First large-scale study reveals important losses of managed honey bee and stingless bee colonies in Latin America.

Over the last quarter century, increasing honey bee colony losses motivated standardized large-scale surveys of managed honey bees (Apis mellifera), particularly in Europe and the United States. Here we present the first large-scale standardized survey of colony losses of managed honey bees and stingless bees across Latin America. Overall, 1736 beekeepers and 165 meliponiculturists participated in the 2-year survey (2016-2017 and 2017-2018). On average, 30.4% of honey bee colonies and 39.6% of stingless bee colonies were lost per year across the region. Summer losses were higher than winter losses in stingless bees (30.9% and 22.2%, respectively) but not in honey bees (18.8% and 20.6%, respectively). Colony loss increased with operation size during the summer in both honey bees and stingless bees and decreased with operation size during the winter in stingless bees. Furthermore, losses differed significantly between countries and across years for both beekeepers and meliponiculturists. Overall, winter losses of honey bee colonies in Latin America (20.6%) position this region between Europe (12.5%) and the United States (40.4%). These results highlight the magnitude of bee colony losses occurring in the region and suggest difficulties in maintaining overall colony health and economic survival for beekeepers and meliponiculturists.

Short communication: Varroa destructor re-invasion dynamics during autumn and winter in Apis mellifera colonies from a temperate climate.

Infestation with Varroa destructor compromises the survival of Apis mellifera colonies, especially during the winter season. In order to prevent colony losses due to the presence of Varroa mites, determination of the infestation level and the application of autumn treatment is highly recommended. However, autumn reinvasion by inter-apiary and intra-apiary horizontal transmission might threaten the control strategies. Drivers like the infestation level of untreated colonies, the presence of bee brood during late autumn and the colonies disposition within the apiary might explain at least partially the re-invasion rates of treated colonies during autumn and winter. A total of six apiaries with five colonies were distributed within a 30 km radius. Colonies were arranged within each apiary following a circular or a lineal disposition. Twenty-four of the thirty colonies were treated during late summer against V. destructor and one colony per apiary remained untreated. The phoretic Varroa infestation and the adult bee population, brood area and nutritional reserves in the colonies was estimated before and after treatment and then for the next four months (late autumn and winter season). In this study, a model for the V. destructor re-invasion dynamics during autumn and winter in honey bee colonies from a temperate climate was fitted. According to these results, the re-invasion probability is higher for lineal disposition of the colonies that also present a higher amount of brood availability during late autumn and winter. Beekeeping practical implications are discussed in the context of an Integrated Pest management approach.

Field evaluation of Varroa-resistance traits in surviving Apis mellifera colonies in Argentina.

Varroa destructor is one of the most important sanitary threats for the beekeeping industry and so far disease control is based mainly on chemical treatment. However, a long-term solution may arise from studying natural surviving colonies of Apis mellifera. We compared the Varroa infestation rate in six commercial colonies that received annual treatment against mites and six non-treated colonies that survived in absence of any treatment for the last 6 years. In addition, we evaluated two potential mechanisms that might be involved in colony survival: hygienic (HYG) and Varroa-sensitive hygiene behavior (VSH) by means of pin-killed and mite artificial infestation, respectively. HYG and VSH were negatively correlated with mite infestation independently of the colony group (treated or non-treated). Furthermore, colonies expressing high levels of pupae removal (≥ 80%) showed higher %HYG and lower mite infestation compared to colonies showing low pupae removal (< 80%). The analysis of reproductive status of mites from the non-removed infested cells evidenced that more infertile mites are found in colonies with more than 80% of pupae removal. To study non-treated colonies that survive for several years, it is a suitable approach for identifying the underlying mechanisms related to Varroa-resistance.

Potential associations between the mite Varroa destructor and other stressors in honeybee colonies (Apis mellifera L.) in temperate and subtropical climate from Argentina.

