Contaminant dynamics in honey bees and hive products of apiaries from environmentally contrasting Argentinean regions.

Argentina is a leading honey producer and honey bees are also critical for pollination services and wild plants. At the same time, it is a major crop producer with significant use of insecticides, posing risks to bees. Therefore, the presence of the highly toxic insecticide chlorpyrifos, and forbidden contaminants (organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs)) was investigated in honey bee, beebread, wax and honey samples in apiaries from three contrasting regions of Argentina. Chlorpyrifos was detected in all samples with higher levels during period 1 (spring) in contrast to period 2 (fall), agreeing with its season-wise use in different crops, reaching 3.05 ng/g in honey bees. A subsequent first-tier pesticide hazard analysis revealed that it was relevant to honey bee health, mainly due to the high concentrations found in wax samples from two sites, reaching 132.4 ng/g. In addition, wax was found to be the most contaminated matrix with a prevalence of OCPs (∑OCPs 58.23-172.99 ng/g). Beebread samples showed the highest concentrations and diversity of pesticide residues during period 1 (higher temperatures). A predominance of the endosulfan group was registered in most samples, consistent with its intensive past use, especially in Central Patagonia before its prohibition. Among the industrial compounds, lighter PCB congeners dominated, suggesting the importance of atmospheric transport. The spatio-temporal distribution of pesticides shows a congruence with the environmental characteristics of the areas where the fields are located (i.e., land use, type of productive activities and climatic conditions). Sustained monitoring of different pollutants in beekeeping matrices is recommended to characterize chemical risks, assess the health status of honey bee hives and the pollution levels of different agroecosystems. This knowledge will set a precedent for South America and be helpful for actions focused on the conservation of pollination services, apiculture and ecosystems in Argentina.

Lethal concentrations of Cymbopogon nardus essential oils and their main component citronellal on Varroa destructor and Apis mellifera.

Varroosis is a disease caused by the mite Varroa destructor, and it is considered one of the biggest threats to honey bee populations globally. Mite control is centered on the use of synthetic acaricides, such as amitraz and flumethrine. However, high usage of these chemicals is associated with a wide variety of undesirable effects on bee colonies, including the development of resistance and persistence of harmful residues of acaricides in hive products used by humans. Botanical extracts have been identified as a potentially suitable organic alternative to synthetic acaricides. Essential oils, such as clove, eucalyptus, lemongrass, and oregano, have been found to exhibit acaricidal activity against V. destructor. The main goal of this work was to assess the bioactivity of the Cymbopogon nardus essential oil from two different locations (Argentina and India), and the activity of its major component the monoterpene citronellal. According to our results, complete essential oil from India is more effective in controlling parasitosis than the isolated citronellal component. The essential oil of C. nardus from Argentina demonstrated promise for the control of varroosis, as well as exhibiting low toxicity against bees (LC50 = 11.84 µL/mL). In addition, this essential oil may avoid the problems caused by synthetic acaricides, such as the emergence of resistance foci in Varroa and residues in hive products. Future research needs to investigate the delivery of volatile essentials oils to target mite populations.

High-resolution (mtDNA) melting analysis for simple and efficient characterization of Africanized honey bee Apis mellifera (Hymenoptera:Apidae).

Analysis of the mtDNA variation in Apis mellifera L. has allowed distinguishing subspecies and evolutionary lineages by means of different molecular methods; from RFLP, to PCR-RFLP and direct sequencing. Likewise, geometric morphometrics (GM) has been used to distinguish Africanized honey bees with a high degree of consistency with studies using molecular information. High-resolution fusion analysis (HRM) allows one to quickly identify sequence polymorphisms by comparing DNA melting curves in short amplicons generated by real-time PCR (qPCR). The objective of this work was to implement the HRM technique in the diagnosis of Africanization of colonies of A. mellifera from Argentina, using GM as a validation method. DNA was extracted from 60 A. mellifera colonies for mitotype identification. Samples were initially analyzed by HRM, through qPCRs of two regions (485 bp/385 bp) of the mitochondrial cytochrome b gene (cytb). This technique was then optimizing to amplify a smaller PCR product (207 bp) for the HRM diagnosis for the Africanization of colonies. Of the 60 colony samples analyzed, 41 were classified as colonies of European origin whereas 19 revealed African origin. All the samples classified by HRM were correctly validated by GM, demonstrating that this technique could be implemented for a rapid identification of African mitotypes in Apis mellifera samples.

