Mulbry, W.; Karns, J.
Insecticide wastes generated from livestock dipping operations are well suited for biodegradation processes since these wastes are concentrated, contained, and have no other significant toxic components. About 400,000 L of cattle dip wastes containing approximately 1500 mg/L of the organophosphate coumaphos are generated yearly along the Mexican border from a USDA program designed to control disease carrying cattle ticks. Use of unlined evaporation pits for the disposal of these wastes has resulted in highly contaminated soils underlying these sites. Previous work has shown that microbial consortia present in selected dip wastes can be induced to mineralize coumaphos. Our laboratory results show that these consortia are able to colonize plastic fibers in trickling biofilters and can be used in these filters to quickly metabolize coumaphos from dip wastes. A field scale biofilter capable of treating 15,000 litre batches of dip waste was used to reduce the coumaphos concentration in two successive 11,000 litre batch trials from 2000 mg/L to 10 mg/L in approximately 14 d. (author)
Osman, A.Z.; Zayed, S.M.A.D.; Hazzaa, N.I.
Our study was initiated to determine the residues of 14 C-coumaphos and its metabolites which might appear in milk, meat and other different organs of lactating goats subjected to coumaphos treatment. 14 C-coumaohos (Asunto) was synthesized and applied dermally on two lactating goats. Two equal dermal applications, with 2 weeks interval, were applied on each goat. One goat was sacrificed 24 hours after the second application, and the second animal after a withdrawal period of two weeks. The results indicate that coumaphos has a slow absorption rate when dermally applied to lactating goats. After the withdrawal period, liver, kidney, skin and subcutaneous fat were found to contain considerable amounts of coumaphos residues in comparison with other different organs and milk. Thin layer chromatography showed the presence of coumaphos, its oxygen analogue, and two unknown metabolites.3 fig.,2 tab
Calumpang, S.M.F.; Medina, M.J.B.; Tejada, A.W.
A survey of cattle dip facilities and current practices employed was done. Coumaphos and ethion were the commonly used acaricides in the four respondent stock farms. The behavior of coumaphos in a simulated model cattle dip was monitored using radiotracer techniques. Degradation was rapid, resulting in the formation of potasan metabolite and bound residues in the sediment. A rapid field method for the detection of organophosphate pesticides was used in monitoring the degradation of coumaphos in a cattle dip. The sensitivity of the method is comparable to the conventional HPLC method employed. This rapid field method can easily be used by cattle ranch owners to monitor coumaphos content of the vat facility so that recharging could be made in order to prevent the onset of resistance development in cattle tick. (author)
Espinosa Gonzalez, J.; Rodriguez, F.; Barrera, R.
The concentration of coumaphos in four field cattle dipping vats and its distribution at different depths in the suspension and in the sediment was monitored over 12 to 48 weeks. The residual concentration of coumaphos and degradation products was determined by HPLC. Coumaphos was extracted from the suspension by shaking with equal volume of methanol and 90% of coumaphos was extracted by this method. A reverse phase C-18 column (25 cm x 0.4 cm) was used in the HPLC and the eluent was a mixture of methanol+ water (80+20,v/v). The initial concentration of coumaphos was 200 mg/L. However, it steadily decreased in all four vats with time. The final concentration was reduced to 39% of the concentration at zero time in vat number 1 after 12 week 17% after 18 weeks in vat number 2, 29% after 19 weeks in vat number 3 and 23% after 48 weeks in vat number 4. The concentration in the sediment increased from 165 mg/kg at zero time to 1960 mg/kg after 18 weeks in vat number 1 and 152 mg/kg to 2020 mg/kg after 48 weeks in vat number 4. The concentration of coumaphos in the suspension ranged between 28 to 81 mg/L at the surface, 46 to 115 mg/L at 20 cm and 86 to 147 mg/L at 100 cm depth. (author)
Jindal, T.; Singh, D.K.; Agarwal, H.C.
The stability of coumaphos was studied in model dipping vats under field conditions using 14 C-labelled and unlabelled coumaphos, with or without additives. Four vats were used each containing 50 litre of water treated with 3.7 MBq 14 C and 10 g (AI) unlabelled coumaphos in 25 mL acetone. Vat 1 was control. Vat 2 and 3 were maintained at pH 5 by addition of superphosphate initially and at regular intervals as required. Vat 3 and 4 were treated with bacteriostat copper sulphate at 0.01 g per litre. The pH of vat 1 and 4 gradually increased with time from initial value of 7 to 8.47 and 7.57 respectively. In vat 1 and 4 the concentration of coumaphos declined from about initial concentration of 200 μg/mL to about 9 μg/mL in 255 days after initial treatment, whereas the persistence of coumaphos was more in vats 2 and 3. The stability of coumaphos in model vats increased significantly by maintaining a pH of 5 by addition of superphosphate. The pesticide residues consisted of 80 % or more of unchanged coumaphos. In addition potasan, chlorferon and 4 - methylumbelliferone were detected in small proportions. In another experiment the effect of pH was studied. Coumaphos was most stable at pH 5 as also observed in model vats under field conditions. Coumaphos did not leach below 10 cm in all the four cases. Dissipation, degradation and bound residue formation was more in case of alkali treated coumaphos than normal coumaphos. Aged residues of vat 3 were reasonably stable as copper sulphate inhibited the degradation of coumaphos in soil by microorganisms. Coumaphos along with the the above three metabolises were detected in the extractable insecticide residues from the soil, though the proportion of metabolises was much more than found in model vats. Again treatment of coumaphos with alkali further increased its metabolism and the soil bound residues were double as compared to untreated coumaphos
Khan, S.U.; Behki, R.
