Previous infection protects house finches from re-infection with St. Louis encephalitis virus

Antibody titers against St. Louis encephalitis virus (SLE) measured by a plaque reduction neutralization test (PRNT) decreased rapidly in house finches (Capodacus mexicanus) after initial infection, whereas antibodies measured by enzyme immunoassay (EIA) remained detectable in all birds for the length of the experiment, indicating long-term persistence and greater assay sensitivity of the EIA. After 52 wk, birds were challenged by subcutaneous inoculation with the same strain of SLE virus. Virus was not detected for 1-4 d postchallenge in blood samples tested by plaque assay and RT-PCR or by xenodiagnosis in Culex tarsalis fed concurrently and then held for 11 d at 26 degrees C. Virus was detected by all three methods in control birds infected concurrently for the first time. Challenge with SLE produced a rapid and marked ananmestic rise in both neutralizing and EIA antibody titers that exceeded the primary response in the same birds or in concurrently inoculated control birds. At necropsy 4 wk postchallenge, 3 of 7 challenged and 1 of 2 positive control birds were chronically infected, with viral RNA detected by RT-PCR in brain, spleen, lung, and/or kidney tissues. Our results indicated that persistence of protective antibody prevents reinfection during the following season and may prevent the recrudescence of infectious virus in chronically infected birds.     

Characterization of unilateral conjunctival inoculation with Mycoplasma gallisepticum in house finches

House finches in much of the continental United States experience annual epidemics of mycoplasmal conjunctivitis, caused by the bacterial pathogen Mycoplasma gallisepticum (MG). Although evidence suggests that natural infections typically begin unilaterally, experimental inoculations of songbirds with MG to date have all been administered bilaterally. Furthermore, studies of free-living finches find more severe clinical signs of mycoplasmal conjunctivitis in left versus right eyes, but the mechanisms underlying this side bias remain unknown. Here, we characterized unilateral inoculation of house finches with MG, and tested whether differential susceptibility of left versus right conjunctiva explains the side bias in disease severity of free-living finches. We directly inoculated house finches in either the left or right conjunctiva and characterized resulting disease severity and pathogen load throughout the course of infection. As expected, unilateral inoculation resulted in significantly more severe conjunctivitis, as well as higher conjunctival bacterial loads, on whichever side (left or right) birds were directly inoculated. However, in 55% of cases, unilateral inoculations resulted in bilateral disease, and in 85% cases there was evidence of bilateral infection. The overall severity of disease did not differ for birds inoculated in the left versus right conjunctiva, suggesting that physiological differences between the conjunctivae cannot explain the side bias in disease severity of free-living birds. Instead, laterality in exposure, perhaps due to feeding handedness, likely explains the detected field patterns.

RESEARCH HIGHLIGHTS

• House finches show more severe disease in the directly inoculated conjunctiva.

• Unilateral inoculations lead to high rates of bilateral infection and disease.

• Overall disease severity does not differ for the left- or right-inoculated conjunctiva.

• Laterality in exposure likely explains the left-side bias in natural infections.

     

Response of house finches to infection with sympatric and allopatric strains of western equine encephalomyelitis and St. Louis encephalitis viruses from California

Adult house finches from Kern County were inoculated subcutaneously with recent sympatric and allopatric isolates of western equine encephalomyelitis and St. Louis encephalitis (SLE) viruses made from Culex tarsalis Coquillett collected in Kern County and Coachella Valley, CA, respectively. Virulence, as measured by the amplitude of the viremia response during days 1 and 2 postinfection, varied significantly among strains, but independently of geographic origin. The intensity of the immune response, as measured by an enzyme immunoassay and a plaque reduction neutralization test, seemed to be independent of virulence, especially for SLE where some strains failed to produce a detectable viremia but elicited a strong antibody response. Our preliminary data indicated that strain virulence may be associated with the level of enzootic activity during the year of isolation.     