Viruses that affect Argentinian honey bees (Apis mellifera).

Beekeeping is a widespread activity in Argentina, mainly producing honey that has gained both national and international recognition. There are more than 3,000,000 hives in the country, mainly concentrated in Buenos Aires Province (approximately 1,000,000 hives). In recent decades, worrying rates of hive loss have been observed in many countries around the world. In Latin America, the estimated loss of hives is between 13% (Peru and Ecuador) and 53% (Chile). Argentina had annual losses of 34% for the period of October 1, 2016 to October 1, 2017. The causes of these losses are not clear but probably involve multiple stressors that can act simultaneously. One of the main causes of loss of bee colonies worldwide is infestation by the ectoparasitic mite Varroa destructor in combination with viral infections. To date, 10 viruses have been detected that affect honey bees (Apis mellifera) in Argentina. Of these, deformed wing virus, sacbrood virus, acute bee paralysis virus, chronic bee paralysis virus, and Israeli acute bee paralysis can be transmitted by mites. Deformed wing virus and the AIK complex are the viruses most often associated with loss of hives worldwide. Considering that bee viruses have been detected in Argentina in several hymenopteran and non-hymenopteran insects, these hosts could act as important natural reservoirs for viruses and play an important role in their dispersal in the environment. Further studies to investigate the different mechanisms by which viruses spread in the environment will enable us to develop various strategies for the control of infected colonies and the spread of viruses in the habitat where they are found.

Fragrance addition improves visitation by honeybees and fruit quality in kiwifruit (Actinidia deliciosa).

BACKGROUND: Kiwifruit is an important horticultural crop all over the world and its development is important in Argentina. This dioecious crop has a short blooming period with nectarless flowers, and its fruit production depends on cross-pollination. Here, we tested whether kiwifruit quality increases by using honeybees exposed to female flowers treated with an artificial fragrance. The three experimental treatments were: A, sprinkled female flowers with 1:1 sugar syrup + Lavandula hybrida extract solution (a new attractant substance especially developed for this study named Lavandin Grosso); B, sprinkled female flowers with 1:1 water + sugar syrup (female flowers with additional sugar syrup reward); C (control; female flowers exposed to honeybees). RESULTS: The results showed a higher number of visits of honeybees to the female flowers sprinkled with the attractant substance, Lavandin Grosso, as well as higher fruit quality (weight, number of seeds, regularity in fruit size). CONCLUSION: Our study demonstrates the potential of fragrance-treated flowers to improve yield production in kiwifruit. © 2021 Society of Chemical Industry.

Chronic bee paralysis virus (CBPV) in South American non-Apis bees.

Chronic bee paralysis virus (CBPV) is a positive single-stranded RNA virus that exhibits a worldwide distribution. Although the effects of this virus on honeybees' health are well known, its presence in other bee species has not been fully studied. In this work, CBPV was detected in several native bees from Argentina, including Bombus pauloensis, Halictillus amplilobus, Peponapis fervens, and members of the genus Xylocopa. Here, we report for the first time the presence of CBPV in native bees from South America.

Impacts of dietary supplementation with p-coumaric acid and indole-3-acetic acid on survival and biochemical response of honey bees treated with tau-fluvalinate.