Dissipation and degradation of 14 C-labelled coumaphos in suspension and binding to sediment was studied in a model laboratory glass vat was studied. The water in the vat was treated with a mixture of 14 C-labelled and unlabelled coumaphos and initial concentration determined. Two weeks later 50 g soil was added to the suspension and 3 x 25 mL samples taken and analysis. The procedure was repeated every two weeks for 44 weeks. pH was also recorded at each sampling time. At the end of the experiment soil was extracted by using Soxhlet apparatus and by supercritical fluid extraction and concentrations of free and bound coumaphos and metabolites were determined. Very little dissipation of total 14 C occurred and 88% of the initial concentration was still present at the end of 44 weeks. The 14 C content in the filtrate declined to 20% but there was a corresponding increase in the 14 C content of the sediment mixed with emulsion. The pH of the suspension steadily decreased with time. HPLC analysis of the filtrate, soil wash and Soxhlet extract indicated the presence of coumaphos and three metabolites chlorferon, potasan and oxygen analogue of coumaphos. Analysis of supercritical fluid extract of the sediment-bound radioactivity showed the presence of coumaphos but no metabolites. In another study filtered and washed soil from the tank was incubated with an EPTC-degrading bacterial strain, Rhodococcus TE1 to study the effect on the metabolism of coumaphos. About 10.9% of 14 C was released in the incubation mixture and it contained mostly chlorferon with some oxygen analogue of coumaphos. Exposure of 14 C-labelled coumaphos to U.V. and sun lamp, to study photolysis, resulted in the formation of a brown precipitate. While the 14 C content in the filtrate decreased, its concentration in the precipitate increased. The filtrate contained primarily coumaphos, whereas, the precipitate contained coumaphos, chlorferon and potasan. These findings suggest that in the field vat the
Valdovinos-Flores, Cesar; Gaspar-Ramírez, Octavio; Heras–Ramírez, María Elena; Dorantes-Ugalde, José Antonio; Saldaña-Loza, Luz María
In the search of alternatives for controlling Aethina tumida Murray, we recently proposed the BAA trap which uses boric acid and an attractant which mimics the process of fermentation caused by Kodamaea ohmeri in the hive. This yeast is excreted in the feces of A. tumida causing the fermentation of pollen and honey of infested hives and releasing compounds that function as aggregation pheromones to A. tumida. Since the boron is the toxic element in boric acid, the aim of this article is to assess the amount of boron residues in honey and beeswax from hives treated with the BAA trap. For this aim, the amount of bioaccumulated boron in products of untreated hives was first determined and then compared with the amount of boron of products from hives treated with the BAA trap in two distinct climatic and soil conditions. The study was conducted in the cities of Padilla, Tamaulipas, and Valladolid, Yucatan (Mexico) from August 2014 to March 2015. The quantity of boron in honey was significantly less in Yucatan than in Tamaulipas; this agrees with the boron deficiency among Luvisol and Leptosol soils found in Yucatan compared to the Vertisol soil found in Tamaulipas. In fact, the honey from Yucatan has lower boron levels than those reported in the literature. The BAA treatment was applied for four months, results show that the BAA trap does not have any residual effect in either honey or wax; i.e., there is no significant difference in boron content before and after treatment. On the other hand, the organophosphate pesticide coumaphos was found in 100% of wax samples and in 64% of honey samples collected from Yucatan. The concentration of coumaphos in honey ranges from 0.005 to 0.040 mg/kg, which are below Maximum Residue Limit (MRL) allowed in the European Union (0.1 mg/kg) but 7.14% of samples exceeded the MRL allowed in Canada (0.02 mg/kg). PMID:27092938
Full Text Available In the search of alternatives for controlling Aethina tumida Murray, we recently proposed the BAA trap which uses boric acid and an attractant which mimics the process of fermentation caused by Kodamaea ohmeri in the hive. This yeast is excreted in the feces of A. tumida causing the fermentation of pollen and honey of infested hives and releasing compounds that function as aggregation pheromones to A. tumida. Since the boron is the toxic element in boric acid, the aim of this article is to assess the amount of boron residues in honey and beeswax from hives treated with the BAA trap. For this aim, the amount of bioaccumulated boron in products of untreated hives was first determined and then compared with the amount of boron of products from hives treated with the BAA trap in two distinct climatic and soil conditions. The study was conducted in the cities of Padilla, Tamaulipas, and Valladolid, Yucatan (Mexico from August 2014 to March 2015. The quantity of boron in honey was significantly less in Yucatan than in Tamaulipas; this agrees with the boron deficiency among Luvisol and Leptosol soils found in Yucatan compared to the Vertisol soil found in Tamaulipas. In fact, the honey from Yucatan has lower boron levels than those reported in the literature. The BAA treatment was applied for four months, results show that the BAA trap does not have any residual effect in either honey or wax; i.e., there is no significant difference in boron content before and after treatment. On the other hand, the organophosphate pesticide coumaphos was found in 100% of wax samples and in 64% of honey samples collected from Yucatan. The concentration of coumaphos in honey ranges from 0.005 to 0.040 mg/kg, which are below Maximum Residue Limit (MRL allowed in the European Union (0.1 mg/kg but 7.14% of samples exceeded the MRL allowed in Canada (0.02 mg/kg.
Williamson, Sally M; Baker, Daniel D; Wright, Geraldine A
The decline of honeybees and other pollinating insects is a current cause for concern. A major factor implicated in their decline is exposure to agricultural chemicals, in particular the neonicotinoid insecticides such as imidacloprid. Honeybees are also subjected to additional chemical exposure when beekeepers treat hives with acaricides to combat the mite Varroa destructor. Here, we assess the effects of acute sublethal doses of the neonicotinoid imidacloprid, and the organophosphate acaricide coumaphos, on honey bee learning and memory. Imidacloprid had little effect on performance in a six-trial olfactory conditioning assay, while coumaphos caused a modest impairment. We report a surprising lack of additive adverse effects when both compounds were administered simultaneously, which instead produced a modest improvement in learning and memory.
Chaimanee, Veeranan; Evans, Jay D; Chen, Yanping; Jackson, Caitlin; Pettis, Jeffery S
Honey bee population declines are of global concern. Numerous factors appear to cause these declines including parasites, pathogens, malnutrition and pesticides. Residues of the organophosphate acaricide coumaphos and the neonicotinoid insecticide imidacloprid, widely used to combat Varroa mites and for crop protection in agriculture, respectively, have been detected in wax, pollen and comb samples. Here, we assess the effects of these compounds at different doses on the viability of sperm stored in the honey bee queens' spermatheca. Our results demonstrate that sub-lethal doses of imidacloprid (0.02ppm) decreased sperm viability by 50%, 7days after treatment. Sperm viability was a downward trend (about 33%) in queens treated with high doses of coumaphos (100ppm), but there was not significant difference. The expression of genes that are involved in development, immune responses and detoxification in honey bee queens and workers exposed to chemicals was measured by qPCR analysis. The data showed that expression levels of specific genes were triggered 1day after treatment. The expression levels of P450 subfamily genes, CYP306A1, CYP4G11 and CYP6AS14 were decreased in honey bee queens treated with low doses of coumaphos (5ppm) and imidacloprid (0.02ppm). Moreover, these two compounds suppressed the expression of genes related to antioxidation, immunity and development in queens at day 1. Up-regulation of antioxidants by these compounds in worker bees was observed at day 1. Coumaphos also caused a repression of CYP306A1 and CYP4G11 in workers. Antioxidants appear to prevent chemical damage to honey bees. We also found that DWV replication increased in workers treated with imidacloprid. This research clearly demonstrates that chemical exposure can affect sperm viability in queen honey bees. Copyright © 2016 Elsevier Ltd. All rights reserved.
Cizelj, Ivanka; Glavan, Gordana; Božič, Janko; Oven, Irena; Mrak, Vesna; Narat, Mojca
The Carniolan honey bee, Apis mellifera carnica, is a Slovenian autochthonous subspecies of honey bee. In recent years, the country has recorded an annual loss of bee colonies through mortality of up to 35%. One possible reason for such high mortality could be the exposure of honey bees to xenobiotic residues that have been found in honey bee and beehive products. Acaricides are applied by beekeepers to control varroosis, while the most abundant common agricultural chemicals found in honey bee and beehive products are fungicides, which may enter the system when applied to nearby flowering crops and fruit plants. Acaricides and fungicides are not intrinsically highly toxic to bees but their action in combination might lead to higher honey bee sensitivity or mortality. In the present study we investigated the molecular immune response of honey bee workers at different developmental stages (prepupa, white-eyed pupa, adult) exposed to the acaricide coumaphos and the fungicide prochloraz individually and in combination. Expression of 17 immune-related genes was examined by quantitative RT-PCR. In treated prepupae downregulation of most immune-related genes was observed in all treatments, while in adults upregulation of most of the genes was recorded. Our study shows for the first time that negative impacts of prochloraz and a combination of coumaphos and prochloraz differ among the different developmental stages of honey bees. The main effect of the xenobiotic combination was found to be upregulation of the antimicrobial peptide genes abaecin and defensin-1 in adult honey bees. Changes in immune-related gene expression could result in depressed immunity of honey bees and their increased susceptibility to various pathogens. Copyright © 2015 Elsevier Inc. All rights reserved.
Malek, M.A.; Rahman, M.M.; Amin, M.R.; Rahman, M.S.; Khatoon, J.
