Electrotarsogram responses to synthetic odorants by Varroa destructor, a primary parasite of western honey bees (Apis mellifera).

Olfaction is a key sensory modality for many arthropods and could be used as a tool in pest management through manipulation of pest behavior. Management of Varroa destructor, important parasitic mites of honey bees, could be improved through better understanding of the chemical ecology of this host-parasite relationship. We refined techniques of mounting mites to obtain electrophysiological recordings (electrotarsograms) of their responses to synthetic odor stimuli. Results of 271 electrotarsogram recordings from V. destructor revealed responses to 10 odorants relative to solvent controls. Electrotarsogram responses to methyl palmitate, ethyl palmitate, and 2-heptanol were highest at the lowest stimulus loading (10 ng) we tested, suggesting that V. destructor may have acute sensitivity to low concentrations of some odors. Results suggest that odorant origin (e.g., methyl oleate from honey bee larvae, geraniol from adult honey bee alarm pheromone, and α-terpineol, a plant secondary metabolite) can influence the degree of electrophysiological response. Varroa destructor tended to be more responsive to known attractants and repellents relative to previously unexplored odorants and some repellent terpenes. Electrotarsograms offer the potential for screening odors to determine their importance in V. destructor host detection.

Varroa destructor mite electrophysiological responses to honey bee (Apis mellifera) colony volatiles.

Detection and interpretation of chemical cues is essential for Varroa destructor Anderson and Trueman, an important parasite of honey bees (Apis mellifera L.), to complete its life cycle. We collected volatiles from honey bee brood at various developmental stages and screened for V. destructor electrophysiological responses to these with gas chromatography-linked electrotarsal detection. Volatile collections contained several methyl-alkanes that evoked electrophysiological responses from V. destructor. Moreover, odors in honey bee colonies that regulate honey bee colony structure and function were also detected by V. destructor. Collections from mid- to late-stage larvae had detectable levels of low-volatility odors identified as components of the honey bee brood pheromone and branched alkanes likely originating from brood cuticle. Among these, several mid- to heavy-molecular weight compounds elicited high proportional electrophysiological responses by V. destructor relative to their abundance but could not be identified using chemical standards of previously documented honey bee brood odors. We suggest further investigation of these unknown volatiles and future behavioral assays to determine attractiveness/repellency (valence) of those identified through chemical standards.

A range-wide domino effect and resetting of the annual cycle in a migratory songbird.

Latitudinal differences in timing of breeding are well documented but how such differences carry over to influence timing of events in the annual cycle of migratory birds is not well understood. We examined geographical variation in timing of events throughout the year using light-level geolocator tracking data from 133 migratory tree swallows ( Tachycineta bicolor) originating from 12 North American breeding populations. A swallow's breeding latitude influenced timing of breeding, which then carried over to affect breeding ground departure. This resulted in subsequent effects on the arrival and departure schedules at autumn stopover locations and timing of arrival at non-breeding locations. This 'domino effect' between timing events was no longer apparent by the time individuals departed for spring migration. Our range-wide analysis demonstrates the lasting impact breeding latitude can have on migration schedules but also highlights how such timing relationships can reset when individuals reside at non-breeding sites for extended periods of time.

Relationships between blood mercury levels, reproduction, and return rate in a small seabird.

Mercury (Hg) is a ubiquitous heavy metal that occurs naturally in the environment, but its levels have been supplemented for decades by a variety of human activities. Mercury can have serious deleterious effects on a variety of organisms, with top predators being particularly susceptible because methylmercury bioaccumulates and biomagnifies in food webs. Among birds, seabirds can have especially high levels of Hg contamination and Leach's storm-petrels (Oceanodroma leucorhoa), in particular, have amongst the highest known levels. Several populations of Leach's storm-petrels have declined recently in the Northwest Atlantic. The causes of these declines remain uncertain, but the toxic effects of Hg could be a potential factor in this decline. Here, we tested for relationships between adult blood total Hg (THg) concentration and several offspring development parameters, and adult return rate of Leach's storm-petrels breeding on Bon Portage Island (43° 28' N, 65° 44' W), Nova Scotia, Canada, between 2011 and 2015 (blood samples n = 20, 36, 6, 15, and 13 for each year, respectively). Overall, THg levels were elevated (0.78 ± 0.43 µg/g wet wt.) compared to other species of seabirds in this region, and varied significantly among years. However, we found no associations between THg levels and reproductive parameters or adult return rate. Our results indicate that levels of mercury observed in Leach's storm-petrel blood, although elevated, appear not to adversely affect their offspring development or adult return rate on Bon Portage Island.