Pollinator populations are in decline worldwide. Multiple factors have been cited as potential causes to these declines. In honey bees, a combination of stressors is known to cause colony losses. Adequate nutrition is a key factor for honey bee growth and colony development. Several studies show that the nutritional quality of the diet is directly proportional to the ability of the bee to face challenges or stressors. We explored the effect of p-coumaric (600 µM) and indole-3-acetic acid (2, 20 or 200 µM) supplementation on the survival and activity of key detoxification enzymes of honey bees exposed to tau-fluvalinate. The dietary supplementation with p-coumaric and indole-3-acetic acids (20 µM) enhanced the survival of bees exposed to tau-fluvalinate (approximately 20%). We also showed that dietary p-coumaric acid increased the levels of cytochrome P450 and glutathione reductase activity in bees treated with tau-fluvalinate, as well as in the untreated controls, while glutathione-S-transferase activity was lower in treated bees than in untreated. In bees fed with indole-3-acetic acid, cytochrome P450 showed increased levels, however, glutathione-S-transferase showed the lowest activity. Moreover, the results showed that supplementation with p-coumaric and indole-3-acetic acids did not alter acetyl cholinesterase activity, nor did treatment with tau-fluvalinate. Altogether, the enzymatic changes related to the detoxification mechanisms observed in bees that were fed with p-coumaric and indole-3-acetic acids could be responsible for the increased survival of bees treated with tau-fluvalinate compared to those that received a control diet. The results presented in this study, together with previous studies, provide evidence of the importance of dietary phytochemicals in the response of honey bees to pesticide exposure. Moreover, these results are the first report of the beneficial effect of the phytohormone indole-3-acetic acid on the survival of honey bees treated with tau-fluvalinate.

Abscisic acid enhances cold tolerance in honeybee larvae.

The natural composition of nutrients present in food is a key factor determining the immune function and stress responses in the honeybee (Apis mellifera). We previously demonstrated that a supplement of abscisic acid (ABA), a natural component of nectar, pollen, and honey, increases honeybee colony survival overwinter. Here we further explored the role of ABA in in vitro-reared larvae exposed to low temperatures. Four-day-old larvae (L4) exposed to 25°C for 3 days showed lower survival rates and delayed development compared to individuals growing at a standard temperature (34°C). Cold-stressed larvae maintained higher levels of ABA for longer than do larvae reared at 34°C, suggesting a biological significance for ABA. Larvae fed with an ABA-supplemented diet completely prevent the low survival rate due to cold stress and accelerate adult emergence. ABA modulates the expression of genes involved in metabolic adjustments and stress responses: Hexamerin 70b, Insulin Receptor Substrate, Vitellogenin, and Heat Shock Proteins 70. AmLANCL2, the honeybee ABA receptor, is also regulated by cold stress and ABA. These results support a role for ABA increasing the tolerance of honeybee larvae to low temperatures through priming effects.

Laurel leaf extracts for honeybee pest and disease management: antimicrobial, microsporicidal, and acaricidal activity.

A diverse set of parasites and pathogens affects productivity and survival of Apis mellifera honeybees. In beekeeping, traditional control by antibiotics and molecules of synthesis has caused problems with contamination and resistant pathogens. In this research, different Laurus nobilis extracts are tested against the main honeybee pests through an integrated point of view. In vivo effects on bee survival are also evaluated. The ethanol extract showed minimal inhibitory concentration (MIC) values of 208 to 416 µg/mL, having the best antimicrobial effect on Paenibacillus larvae among all substances tested. Similarly, this leaf extract showed a significant antiparasitic activity on Varroa destructor, killing 50 % of mites 24 h after a 30-s exposure, and on Nosema ceranae, inhibiting the spore development in the midgut of adult bees ingesting 1 × 10(4) µg/mL of extract solution. Both ethanol extract and volatile extracts (essential oil, hydrolate, and its main component) did not cause lethal effects on adult honeybees. Thus, the absence of topical and oral toxicity of the ethanol extract on bees and the strong antimicrobial, microsporicidal, and miticidal effects registered in this study place this laurel extract as a promising integrated treatment of bee diseases and stimulates the search for other bioactive phytochemicals from plants.