The study was undertaken to evaluate the stability of coumaphos, 14 C-labelled lindane and chlorpyrifos insecticides/acaricides in a simulated cattle dipping vat. During the period of the study the pH of the suspension in vat increased due to addition of soil from the vacinity of cattle barn as well as due to standing in cement-concrete of the vat. The residual amounts of lindane and chiorpyrifos were estimated by counting the corresponding radioactivity in the suspension. Lindane dissipated rapidly and its half-life during the first phase was 4 d only. After standing for 3 weeks lindane residues were equivalent to 3.5% of the original amount in the suspension and 9.6% in the supernatent. Similarly, 97% of the original amount of lindane bad dissipated in the soil sediment at the end of 3 weeks, indicating that dissipation of lindane was primarily due to volatilization. Chlorpyrifos was more stable under the experimental conditions and its half life was calculated to be 22 d during the first phase of dissipation. After 3 weeks the concentration of chlorpyriphos in the suspension remained almost unchanged. However, in the supernatant its concentration increased to 115% of the initial amount. The concentration of soil-bound residues of chlopyrofos remained less than 5% of the total amount applied through out the period of study
... follows: Commodity Parts per million Cattle, fat 1.0 Cattle, meat 1.0 Cattle, meat byproducts 1.0 Goat, fat 1.0 Goat, meat 1.0 Goat, meat byproducts 1.0 Hog, fat 1.0 Hog, meat 1.0 Hog, meat byproducts 1.0 Honey 0.15 Honeycomb 45.0 Horse, fat 1.0 Horse, meat 1.0 Horse, meat byproducts 1.0 Milk, fat (=n in...
Gregorc, Aleš; Alburaki, Mohamed; Sampson, Blair; Knight, Patricia R; Adamczyk, John
The efficacies of various acaricides in order to control a parasitic mite, the Varroa mite, Varroa destructor , of honey bees, were measured in two different settings, namely, in laboratory caged honey bees and in queen-right honey bee colonies. The Varroa infestation levels before, during, and after the acaricide treatments were determined in two ways, namely: (1) using the sugar shake protocol to count mites on bees and (2) directly counting the dead mites on the hive bottom inserts. The acaricides that were evaluated were coumaphos, tau-fluvalinate, amitraz, thymol, and natural plant compounds (hop acids), which were the active ingredients. The acaricide efficacies in the colonies were evaluated in conjunction with the final coumaphos applications. All of the tested acaricides significantly increased the overall Varroa mortality in the laboratory experiment. Their highest efficiencies were recorded at 6 h post-treatment, except for coumaphos and thymol, which exhibited longer and more consistent activity. In the honey bee colonies, a higher Varroa mortality was recorded in all of the treatments, compared with the natural Varroa mortality during the pretreatment period. The acaricide toxicity to the Varroa mites was consistent in both the caged adult honey bees and workers in the queen-right colonies, although, two of these acaricides, coumaphos at the highest doses and hop acids, were comparatively more toxic to the worker bees.
... [Docket No. FDA-2010-N-0002] Animal Drugs, Feeds, and Related Products; Withdrawal of Approval of a New Animal Drug Application; Buquinolate; Coumaphos AGENCY: Food and Drug Administration, HHS. ACTION: Final rule. SUMMARY: The Food and Drug Administration (FDA) is amending the animal drug regulations by...
Williamson, Sally M; Wright, Geraldine A
Pesticides are important agricultural tools often used in combination to avoid resistance in target pest species, but there is growing concern that their widespread use contributes to the decline of pollinator populations. Pollinators perform sophisticated behaviours while foraging that require them to learn and remember floral traits associated with food, but we know relatively little about the way that combined exposure to multiple pesticides affects neural function and behaviour. The experiments reported here show that prolonged exposure to field-realistic concentrations of the neonicotinoid imidacloprid and the organophosphate acetylcholinesterase inhibitor coumaphos and their combination impairs olfactory learning and memory formation in the honeybee. Using a method for classical conditioning of proboscis extension, honeybees were trained in either a massed or spaced conditioning protocol to examine how these pesticides affected performance during learning and short- and long-term memory tasks. We found that bees exposed to imidacloprid, coumaphos, or a combination of these compounds, were less likely to express conditioned proboscis extension towards an odor associated with reward. Bees exposed to imidacloprid were less likely to form a long-term memory, whereas bees exposed to coumaphos were only less likely to respond during the short-term memory test after massed conditioning. Imidacloprid, coumaphos and a combination of the two compounds impaired the bees' ability to differentiate the conditioned odour from a novel odour during the memory test. Our results demonstrate that exposure to sublethal doses of combined cholinergic pesticides significantly impairs important behaviours involved in foraging, implying that pollinator population decline could be the result of a failure of neural function of bees exposed to pesticides in agricultural landscapes.
Harriet, Jorge; Campá, Juan Pablo; Grajales, Mauricio; Lhéritier, Christophe; Gómez Pajuelo, Antonio; Mendoza-Spina, Yamandú; Carrasco-Letelier, Leonidas
Over the last decade, Uruguay has expanded and intensified its rainfed crop production. This process has affected beekeeping in several ways: for example, by reducing the space available. This has increased the density of apiaries, the risk of varroosis and acaricide use. Additionally, the dominance of no-tillage crops has increased the frequencies of application and of loads of pesticides in regions where such crops share the land with beekeeping and honey production. Therefore, the exposure of bees to xenobiotics (agricultural pesticides and veterinary products) has increased in line with pollution of hives and their products. To document pollution from hive exposure to pesticides, we surveyed the presence of 30 xenobiotics normally used in Uruguay, in recycled beeswax (RB) and in honey cappings (HC) from the main Uruguayan beekeeping regions. There was contamination of all the analyzed samples (RB and HC) with the herbicide atrazine at a range of 1-2 ng g -1 . At least three or four additional xenobiotics were detected: insecticides (chlorpyrifos-ethyl and thiacloprid); fungicides (azoxystrobin and tebuconazole); and veterinary products (coumaphos, ethion, and tau-fluvalinate). The frequency of detection of chlorpyrifos-ethyl and coumaphos in RB samples was higher than in those of HC. Moreover, the concentrations of azoxystrobin, coumaphos, and tebuconazole in RB samples were higher than in HC samples. Therefore, we suggest the use of HC to produce recycled printed beeswax films for use in hives to minimize pollution transfer. Copyright © 2017 Elsevier Ltd. All rights reserved.
Full Text Available Recently, the widespread distribution of pesticides detected in the hive has raised serious concerns about pesticide exposure on honey bee (Apis mellifera L. health. A larval rearing method was adapted to assess the chronic oral toxicity to honey bee larvae of the four most common pesticides detected in pollen and wax--fluvalinate, coumaphos, chlorothalonil, and chloropyrifos--tested alone and in all combinations. All pesticides at hive-residue levels triggered a significant increase in larval mortality compared to untreated larvae by over two fold, with a strong increase after 3 days of exposure. Among these four pesticides, honey bee larvae were most sensitive to chlorothalonil compared to adults. Synergistic toxicity was observed in the binary mixture of chlorothalonil with fluvalinate at the concentrations of 34 mg/L and 3 mg/L, respectively; whereas, when diluted by 10 fold, the interaction switched to antagonism. Chlorothalonil at 34 mg/L was also found to synergize the miticide coumaphos at 8 mg/L. The addition of coumaphos significantly reduced the toxicity of the fluvalinate and chlorothalonil mixture, the only significant non-additive effect in all tested ternary mixtures. We also tested the common 'inert' ingredient N-methyl-2-pyrrolidone at seven concentrations, and documented its high toxicity to larval bees. We have shown that chronic dietary exposure to a fungicide, pesticide mixtures, and a formulation solvent have the potential to impact honey bee populations, and warrants further investigation. We suggest that pesticide mixtures in pollen be evaluated by adding their toxicities together, until complete data on interactions can be accumulated.
