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Canvasback ducks wintering on Chesapeake Bay had a 6% incidence of Leucocytozoon sirnondi and 2% incidence of Haemoproteus. Sub-inoculation of whole blood into Pekin ducklings produced a Plasmodium infection rate of 31%. Females were more frequently infected (12/22) than males (15/68). The parasite was identified as P. circumflexum.
Hawaiʻi’s avifauna has undergone profound changes over the past 1,000 years, with many endemic bird species driven to extinction and non-native introductions reshaping forest ecosystems. On the island of Molokaʻi, habitat degradation and the introduction of mosquito-borne disease, mainly avian malaria ( Plasmodium relictum ), have caused extinctions and severe declines in native forest bird populations. To assess these changes, point-transect distance sampling surveys were initiated in 1979 and 1980 and repeated six times, most recently in 2021, covering a 3,527 ha area. This study analyzes species composition, population densities, abundances, and long-term trends for species with sufficient detections using log-linear regression. Since 1979–1980, a total of 17 non-native and 3 native species were detected across all six surveys. The non-native Warbling White-eye ( Zosterops japonicus ) had the highest density and abundance in every survey, while abundance of Red-billed Leiothrix ( Leiothrix lutea ) significantly trended upward. The Japanese Bush Warbler ( Horornis diphone ) irrupted in the survey area, increasing in abundance by more than 99% since 1979–1980. Among native species, ʻApapane ( Himatione sanguinea ) populations increased by 33%, while Hawaiʻi ʻAmakihi ( Chlorodrepanis virens ) declined by 78% from their peak in 1995. Federally threatened ʻIʻiwi ( Drepanis coccinea ), highly susceptible to avian malaria, have not been recorded on Molokaʻi since 2010, and the endemic Olomaʻo ( Myadestes lanaiensis ) has not been detected since 1980 and is possibly extinct. Our results underscore the ongoing conservation crisis for Molokaʻi’s forest birds and the continued vulnerability of native birds to disease and habitat loss.
Introduced mosquito-borne avian disease is a major limiting factor in the recovery and restoration of native Hawaiian forest birds. Annual epizootics of avian pox (Avipoxvirus) and avian malaria (Plasmodium relictum) likely led to the extinction of some species and continue to impact populations of susceptible Hawaiian honeycreepers (Drepanidinae). The introduction of a novel pathogen, such as West Nile virus (WNV), could result in further population declines and extinctions. During September and October 2004, we infected Hawai'i' Amakihi (Hemignathus virens) with a North American isolate of WNV by needle inoculation and mosquito bite to observe susceptibility, mortality, and illness in this endemic passerine, and to determine the vector competence of the co-occurring, introduced mosquito Culex quinquefasciatus. All experimentally infected Hawai'i ;Amakihi became viremic, with a mean titer >10(5) plaque-forming units (PFU)/ml, and they experienced clinical signs ranging from anorexia and lethargy to ataxia. The fatality rate among needle-inoculated Hawai'i' Amakihi (n=16) was 31.3%, but mortality in free-ranging birds is likely to increase due to predation, starvation, thermal stress, and concomitant infections of avian malaria and pox. Surviving Hawai'i' Amakihi seem to clear WNV from the peripheral blood by 7-10 days postinfection (DPI), and neutralizing antibodies were detected from 9 to 46 DPI. In transmission trials, Hawaiian Cx. quinquefasciatus proved to be a competent vector and Hawai'i Amakihi an adequate amplification host of WNV, suggesting that epizootic WNV could readily become an additional limiting factor of some native Hawaiian bird populations.
Several polymerase chain reaction (PCR)-based methods have recently been developed for diagnosing malarial infections in both birds and reptiles, but a critical evaluation of their sensitivity in experimentally-infected hosts has not been done. This study compares the sensitivity of several PCR-based methods for diagnosing avian malaria ( Plasmodium relictum ) in captive Hawaiian honeycreepers using microscopy and a recently developed immunoblotting technique. Sequential blood samples were collected over periods of up to 4.4 yr after experimental infection and rechallenge to determine both the duration and detectability of chronic infections. Two new nested PCR approaches for detecting circulating parasites based on P. relictum 18S rRNA genes and the thrombospondin-related anonymous protein (TRAP) gene are described. The blood smear and the PCR tests were less sensitive than serological methods for detecting chronic malarial infections. Individually, none of the diagnostic methods was 100% accurate in detecting subpatent infections, although serological methods were significantly more sensitive (97%) than either nested PCR (61–84%) or microscopy (27%). Circulating parasites in chronically infected birds either disappear completely from circulation or to drop to intensities below detectability by nested PCR. Thus, the use of PCR as a sole means of detection of circulating parasites may significantly underestimate true prevalence.
