The frequency of occurrence and relative concentration of 44 pesticides in apicultural (Apis mellifera) matrices collected from five French locations (24 apiaries) were assessed from 2002 to 2005. The number and nature of the pesticides investigated varied with the matrices examined-living honeybees, pollen loads, honey, and beeswax. Pollen loads and beeswax had the highest frequency of pesticide occurrence among the apiary matrices examined in the present study, whereas honey samples had the lowest. The imidacloprid group and the fipronil group were detected in sufficient amounts in all matrices to allow statistical comparisons. Some seasonal variation was shown when residues were identified in pollen loads. Given the results (highest frequency of presence) and practical aspects (easy to collect; matrix with no turnover, unlike with bees that are naturally renewed), pollen loads were the best matrix for assessing the presence of pesticide residues in the environment in our given conditions.

In 2002, a field survey was initiated on French apiaries to monitor weakness of honey bee, Apis mellifera L., colonies. Apiaries were evenly distributed in five sites located on continental France. Five colonies were randomly selected in each apiary, leading to a total of 125 studied honey bee colonies. For 3 yr (starting in autumn 2002), colonies were visited four times per year: after winter, before summer, during summer, and before winter. Pollen loads from traps were collected at each visit. Multiresidue analyses were performed in pollen to search residues of 36 different molecules. Specific analyses were conducted to search fipronil and metabolites and also imidacloprid and metabolites. Residues of 19 searched compounds were found in samples. Contamination by pesticides ranged from 50 to 0%. Coumaphos and tau-fluvalinate residues were the most concentrated of all residues (mean concentrations were 925.0 and 487.2 microg/kg, respectively). Fipronil and metabolite contents were superior to the limit of detection in 16 samples. Residues of fipronil were found in 10 samples. Nine samples contained the sulfone compound, and three samples contained the desulfinyl compound. Residues of imidacloprid and 6-chloronicotinic acid were found in 69% of samples. Imidacloprid contents were quantified in 11 samples with values ranging from 1.1 to 5.7 microg/kg. 6-Chloronicotinic acid content was superior to the limit of quantification in 28 samples with values ranging from 0.6 to 9.3 microg/kg. Statistical tests showed no difference between places of sampling with the exception of fipronil. Possible origins of these contaminations, concentration and toxicity of pesticides, and the possible consequences for bees are discussed.

A 3-yr field survey was carried out in France, from 2002 to 2005, to study honey bee (Apis mellifera L.) colony health in relation to pesticide residues found in the colonies. This study was motivated by recent massive losses of honey bee colonies, and our objective was to examine the possible relationship between low levels of pesticide residues in apicultural matrices (honey, pollen collected by honey bees, beeswax) and colony health as measured by colony mortality and adult and brood population abundance. When all apicultural matrices were pooled together, the number of pesticide residue detected per sampling period (four sampling periods per year) and per apiary ranged from 0 to 9, with the most frequent being two (29.6%). No pesticide residues were detected during 12.7% of the sampling periods. Residues of imidacloprid and 6- chloronicotinic acid were the most frequently detected in pollen loads, honey, and honey bee matrices. Several pairs of active ingredients were present concurrently within honey bees and in pollen loads but not in beeswax and honey samples. No statistical relationship was found between colony mortality and pesticide residues. When pesticide residues from all matrices were pooled together, a mixed model analysis did not show a significant relationship between the presence of pesticide residues and the abundance of brood and adults, and no statistical relationship was found between colony mortality and pesticide residues. Thus, although certain pesticide residues were detected in apicultural matrices and occasionally with another pesticide residual, more work is needed to determine the role these residues play in affecting colony health.

