Volatile and semivolatile compounds in gray catbird uropygial secretions vary with age and between breeding and wintering grounds.

The uropygial secretions of some bird species contain volatile and semivolatile compounds that are hypothesized to serve as chemical signals. The abundance of secretion components varies with age and season, although these effects have not been investigated in many species. We used solid-phase microextraction headspace sampling and solvent extraction coupled with gas chromatography-mass spectrometry to detect and identify volatile and semivolatile chemical compounds in uropygial secretions of gray catbirds (Dumetella carolinensis). We identified linear and branched saturated carboxylic acids from acetic (C2) through hexacosanoic (C26); linear alcohols from decanol (C10) through docosanol (C22); one aromatic aldehyde; one monounsaturated carboxylic acid; two methyl ketones; and a C28 ester. We tested for the effect of age on signal strength and found that juvenile birds produced greater amounts of volatile C4 through C7 acids and semivolatile C20 through C26 acids, although the variation among individuals was large. Adult birds displayed small concentrations and minimal individual variation among volatile compounds, but produced significantly higher levels of long-chain linear alcohols than juvenile birds. We tested for the effects of season/location by sampling adult catbirds at their Ohio breeding grounds and at their Florida wintering grounds and found that the heaviest carboxylic acids are significantly more abundant in secretions from birds sampled during winter at the Florida site, whereas methyl ketones are more abundant in birds sampled during summer on the Ohio breeding grounds. We observed no effect of sex on semivolatile compounds, but we found a significant effect of sex on levels of carboxylic acids (C4 through C7) for juvenile birds only.

Test of recrudescence hypothesis for overwintering of eastern equine encephalomyelitis virus in gray catbirds.

Eastern equine encephalitis virus (EEEV; family Togaviridae, genus Alphavirus) epizootics are infrequent, but they can lead to high mortality in infected horses and humans. Despite the importance of EEEV to human and animal health, little is known about how the virus overwinters and reinitiates transmission each spring, particularly in temperate regions where infected adult mosquitoes are unlikely to survive through the winter. One hypothesis to explain the mechanism by which this virus persists from year to year is the spring recrudescence of latent virus in avian reservoir hosts. In this study, we tested the recrudescence hypothesis with gray catbirds (Dumatella carolinensis) captured in northern Ohio (July-August 2007). Birds were experimentally infected with EEEV on 1 October 2007. In January 2008, they were then exposed to exogenous testosterone and/or extended photoperiod to initiate reactivation of latent EEEV infection. All birds became viremic with EEEV, with mean viremia of 6.0 log10 plaque-forming units/ml serum occurring at 1 d postinoculation. One male in the testosterone, long-day treatment group had EEEV viral RNA in a cloacal swab collected on 18 January 2008. Otherwise, no other catbirds exhibited reactivated infections in cloacal swabs or blood. Antibody titers fluctuated over the course of the study, with lowest titers observed in January 2008, which corresponded with the lowest mean weight of the birds. No EEEV viral RNA was detected in the blood, kidney, spleen, brain, liver, and lower intestine upon necropsy at 19 wk postinfection.

A survey of trace metal burdens in increment cores from eastern cottonwood (Populus deltoides) across a childhood cancer cluster, Sandusky County, OH, USA.

A dendrochemical study of cottonwood trees (Populus deltoides) was conducted across a childhood cancer cluster in eastern Sandusky County (Ohio, USA). The justification for this study was that no satisfactory explanation has yet been put forward, despite extensive local surveys of aerosols, groundwater, and soil. Concentrations of eight trace metals were measured by ICP-MS in microwave-digested 5-year sections of increment cores, collected during 2012 and 2013. To determine whether the onset of the first cancer cases could be connected to an emergence of any of these contaminants, cores spanning the period 1970-2009 were taken from 51 trees of similar age, inside the cluster and in a control area to the west. The abundance of metals in cottonwood tree annual rings served as a proxy for their long-term, low-level accumulation from the same sources whereby exposure of the children may have occurred. A spatial analysis of cumulative metal burdens (lifetime accumulation in the tree) was performed to search for significant 'hotspots', employing a scan statistic with a mask of variable radius and center. For Cd, Cr, and Ni, circular hotspots were found that nearly coincide with the cancer cluster and are similar in size. No hotspots were found for Co, Cu, and Pb, while As and V were largely below method detection limits. Whereas our results do not implicate exposure to metals as a causative factor, we conclude that, after 1970, cottonwood trees have accumulated more Cd, Cr, and Ni, inside the childhood cancer cluster than elsewhere in Sandusky County.