The presence of Varroa destructor in colonies of Apis mellifera is explained by the interaction among a number of factors including beekeeping practices and surrounding environment features. The aim of this study was to evaluate the relative impact of environment geographical region and beekeeping management on Varroa infestation levels throughout a year. A monitoring study was carried out during 2015 in north-central regions from Argentina, consisting of three sampling dates: 1) autumn survey before autumn acaricide treatment; 2) autumn survey after autumn acaricide treatment and 3) spring survey. During these visits, we collected samples for Varroa mites and Nosema sp. presence assessment and information concerning the apiary management practices during each period. Both regional location and beekeeping practices impact on V. destructor infestation level during the course of the year, but relative importance depend partially on the time of year when this was observed. Varroa infestation level is driven simultaneously by a wide-range of environmental factors (regional effect) and honeybee population dynamics. Additionally, colony life histories are also strongly affected by the management practices employed by beekeepers, especially regarding the Varroa mites control and the supplementary feeding. Complexity involving multiple factors interaction in socio-ecological systems like beekeeping is discussed.

Prevalence of honey bee (Apis mellifera) viruses in temperate and subtropical regions from Argentina / Prevalencia de virus en abejas melíferas (Apis mellifera) en ambientes templados y subtropicales de Argentina

In Argentina, bee virus studies are still incipient, and there are no studies regarding the climatic effect. The aim of this study was to assess and compare the presence of honeybee viruses in different climatic regions from Argentina. A total of 385 colonies distributed in five Argentinean eco-regions were examined to evaluate the percentage of infestation with Varroa destructor and the presence of seven virus species (Deformed wing virus, DWV; Acute bee paralysis virus, ABPV; Chronic bee paralysis virus, CBPV; Black queen cell virus, BQCV; Kashmer bee virus, KBV; Israeli acute bee paralysis virus, IAPV; and Sacbrood bee virus, SBV) after honey yield. Two viruses, KBV and IAPV, were not detected. The other five viruses were found in different prevalences: DWV (35%), ABPV (21.5%), BQCV (8.0%), CBPV (2.2%), and SBV (1.1%). We found double and triple viral associations in approximately 25% of the sampled colonies. The mean V. destructor infestation in the colonies prior to the acaricide treatment was 7.12% ± 8.7%. The knowledge of the prevalence of these viruses in the region and their relation with the mite and other possible influencing factors is important for preventing colony losses. Further studies are necessary to identify the risk factors associated with virus presence and its relationship with other pathogens such as V. destructor.

En Argentina, los estudios sobre prevalencia de virus en abejas continúan siendo incipientes y no existen reportes acerca de cómo inciden sobre dicha prevalencia las variables climáticas. El objetivo de este estudio fue evaluar y comparar la presencia de virus en abejas melíferas en diferentes regiones agroecológicas de Argentina. A tal fin se evaluaron 385 colmenas distribuidas en 5 regiones agroecológicas de las provincias de Chaco y Santa Fe; en ellas se analizó el porcentaje de infestación con Varroa destructor (ácaro patógeno de abejas) y la presencia de 7 especies de virus (DWV, virus de las alas deformadas; ABPV, virus de la parálisis aguda de la abeja; CBPV, virus de la parálisis crónica de la abeja; BQCV, virus de celda negra de la reina; KBV, virus de la abeja de Cachemira; IAPV, virus israelí de la parálisis aguda y SBV, virus de la cría ensacada). luego de la cosecha de miel. Dos virus (KBV y IAPV) no fueron detectados. Las otras 5 especies de virus se encontraron con prevalencias variables: DWV (35%), ABPV (21,5%), BQCV (8%), CBPV (2,2%) y SBV (1,1%). Fue posible identificar la presencia de 3 y hasta 3 virus simultáneamente en el 25% de las colmenas evaluadas. El promedio de infestación por V. destructor en las colmenas luego de la cosecha de miel y antes del tratamiento con acaricidas fue de 7,12% (±8,7). Conocer la prevalencia de virus en las diferentes regiones agroecológicas y su relación con la presencia del ácaro V. destructor e identificar otros posibles factores que podrían influir en su presencia es relevante para definir estrategias que reduzcan la mortandad de colmenas. Es necesario realizar estudios adicionales para identificar los factores de riesgo asociados a la presencia de virus en las colmenas y su relación con otros patógenos, como V. destructor.