Introduced mosquito-borne pathogens avian malaria (Plasmodium relictum Grassi and Feletti) and avian pox virus (Avipoxvirus) have been implicated in the past extinctions and declines of Hawaiian avifauna and remain significant obstacles to the recovery and restoration of endemic Hawaiian birds. Effective management of avian disease will require extensive mosquito control efforts that are guided by the local ecology of the vector Culex quinquefasciatus Say (Diptera: Culicidae). During October and November 1997 and September through November 1998 five mark-release-recapture experiments with laboratory-reared Cx. quinquefasciatus were conducted in a native rain forest on Hawaii Island. Of the overall 66,047 fluorescent dye-marked and released females, 1,192 (1.8%) were recaptured in 43-52 CO2-baited traps operated for 10-12-d trapping periods. Recaptured mosquitoes were trapped in all directions and at distances up to 3 km from the release site. The cumulative mean distance traveled (MDTs) over the trapping period ranged from a high of 1.89 km after 11 d (September 1998) to a low of 0.81 km after 11 d (November 1998). Released mosquitoes moved predominately in a downwind direction and they seemed to use forestry roads as dispersal corridors. Applying an estimated MDT of 1.6 km to a geographical information system-generated map of the Hakalau Forest National Wildlife Refuge clearly demonstrated that the effective refuge area could be reduced 60% by mosquitoes infiltrating into managed refuge lands. These findings should have significant implications for the design of future refuges and development of effective mosquito-borne avian disease control strategies.
Plasmodium parasites infect thousands of species and provide an exceptional system for studying host-pathogen dynamics, especially for multi-host pathogens. However, understanding these interactions requires an accurate assay of infection. Assessing Plasmodium infections using microscopy on blood smears often misses infections with low parasitemias (the fractions of cells infected), and biases in malaria prevalence estimates will differ among hosts that differ in mean parasitemias. We examined Plasmodium relictum infection and parasitemia using both microscopy of blood smears and quantitative polymerase chain reaction (qPCR) on 299 samples from multiple bird species in Hawai'i and fit models to predict parasitemias from qPCR cycle threshold (Ct) values. We used these models to quantify the extent to which microscopy underestimated infection prevalence and to more accurately estimate infection patterns for each species for a large historical study done by microscopy. We found that most qPCR-positive wild-caught birds in Hawaii had low parasitemias (Ct scores ≥35), which were rarely detected by microscopy. The fraction of infections missed by microscopy differed substantially among eight species due to differences in species’ parasitemia levels. Infection prevalence was likely 4–5-fold higher than previous microscopy estimates for three introduced species, including Zosterops japonicus , Hawaii’s most abundant forest bird, which had low average parasitemias. In contrast, prevalence was likely only 1.5–2.3-fold higher than previous estimates for Himatione sanguinea and Chlorodrepanis virens , two native species with high average parasitemias. Our results indicate that relative patterns of infection among species differ substantially from those observed in previous microscopy studies, and that differences depend on variation in parasitemias among species. Although microscopy of blood smears is useful for estimating the frequency of different Plasmodium stages and host attributes, more sensitive quantitative methods, including qPCR, are needed to accurately estimate and compare infection prevalence among host species.
An epizoological study of Plasmodium infections in wild birds of Kern County, California, in the years 1946 through 1951 greatly extended knowledge of the occurrence of these parasites and their behavior in nature. Examination of 10,459 blood smears from 8,674 birds representing 73 species resulted in the observation of Plasmodium spp. in 1,094 smears representing 888 individual birds of 27 species. Seven species of Plasmodium were found: relictum, elongatum, hexamerium, nucleophilum, polare, rouxi and vaughani. Plasmodium relictum was by far the most frequently observed species, occurring in at least 79 per cent of the infected birds. Twelve new host species are recorded for this parasite. Sufficient morphological variation was observed to indicate that two strains of this species probably exist in nature. Numerous new host records were made of plasmodia with elongate gametocytes. The finding of parasites believed to be P. rouxi in two new host species represents the first record of the occurrence of this Plasmodium outside of Algeria. Multiple smears were obtained from a number of individual birds over varying time periods. Evidence of prolonged parasitemia was unusual, but some individuals had parasitemia on consecutive months and even for three successive years. In most individuals, parasitemias were of short duration. The inoculation of blood from wild birds into canaries led to the demonstration of many infections not observed on blood smear examination of donors. Use of these two complementary techniques led to more complete host records and a truer picture of the prevalence of infection. Three age classes of birds were studied--nestling, immature (less than 1 year of age) and adult. Parasites were observed in all three groups but infections in the younger individuals were most susceptible to interpretation. As to time of onset, numerous records were obtained of infection in nestling birds. Prevalence rates in immature birds after a single season's exposure ranged from 64 to 100 per cent in the house finch and 17 to 68 per cent in the English sparrow in different areas and years. Marked differences were found in the prevalence rates in different summer months, years and areas. It is believed these differences reflect variation in a number of environmental factors. This study indicates the extensive distribution of Plasmodium infection in a wide range of wild avian hosts. The observations are of possible importance in epidemiological studies of other arthropod-borne diseases such as the viral encephalitides for which these birds serve as hosts.