Attraction of Culex pipiens to House Sparrows Is Influenced by Host Age but Not Uropygial Gland Secretions.

Culex pipiens serves as the endemic vector of West Nile virus (WNV) in eastern North America, where house sparrows (HOSP, Passer domesticus) serve as a reservoir host. We tested the hypotheses that: (1) Attraction of Cx. pipiens to HOSP is influenced by bird age and (2) that age-specific variation in chemical profiles of bird uropygial gland secretions informs this choice. We conducted mosquito choice trials in an olfactometer and found that Cx. pipiens were more often attracted to adult sparrows over nestlings, however, they demonstrated no preference for adults over fledglings. Using gas chromatography-mass spectrometry we observed age-specific differences in the semi-volatile chemical profiles of house sparrow uropygial gland secretions. Contrary to our hypothesis, we found no significant difference in mosquito feeding preference between the secretions of adults and those of either nestlings or fledglings. We suggest that other chemical cues influence the feeding preference of Cx. pipiens, either independently of uropygial gland secretions, or synergistically with them.

Attraction of Culex pipiens to uropygial gland secretions does not explain feeding preference for American robins.

Culex pipiens, the endemic mosquito vector of West Nile virus in eastern North America, is responsible for maintenance of the virus in avian reservoir hosts, the most important of which appears to be the American robin. One reason for the greater involvement of robins is believed to be the feeding preference of Cx. pipiens, however, the basis of this preference is not understood. We tested the hypothesis that the species-specific chemical profile of avian uropygial gland secretions are used by Cx. pipiens as cues to locate birds and, therefore, may contribute to the observed feeding preferences. We used gas chromatography-mass spectrometry to identify the semi-volatile components of the uropygial gland secretions of American robins and two other common reservoir host species, the house sparrow and European starling. We found that the chemical composition of the robin secretions was different from those of the sparrows and starlings. Through behavioral choice trials conducted in a dual-port olfactometer, we also found that Cx. pipiens did not prefer the secretions of robins over the other two species. Surprisingly, however, we found that Cx pipiens were more often attracted to live starlings over robins and to the secretions of starlings over those of robins.

West Nile virus in mosquitoes of northern Ohio, 2003.

From June 19, 2003 to August 18, 2003, we surveyed the mosquitoes of Oberlin, OH, for West Nile Virus (WNV) infection using reverse transcriptase-polymerase chain reaction. A total of 12,055 mosquitoes, representing 17 species or species groups and 4 genera, were collected in gravid traps at seven sites throughout the city, with Culex pipiens/restuans being the most abundant and showing the highest minimum infection rate (MIR) of 0.78. This represents a decrease in WNV enzootic activity from the previous year. Both Cx. pipiens/restuans abundance and MIR increased significantly with date. However, we found no correlation between Cx. pipiens/restuans abundance and MIR.

Survey of Aedes triseriatus (Diptera: Culicidae) for Lacrosse encephalitis virus and West Nile virus in Lorain County, Ohio.

From June through September 2003, we conducted a survey of female Aedes triseriatus (Say) for infection with La Crosse encephalitis virus (family Bunyaviridae, genus Orthobunyavirus, LACV) and West Nile virus (family Flaviviridae, genus Flavivirus, WNV) at three locations in Lorain County, Ohio. To determine infection rate and seasonal variation of both viruses in the Ae. triseriatus population, Ae. triseriatus were collected weekly by using gravid traps and CO2-baited CDC light traps and tested for virus by using reverse transcriptase polymerase chain reaction. In total, 170 pools comprised of 2,143 females were tested for LACV, of which seven were positive; the maximum likelihood estimate of infection rate combined throughout the season was 3.22/1,000. None of 170 pools comprised of 2,158 females tested for WNV were positive. LACV-positive pools were detected between late July and early September.

Hormone levels and infection of Haemoproteus danilewskyi in free-ranging blue jays (Cyanocitta cristata).

Annual spring relapse of blood parasite infections in birds is believed to be the result of hormonal changes associated with breeding. As part of a larger study on the epizootiology of Haemoproteus danilewskyi in blue jays in south-central Florida, we studied the relationship between H. danilewskyi infections and levels of luteinizing hormone, prolactin, progesterone, testosterone, estradiol, and corticosterone. We found a positive association between intensity of H. danilewskyi infection and corticosterone levels in females but not in males. We also found no association between infection and levels of prolactin, luteinizing hormone, progesterone, testosterone, or estradiol in males or females. In addition, we found a positive relationship between levels of corticosterone and handling time and between corticosterone and testosterone levels. We suggest a possible influence of corticosterone on spring relapse of Haemoproteus spp. infections in birds but provide no support for the influence of breeding hormones on relapse of these parasites.