Risk factors for the presence of Deformed wing virus and Acute bee paralysis virus under temperate and subtropical climate in Argentinian bee colonies.

Beekeepers all across the world are suffering important losses of their colonies, and the parasitic mites Varroa destructor and Nosema sp, as well as several bee viruses, are being pointed out as the possible causes of these losses, generally associated with environmental and management factors. The objective of the present study was to evaluate the presence of seven virus species (Deformed wing virus -DWV-, Acute bee paralysis virus -ABPV-, Chronic bee paralysis virus -CBPV-, Black queen cell virus -BQCV-, Kashmir bee virus -KBV-, Israeli acute bee paralysis virus -IAPV-, and Sacbrood bee virus -SBV), as well as the prevalence of Nosema sp. and Varroa destructor, and their possible associated factors, under temperate and subtropical climate conditions in Argentinean colonies. A total of 385 colonies distributed in five Argentinean eco-regions were examined after honey harvest. The final multivariable model revealed only one variable associated with the presence of DWV and two with the presence of ABPV. The apiary random effect was significant in both cases (P=0.018; P=0.006, respectively). Colonies with a Varroa infestation rate >3% showed higher presence of DWV than colonies with <3% of Varroa infestation level (OR=1.91; 95% CI: 1.02-3.57; P<0.044). The same pattern was observed for the presence of ABPV (OR=2.23; 95% CI: 1.04-4.77; P<0.039). Also, colonies where replacement of old combs was not a common practice had higher presence of ABPV (OR=6.02; 95% CI: 1.16-31.25; P<0.033). Regardless of the location of the colonies, virus presence was strongly associated with V. destructor level. Therefore, all the factors that directly or indirectly influence the levels of mites will be also influencing the presence of the viruses.

Prevalence of honey bee (Apis mellifera) viruses in temperate and subtropical regions from Argentina.

In Argentina, bee virus studies are still incipient, and there are no studies regarding the climatic effect. The aim of this study was to assess and compare the presence of honeybee viruses in different climatic regions from Argentina. A total of 385 colonies distributed in five Argentinean eco-regions were examined to evaluate the percentage of infestation with Varroa destructor and the presence of seven virus species (Deformed wing virus, DWV; Acute bee paralysis virus, ABPV; Chronic bee paralysis virus, CBPV; Black queen cell virus, BQCV; Kashmer bee virus, KBV; Israeli acute bee paralysis virus, IAPV; and Sacbrood bee virus, SBV) after honey yield. Two viruses, KBV and IAPV, were not detected. The other five viruses were found in different prevalences: DWV (35%), ABPV (21.5%), BQCV (8.0%), CBPV (2.2%), and SBV (1.1%). We found double and triple viral associations in approximately 25% of the sampled colonies. The mean V. destructor infestation in the colonies prior to the acaricide treatment was 7.12%±8.7%. The knowledge of the prevalence of these viruses in the region and their relation with the mite and other possible influencing factors is important for preventing colony losses. Further studies are necessary to identify the risk factors associated with virus presence and its relationship with other pathogens such as V. destructor.

Environment or beekeeping management: What explains better the prevalence of honey bee colonies with high levels of Varroa destructor?

Varroa destructor is one of the major threats to honey bee colonies. The mite abundance in the colonies is affected by environmental conditions as well as by beekeeping management. The aim of this study was to recognize the main drivers associated with autumn V. destructor infestation in honey bee colonies when different regions from Argentina are compared. A total of 361 colonies distributed in five Argentinean eco-regions were examined to evaluate Varroa mite infestation rate during autumn and Nosema sp. presence. Regions were different regarding annual temperature, precipitation and especially vegetation landscape. In addition, beekeeping management practices were obtained from a checklist questionnaire answered by the beekeepers. The prevalence of colonies with high infestation level was lower in semi-arid Chaco followed by humid and transition Chaco regions. Also, colonies that were positive for Nosema sp. showed a higher Varroa infestation rate. The "environmental" effect was stronger compared with the influence of secondary drivers associated with beekeeping activities. As well, a significant association between V. destructor infestation rates and Nosema presence was identified. Under contrasting natural conditions, environment seems a predominant driver on Varroa destructor infestation level in honey bee colonies.