Zhu, Wanyi; Schmehl, Daniel R; Mullin, Christopher A; Frazier, James L
Recently, the widespread distribution of pesticides detected in the hive has raised serious concerns about pesticide exposure on honey bee (Apis mellifera L.) health. A larval rearing method was adapted to assess the chronic oral toxicity to honey bee larvae of the four most common pesticides detected in pollen and wax--fluvalinate, coumaphos, chlorothalonil, and chloropyrifos--tested alone and in all combinations. All pesticides at hive-residue levels triggered a significant increase in larval mortality compared to untreated larvae by over two fold, with a strong increase after 3 days of exposure. Among these four pesticides, honey bee larvae were most sensitive to chlorothalonil compared to adults. Synergistic toxicity was observed in the binary mixture of chlorothalonil with fluvalinate at the concentrations of 34 mg/L and 3 mg/L, respectively; whereas, when diluted by 10 fold, the interaction switched to antagonism. Chlorothalonil at 34 mg/L was also found to synergize the miticide coumaphos at 8 mg/L. The addition of coumaphos significantly reduced the toxicity of the fluvalinate and chlorothalonil mixture, the only significant non-additive effect in all tested ternary mixtures. We also tested the common 'inert' ingredient N-methyl-2-pyrrolidone at seven concentrations, and documented its high toxicity to larval bees. We have shown that chronic dietary exposure to a fungicide, pesticide mixtures, and a formulation solvent have the potential to impact honey bee populations, and warrants further investigation. We suggest that pesticide mixtures in pollen be evaluated by adding their toxicities together, until complete data on interactions can be accumulated.
Balayiannis, George; Balayiannis, Panos
The pollution of six agricultural areas of Greece (north, central, south) by insecticides used in crop protection has been investigated utilizing, as a bioindicator, bee honey produced in those areas. Honey samples collected randomly from apiaries located in those areas were analyzed for pesticide residues with a multianalytical method, able to determine simultaneously up to 10 organophosphorous insecticides from the same honey extract. Findings concerning the acaricide coumaphos were also included, even though it is not used in crop protection. Coumaphos is used to control the mite Varroa destructor, an external parasite of the honeybee. The above areas are cultivated in large extent with citrus trees or cotton or sunflower crops, which are good forages for honeybees. The main pests of those crops are insects; hence, insecticides are used on a large scale for crop protection. The most contaminated samples originated from citrus groves; 16 out of 19 had pesticide residues: 4 samples had chlorfenvinphos (21.05%), 10 had chlorpyrifos (52.63%) and 2 had phorate (10.53%). Out of 17 samples from cotton fields, residues were found in 8, phorate in 6 (35.29%), chlorfenvinphos in 1 (5.88%), and chlorpyrifos in 1 (5.88%). Out of nine samples from fields of sunflower, four had phorate residues (44.44%). In brief, from the 50 analyzed samples, residues of chlorfenvinphos were detected in 5 samples (10%), residues of chlorpyrifos in 11 samples (22%), and residues of phorate in 12 samples (24%). Their levels ranged between 0.70 and 0.89 microg/kg. Coumaphos residues ranged from 0.10 up to 4.80 microg/kg and were derived exclusively from beehives treated with Perizin (the commercial formulation of coumaphos) for Varroa control. This study indicates that in agricultural areas with developed apiculture, useful information about the occurrence and the distribution of pesticide residues due to crop protection treatments can be derived from the analysis of randomly collected honey
Full Text Available The influence of insecticides commonly used for agricultural purposes on beehive depopulation in Uruguay was investigated. Honeycombs, bees, honey and propolis from depopulated hives were analyzed for pesticide residues, whereas from active beehives only honey and propolis were evaluated. A total of 37 samples were analyzed, representing 14,800 beehives. In depopulated beehives only imidacloprid and fipronil were detected and in active beehives endosulfan, coumaphos, cypermethrin, ethion and chlorpyrifos were found. Coumaphos was present in the highest concentrations, around 1,000 µg/kg, in all the propolis samples from active beehives. Regarding depopulated beehives, the mean levels of imidacloprid found in honeycomb (377 µg/kg, Standard Deviation: 118 and propolis (60 µg/kg, Standard Deviation: 57 are higher than those described to produce bee disorientation and fipronil levels detected in bees (150 and 170 µg/kg are toxic per se. The other insecticides found can affect the global fitness of the bees causing weakness and a decrease in their overall productivity. These preliminary results suggest that bees exposed to pesticides or its residues can lead them in different ways to the beehive.
Pareja, Lucía; Colazzo, Marcos; Pérez-Parada, Andrés; Niell, Silvina; Carrasco-Letelier, Leonidas; Besil, Natalia; Cesio, María Verónica; Heinzen, Horacio
The influence of insecticides commonly used for agricultural purposes on beehive depopulation in Uruguay was investigated. Honeycombs, bees, honey and propolis from depopulated hives were analyzed for pesticide residues, whereas from active beehives only honey and propolis were evaluated. A total of 37 samples were analyzed, representing 14,800 beehives. In depopulated beehives only imidacloprid and fipronil were detected and in active beehives endosulfan, coumaphos, cypermethrin, ethion and chlorpyrifos were found. Coumaphos was present in the highest concentrations, around 1,000 μg/kg, in all the propolis samples from active beehives. Regarding depopulated beehives, the mean levels of imidacloprid found in honeycomb (377 μg/kg, Standard Deviation: 118) and propolis (60 μg/kg, Standard Deviation: 57) are higher than those described to produce bee disorientation and fipronil levels detected in bees (150 and 170 μg/kg) are toxic per se. The other insecticides found can affect the global fitness of the bees causing weakness and a decrease in their overall productivity. These preliminary results suggest that bees exposed to pesticides or its residues can lead them in different ways to the beehive.
Mullin, Christopher A; Frazier, Maryann; Frazier, James L; Ashcraft, Sara; Simonds, Roger; Vanengelsdorp, Dennis; Pettis, Jeffery S
Recent declines in honey bees for crop pollination threaten fruit, nut, vegetable and seed production in the United States. A broad survey of pesticide residues was conducted on samples from migratory and other beekeepers across 23 states, one Canadian province and several agricultural cropping systems during the 2007-08 growing seasons. We have used LC/MS-MS and GC/MS to analyze bees and hive matrices for pesticide residues utilizing a modified QuEChERS method. We have found 121 different pesticides and metabolites within 887 wax, pollen, bee and associated hive samples. Almost 60% of the 259 wax and 350 pollen samples contained at least one systemic pesticide, and over 47% had both in-hive acaricides fluvalinate and coumaphos, and chlorothalonil, a widely-used fungicide. In bee pollen were found chlorothalonil at levels up to 99 ppm and the insecticides aldicarb, carbaryl, chlorpyrifos and imidacloprid, fungicides boscalid, captan and myclobutanil, and herbicide pendimethalin at 1 ppm levels. Almost all comb and foundation wax samples (98%) were contaminated with up to 204 and 94 ppm, respectively, of fluvalinate and coumaphos, and lower amounts of amitraz degradates and chlorothalonil, with an average of 6 pesticide detections per sample and a high of 39. There were fewer pesticides found in adults and brood except for those linked with bee kills by permethrin (20 ppm) and fipronil (3.1 ppm). The 98 pesticides and metabolites detected in mixtures up to 214 ppm in bee pollen alone represents a remarkably high level for toxicants in the brood and adult food of this primary pollinator. This represents over half of the maximum individual pesticide incidences ever reported for apiaries. While exposure to many of these neurotoxicants elicits acute and sublethal reductions in honey bee fitness, the effects of these materials in combinations and their direct association with CCD or declining bee health remains to be determined.
David J Hawthorne
Full Text Available BACKGROUND: Honey bees (Apis mellifera have recently experienced higher than normal overwintering colony losses. Many factors have been evoked to explain the losses, among which are the presence of residues of pesticides and veterinary products in hives. Multiple residues are present at the same time, though most often in low concentrations so that no single product has yet been associated with losses. Involvement of a combination of residues to losses may however not be excluded. To understand the impact of an exposure to combined residues on honey bees, we propose a mechanism-based strategy, focusing here on Multi-Drug Resistance (MDR transporters as mediators of those interactions. METHODOLOGY/PRINCIPAL FINDINGS: Using whole-animal bioassays, we demonstrate through inhibition by verapamil that the widely used organophosphate and pyrethroid acaricides coumaphos and τ-fluvalinate, and three neonicotinoid insecticides: imidacloprid, acetamiprid and thiacloprid are substrates of one or more MDR transporters. Among the candidate inhibitors of honey bee MDR transporters is the in-hive antibiotic oxytetracycline. Bees prefed oxytetracycline were significantly sensitized to the acaricides coumaphos and τ-fluvalinate, suggesting that the antibiotic may interfere with the normal excretion or metabolism of these pesticides. CONCLUSIONS/SIGNIFICANCE: Many bee hives receive regular treatments of oxytetracycline and acaricides for prevention and treatment of disease and parasites. Our results suggest that seasonal co-application of these medicines to bee hives could increase the adverse effects of these and perhaps other pesticides. Our results also demonstrate the utility of a mechanism-based strategy. By identifying pesticides and apicultural medicines that are substrates and inhibitors of xenobiotic transporters we prioritize the testing of those chemical combinations most likely to result in adverse interactions.