Role of nestling mourning doves and house finches as amplifying hosts of St. Louis encephalitis virus

Nestling mourning doves and house finches produced elevated viremias after inoculation with 2-3 log10 plaque-forming units (PFU) of St Louis encephalitis (SLE) virus and infected 67 and 70% of Culex tarsalis Coquillett that engorged upon them, respectively. Mosquito infection rates as well as the quantity of virus produced after extrinsic incubation increased as a function of the quantity of virus ingested and peaked during days 3-5 postinoculation in mourning doves and days 2-4 in house finches. Only female Cx. tarsalis with body titers > or = 4.6 log10 PFU were capable of transmitting virus. Overall, 38% of females infected by feeding on mourning doves and 22% feeding on house finches were capable of transmission. The quantity of virus expectorated was variable, ranging from 0.8 to 3.4 log10 PFU and was greatest during periods when avian viremias were elevated. Our data indicated that nestling mourning doves and house finches were competent hosts for SLE virus and that the quantity of virus ingested from a viremic avian host varies during the course of the infection and determines transmission rates by the mosquito vector.     

Experimental evidence for transmission of Mycoplasma gallisepticum in house finches by fomites.

Ever since Mycoplasma gallisepticum emerged among house finches in North America, it has been suggested that bird aggregations at feeders are an important cause of the epidemic of mycoplasmal conjunctivitis because diseased birds could deposit droplets of pathogen onto the feeders and thereby promote indirect transmission by fomites. In this paper we bring the first experimental evidence that such transmission (bird-to-feeder-to-bird) does actually take place. House finches infected via this route, however, developed only mild disease and recovered much more rapidly than birds infected from the same source birds but directly into the conjunctiva. While it is certainly probable that house finch aggregations at artificial feeders enhance pathogen transmission, to some degree transmission of M. gallisepticum by fomites may serve to immunize birds against developing more severe infections. Some such birds develop M. gallisepticum antibodies, providing indication of an immune response, although no direct evidence of protection.     

Effects of route of inoculation on Mycoplasma gallisepticum infection in captive house finches.

The routes by which Mycoplasma gallisepticum initiates infection during outbreaks of conjunctivitis in house finches remain uncertain. As M. gallisepticum recovered from the cloaca of chickens remains viable for up to 3 days in chicken faeces, the possibility of spread via faecal contamination has been suggested. To test the hypothesis that food or water contaminated with M. gallisepticum may initiate infection, 20 house finches were experimentally inoculated by the oral or the conjunctival route. Clinical and immunological responses were compared. All inoculated birds seroconverted, thus demonstrating infection. Only two of the birds inoculated via the oral route developed very mild unilateral conjunctivitis while all 10 of those infected by eye-drop inoculation developed severe bilateral conjunctivitis. The orally inoculated birds had reduced levels of activity for only a few days, while those infected by conjunctival inoculation had reduced activity for several weeks. M. gallisepticum DNA was detected in conjunctival swabs by polymerase chain reaction in only three orally inoculated birds but in all birds in the conjunctivally inoculated group. Antibodies developed more slowly after oral inoculation than after conjunctival inoculation. We showed that oral exposure to M. gallisepticum can initiate infection, disease, and a serological response, which suggests that food or water contaminated with secretions or excretions may be a route of transmission between house finches.     

Virologic and serologic studies of zoo birds for Marek's disease virus infection.

One hundred and eleven zoo birds representing 49 species in 14 orders were examined for Marek's disease (MD) herpesvirus (MDHV) infection. MDHV was isolated from 10 birds, all belonging to genus Gallus. The precipitating antibodies against MDHV were demonstrated only in the Gallus birds, when 51 selected birds including 34 Galliformes and 17 other birds representing 12 species from nine orders were examined. The 10 MDHV isolates all induced morphologically similar plaques in cell cultures closely resembling those of HN strain, a low pathogenic isolate of MDHV. Six of the 10 isolates, when inoculated into an experimental line of chickens highly susceptible to MD, caused only a minimal degree of histologic lesions without causing clinical MD, gross MD lesions, or deaths from MD. Natural hosts of MD are probably Galliformes, primarily affecting Gallus and less often other genera of Galliformes.     

Hepatic iron overload in birds: Analytical and morphological studies

Hepatic iron content was determined in post mortem specimens from a wide range of avian species collected over a 12-month period. The majority (> 90%) of these specimens (n = 40) showed high iron content (up to 12 mg Fe/g tissue). The highest concentrations were associated with fibrosis and regenerative nodules. Dietary analysis indicated that the iron intake was not excessive, suggesting that iron-loading was due to enhanced intestinal absorption.     