Blood parasites of Nearctic-Neotropical migrant passerine birds during spring trans-Gulf migration: impact on host body condition.

To test the hypothesis that migrants infected with blood parasites arrive on the northern coast of the Gulf of Mexico in poorer condition than uninfected birds, we examined 1705 migrant passerine birds representing 54 species of 11 families from 2 Gulf Coast sites for blood parasites. Three hundred and sixty (21.1%) were infected with 1 or more species of 4 genera of blood parasites. The prevalence of parasites was as follows: Haemoproteus spp. (11.7%), Plasmodium spp. (6.7%), Leucocytozoon spp. (1.3%), and Trypanosoma spp. (1.2%). Both prevalence and density of Haemoproteus spp. infection varied among species. We found no relationship of gender or age with the prevalence of Haemoproteus spp. infection or Plasmodium spp. infection, with the exception of the orchard oriole (Icterus spurius) for which older birds were more likely to be infected with Haemoproteus spp. than younger birds. We also found that scarlet tanagers and summer tanagers infected with species of Haemoproteus have lower fat scores than uninfected individuals and that rose-breasted grosbeaks and Baltimore orioles infected with Haemoproteus spp. have a smaller mean body mass than uninfected individuals. Blood parasites do seem to pose a physiological cost for Neotropical migrant passerines and may be important components of the ecology of these species.

Patterns of West Nile virus infection in Ohio blue jays: implications for initiation of the annual cycle.

West Nile virus (WNV) was first detected in North America in New York City in 1999 and rapidly moved westward. Understanding the mechanisms by which the amplification cycle is reinitiated each year increases our ability to predict epizootics and geographic expansion of the disease. Such understanding is enhanced by knowledge of the patterns of infection in the vertebrate reservoir hosts. Blue jays (Cyanocitta cristata) may serve as reservoir hosts for WNV. We examined the influence of age and date on the prevalence of WNV in jay carcasses in Ohio during May-August 2002. Percent of carcasses that were infected increased significantly with time from 3% in May to more than 90% by August. We found no difference in prevalence between juvenile (nestlings and fledglings) and adult jays early in the season, which contradicts the expected pattern if the majority of the adults sampled in 2002 had been exposed to the virus in 2001. Therefore, jays infected in 2001 were unlikely to have been important in initiating the 2002 virus cycle in Ohio.

West Nile virus in mosquitoes of Northern Ohio, 2001-2002.

From June 14 to August 30, 2001 and June 10 to August 22, 2002, the mosquitoes of Oberlin, Ohio were surveyed and tested for West Nile virus (WNV). Mosquitoes were trapped weekly using gravid traps and CO(2)-baited Centers for Disease Control light traps at seven sites in 2001 and eight sites in 2002 in woodlots within the city limits. A total of 12,151 mosquitoes, representing 14 species and one species group, were collected in gravid traps, with Culex pipiens/restuans being the most abundant, followed by Ochlerotatus triseriatus. In light traps, 12,510 mosquitoes were collected, with Aedes vexans being the most abundant, followed by Culex pipiens/restuans. All gravid trap collections were tested for WNV via reverse transcriptase-polymerase chain reaction. West Nile virus was first detected in July 2002. Of all species tested, the Cx. pipiens/restuans species group exhibited the highest minimum infection rate (MIR) and the greatest percentage of positive pools. Both the MIR and percent of positive pools increased significantly with the date, although abundance of Cx. pipiens/restuans in gravid trap collections did not.

Arboviral infection in two species of wild jays (Aves: Corvidae): evidence for population impacts.

We examined the prevalence of antibodies to three mosquito-borne arboviruses in blue jays, Cyanocitta cristata, and Florida scrub-jays, Aphelocoma coerulescens, to identify the effects on host survival, the influence of sex and age on infection, and the temporal patterns of antibody prevalence. Blood samples from 306 blue jays and 219 Florida scrub-jays were collected at Archbold Biological Station (Lake Placid, FL) from April 1994 through December 1995. Sera were analyzed for hemagglutination-inhibition antibody to eastern equine encephalitis (EEE) and St. Louis encephalitis (SLE) viruses, and neutralizing antibodies to EEE, Highlands J (HJ), and SLE viruses. Overall, 31.4% of blue jay samples and 22.1% of scrub-jay samples had antibodies to EEE. Antibodies to HJ were detected in slightly >15% of samples in each jay species, and SLE was detected in <3% of the samples in each jay species. A single EEE virus isolation was made from the blood of an 11-d-old scrub-jay nestling. Survival of adult blue jays seropositive to EEE was significantly lower than that of seronegative birds based on resight rates, but infection did not seem to affect survival of adult or juvenile Florida scrub-jays.