Daun Binahong (Andredera cordifolia Steenis Sebagai Alternatif Insektisida Terhadap Miasis yang Disebabkan Lalat Chrysomya bezziana (ANREDERA CORDIFOLIA STEENIS (BINAHONG LEAF AS AN ALTERNATIVE INSECTICIDE AGAINST CHRYSOMYA BEZZIANA CAUSED MYIASIS
Full Text Available Larvae Chrysomya bezziana caused myiasis in most livestock in Indonesia. Drugs of choice for treating myasis are synthetic insecticides. Unfortunately the insecticides has negative effect on animal product, killed insect non target and insect resistenst. The research was conducted on the activity of ethanol extract of binahong leaves against various stages of C. bezziana larvae (L1, L2, and L3. Five treatments group tested were: group without any treatment (negative control, groups treated with ethanol extracts of binahong leaves 0.5%, 1%, and 2%, respectively, positive control were given coumaphos. The treatments were applied with five replications. Number of living larvae and pupae, pupae weight and number of imago were observed. The results demonstrated that 0.5% of the extract effective consentration which was able to kill the larvae (L1 and L2. Finally 2% of the extract was the most effective concentration which was able to kill larvae (L3 and decrease the pupae weight. Ethanol extract of binahong leaves was significantly able to reduce the growth of C. bezziana larvae due to contact and digestive effect of the active compounds contained in Anredera cordifolia leaf. ABSTRAK Miasis merupakan infestasi larva lalat yang terdapat pada jaringan hidup. Penyakit ini umum menyerang hewan maupun manusia. Obat yang digunakan sebagai pengendalian kasus miasis di lapangan sampai saat ini sangat terbatas dan menggunakan insektisida sintetik. Penelitian ini bertujuan melihat aktivitas ekstrak etanol daun binahong terhadap larva lalat Chrysomya bezziana pada stadium L1, L2, dan L3. Penelitian dibagi menjadi lima kelompok perlakuan dengan lima ulangan. Kelompok perlakuan terdiri dari kontrol negatif (tanpa pemberian obat, kelompok perlakuan yang diberikan ekstrak binahong dengan konsentrasi berturut-turut 0,5%, 1%, 2%, dan kelompok yang diberikan coumaphos (kontrol positif. Peubah yang diamati adalah jumlah larva yang hidup dan berkembang menjadi pupa
Mao, Wenfu; Schuler, Mary A; Berenbaum, May R
As a managed pollinator, the honey bee Apis mellifera is critical to the American agricultural enterprise. Recent colony losses have thus raised concerns; possible explanations for bee decline include nutritional deficiencies and exposures to pesticides and pathogens. We determined that constituents found in honey, including p-coumaric acid, pinocembrin, and pinobanksin 5-methyl ether, specifically induce detoxification genes. These inducers are primarily found not in nectar but in pollen in the case of p-coumaric acid (a monomer of sporopollenin, the principal constituent of pollen cell walls) and propolis, a resinous material gathered and processed by bees to line wax cells. RNA-seq analysis (massively parallel RNA sequencing) revealed that p-coumaric acid specifically up-regulates all classes of detoxification genes as well as select antimicrobial peptide genes. This up-regulation has functional significance in that that adding p-coumaric acid to a diet of sucrose increases midgut metabolism of coumaphos, a widely used in-hive acaricide, by ∼60%. As a major component of pollen grains, p-coumaric acid is ubiquitous in the natural diet of honey bees and may function as a nutraceutical regulating immune and detoxification processes. The widespread apicultural use of honey substitutes, including high-fructose corn syrup, may thus compromise the ability of honey bees to cope with pesticides and pathogens and contribute to colony losses.
Forkpah, Cordelia; Dixon, Luke R; Fahrbach, Susan E; Rueppell, Olav
The causes of the current global decline in honey bee health are unknown. One major group of hypotheses invokes the pesticides and other xenobiotics to which this important pollinator species is often exposed. Most studies have focused on mortality or behavioral deficiencies in exposed honey bees while neglecting other biological functions and target organs. The midgut epithelium of honey bees presents an important interface between the insect and its environment. It is maintained by proliferation of intestinal stem cells throughout the adult life of honey bees. We used caged honey bees to test multiple xenobiotics for effects on the replicative activity of the intestinal stem cells under laboratory conditions. Most of the tested compounds did not alter the replicative activity of intestinal stem cells. However, colchicine, methoxyfenozide, tetracycline, and a combination of coumaphos and tau-fluvalinate significantly affected proliferation rate. All substances except methoxyfenozide decreased proliferation rate. Thus, the results indicate that some xenobiotics frequently used in apiculture and known to accumulate in honey bee hives may have hitherto unknown physiological effects. The nutritional status and the susceptibility to pathogens of honey bees could be compromised by the impacts of xenobiotics on the maintenance of the midgut epithelium. This study contributes to a growing body of evidence that more comprehensive testing of xenobiotics may be required before novel or existing compounds can be considered safe for honey bees and other non-target species.
Chernetsova, Elena S; Revelsky, Alexander I; Morlock, Gertrud E
The present study is a first step towards the unexplored capabilities of Direct Analysis in Real Time (DART) mass spectrometry (MS) arising from the possibility of the desorption at an angle: scanning analysis of surfaces, including the coupling of thin-layer chromatography (TLC) with DART-MS, and a more sensitive analysis due to the preliminary concentration of analytes dissolved in large volumes of liquids on glass surfaces. In order to select the most favorable conditions for DART-MS analysis, proper positioning of samples is important. Therefore, a simple and cheap technique for the visualization of the impact region of the DART gas stream onto a substrate was developed. A filter paper or TLC plate, previously loaded with the analyte, was immersed in a derivatization solution. On this substrate, owing to the impact of the hot DART gas, reaction of the analyte to a colored product occurred. An improved capability of detection of DART-MS for the analysis of liquids was demonstrated by applying large volumes of model solutions of coumaphos into small glass vessels and drying these solutions prior to DART-MS analysis under ambient conditions. This allowed the introduction of, by up to more than two orders of magnitude, increased quantities of analyte compared with the conventional DART-MS analysis of liquids. Through this improved detectability, the capabilities of DART-MS in trace analysis could be strengthened. Copyright © 2011 John Wiley & Sons, Ltd.
Full Text Available BACKGROUND: Over the last two winters, there have been large-scale, unexplained losses of managed honey bee (Apis mellifera L. colonies in the United States. In the absence of a known cause, this syndrome was named Colony Collapse Disorder (CCD because the main trait was a rapid loss of adult worker bees. We initiated a descriptive epizootiological study in order to better characterize CCD and compare risk factor exposure between populations afflicted by and not afflicted by CCD. METHODS AND PRINCIPAL FINDINGS: Of 61 quantified variables (including adult bee physiology, pathogen loads, and pesticide levels, no single measure emerged as a most-likely cause of CCD. Bees in CCD colonies had higher pathogen loads and were co-infected with a greater number of pathogens than control populations, suggesting either an increased exposure to pathogens or a reduced resistance of bees toward pathogens. Levels of the synthetic acaricide coumaphos (used by beekeepers to control the parasitic mite Varroa destructor were higher in control colonies than CCD-affected colonies. CONCLUSIONS/SIGNIFICANCE: This is the first comprehensive survey of CCD-affected bee populations that suggests CCD involves an interaction between pathogens and other stress factors. We present evidence that this condition is contagious or the result of exposure to a common risk factor. Potentially important areas for future hypothesis-driven research, including the possible legacy effect of mite parasitism and the role of honey bee resistance to pesticides, are highlighted.