Distribution and prevalence of Nosema apis and N. ceranae in temperate and subtropical eco-regions of Argentina.

A total of 361 colonies from 59 apiaries located in two temperate and three subtropical eco-regions were examined during the post-harvest period to determine distribution and prevalence of Nosema spp. Apiaries from subtropical eco-regions showed a lower spore count than those from temperate eco-regions. Pure N. ceranae and co-infection were detected in apiaries from all regions. In contrast, pure N. apis infection was exclusively observed in the subtropical study region. The predominant detection of N. apis in a subtropical region joining a southern temperate region where mainly co-infected apiaries were identified is in contrast to previous reports.

Key management practices to prevent high infestation levels of Varroa destructor in honey bee colonies at the beginning of the honey yield season.

Varroa destructor is considered one of the main threats to worldwide apiculture causing a variety of physiological effects at individual and colony level. Also, Varroa mites are often associated with several honey bee viruses presence. Relatively low levels of Varroa during the spring, at the beginning of the honey yield season, can have a significant economic impact on honey production and colony health. Winter treatments against Varroa and certain management practices may delay mite population growth during following spring and summer improving colonies performance during the honey yield season. The aim of this study was to identify risk factors associated with the presence of Varroa destructor in late spring in apiaries from temperate climate. A longitudinal study was carried out in 48 apiaries, randomly selected to evaluate V. destructor infestation level throughout the year. The percentage of infestation with V. destructor was assessed four times during one year and the beekeepers answered a survey concerning all management practices applied in the colonies. We used a generalized linear mixed model to determine association between risk of achieving 2% infestation on adult bees at the beginning of the honey yield season and all potential explanatory variables. The complete dataset was scanned to identify colonies clusters with a higher probability of achieving damage thresholds throughout the year. Colonies that achieved ≥2% of infestation with V. destructor during spring were owned by less experienced beekeepers. Moreover, as Varroa populations increase exponentially during spring and summer, if the spring sampling time is later this growth remains unobserved. Monitoring and winter treatment can be critical for controlling mite population during the honey production cycle. Spatial distribution of colonies with a higher risk of achieving high Varroa levels seems to be better explained by management practices than a geographical condition.

Varroa destructor and viruses association in honey bee colonies under different climatic conditions.

Honey bee colonies are threatened by multiple factors including complex interactions between environmental and diseases such as parasitic mites and viruses. We compared the presence of honeybee-pathogenic viruses and Varroa infestation rate in four apiaries: commercial colonies that received treatment against Varroa and non-treated colonies that did not received any treatment for the last 4 years located in temperate and subtropical climate. In addition, we evaluated the effect of climate and Varroa treatment on deformed wing virus (DWV) amounts. In both climates, DWV was the most prevalent virus, being the only present virus in subtropical colonies. Moreover, colonies from subtropical climate also showed reduced DWV amounts and lower Varroa infestation rates than colonies from temperate climate. Nevertheless, non-treated colonies in both climate conditions are able to survive several years. Environment appears as a key factor interacting with local bee populations and influencing colony survival beyond Varroa and virus presence.

Critical appraisal of the MTT assay in the presence of rottlerin and uncouplers.

Rottlerin is a natural product isolated from Mallotus philippinensis. This polyphenolic compound, originally described as a selective inhibitor of PKCδ, can inhibit many other PKC-unrelated kinases and has a number of biological actions, including mitochondrial uncoupling effects. We recently found that Rottlerin inhibits the transcription factor nuclear factor κB in different cell types, causing downregulation of cyclin D1 and growth arrest. The present study was carried out to clarify the surprising lack of effect of Rottlerin on MCF-7 cell viability, assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) test. We found that Rottlerin causes overestimation of the MTT test, leading to inconsistent results between cell number and cell viability. Rottlerin, however, strongly differs from other antioxidant polyphenols, which directly reduce tetrazolium salts, since it does not exhibit any reactivity toward the tetrazolium salts in vitro nor does it modulate lactate dehydrogenase activity. The interference in the MTT assay occurred only in cultured cells, concomitantly with a decrease in the energy charge. Because the same MTT overestimation was observed in the presence of uncoupling agents, we conclude that the Rottlerin artifact is linked to its uncoupling action that, by accelerating oxidative chain, accidentally results in enhanced MTT reduction. These results suggest caution in the use of the MTT assay in the presence of Rottlerin and uncouplers in general.