Effects of fluvalinate on honey bee learning, memory, responsiveness to sucrose, and survival.

Contaminants can affect organisms' behaviour and, as a consequence, survival. Tau-fluvalinate (hereafter fluvalinate) is the active ingredient in a pesticide commonly used in North America to control Varroa destructor mites in honey bee (Apis mellifera) colonies. Fluvalinate's effects on honey bees are not well known. Honey bee cognitive and neural function can be assessed using the proboscis extension reflex (PER), which applies Pavlovian conditioning techniques. This study used PER to evaluate effects of fluvalinate on honey bee acquisition learning, (long-term) memory recall, responsiveness to sucrose, and mortality. We also evaluated how exclusion criteria for honey bees that did not exhibit PER during training and memory trials affected interpretation of results. Fluvalinate was administered both orally and dermally at high and low doses to mimic routes by which honey bees are exposed. We found negative effects of fluvalinate on honey bee learning, memory, responsiveness to sucrose, and survival, especially in high oral doses. We also found significant consequences to interpretation of results using different exclusion criteria. For example, almost 50% of individuals that failed to show evidence of learning subsequently showed evidence of memory. The latter results have important implications regarding traditional assessment of PER-based learning and memory; the former results suggest that evaluation of honey bee exposure to fluvalinate and attendant consequences warrants further investigation.

Fate of dermally applied miticides fluvalinate and amitraz within honey bee (Hymenoptera: Apidae) bodies.

Varroa mites, Varroa destructor Anderson & Trueman, are economically important pests of honey bees. Varroa mites are principally controlled within honey bee colonies using miticides. However, despite their importance in managing mite populations for apiculture, potential effects of miticides on honey bees are poorly understood. Using gas chromatography-flame ionization detection, we investigated concentrations, over variable time frames and within different body regions, of two commonly used miticides, tau-fluvalinate and amitraz, after dermal exposure to honey bees. We also quantified mortality of honey bees exposed to each miticide at both a low and high dose. Significant differences were observed in distributions of miticides among body regions. Within honey bee body parts, tau-fluvalinate was more readily absorbed and decreased in concentration more rapidly than amitraz. Mortality increased with higher dosages of miticides, and at higher dosages mortality was greater from fluvalinate than from amitraz. For individual honey bees, our results for rate of breakdown suggest that fluvalinate may be the preferred miticide for apiculturists, whereas our mortality results suggest that amitraz may be preferable. Either choice must be weighed against geographic variation in varroa resistance to each pesticide and attendant costs of parasitism.

Factors influencing mercury levels in Leach's storm-petrels at northwest Atlantic colonies.

Mercury (Hg) is a globally distributed heavy metal, with negative effects on wildlife. Its most toxic form, methylmercury (MeHg), predominates in aquatic systems. Levels of MeHg in marine predators can vary widely among individuals and populations. Leach's storm-petrels (Hydrobates leucorhous) have elevated levels of Hg but the role of Hg in storm-petrel population declines is unknown. In this study, we used egg and blood samples to study variation in Hg exposure among several northwest Atlantic colonies during breeding seasons, thereby evaluating relative toxicity risk within and among colonies. Total mercury (THg) concentrations were higher with increasing colony latitude, and were more pronounced in blood than in eggs. THg concentrations in blood were mostly associated with low toxicity risk in birds from the southern colonies and moderate risks in birds from the northern colonies; however, those values did not affect hatching or fledging success. THg concentrations in both eggs and blood were positively correlated with δ34S, emphasizing the role of sulfate-reducing bacteria in methylation of THg acquired through marine food webs, which is consistent with enriched δ34S profiles. By associating tracking data from foraging trips with THg from blood, we determined that blood THg levels were higher when storm-petrel's intensive search locations were over deeper waters. We conclude that spatial variation in THg concentrations in Leach's storm-petrels is attributable to differences in ocean depth at foraging locations, both at individual and colony levels. Differences in diet among colonies observed previously are the most likely cause for observed blood THg differences. As one of the few pelagic seabird species breeding in Atlantic Canada, with limited overlap in core foraging areas among colonies, Leach's storm-petrels can be used as biomonitors for less sampled offshore pelagic regions. The global trend in Hg emissions combined with legacy levels warrant continued monitoring for toxicity effects in seabirds.

The proboscis extension reflex to evaluate learning and memory in honeybees (Apis mellifera): some caveats.