Lisa A. De Stefano
Full Text Available This paper describes a mathematical model of the learning process suitable for studies of conditioning using the proboscis extension reflex (PER in honey bees when bees are exposed to agrochemicals. Although procedural variations exist in the way laboratories use the PER paradigm, proboscis conditioning is widely used to investigate the influence of pesticides and repellents on honey bee learning. Despite the availability of several mathematical models of the learning process, no attempts have been made to apply a mathematical model to the learning curve in honey bees exposed to agrochemicals. Our model is based on the standard transfer function in the form Y=B3 e-B2 (X-1 +B4(1-e-B2 (X-1 where X is the trial number, Y is the proportion of correct responses, B2 is the learning rate, B3 is readiness to learn, and B4 is ability to learn. We reanalyze previously published data on the effect of several classes of agrochemicals including: (1 those that are considered harmless to bees (e.g., pymetrozine, essential oils, dicofol; (2 sublethal exposure to pesticides known to harm honey bees (e.g., coumaphos, cyfluthrin, fluvalinate, permethrin; and (3 putative repellents of honey bees (e.g., butyric acid, citronella. The model revealed additional effects not detected with standard statistical tests of significance.
Assessing the Developmental Neurotoxicity of 27 Organophosphorus Pesticides Using a Zebrafish Behavioral Assay, Waalkes, M., Hunter, D.L., Jarema, K., Mundy, W., and S. Padilla. The U.S. Environmental Protection Agency is evaluating methods to screen and prioritize organophosphorus pesticides for developmental neurotoxicity. As such, we are exploring a behavioral testing paradigm that can assess the effects of sublethal and subteratogenic concentrations of developmental neurotoxicants on zebrafish (Danio rerio). This in vivo assay quantifies the locomotor response to light stimuli under tandem light and dark conditions in a 96-well plate using a video tracking system on 6 day post fertilization zebrafish larvae. Each of twenty-seven organophosphorus pesticides was tested for their developmental neurotoxic potential by exposing zebrafish embryos/larvae to the pesticide at several concentrations (≤ 100 μM nominal concentration) during the first five days of development, followed by 24 hours of depuration and then behavioral testing. Approximately 22% of the chemicals (Acephate, Dichlorvos, Diazoxon, Bensulide,Tribufos, Tebupirimfos) did not produce any behavioral changes after developmental exposure, while many (Malaoxon Fosthiazate, Dimethoate, Dicrotophos, Ethoprop, Malathion, Naled, Diazinon, Methamidophos, Terbufos, Trichlorfon, Phorate, Pirimiphos-methyl, Profenofos, Z-Tetrachlorvinphos, Chlorpyrifos, Coumaphos, Phosmet, Omethoate) produced changes in swi
Burley, Lisa M; Fell, Richard D; Saacke, Richard G
We conducted research to examine the potential impacts ofcoumaphos, fluvalinate, and Apilife VAR (Thymol) on drone honey bee, Apis mellifera L. (Hymenoptera: Apidae), sperm viability over time. Drones were reared in colonies that had been treated with each miticide by using the dose recommended on the label. Drones from each miticide treatment were collected, and semen samples were pooled. The pooled samples from each treatment were subdivided and analyzed for periods of up to 6 wk. Random samples were taken from each treatment (n = 6 pools) over the 6-wk period. Sperm viability was measured using dual-fluorescent staining techniques. The exposure of drones to coumaphos during development and sexual maturation significantly reduced sperm viability for all 6 wk. Sperm viability significantly decreased from the initial sample to week 1 in control colonies, and a significant decrease in sperm viability was observed from week 5 to week 6 in all treatments and control. The potential impacts of these results on queen performance and failure are discussed.
De Roos, A J; Zahm, S H; Cantor, K P; Weisenburger, D D; Holmes, F F; Burmeister, L F; Blair, A
An increased rate of non-Hodgkin's lymphoma (NHL) has been repeatedly observed among farmers, but identification of specific exposures that explain this observation has proven difficult. During the 1980s, the National Cancer Institute conducted three case-control studies of NHL in the midwestern United States. These pooled data were used to examine pesticide exposures in farming as risk factors for NHL in men. The large sample size (n = 3417) allowed analysis of 47 pesticides simultaneously, controlling for potential confounding by other pesticides in the model, and adjusting the estimates based on a prespecified variance to make them more stable. Reported use of several individual pesticides was associated with increased NHL incidence, including organophosphate insecticides coumaphos, diazinon, and fonofos, insecticides chlordane, dieldrin, and copper acetoarsenite, and herbicides atrazine, glyphosate, and sodium chlorate. A subanalysis of these "potentially carcinogenic" pesticides suggested a positive trend of risk with exposure to increasing numbers. Consideration of multiple exposures is important in accurately estimating specific effects and in evaluating realistic exposure scenarios.
Full Text Available The honey bee, Apis mellifera, is frequently used as a sentinel to monitor environmental pollution. In parallel, general weakening and unprecedented colony losses have been reported in Europe and the USA, and many factors are suspected to play a central role in these problems, including infection by pathogens, nutritional stress and pesticide poisoning. Honey bee, honey and pollen samples collected from eighteen apiaries of western France from four different landscape contexts during four different periods in 2008 and in 2009 were analyzed to evaluate the presence of pesticides and veterinary drug residues.A multi-residue analysis of 80 compounds was performed using a modified QuEChERS method, followed by GC-ToF and LC-MS/MS. The analysis revealed that 95.7%, 72.3% and 58.6% of the honey, honey bee and pollen samples, respectively, were contaminated by at least one compound. The frequency of detection was higher in the honey samples (n = 28 than in the pollen (n = 23 or honey bee (n = 20 samples, but the highest concentrations were found in pollen. Although most compounds were rarely found, some of the contaminants reached high concentrations that might lead to adverse effects on bee health. The three most frequent residues were the widely used fungicide carbendazim and two acaricides, amitraz and coumaphos, that are used by beekeepers to control Varroa destructor. Apiaries in rural-cultivated landscapes were more contaminated than those in other landscape contexts, but the differences were not significant. The contamination of the different matrices was shown to be higher in early spring than in all other periods.Honey bees, honeys and pollens are appropriate sentinels for monitoring pesticide and veterinary drug environmental pollution. This study revealed the widespread occurrence of multiple residues in beehive matrices and suggests a potential issue with the effects of these residues alone or in combination on honey bee health.