Evaluation of the Protoscolicidal Activity of Humulus lupulus Methanolic Extracts on Echinococcus granulosus Sensu Stricto.

The larval stage of the parasite Echinococcus granulosus sensu lato (s.l) is responsible for cystic echinococcosis (CE), a long-term infection affecting humans and animals worldwide, and constitutes a serious public health concern. If left untreated, CE can cause serious damage to multiple organs, especially the liver and lungs. Regarding the treatment, in the last few years, the use of pharmacological treatment has increased, suggesting that in the future, drug therapy may replace surgery for uncomplicated cysts. However, the only available anthelmintic drug to treat this infection is the albendazole, which has an efficacy that does not exceed 50%. On the basis of the above-mentioned evidence, new and improved alternative treatments are urgently needed. The use of natural products and their active fractions and components holds great promise as a valuable resource for the development of novel and effective therapies. Hop (Humulus lupulus L.) is a bittering agent in the brewing industry for which the sedative, digestive, anti-inflammatory, and antimicrobial effects have been reported. The purpose of this study was to assess the in vitro efficacy of methanolic extracts from the leaves of hop varieties against E. granulosus sensu stricto (s.s) protoscoleces. Varieties Mapuche and Victoria caused a stronger protoscolicidal effect compared to the Bullion, Cascade, and Traful varieties (P < 0.01), coinciding with their highest content of flavonoids, total polyphenols, and saponins. The viability of protoscoleces treated with the varieties Mapuche and Victoria decreased to approximately 50% at days 5 y 8, respectively, showing alterations such as soma contraction and impaired microtriches. After 18 days of treatment with both varieties, protoscoleces were completely altered both structurally and ultrastructurally. In conclusion, the methanolic extracts of the H. lupulus varieties Mapuche and Victoria demonstrated a marked in vitro effect against E. granulosus s.s. protoscoleces. The beer-making industry exclusively uses hop cones, leaving behind large amounts of hop leaves as an agricultural by-product that is not being utilized. On the basis of our study, we propose that hop leaves could also be used as a source of secondary metabolites with anthelmintic activity.

Occurrence of chlorpyrifos and organochlorine pesticides in a native bumblebee (Bombus pauloensis) living under different land uses in the southeastern Pampas, Argentina.

Pollinators such as Apidae bees are vital for ecosystems and food security. Unfortunately, their populations have declined due to several factors including pesticide use. Among them, the organophosphate insecticide chlorpyrifos, poses a global threat, while legacy compounds like organochlorine pesticides (OCPs) easily bioaccumulate, increasing the concern. Bombus pauloensis, a widely distributed native bee in Argentina, is used for commercial pollination; however, information regarding their health status is scarce. This study assessed chlorpyrifos and OCP levels in B. pauloensis (workers and males) and related environmental matrices living from three different land uses schemes, by means of GC-ECD and GC-MS. The ornamental horticulture field (OP) showed the highest total pesticide concentrations in workers (13.1 ng/g), flowers and soils, whereas the organic agriculture field (OA) exhibited the lowest. Chlorpyrifos was the most abundant compound, accounting for at least 20 % of pesticide load across all matrices. The food production horticulture field (FH) had the highest chlorpyrifos concentration in workers, males and soils (5.0, 4.4 and 3.3 ng/g, respectively), suggesting a local greater usage, whereas OA showed the lowest. Regarding OCPs groups, Drins and DDTs were predominant in most matrices, with FH males registering the highest levels (4.0 and 2.5 ng/g, respectively), closely followed by OP. However, metabolites' contribution indicated historical use and atmospheric inputs in all sites. Multivariate analyses confirmed the significance of site and bumblebee sex to explain pesticide composition. Males from all sites exhibited higher chlorpyrifos levels than workers and this trend was similar for some OCP groups. Overall, OA differed from FH and OP, indicating a correlation between production modes and pesticide profiles. This study demonstrates the value of B. pauloensis as a pesticide biomonitor but also offers insights into its populations' health in the area. In this sense, this information could be useful towards the preservation of this crucial pollinator.