The proboscis extension reflex (PER) is widely used in a classical conditioning (Pavlovian) context to evaluate learning and memory of a variety of insect species. The literature is particularly prodigious for honeybees (Apis mellifera) with more than a thousand publications. Imagination appears to be the only limit to the types of challenges to which researchers subject honeybees, including all the sensory modalities and a broad diversity of environmental treatments. Accordingly, some remarkable insights have been achieved using PER. However, there are several challenges to evaluating the PER literature that warrant a careful and thorough review. We assess here variation in methods that makes interpretation of studies, even those researching the same question, tenuous. We suggest that the numerous variables that might influence experimental outcomes from PER be thoroughly detailed by researchers. Moreover, the influence of individual variables on results needs to carefully evaluated, as well as among two or more variables. Our intent is to encourage investigation of the influence of numerous variables on PER results.

Manipulation by Plasmodium Parasites of Anopheles Mosquito Behavior and Human Odors.

PURPOSE: The phenomenon of parasites manipulating host phenotypes is well documented; the best-known examples are manipulations of host behavior. More recently, there has been interest in whether parasites can manipulate host odor phenotypes to enhance their attractiveness to vectors. We review here evidence that Plasmodium-infected mosquitoes have enhanced attraction to human hosts, especially when the parasite is sufficiently developed to be transmissible. We also review evidence suggesting that malaria-infected host odors elicit greater mosquito attraction compared to uninfected controls. METHODS: We reviewed and summarized the relevant literature. RESULTS: Though evidence is mounting that supports both premises we reviewed, there are several confounds that complicate interpretation. These include differences in Plasmodium and mosquito species studied, stage of infection tested, age of human participants in trials, and methods used to quantify volatiles. In addition, a key requirement to support the hypothesis of manipulation by parasites is that costs of manipulation be identified, and ideally, quantified. CONCLUSIONS: Substantial progress has been made to unlock the importance of odor for enhancing transmission of Plasmodium. However, there needs to be more replication using similar methods to better define the odor parameters involved in this enhancement.

Sexual selection: when to expect trade-offs.

Empirical evidence is mixed for interspecific trade-offs in investment among sexually selected traits. One important reason may be the way resources are allocated among species. Consider a set of species that obtains the same fitness pay-off for investment in song or plumage. Simulations where resources were normally distributed among species revealed significant trade-offs between song and plumage ( ± s.d. of r = -0.54 ± 0.06). However, simulations where resources were distributed in a negative binomial fashion usually produced positive correlations (r = 0.11 ± 0.09). Repeating simulations on three published studies that concomitantly quantified elaboration of song and plumage indicated that trade-offs are likely, although these analyses make assumptions that require further evaluation. Moreover, there are currently too few empirical distributions to make generalizations about the likelihood of interspecific trade-offs in sexually selected traits.

Detection of Borrelia spp., Ehrlichia canis, Anaplasma phagocytophilum, and Dirofilaria immitis in Eastern Coyotes (Canis latrans) in Nova Scotia, Canada.

Borrelia burgdorferi and Borrelia miyamotoi are tickborne zoonotic pathogens in Canada. Both bacteria are vectored by ticks, Ixodes scapularis in Atlantic Canada, but require wildlife reservoir species to maintain the bacteria for retransmission to future generations of ticks. Coyotes (Canis latrans) are opportunistic feeders, resulting in frequent contact with other animals and with ticks. Because coyotes are closely related to domestic dogs (Canis lupus familiaris), it is probable that coyote susceptibility to Borrelia infection is similar to that of dogs. We collected livers and kidneys of eastern coyotes from licensed harvesters in Nova Scotia, Canada, and tested them using nested PCR for the presence of B. burgdorferi, B. miyamotoi, and Dirofilaria immitis. Blood obtained from coyote livers was also tested serologically for antibodies to B. burgdorferi, Ehrlichia canis, Anaplasma phagocytophilum, and D. immitis. Borrelia burgdorferi and D. immitis were detected by both nested PCR and serology tests. Seroreactivity to A. phagocytophilum was also found. Borrelia miyamotoi and E. canis were not detected. Our results show that coyotes in Nova Scotia have been exposed to a number of vectorborne pathogens.

New geographic records for Echinococcus canadensis in coyotes and moose from Nova Scotia, Canada.