Schmehl, Daniel R; Teal, Peter E A; Frazier, James L; Grozinger, Christina M
Populations of pollinators are in decline worldwide. These declines are best documented in honey bees and are due to a combination of stressors. In particular, pesticides have been linked to decreased longevity and performance in honey bees; however, the molecular and physiological pathways mediating sensitivity and resistance to pesticides are not well characterized. We explored the impact of coumaphos and fluvalinate, the two most abundant and frequently detected pesticides in the hive, on genome-wide gene expression patterns of honey bee workers. We found significant changes in 1118 transcripts, including genes involved in detoxification, behavioral maturation, immunity, and nutrition. Since behavioral maturation is regulated by juvenile hormone III (JH), we examined effects of these miticides on hormone titers; while JH titers were unaffected, titers of methyl farnesoate (MF), the precursor to JH, were decreased. We further explored the association between nutrition- and pesticide-regulated gene expression patterns and demonstrated that bees fed a pollen-based diet exhibit reduced sensitivity to a third pesticide, chlorpyrifos. Finally, we demonstrated that expression levels of several of the putative pesticide detoxification genes identified in our study and previous studies are also upregulated in response to pollen feeding, suggesting that these pesticides and components in pollen modulate similar molecular response pathways. Our results demonstrate that pesticide exposure can substantially impact expression of genes involved in several core physiological pathways in honey bee workers. Additionally, there is substantial overlap in responses to pesticides and pollen-containing diets at the transcriptional level, and subsequent analyses demonstrated that pollen-based diets reduce workers' pesticide sensitivity. Thus, providing honey bees and other pollinators with high quality nutrition may improve resistance to pesticides. Copyright © 2014 Elsevier Ltd. All
Al Naggar, Yahya; Codling, Garry; Vogt, Anja; Naiem, Elsaied; Mona, Mohamed; Seif, Amal; Giesy, John P
There is no clear single factor to date that explains colony loss in bees, but one factor proposed is the wide-spread application of agrochemicals. Concentrations of 14 organophosphorous insecticides (OPs) in honey bees (Apis mellifera) and hive matrices (honey and pollen) were measured to assess their hazard to honey bees. Samples were collected during spring and summer of 2013, from 5 provinces in the middle delta of Egypt. LC/MS-MS was used to identify and quantify individual OPs by use of a modified Quick Easy Cheap Effective Rugged Safe (QuEChERS) method. Pesticides were detected more frequently in samples collected during summer. Pollen contained the greatest concentrations of OPs. Profenofos, chlorpyrifos, malation and diazinon were the most frequently detected OPs. In contrast, ethoprop, phorate, coumaphos and chlorpyrifos-oxon were not detected. A toxic units approach, with lethality as the endpoint was used in an additive model to assess the cumulative potential for adverse effects posed by OPs. Hazard quotients (HQs) in honey and pollen ranged from 0.01-0.05 during spring and from 0.02-0.08 during summer, respectively. HQs based on lethality due to direct exposure of adult worker bees to OPs during spring and summer ranged from 0.04 to 0.1 for best and worst case respectively. It is concluded that direct exposure and/or dietary exposure to OPs in honey and pollen pose little threat due to lethality of bees in Egypt. Copyright © 2014 Elsevier Inc. All rights reserved.
Berry, Jennifer A; Hood, W Michael; Pietravalle, Stéphane; Delaplane, Keith S
In a study replicated across two states and two years, we tested the sublethal effects on honey bees of the miticides Apistan (tau fluvalinate) and Check Mite+ (coumaphos) and the wood preservative copper naphthenate applied at label rates in field conditions. A continuous covariate, a colony Varroa mite index, helped us disambiguate the effects of the chemicals on bees while adjusting for a presumed benefit of controlling mites. Mite levels in colonies treated with Apistan or Check Mite+ were not different from levels in non-treated controls. Experimental chemicals significantly decreased 3-day brood survivorship and increased construction of queen supercedure cells compared to non-treated controls. Bees exposed to Check Mite+ as immatures had higher legacy mortality as adults relative to non-treated controls, whereas bees exposed to Apistan had improved legacy mortality relative to non-treated controls. Relative to non-treated controls, Check Mite+ increased adult emergence weight. Although there was a treatment effect on a test of associative learning, it was not possible to statistically separate the treatment means, but bees treated with Apistan performed comparatively well. And finally, there were no detected effects of bee hive chemical on colony bee population, amount of brood, amount of honey, foraging rate, time required for marked released bees to return to their nest, percentage of released bees that return to the nest, and colony Nosema spore loads. To our knowledge, this is the first study to examine sublethal effects of bee hive chemicals applied at label rates under field conditions while disambiguating the results from mite control benefits realized from the chemicals. Given the poor performance of the miticides at reducing mites and their inconsistent effects on the host, these results defend the use of bee health management practices that minimize use of exotic hive chemicals.
Kakumanu, Madhavi L; Reeves, Alison M; Anderson, Troy D; Rodrigues, Richard R; Williams, Mark A
Honey bees (Apis mellifera) are the primary pollinators of major horticultural crops. Over the last few decades, a substantial decline in honey bees and their colonies have been reported. While a plethora of factors could contribute to the putative decline, pathogens, and pesticides are common concerns that draw attention. In addition to potential direct effects on honey bees, indirect pesticide effects could include alteration of essential gut microbial communities and symbionts that are important to honey bee health (e.g., immune system). The primary objective of this study was to determine the microbiome associated with honey bees exposed to commonly used in-hive pesticides: coumaphos, tau-fluvalinate, and chlorothalonil. Treatments were replicated at three independent locations near Blacksburg Virginia, and included a no-pesticide amended control at each location. The microbiome was characterized through pyrosequencing of V2-V3 regions of the bacterial 16S rRNA gene and fungal ITS region. Pesticide exposure significantly affected the structure of bacterial but not fungal communities. The bee bacteriome, similar to other studies, was dominated by sequences derived from Bacilli, Actinobacteria, α-, β-, γ-proteobacteria. The fungal community sequences were dominated by Ascomycetes and Basidiomycetes. The Multi-response permutation procedures (MRPP) and subsequent Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) analysis indicated that chlorothalonil caused significant change to the structure and functional potential of the honey bee gut bacterial community relative to control. Putative genes for oxidative phosphorylation, for example, increased while sugar metabolism and peptidase potential declined in the microbiome of chlorothalonil exposed bees. The results of this field-based study suggest the potential for pesticide induced changes to the honey bee gut microbiome that warrant further investigation.
Full Text Available In Italy a nation-wide monitoring network was established in 2009 in response to significant honey bee colony mortality reported during 2008. The network comprised of approximately 100 apiaries located across Italy. Colonies were sampled four times per year, in order to assess the health status and to collect samples for pathogen, chemical and pollen analyses. The prevalence of Nosema ceranae ranged, on average, from 47-69% in 2009 and from 30-60% in 2010, with strong seasonal variation. Virus prevalence was higher in 2010 than in 2009. The most widespread viruses were BQCV, DWV and SBV. The most frequent pesticides in all hive contents were organophosphates and pyrethroids such as coumaphos and tau-fluvalinate. Beeswax was the most frequently contaminated hive product, with 40% of samples positive and 13% having multiple residues, while 27% of bee-bread and 12% of honey bee samples were contaminated. Colony losses in 2009/10 were on average 19%, with no major differences between regions of Italy. In 2009, the presence of DWV in autumn was positively correlated with colony losses. Similarly, hive mortality was higher in BQCV infected colonies in the first and second visits of the year. In 2010, colony losses were significantly related to the presence of pesticides in honey bees during the second sampling period. Honey bee exposure to poisons in spring could have a negative impact at the colony level, contributing to increase colony mortality during the beekeeping season. In both 2009 and 2010, colony mortality rates were positively related to the percentage of agricultural land surrounding apiaries, supporting the importance of land use for honey bee health.
Full Text Available The human paraoxonase 2 (PON2 has been described as a highly specific lactonase hydrolysing the quorum sensing molecule N-(3-oxododecanoyl-L-homoserine lactone (3OC12-HSL and having secondary esterase but not phosphotriesterase activity, in contrast with the related enzymes PON1 and PON3. It has been suggested that PON2 enzyme activity is dependent on glycosylation and its N-terminal region has been recently demonstrated to be a transmembrane domain mediating association to membranes. In the present study we describe a mutated form of PON2, lacking the above N-terminal region, which has been further stabilized by the insertion of six amino acidic substitutions. The engineered version, hence forth called rPON2, has been over-expressed in E.coli, refolded from inclusion bodies and purified, yielding an enzyme with the same characteristics as the full length enzyme. Therefore the first conclusion of this work was that the catalytic activity is independent from the N-terminus and protein glycosylation. The kinetic characterization confirmed the primary activity on 3OC12-HSL; accordingly, in vitro experiments of inhibition of the biofilm formed by Pseudomonas aeruginosa (PAO1 have demonstrated that rPON2 is more effective than PON1. In addition, we observed small but significant activity against organophosphorothiotes pesticides, m-parathion, coumaphos and malathion.The availability of fair amount of active protein allowed to pinpoint, by mass-spectrometry, ubiquitination of Lys 168 induced in rPON2 by HeLa extract and to correlate such post-translational modification to the modulation of catalytic activity. A mutational analysis of the modified residue confirmed the result.