The gut microbiome of solitary bees is mainly affected by pathogen assemblage and partially by land use.

Pollinators, including solitary bees, are drastically declining worldwide. Among the factors contributing to this decline, bee pathogens and different land uses are of relevance. The link between the gut microbiome composition and host health has been recently studied for social pollinators (e.g. honeybees), whereas the information related to solitary bees is sparse. This work aimed at the characterization of the gut microbiome of the solitary bees Xylocopa augusti, Eucera fervens and Lasioglossum and attempted to correlate the gut microbial composition with the presence and load of different pathogens and land uses. Solitary bees were sampled in different sites (i.e. a farm, a natural reserve, and an urban plant nursery) showing different land uses. DNA was extracted from the gut, 16S rRNA gene amplified and sequenced. Eight pathogens, known for spillover from managed bees to wild ones, were quantified with qPCR. The results showed that the core microbiome profile of the three solitary bees significantly varied in the different species. Pseudomonas was found as the major core taxa in all solitary bees analyzed, whereas Lactobacillus, Spiroplasma and Sodalis were the second most abundant taxa in X. augusti, E. fervens and Lasioglossum, respectively. The main pathogens detected with qPCR were Nosema ceranae, Nosema bombi and Crithidia bombi, although differently abundant in the different bee species and sampling sites. Most microbial taxa did not show any correlation with the land use, apart from Snodgrassella and Nocardioides, showing higher abundances on less anthropized sites. Conversely, the pathogens species and load strongly affected the gut microbial composition, with Bifidobacterium, Apibacter, Serratia, Snodgrassella and Sodalis abundance that positively or negatively correlated with the detected pathogens load. Therefore, pathogens presence and load appear to be the main factor shaping the gut microbiome of solitary bees in Argentina.

Body size variability of Varroa destructor and its role in acaricide tolerance.

Phenotypic plasticity has been defined as the ability of a genotype to produce different phenotypes when exposed to distinct environments throughout its ontogeny. Morphological variability of individuals is an example of this plasticity. Taking into account that several studies have reported a wide morphological variability in Varroa destructor populations, we evaluated if the body size plasticity of the parasite constituted a key factor able to modulate mites survival when they were exposed to a drug bioassays. Drug bioassays against mites were conducted using three different Syzygium aromaticum essential oil concentrations (0.5, 1, and 5 µl/capsule) and controls. After 4 h of exposition, mite mortality was registered. The width (WS) and length (LS) of the dorsal shield were measured in dead mites. General lineal models were carried to determine if V. destructor survival to acaricides was related to the explanatory variables. Data modelling confirmed that WS and LS variables, together with time interaction, were significantly related to V. destructor survival when the parasites were exposed to acaricides. The models proposed demonstrated that for the smaller S. aromaticum essential oil concentration, the larger the parasite body, the greater the probability that it remains alive at the end of the bioassay. Such relationship was inverse for the other two concentrations tested. Possible causes explaining the body size variability in V. destructor individuals were discussed.

Africanized honeybee population (Apis mellifera L.) in Nicaragua: Forewing length and mitotype lineages.