Epizootiology of Haemoproteus danilewskyi (Haemosporina: Haemoproteidae) in blue jays (Cyanocitta cristata) in southcentral Florida.

Prevalence and density of Haemoproteus danilewskyi was studied in a population of free-ranging blue jays (Cyanocitta cristata) in southcentral Florida (USA) from May 1992 to December 1995. Prevalence of infection was 27% for data combined over years, seasons, ages, and sexes. Prevalence did not vary between sexes or among years, but increased with age and varied with season, being highest in June-July and lowest in November-January. Parasite density did not vary between sexes or among seasons, but was higher in younger birds when controlling for season. To determine periods of natural transmission, seasonal patterns of infection were compared with previous month abundance of the biting fly vectors. Mean monthly prevalence of H. danilewskyi in older jays was positively correlated with previous month abundance of Culicoides edeni and C. arboricola, both capable of sporogonic development of H. danilewskyi.

Ecology of Culicoides (Diptera: Ceratopogonidae) in southcentral Florida and experimental Culicoides vectors of the avian hematozoan Haemoproteus danilewskyi Kruse.

To determine the vectors of Haemoproteus danilewskyi in blue jays (Cyanocitta cristata) in southcentral Florida (USA), we conducted a 2 yr study from January 1993 to December 1995 of the presence and seasonal abundance of Culicoides spp. Of the 14 species of Culicoides captured in Centers for Disease Control light traps, 10 were ornithophilic. Of these, C. edeni, C. knowltoni, C. stellifer, C. beckae, and C. arboricola were most abundant, representing 46% of the total collection and 99% of the ornithophilic collection. The presence of C. stellifer in Bennett trap collections represents a new biting record for this species on passerine birds. We experimentally challenged the most abundant ornithophilic species to determine which were capable of supporting sporogonic development of H. danilewskyi. Culicoides edeni, C. knowltoni, and C. arboricola supported sporogonic development of H. danilewskyi.

Pathogenicity of Haemoproteus danilewskyi, Kruse, 1890, in blue jays (Cyanocitta cristata).

Although the impact of blood parasite infections on passerine birds is potentially great, little is known of their pathologic effects. We studied Haemoproteus danilewskyi in experimentally infected captive and naturally infected free-ranging blue jays (Cyanocitta cristata) to determine patterns of infection and examine the pathologic effects of the parasite on the host. Physiologic changes, such as elevated numbers of lymphocytes, heterophils, basophils, eosinophils, and monocytes and decreased packed cell volume in the peripheral blood were associated with the erythrocytic phase of experimental infections of captive juvenile jays. Sublethal pathologic changes associated with the pre-erythrocytic phase of infections were observed in the liver, lung, and spleen. Schizonts were observed in the pulmonary capillaries of a 1 yr old jay necropsied 31 days post-inoculation, but not in 20 juvenile jays necropsied 57 days post-inoculation. In free-ranging naturally infected jays plasma protein concentration increased with density of natural infections.

Prevalence of hematozoa in overwintering American redstarts (Setophaga ruticilla): no evidence for local transmission.

We examined American redstarts (Setophaga ruticilla) for protozoan blood parasites on their wintering grounds to determine whether transmission of these parasites occurs prior to spring migration. A total of 73 blood smears from 37 birds were examined for presence and intensity of infection. Thirty-six birds were sampled in the fall, soon after arriving from northern breeding grounds, and the spring prior to departure. Two (5%) of the samples collected in the fall were positive for Haemoproteus fringillae and one (3%) had detectable infections of Trypanosoma avium. Individuals infected with H. fringillae were hatching year redstarts sampled in September and October. Intensity of infection was 78 and < 1 infected erythrocytes per 10,000 erythrocytes, respectively. None of the birds had detectable infections when resampled prior to spring migration the following March.

Patterns of Haemoproteus beckeri parasitism in the gray catbird (Dumatella carolinensis) during the breeding season.