Echinococcus spp. tapeworms can cause serious diseases in mammals, including humans. Within the E. granulosus species complex, metacestodes produce unilocular cysts that are responsible for cystic echinococcosis in animal intermediate hosts. Canids are definitive hosts, harbouring adult cestodes in their intestines. Adult E. canadensis were recovered from the small intestine of 1 of 262 coyotes (Canis latrans) from Nova Scotia, Canada. Subsequently, we found unilocular cysts in lungs and livers of 4 of 8 sympatric moose (Alces alces) from Cape Breton Island. DNA was extracted from three cysts using the Qiagen DNeasy Blood and Tissue kit and assayed by polymerase chain reaction (PCR) with primers (cest4 and cest5) for a 117-bp region of the small subunit of ribosomal RNA of E. granulosus sensu lato, and further validated as E. canadensis G8 using primers targeting nicotinamide adenosine dinucleotide dehydrogenase subunit 1 (ND1) and cytochrome c oxidase subunit 1 (CO1) mitochondrial genes. These are the first records of E. canadensis in any of the three Maritime provinces, which include Nova Scotia, New Brunswick, and Prince Edward Island. The parasite was thought to be absent in this region due to extirpation of wolves (Canis spp.) in the 1800s. These findings suggest that further wildlife surveillance and risk assessment is warranted.

Effects at Nearctic north-temperate latitudes of indoor versus outdoor overwintering on the microsporidium Nosema ceranae and western honey bees (Apis mellifera).

In northern temperate climates, western honey bee (Apis mellifera) colonies can be wintered outdoors exposed to ambient conditions, or indoors in a controlled setting. Because very little is known about how this affects the recently-detected microsporidium Nosema ceranae, we investigated effects of indoor versus outdoor overwintering on spring N. ceranae intensity (spores per bee), and on winter and spring colony mortality. For colonies medicated with Fumagilin-B(R) to control N. ceranae, overwintering treatment did not affect N. ceranae intensity, despite outdoor-wintered colonies having significantly greater mortality. These findings suggest that N. ceranae may not always pose the most significant threat to western honey bees, and that indoor-wintering may ensure that a greater number of colonies are available for honey production and pollination services during the summer.

Relationships between circulating leucocytes and Leucocytozoon simondi in mallard, Anas platyrhynchos, ducklings.

Leucocytozoon simondi is an apicomplexan blood parasite of waterfowl that frequently causes significant mortality, and thus is expected to provoke a significant immune response in hosts. Using blood smears collected in consecutive weeks from 30 wild-stock mallard, Anas platyrhynchos, ducklings, we tested with repeated measures analyses for associations between leucocyte profiles and L. simondi infection intensities. With each of the five weeks of leucocyte profiles as response variables, we found evidence of fewer circulating heterophils and more circulating lymphocytes in the third week of infection associated with more intense L. simondi infections from the second week, but no significant relationships between leucocytes and the other four weeks of L. simondi infection. With each of the five weeks of L. simondi infection intensities as response variables, we found no associations with leucocyte profiles. Collectively, our results did not reveal tight links between leucocyte profiles and parasitism by L. simondi. Our data suggest that L. simondi was relatively benign to our wild ducks.

Deformed wing virus in western honey bees (Apis mellifera) from Atlantic Canada and the first description of an overtly-infected emerging queen.

Deformed wing virus (DWV) in western honey bees (Apis mellifera) often remains asymptomatic in workers and drones, and symptoms have never been described from queens. However, intense infections linked to parasitism by the mite Varroa destructor can cause worker wing deformity and death within 67 h of emergence. Ten workers (eight with deformed wings and two with normal wings) and three drones (two with deformed wings and one with normal wings) from two colonies infected with V. destructor from Nova Scotia, Canada, and two newly-emerged queens (one with deformed wings and one with normal wings) from two colonies infected with V. destructor from Prince Edward Island, Canada, were genetically analyzed for DWV. We detected DWV in all workers and drones, regardless of wing morphology, but only in the deformed-winged queen. This is the first report of DWV from Atlantic Canada and the first detection of a symptomatic queen with DWV from anywhere.

Splenic mass of masked shrews, Sorex cinereus, in relation to body mass, sex, age, day of the year, and bladder nematode, Liniscus (=Capillaria) maseri, infection.

The spleen is an important organ of vertebrates. Splenic mass can change in response to a variety of factors. We tested whether splenic mass of masked shrews, Sorex cinereus, was related to sex, age, time of the year, or intensity of bladder nematode (Liniscus [=Capillaria] maseri) infection, after controlling for host body mass. For females, body mass was a strong predictor of splenic mass. For males, splenic masses were greater later in the year and in more heavily infected males. The latter appeared to represent a threshold response wherein only the most heavily infected individuals had enlarged spleens.

Cophylogeny of Nosema (Microsporidia: Nosematidae) and bees (Hymenoptera: Apidae) suggests both cospeciation and a host-switch.

