passim currently appears to be the predominant trypanosome in honey bees worldwide. Although, the specific effects of L. passim by single or mixed with other gut parasites such as Nosema ceranae on honey bees' health is still unclear. We consequently measured bees' survival, parasite loads, the expression of antimicrobial peptides (AMPs) and vitellogenin gene. Thus, (1) bees naturally infected with L. passim, (2) healthy bees inoculated with Nosema ceranae, (3) bees naturally infected with L. passim and inoculated with N. ceranae and (4) healthy bees (control) were maintained under controlled conditions. Honey bees infected with N. ceranae or with mixed infections of L. passim and N. ceranae had significantly lower survival rates than the control group at 20 days post-inoculation (dpi). A competitive suppression was also detected, provided that the L. passim load was significantly affected by the presence of N. ceranae at 15 dpi. Expressions of the AMPs defensin and hymenoptaecin rapidly (two hours post-inoculation) increased in bees infected with N. ceranae and mixed infections. However, this effect was not continuous. In fact, expressions of abaecin, defensin, hymenoptaecin and vitellogenin decreased drastically at 15 dpi in bees with both single and mixed infections. The decrease in the expression of AMPs and vitellogenin throughout this period was consistent with the reduced survivals observed in this study, indicating that mixed infections of L. passim and N. ceranae, and even into a scenario of competition between them, may have a synergic effect on the survival and immune-related gene expressions (biomarkers) of worker bees.

PCR-specific detection of recently described Lotmaria passim (Trypanosomatidae) in Chilean apiaries.

The recently described trypanosome Lotmaria passim is currently considered the most predominant trypanosomatid in honey bees worldwide and could be a factor in honey bee declines. For a specific and quick detection of this pathogen, we developed primers based on the SSU rRNA and gGAPDH genes for the detection of L. passim in Chilean honey beehives. PCR products amplified and sequenced for these primers shared 99-100% identity with other sequences of L. passim. The designed primers were specific and we were able to detect a high prevalence (40-90%) of L. passim in bee hives distributed throughout Chile. Our described PCR-based method offers a feasible and specific detection of L. passim in any honey bee samples.

Molecular differentiation of recently described Neozygites osornensis (Neozygitales: Neozygitaceae) from two morphologically similar species.

Neozygites osornensis is a recently described aphid pathogen characterized by morphological criteria and compared with two morphologically close taxa, Neozygites cinarae and Neozygites turbinata. Neozygites species are traditionally classified based on morphological characteristics and molecular data that would confirm these taxa and permit investigation of their phylogenetic relationships are scarce. In this study, we successfully evaluated a genetic variation within partial 18S rDNA sequences among the three Neozygites species and supported the recent erection of N. osornensis as a new species. A phylogenic analysis was also included to support our results.

DAPI staining and fluorescence microscopy techniques for phytoplasmas.

The 4',6-diamidino-2-phenylindole (DAPI) stain technique is a simple method that was developed for confirming the presence of phytoplasmas in hand-cut or freezing microtome sections of infected tissues. DAPI binds AT-rich DNA preferentially, so that phytoplasmas, localized among phloem cells, can be visualized in a fluorescence microscope. The procedure is quick, easy to use, inexpensive, and can be used as a preliminary or quantitative method to detect or quantify phytoplasma-like bodies in infected plants.

Evaluación del color y la posición de trampas en la captura de cicadélidos en Gaultheria phillyreifolia (Ericaceae) afectadas por fitoplasmas / Evaluation of trap color and position on the capture of cicadellids in Gaultheria phillyreifolia (Ericaceae) affected by phytoplasmas

Color sticky traps are one of the main alternatives to collect insect vectors, as they are easy to handle and are inexpensive. We aimed to compare the effect of color and height of the traps on the attractiveness to potential cicadellids vectors of plant pathogens. Yellow and green colored stick traps were placed at two different heights in plant of Gaultheria phillyreifolia. Seventeen leafhopper species were identified, with Ribautiana tenerrima Herrich-Shãffer (49 percent), Carelmapu ramosi Linnavuori & DeLong (33 percent), Carelmapu aurionitens Linnavuori (5 percent) and Atanus sp. (6 percent) being the most common. All these species were significantly attracted by yellow sticky traps. Ribautiana tenerrima was the only species affected by the height of the sticky traps. However, this was also dependent on the sampling season. The phytoplasma vector candidate, C. ramosi, showed two population peaks in early and late summer, which may indicate two different generations. Males of this species were more abundant than females on sticky traps, but were both similarly attracted to yellow sticky traps. No differences were detected in the capture efficiency of both sexes at any height of the traps. The high proportion of C. ramosi captured suggests that the yellow sticky traps can be an important element for monitoring this species.

Las trampas pegajosas de colores son una de las principales alternativas en la captura de insectos vectores, debido a su bajo costo y su fácil implementación. El objetivo fue comparar las preferencias a los colores y la altura de colocación de trampas para los posibles cicadélidos vectores de fitopatógenos. Trampas pegajosas amarillas y verdes fueron colocadas en plantas de Gaultheria phillyreifolia a dos distintas alturas. Se diferenciaron 17 especies de cicadélidos, siendo Ribautiana tenerrima Herrich-Shãffer (49 por ciento), Carelmapu ramosi Linnavuori & DeLong, Carelmapu aureonitens Linnavuori (33 y 5 por ciento), Atanus sp. (6 por ciento) las especies más comunes. Todas estas especies se vieron significativamente atraídas por trampas de color amarillo. Ribautiana tenerrima fue la única especie afectada por la altura de las trampas pegajosas, aunque este efecto fue influenciado por la temporada de colecta. La especie candidato a vector, C. ramosi, presentó dos curvas de crecimiento al inicio y finales del verano, lo cual puede representar la emergencia de dos generaciones distintas. Machos de esta especie fueron más abundantes que las hembras en las trampas pegajosas, pero ambos con una similar preferencia hacia el color amarillo. Además, no se detectaron diferencias en la posición de las trampas en la captura de ambos sexos. La alta proporción capturada de C. ramosi, sugiere que trampas de color amarillo pueden ser um elemento importante en el monitoreo de esta especie.

[Evaluation of trap color and position on the capture of cicadellids in Gaultheria phillyreifolia (Ericaceae) affected by phytoplasmas]. / Evaluación del color y la posición de trampas en la captura de cicadélidos en Gaultheria phillyreifolia (Ericaceae) afectadas por fitoplasmas.

Color sticky traps are one of the main alternatives to collect insect vectors, as they are easy to handle and are inexpensive. We aimed to compare the effect of color and height of the traps on the attractiveness to potential cicadellids vectors of plant pathogens. Yellow and green colored stick traps were placed at two different heights in plant of Gaultheria phillyreifolia. Seventeen leafhopper species were identified, with Ribautiana tenerrima Herrich-Shäffer (49%), Carelmapu ramosi Linnavuori & DeLong (33%), Carelmapu aurionitens Linnavuori (5%) and Atanus sp. (6%) being the most common. All these species were significantly attracted by yellow sticky traps. Ribautiana tenerrima was the only species affected by the height of the sticky traps. However, this was also dependent on the sampling season. The phytoplasma vector candidate, C. ramosi, showed two population peaks in early and late summer, which may indicate two different generations. Males of this species were more abundant than females on sticky traps, but were both similarly attracted to yellow sticky traps. No differences were detected in the capture efficiency of both sexes at any height of the traps. The high proportion of C. ramosi captured suggests that the yellow sticky traps can be an important element for monitoring this species.