We determined the prevalence and intensity of blood parasites in breeding gray catbirds (Dumatella carolinensis) at Killbuck Wildlife Area in Wayne and Holmes Counties, Ohio (USA) from June through August 2000. Of 98 catbirds sampled, 40 (40.8%) had detectable infections of Haemoproteus beckeri. Overall prevalence of H. beckeri in this population is high relative to that reported in earlier blood parasite surveys of both breeding and migrant catbirds. Mean intensity of H. beckeri infection did not vary significantly between young and old birds or among sampling periods. We found no effect of age on prevalence or intensity of H. beckeri infection. Older birds were not more likely to be infected than younger birds, despite longer exposure to arthropod vectors. Prevalence varied significantly with season and was highest in June and lowest in August. This pattern also was observed in older birds sampled repeatedly. This seasonal variation may reflect both newly acquired infections and chronic infections relapsing in response to hormonal changes associated with breeding. Evidence of transmission was observed in the single hatching year bird that lacked detectable infection in early summer, but demonstrated a very high intensity infection in late summer. These observations provide supportive evidence that hematozoa infections are acquired on the breeding grounds during the first year of life and relapse during the breeding season in subsequent years.

Abundance and temporal distribution of Ornithonyssus sylviarum Canestrini and Fanzago (Acarina: Mesostigmata) in gray catbird (Dumatella carolinensis) nests.

The northern fowl mite, Ornithonyssus sylviarum Canestrini and Fanzago, is a common ectoparasite of wild birds. Despite its ability to transmit eastern equine encephalitis (EEE) virus under laboratory conditions and potential for involvement in the natural EEE virus cycle, we know little about its abundance or temporal distribution in nature. From June to August 2000, we studied the abundance of O. sylviarum in the nests of gray catbirds (Dumatella carolinensis), a reservoir host for EEE virus, at Killbuck Marsh Wildlife Area (KMWA), a known EEE virus focus in Wayne County, Ohio. A total of 7,883 O. sylviarum, including 1,910 adults and 5,973 protonymphs, were recovered from 23 of 26 gray catbird nests collected during various phases of the nesting cycle. We found no association between mite abundance and number of catbird nestlings in successful nests. However, mite abundance increased significantly with date of nest collection and peaked in late July when transmission of EEE virus is likely to occur at KMWA. We therefore suggest that O. sylviarum may contribute to the transmission of EEE virus among gray catbirds at KMWA.

Seroprevalence of eastern equine encephalomyelitis virus in birds and larval survey of Culiseta melanura Coquillett during an interepizootic period in central Ohio.

From June through August in 1999 and 2000, we conducted an avian serosurvey for eastern equine encephalomyelitis (EEE) virus at Killbuck Marsh Wildlife Area (KMWA), a focus of infection in central Ohio. We also monitored abundance of the suspected enzootic vector, Culiseta melanura Coquillett, in Brown's Lake Bog, an adjacent wetland. Of the 363 birds of 30 species sampled in 1999, three gray catbirds (Dumetella carolinensis) were positive for antibodies to EEE virus, representing 1.2% of the avian samples and 4.2% of the gray catbirds sampled. Given these results and the abundance of gray catbirds at this site, this species became the focus of our sampling efforts in 2000. However, none of the 109 samples collected from 98 catbirds in 2000 was positive for the virus. Culiseta melanura adults were monitored using resting boxes and CDC CO, light traps at both sites in 1999. Culiseta melanura larvae were monitored in 1999 and 2000 at Brown's Lake Bog, the closest known source of this species, approximately 5km from the avian serosurvey site. We suggest that dry conditions reduced the breeding and abundance of Cs. melanura in 2000 and possibly the transmission of EEE virus at KMWA.

Short-chain carboxylic acids from gray catbird (Dumetella carolinensis) uropygial secretions vary with testosterone levels and photoperiod.

The uropygial gland of birds produces secretions that are important in maintaining the health and structural integrity of feathers. Non-volatile components of uropygial secretions are believed to serve a number of functions including waterproofing and conditioning the feathers. Volatile components have been characterized in fewer species, but are particularly interesting because of their potential importance in olfactory interactions within and across species. We used solid-phase microextraction headspace sampling with gas chromatography-mass spectrometry to detect and identify volatiles in uropygial secretions of gray catbirds (Dumetella carolinensis), a North American migratory bird. We consistently detected the following carboxylic acids: acetic, propanoic, 2-methylpropanoic, butanoic, and 3-methylbutanoic. We tested for the effect of lengthened photoperiod and/or exogenous testosterone on volatile signal strength and found a negative effect of lengthened photoperiod on the signal strength of propanoic, 2-methylpropanoic, and butanoic acids, suggesting a trade-off between their production and heightened night-time activity associated with lengthened photoperiod. Signal strength of propanoic and 2-methylpropanoic acids was lower in birds treated with exogenous testosterone than in birds treated with placebos. Sex did not affect signal strength of any of the volatile compounds.