Various subspecies of Apis mellifera L. were introduced to Central America since colonization 500 years ago. Hybridization increased with the entrance of the Africanized bee in Nicaragua in 1984. Rustic beekeeping activities and numerous feral swarms define the genetic pattern, reflected in phenotypic heterogeneity and remarkable differences in the behaviour of the bee colonies, especially the nest defence. Due to these facts, the question emerge about the degree of Africanization of honeybee colonies in Nicaragua. In this study, we identified Africanized honeybee colonies based on the single character "mean forewing length" and we corroborated our results by determining mitotypes using mtDNA analysis. Morphometric and genetic approaches were realized in three different geographical zones of Nicaragua and related to beehive characteristics and management. Worker bee samples were taken from the inside of 146 hives from 26 apiaries. Abdominal colour as phenotypic character was the first examination, followed by measurement of 1460 right forewings to determine corresponding probability of Africanization. More than 60% of the beehives showed phenotypic heterogeneity and mean forewing length of 8.74 mm (SD 0.16 mm) indicated a high degree of Africanization. Those results provided a selection of 96 worker bees to perform PCR of two worker bees per hive. For mitochondrial DNA analysis 14 samples from sentinel apiaries were added. Three from 61 beehives presented bees with different mtDNA. Throughout, three mitotypes of the African (A) lineage were detected; one mitotype is still unidentified. Mitotype A1 A. mellifera iberiensis was represented by 88 bees and mitotype A4 A. mellifera scutellata by 21 bees. Phylogenetic analysis confirmed the PCR findings. No associations were found between mitotypes, forewing length, beehive characteristics and management. A high degree of Africanization in A. mellifera colonies represented by two predominating mitotypes from the A lineage, prevail in Neotropical Nicaragua, with mitotype A4 predominating at higher altitudes.

Hydrolats from Humulus lupulus and Their Potential Activity as an Organic Control for Varroa destructor.

Varroa destructor is a parasitic mite, which is considered a severe pest for honey bees causing serious losses to beekeeping. Residual hydrolats from steam extraction of hop essential oils, generally considered as a waste product, were tested for their potential use as acaricides on V. destructor. Four hop varieties, namely Cascade, Spalt, Victoria, and Mapuche, showed an interesting performance as feasible products to be used in the beekeeping industry. Some volatile oxidized terpenoids were found in the hydrolats, mainly ß-caryophyllene oxide, ß-linalool, and isogeraniol. These compounds, together with the presence of polyphenols, flavonoids, and saponins, were probably responsible for the promissory LC50 values obtained for mites after hydrolat exposition. Victoria hydrolat was the most toxic for mites (LC50: 16.1 µL/mL), followed by Mapuche (LC50 value equal to 30.1 µL/mL), Spalt (LC50 value equal to 114.3 µL/mL), and finally Cascade (LC50: 117.9 µL/mL). Likewise, Spalt had the highest larval survival, followed by Victoria and Mapuche. Cascade was the variety with the highest larval mortality. In addition, none of the extracts showed mortality higher than 20% in adult bees. The Victoria hydrolat presented the best results, which makes it a good compound with the prospect of an acaricide treatment against V. destructor.

Repellent and acaricidal effects of botanical extracts on Varroa destructor.

Extracts of indigenous plants from South America have shown a broad spectrum of bioactivities. No-contaminant and natural substances have recently resurged as control treatment options for varroosis in honey bee colonies from Argentina. The aim of this work was to evaluate the biological activity of botanical extracts from Baccharis flabellata and Minthostachys verticillata on Varroa destructor and Apis mellifera. The acaricidal and insecticidal activities were assessed by the spraying application method. Both ethanolic extracts showed high levels of toxicity against the mites and were harmless to their host, A. mellifera. During the attractive-repellent test, the olfactory stimulus evoked for the extract from B. flabellata resulted as a repellent for mites. The aromatic stimulus of these extracts would be strong enough to cause disturbance on the behavior of V. destructor. Thus, the repellent effect of these substances plus the toxicity on mites postulate these botanical extracts like promising natural compound to be incorporated for the control of varroosis.

Susceptibility of Varroa destructor (Acari: Varroidae) to synthetic acaricides in Uruguay: Varroa mites' potential to develop acaricide resistance.