Full Text Available Novel hybrid TiO2 particles were developed and assessed as an adsorbent for solid phase extraction (SPE of organophosphorus pesticides (fensulfothion, parathion methyl, coumaphos, and diazinon from spiked water. The sol-gel method was used to synthesize TiO2 particles, which were coated with free-radical polystyrene (PS and poly(3-chloro-2-hydroxypropyl methacrylate (PClHPMA polymers. Particle structures were determined via Fourier transform infrared spectroscopy to confirm that the polymers were successfully anchored to the TiO2 particles. Thermogravimetric analysis was conducted to determine organic and inorganic matter in TiO2-PS and TiO2-PClHPMA particles showing results of 20 : 80 wt/wt% and 23 : 77 wt/wt%, respectively. SEM-EDS and X-ray diffraction test were conducted to determine the morphology and semielemental composition of the particles showing amorphous characteristics. By observing the contact angle, particles coated with PClHPMA were determined to be more hydrophilic than TiO2-PS particles. The pore size distributions obtained from the N2 adsorption-desorption isotherms were 0.150 and 0.168 cm3g−1. The specific surface area (BET was 239.9 m2g−1 for TiO2-PS and 225.7 m2g−1 for TiO2-PClHPMA. The synthesized particles showed relatively high yields of adsorption in SPE. The pesticide recoveries obtained by high performance liquid chromatography ranged from 6 to 26% for TiO2-PClHPMA and 44 to 92% for TiO2-PS.
Hakme, E; Lozano, A; Gómez-Ramos, M M; Hernando, M D; Fernández-Alba, A R
This work presents a non-targeted screening approach for the detection and quantitation of contaminants in bees and pollen, collected from the same hive, by GC-EI-ToF-MS. It consists of a spectral library datasets search using a compound database followed by a manual investigation and analytical standard confirmation together with semi-quantitation purposes. Over 20% of the compounds found automatically by the library search could not be confirmed manually. This number of false positive detections was mainly a consequence of an inadequate ion ratio criterion (±30%), not considered in the automatic searching procedure. Eight compounds were detected in bees and pollen. They include insecticides/acaricides (chlorpyrifos, coumaphos, fluvalinate-tau, chlorfenvinphos, pyridaben, and propyl cresol) at a concentration range of 1-1207 μg kg -1 , herbicides (oxyfluorfen) at a concentration range of 212-1773 μg kg -1 and a growth regulator hormone (methoprene). Some compounds were detected only in pollen; such as herbicides (clomazone), insecticides/acaricides and fungicides used to control Varroa mites as benzylbenzoate, bufencarb, allethrin, permethrin, eugenol and cyprodinil. Additional compounds were detected only in bees: flamprop-methyl, 2-methylphenol (2-49 μg kg -1 ) and naphthalene (1-23 μg kg -1 ). The proposed method presents important advantages as it can avoid the use of an unachievable number of analytical standards considered target compounds "a priori" but not present in the analyzed samples. Copyright © 2017 Elsevier Ltd. All rights reserved.
Jennifer A Berry
Full Text Available In a study replicated across two states and two years, we tested the sublethal effects on honey bees of the miticides Apistan (tau fluvalinate and Check Mite+ (coumaphos and the wood preservative copper naphthenate applied at label rates in field conditions. A continuous covariate, a colony Varroa mite index, helped us disambiguate the effects of the chemicals on bees while adjusting for a presumed benefit of controlling mites. Mite levels in colonies treated with Apistan or Check Mite+ were not different from levels in non-treated controls. Experimental chemicals significantly decreased 3-day brood survivorship and increased construction of queen supercedure cells compared to non-treated controls. Bees exposed to Check Mite+ as immatures had higher legacy mortality as adults relative to non-treated controls, whereas bees exposed to Apistan had improved legacy mortality relative to non-treated controls. Relative to non-treated controls, Check Mite+ increased adult emergence weight. Although there was a treatment effect on a test of associative learning, it was not possible to statistically separate the treatment means, but bees treated with Apistan performed comparatively well. And finally, there were no detected effects of bee hive chemical on colony bee population, amount of brood, amount of honey, foraging rate, time required for marked released bees to return to their nest, percentage of released bees that return to the nest, and colony Nosema spore loads. To our knowledge, this is the first study to examine sublethal effects of bee hive chemicals applied at label rates under field conditions while disambiguating the results from mite control benefits realized from the chemicals. Given the poor performance of the miticides at reducing mites and their inconsistent effects on the host, these results defend the use of bee health management practices that minimize use of exotic hive chemicals.
Notardonato, Ivan; Salimei, Elisabetta; Russo, Mario Vincenzo; Avino, Pasquale
chlorpyrifos-methyl, 9 pirimiphos-methyl, 10 malathion, 11 chlorpyrifos, 12 parathion-ethyl, 13 pirimiphos-ethyl, 14 bromophos, 15 chlorfenvinphos, 16 bromophos-ethyl, 17 stirophos, 18 diethion, 19 coumaphos, A dimethyl phthalate, B diethyl phthalate, C dibutyl phthalate, D butyl cyclohexyl phthalate, E benzyl butyl phthalate, F bis(2-ethylhexyl) phthalate, IS internal standard.
Controle de Rhipicephalus sanguineus (Latreille, 1806 (Acari: Ixodidae no canil da Escola de Veterinária da UFMG, Belo Horizonte, Minas Gerais, Brasil Control of Rhipicephalus sanguineus (Latreille, 1806 (Acari: Ixodidae in the kennel of the UFMG Veterinary School, Belo Horizonte, Minas Gerais, Brazil
Gustavo F. Paz
Full Text Available O presente estudo teve como proposta controlar uma infestação natural de Rhipicephalus sanguineus em um canil pertencente à Escola de Veterinária da UFMG, com uma área de 72 m² e 25 cães de diversas raças. Um teste de sensibilidade a produtos acaricidas foi aplicado em uma amostra de carrapatos capturados no canil. Dos produtos testados: Amitraz 12,5% (Produto 1; uma associação de Alfametrina 2% e Dichlorvós 60% (produto 2; Deltametrina 5% (produto 3 e uma associação de Trichlorfon 77,6%, Coumaphós 1% e Cyfluthrin 1% (produto 4; somente os produtos 1 e 4 apresentaram 100% de eficácia. O Amitraz 12,5% foi utilizado no controle dos estádios de R. sanguineus presentes no ambiente do canil, com um esquema de 4 aplicações a cada 7 dias de intervalo. Para os estádios parasitários, um único tratamento com Flumetrina 1.0% pour-on foi empregado em todos os animais. A inspeção e o monitoramento do ambiente e dos animais foram realizados durante um período de um ano. Estas medidas de controle reduziram significativamente a população de carrapatos dentro do canil. O canil não apresentou infestação pelo carrapato nos seis meses seguintes aos tratamentos. Uma infestação leve foi observada no 7º mês pós-tratamento, quando foi realizada uma nova aplicação de Amitraz 12,5% no ambiente do canil e um novo tratamento com Flumetrina 1.0% pour-on nos animais, com resultados satisfatórios.The present work was aimed at proposing a control measure for Rhipicephalus sanguineus in a naturally infected kennel of 72 m² of the UFMG Veterinary School with 25 dogs of different breeds. A sensitivity test to acaricide products was applied in a sample of R. sanguineus. Out of the acaricide products tested: 12.5% Amitraz (product 1; 2% Alfamethrin and 60% Dichlorvos association (product 2; 5% Deltamethrin (product 3 and the association between 77.6% Trichlorfon, 1.0% Coumaphos and 1.0% Cyfluthrin (product 4; only 1 and 4 products showed 100