Detection of egg drop syndrome 1976 virus by polymerase chain reaction and study of its persistence in experimentally infected layer birds

Polymerase chain reaction (PCR) assay was developed for the detection of Egg drop syndrome 1976 (EDS-76) virus in tissues, namely in the uterus, spleen and buffy coat. It was also used to study the persistence of the virus in tissues of experimentally infected layer birds. The PCR assay could detect as little as 10 fg of purified EDS-76 viral DNA. It also amplified the DNA of Fowl adenovirus serotypes 4 (FAV-4) and 8 (FAV-8). The virus persisted in the uterus up to day 21 post infection (p.i.). Detection of EDS-76 viral DNA in the buffy coat could be useful for studying the occurrence of the respective disease in layer bird flocks.     

Mark-release-recapture studies with Culex mosquitoes (Diptera: Culicidae) in Southern California

Cohorts of Culex adults were marked uniquely with date- and site-specific fluorescent dust colors and were released at centrally located residences and at peripheral breeding sources to study population dispersal, size, additions, and deletions. The recapture rate of Cx. quinquefasciatus females was higher and the mean distance dispersed was lower in residential than in agricultural or park habitats. Dispersal was associated with host-seeking activity and ranged from 0.6 to 1.0 km/d. Survivorship ranged from 0.65 to 0.84 per day, and population density ranged from 36,612 to 671,634 females per km2. The sampling efficiency of CO2-baited traps in residential habitats increased coincidentally with increasing population density. Gravid traps were most effective in residential habitats where there were few competitive oviposition sites. Teneral Cx. stigmatosoma were extremely dispersive, and few marked females were recaptured. Unmarked females were more abundant at CO2-baited traps in residential habitats than at traps near productive peripheral breeding sources. Few Cx. tarsalis were released, and the recapture rate in residential habitats was low when compared with rural sites.     

Junín virus persistence in mice

Newborn mice surviving intracerebral infection with Junín virus (JV) strain XJ showed viral persistence in brain up to 140 days post-infection (p.i.). Mild meningoencephalitis or encephalitis, but not the neutralizing antibody titres (NtAb) correlated with virus presence.     

Defective interfering particles: effects in modulating virus growth and persistence

Defective interfering virus particles (DIP) frequently play an important part in viral persistence in vitro, and may in some instances modify a virus infection in vivo, causing attenuation or persistence of the infection. To explain certain aspects of the growth of these mutants in vitro, other factors have been invoked such as interferon, mutations in the wild-type virus or the infected cells, or other substances released by infected cells that attenuate the infection. We present here a simple model of the growth of DIP in vitro which shows that (a) the observed population dynamics of DIP can readily be explained without invoking such extrinsic factors; (b) the initial multiplicity of infection of DIP is the principal determinant of the outcome of infection in both single- and repeated-passage cultures; and (c) in a long-term culture in vitro, the criterion used to decide the time of virus passage directly determines how long the standard virus, DIP, and cells survive. This model may be used with minor modifications to predict the behavior in vitro of other mutant viruses with a dominantly interfering phenotype.     

Seasonal abundance and survivorship of Culicoides sonorensis (Diptera: Ceratopogonidae) at a southern California dairy, with reference to potential bluetongue virus transmission and persistence

Seasonal abundance and survivorship of Culicoides sonorensis Wirth & Jones were examined at a dairy in southern California from January 1995 to December 1997. Insects were collected one to two times per week using five CDC-type suction traps (without light) baited with CO2 at a constant release rate of 1,000 ml/min. Female and male abundance was greatest during late summer and early fall and was directly correlated with mean monthly air temperature. Parity of females was lowest during late summer and early fall. The gonotrophic cycle was estimated to require 3-4 during hot summer months and up to 14 d during cool winter months. Estimated extrinsic incubation of bluetongue virus (BLU) was 9-10 d during August and September. The estimated daily survival ranged from < 60% in the summer to > 95% in the winter, resulting in an expectation of life of only 2-3 d in summer and > 10 d in winter. The probability of females surviving the extrinsic incubation period for BLU virus, and the expectation of infective life were both lowest during late summer and early fall. During 1997, midge abundance during late summer was not high enough to overcome very low survivorship, and the absolute number of females expected to survive the extrinsic incubation period was relatively low. However, in 1995 and 1996, very high midge abundance compensated for low survivorship during summer and the number of females expected to survive the extrinsic incubation period was relatively high. Although abundance was generally very low during the cool winter and spring, host-seeking females were captured throughout the year, and their winter survival was high. Overwintering of BLU virus by continued transmission of the virus by active midges appears possible.