In 2002, a field survey was initiated on French apiaries to monitor weakness of honey bee, Apis mellifera L., colonies. Apiaries were evenly distributed in five sites located on continental France. Five colonies were randomly selected in each apiary, leading to a total of 125 studied honey bee colonies. For 3 yr (starting in autumn 2002), colonies were visited four times per year: after winter, before summer, during summer, and before winter. Pollen loads from traps were collected at each visit. Multiresidue analyses were performed in pollen to search residues of 36 different molecules. Specific analyses were conducted to search fipronil and metabolites and also imidacloprid and metabolites. Residues of 19 searched compounds were found in samples. Contamination by pesticides ranged from 50 to 0%. Coumaphos and tau-fluvalinate residues were the most concentrated of all residues (mean concentrations were 925.0 and 487.2 microg/kg, respectively). Fipronil and metabolite contents were superior to the limit of detection in 16 samples. Residues of fipronil were found in 10 samples. Nine samples contained the sulfone compound, and three samples contained the desulfinyl compound. Residues of imidacloprid and 6-chloronicotinic acid were found in 69% of samples. Imidacloprid contents were quantified in 11 samples with values ranging from 1.1 to 5.7 microg/kg. 6-Chloronicotinic acid content was superior to the limit of quantification in 28 samples with values ranging from 0.6 to 9.3 microg/kg. Statistical tests showed no difference between places of sampling with the exception of fipronil. Possible origins of these contaminations, concentration and toxicity of pesticides, and the possible consequences for bees are discussed.

32 Short Report 34A large variety of viruses multiply in the honey bee Apis mellifera L. (Allen & Ball, 1996). 35Knowledge of the spreading mechanism of honey bee pathogens within the hive and 36 apiary is essential to our understanding of bee disease dynamics. Among the viruses 37 infecting honey bees, Chronic bee paralysis virus (CBPV) is the causal agent of chronic 38 paralysis known to induce significant losses in honey bee colonies (Ball & Bailey, 1997). 39The pathology is characterized by clusters of trembling, flightless, crawling bees and by 40 individual bees, sometimes hairless, standing at the hive entrance (Bailey et al., 1983). A 41 correlation between chronic paralysis and high viral loads of CBPV was demonstrated 42 particularly in symptomatic bees (Blanchard et al., 2007a). Moderate viral loads were also 43 demonstrated in colonies without symptoms (Blanchard et al., 2007a). To date, CBPV 44 has been detected only in A. mellifera (Allen & Ball, 1996) and the presence of this virus 45 has been observed on every continent (Bailey, 1967 destructor (Ball & Allen, 1988;Tentcheva et al., 2004;Yue & Genersch, 2005). In addition, 50DWV replicative RNA was detected in V. destructor (Yue & Genersch, 2005 63secondly to determine whether the CBPV was able to replicate in these hosts, and thirdly 64to evaluate the genome variability of a partial sequence of CBPV between them. 66Sample collection and preparation, RNA extraction and cDNA synthesis. 67In this study, the honey bees, ants and mites were collected from three apiaries where Five microliters of the cDNA were then used as template for the CBPV TaqMan PCR, or 78 the minus-strand RNA detection. 80Upgrading the CBPV real-time two-step RT-PCR assay. 81A real-time two-step TaqMan RT-PCR assay has recently been developed to quantify the 82 CBPV genomic load in bee samples (Blanchard et al., 2007a). This assay was adapted to 109Unexpectedly, a CBPV genomic load was detected in the mite sample from apiary 1 but 123Given that CBPV is a positive-strand RNA virus, the synthesis of minus-strand CBPV 124RNA is carried out during viral replication. Hence, detection of the minus-strand RNA is 125 indicative of virus replication (Craggs et al., 2001;Yue & Genersch, 2005). A specific RT- 126PCR was developed to assess the presence or absence of minus-strand CBPV RNA in 127 different samples. First strand cDNA was synthesized from the extracted RNA described 128 above, using a minus-strand specific primer. This primer consisted of a tag unrelated to 129 CBPV (lower-case) coupled to a specific primer of CBPV (upper-case) (RT ms CBPV = 130 5'-atcggaatcgcctagcttGCTTGATCTCCTCCTGCTTG-3'). The reverse transcription was 148 Sequence analysis 149The PCR products obtained from the different samples (bees, ants, varroas) were 156 Discussion 157These data show for the first time the presence of CBPV in hosts other than A. mellifera. 158In this study we demonstrated the presence of CBPV in two species of carnivore ants (C. 180This is the first discovery of a bee virus a...