Role of PTHrp and PTHrp-engaged pathways in MCF-7 cells migration/invasion.

In this paper the effect of N-terminal parathyroid hormone-related protein (PTHrp) and PTHrp-engaged pathways on MCF-7 breast cancer cell migration/invasivity and matrix metalloproteinases (MMPs) production were investigated. We found that: a) migration is not affected by PTHrp and Forskolin (FK)-activated PKA, while Phorbol Myristate Acetate (PMA)-activated PKC strongly stimulates MCF-7 cells motility. b) MMPs production was unaffected by PTHrp, but FK reduced membrane-type (MT)-1 MMP expression. Conversely, PMA induced a marked increase of MT1-MMP and MMP-9. c) Chemical activation of PKC is not sufficient, by itself, to confer invasive ability to MCF-7 cells, unless they were provided with additional factors, supplied by fibroblasts. d) Matrix invasion likely occurs through an activation cascade, involving at least three components: pro-MMP-9 and MT-1 MMP (supplied by PMA-stimulated MCF-7 cells) and pro MMP-2 (supplied by fibroblasts). e) The selective chemical inhibition of the adenylylciclase (AC)/PKA and phospholipase C (PLC)/PKC pathways confirmed that MCF-7 cells invasivity is not affected by exogenous PTHrp, which can only modulate their growth. However, the PTHrp responsibility in breast cancer invasion cannot be completely excluded. Indeed, fibroblasts are known to respond to PTHrp (which is a normal product of MCF-7 as well as other breast cancer cells) with enhanced release of MMP-2. On the basis of the documented requirement of fibroblast-derived MMP-2 for MCF-7 cell invasivity, a novel humoral fibroblast-breast cancer cell interaction, mediated by PTHrp, can be recognised.

Parathyroid hormone-related protein in preeclampsia: a linkage between maternal and fetal failures.

Preeclampsia is a disorder associated with pregnancy that affects both the mother and the fetus. Typical features of the disease are maternal hypertension, proteinuria, and edema as well as fetal growth retardation. Although the etiological details are still being debated, a consensus exists that the starting point is deficient placentation in the first half of pregnancy. The crucial early steps are reduced trophoblast invasiveness and enhanced apoptotic death. In the present review, we demonstrate that parathyroid hormone-related protein is involved not only in the maternal and fetal failures but also in the etiological aspects of the disease. We hypothesize that reduced local production of the peptide is a major causative event.

Small HDL form via apo A-I a complex with atrial natriuretic peptide.

The goal of this study was to test the ability of small high density lipoproteins (small HDL) to bind human alpha-atrial natriuretic peptide (alpha-hANP), an amyloidogenic peptide whose involvement in cardiac pathologies is gaining increasing clinical evidence. After incubation of HDL with labeled ANP, the peptide associated to lipoprotein was detectable only in small HDL containing preparations. HDL-associated alpha-[(125)I]hANP was subjected to chromatography, electrophoresis, and autoradiography. The autoradiograph showed two radioactive bands, whose molecular weight was consistent with the chromatographic pattern. Immunoblotting showed the presence of apo A-I in both autoradiographic bands. The proteins of the main band were electroeluted, incubated with labeled ANP, and subjected to two-dimensional electrophoresis followed by autoradiography. The mass spectrometry and molecular weight analyses of the radioactive spot demonstrated the presence of an apo A-I dimer. This finding provided a novel solid evidence that small HDL via apo A-I dimer are involved in the ANP sequestration and thus may play a role in preventing amyloid fibril formation.