The purpose of this study was to estimate the acaricide susceptibility of Varroa destructor populations from Uruguay, which had never been exposed to synthetic acaricides. It was also to determine whether acaricide resistance to coumaphos occurred in apiaries in which acaricide rotation had been applied. Bioassays with acaricides against mite populations that had never been exposed to synthetic acaricides were performed, also against mite populations in which control failures with coumaphos had been reported. Additionally, coumaphos' effectiveness in honeybee colonies was experimentally tested. The lethal concentration that kills 50% of the exposed animals (LC(50)) for susceptible mite populations amounted to 0.15 µg/Petri dish for coumaphos and to less than 0.3 µg/Petri dish for the other acaricides. Coumaphos LC(50) was above 40 µg/Petri dish for resistant mites. The effectiveness of coumaphos in honeybee colonies parasitized by V. destructor ranged from 17.6% to 93.9%. LC(50) for mite populations susceptible to the most commonly applied miticides was determined, and the first case of coumaphos resistance recorded in Uruguay was established.

Parasites and RNA viruses in wild and laboratory reared bumble bees Bombus pauloensis (Hymenoptera: Apidae) from Uruguay.

Bumble bees (Bombus spp.) are important pollinators insects involved in the maintenance of natural ecosystems and food production. Bombus pauloensis is a widely distributed species in South America, that recently began to be managed and commercialized in this region. The movement of colonies within or between countries may favor the dissemination of parasites and pathogens, putting into risk while populations of B. pauloensis and other native species. In this study, wild B. pauloensis queens and workers, and laboratory reared workers were screened for the presence of phoretic mites, internal parasites (microsporidia, protists, nematodes and parasitoids) and RNA viruses (Black queen cell virus (BQCV), Deformed wing virus (DWV), Acute paralysis virus (ABCV) and Sacbrood virus (SBV)). Bumble bee queens showed the highest number of mite species, and it was the only group where Conopidae and S. bombi were detected. In the case of microsporidia, a higher prevalence of N. ceranae was detected in field workers. Finally, the bumble bees presented the four RNA viruses studied for A. mellifera, in proportions similar to those previously reported in this species. Those results highlight the risks of spillover among the different species of pollinators.

Lotmaria passim (Kinetoplastea: Trypanosomatidae) in honey bees from Argentina.

Lotmaria passim (Kinetoplastea) is considered the most prevalent as well as the most virulent trypanosomatid associated to the European honey bee Apis mellifera. We used qPCR to screen for the presence of this parasite in 57 samples from ten Argentinian provinces, and were able to detect its presence throughout most of the country with 41% of the samples testing positive. In a retrospective analysis, we detected L. passim in 73% of honey bee samples from 2006 showing that this flagellate has been widely present in Argentina for at least ~15 years. Additionally, three primer sets for L. passim detection were compared, with the pair that produced smallest PCR product having the best detection capability. Finally, we also found L. passim DNA in 100% (n = 6) of samples of the mite Varroa destructor. The role of this ectoparasite in the lifecycle of Lotmaria, if any, remains unrevealed.

Resistance phenomena to amitraz from populations of the ectoparasitic mite Varroa destructor of Argentina.

In Argentina, Varroa destructor resistance to coumaphos has been previously reported. However, the status of mite susceptibility to other hard acaricides is still unknown. At present, high infestation levels of V. destructor are being detected in colonies of Apis mellifera after treatment with amitraz. The aim of the present study was to determine the LC(50) of amitraz in V. destructor from three apiaries with high mite density after treatment with the acaricide. The LC(50) values were 3.9, 3.5, and 3.7 µg/Petri dish for mites from three different apiaries. Significant LC(50) differences were detected between resistant and susceptible mites. LC(50) increased 35-39-fold when compared to the corresponding baseline, suggesting the development of resistance. These results are the first report of resistance to amitraz in V. destructor in Argentina and extend the knowledge according to the status of acaricides resistance in the country.