Some microsporidian parasites belonging to the genus Nosema infect bees. Previous phylogenies of these parasites have produced alternative, conflicting relationships. We analyzed separately, and in combination, large and small subunit ribosomal DNA sequences of Nosema species infecting bees under neighbor-joining, maximum parsimony, maximum likelihood, and Bayesian frameworks. We observed a sister relationship between Nosema ceranae and Nosema bombi, with Nosema apis as a basal member to this group. When compared to their respective hosts (Apis cerana, Bombus spp., and A. mellifera), 2 plausible evolutionary scenarios emerged. The first hypothesis involves a common ancestor of N. bombi host-switching from a historical Bombus lineage to A. cerana. The second suggests an ancestral N. ceranae host-switching to a species of Bombus. The reported events offer insight into the evolutionary history of these organisms and may explain host specificity and virulence of Nosema in these economically important insects.

Relationships between leukocytes and Hepatozoon spp. In green frogs, Rana clamitans.

There are few published data on amphibian leukocyte profiles, and relationships between amphibian leukocytes and parasites are even less well known. Using counts from 35 pairs of blood smears taken 2 days apart, we tested for correlations between leukocyte proportions and infection intensities of Hepatozoon spp. (either Hepatozoon catesbianae or Hepatozoon clamatae) in green frogs (Rana clamitans). On average (SE), we counted 65.4 (1.7) lymphocytes, 14.0 (1.3) neutrophils, 19.3 (1.6) eosinophils, 0.9 (0.1) monocytes, and 0.4 (0.1) basophils per 100 leukocytes. All frogs harbored Hepatozoon spp. (median seven parasites per 100 leukocytes; range 1-250). Significant relationships were not observed between numbers of leukocytes and infection intensities of Hepatozoon spp. Among the possible explanations for these null results are that Hepatozoon spp. is benign, that Hepatozoon spp. is able to evade detection by the immune system, that Hepatozoon spp. is able to manipulate leukocyte investment, or that other unmeasured or undetected parasites were more important in affecting immune response.

Does fumagillin control the recently detected invasive parasite Nosema ceranae in western honey bees (Apis mellifera)?

Western honey bee (Apis mellifera) colonies in Nova Scotia, Canada were sampled in spring and late summer 2007 to evaluate efficacy of fumagillin dicyclohexylammonium (hereafter, fumagillin) against Nosema ceranae. Colonies treated with fumagillin in September 2006 (n=94) had significantly lower Nosema intensity in spring 2007 than did colonies that received no treatment (n=51), but by late summer 2007 no difference existed between groups. Molecular sequencing of 15 infected colonies identified N. ceranae in 93.3% of cases, suggesting that fumagillin is successful at temporarily reducing this recent invasive parasite in western honey bees.

Foraging areas, offshore habitat use, and colony overlap by incubating Leach's storm-petrels Oceanodroma leucorhoa in the Northwest Atlantic.

Despite their importance in marine food webs, much has yet to be learned about the spatial ecology of small seabirds. This includes the Leach's storm-petrel Oceanodroma leucorhoa, a species that is declining throughout its Northwest Atlantic breeding range. In 2013 and 2014, we used global location sensors to track foraging movements of incubating storm-petrels from 7 eastern Canadian breeding colonies. We determined and compared the foraging trip and at-sea habitat characteristics, analysed spatial overlap among colonies, and determined whether colony foraging ranges intersected with offshore oil and gas operations. Individuals tracked during the incubation period made 4.0 ± 1.4 day foraging trips, travelling to highly pelagic waters over and beyond continental slopes which ranged, on average, 400 to 830 km from colonies. Cumulative travel distances ranged from ~900 to 2,100 km among colonies. While colony size did not influence foraging trip characteristics or the size of areas used at sea, foraging distances tended to be shorter for individuals breeding at the southern end of the range. Core areas did not overlap considerably among colonies, and individuals from all sites except Kent Island in the Bay of Fundy foraged over waters with median depths > 1,950 m and average chlorophyll a concentrations ≤ 0.6 mg/m3. Sea surface temperatures within colony core areas varied considerably (11-23°C), coincident with the birds' use of cold waters of the Labrador Current or warmer waters of the Gulf Stream Current. Offshore oil and gas operations intersected with the foraging ranges of 5 of 7 colonies. Three of these, including Baccalieu Island, Newfoundland, which supports the species' largest population, have experienced substantial declines in the last few decades. Future work should prioritize modelling efforts to incorporate information on relative predation risk at colonies, spatially explicit risks at-sea on the breeding and wintering grounds, effects of climate and marine ecosystem change, as well as lethal and sub-lethal effects of environmental contaminants, to better understand drivers of Leach's storm-petrel populations trends in Atlantic Canada.