Dead Wild Birds Research Articles

Captive Rearing and Release of Bar-headed Geese (Anser indicus) in China: A Possible HPAI H5N1 Virus Infection Route to Wild Birds

In April–June 2005, more than 6,000 birds, including more than 3,000 Barheaded Geese Anser indicus, died at Lake Qinghai, northern China, during an outbreak of highly pathogenic avian influenza (HPAI) H5N1 (Chen et al., 2006). In spring 2006, more waterbirds, again mainly Bar-headed Geese (939 recorded at three Qinghai sites but not counted at others), died at four sites in Qinghai province and three in Tibet. Two virus isolations, from a Bar-headed Goose and a Great Black-headed Gull (Larus ichthyaetus), were reported from Lake Qinghai in 2007 (Li et al., 2010) and in May 2009, more wild birds (species not recorded) died at two sites in Qinghai province (Wang et al., 2008; World Organisation for Animal Health [OIE], 2010). In May 2010, dead wild birds, including Barheaded Geese, were reported from Nagqu province, Tibet (OIE, 2010). In 2005, 2006, 2009, and 2010 outbreaks on the Qinghai-Tibet plateau were contemporaneous with or followed by outbreaks in wild birds in Mongolia and southern Russia (OIE, 2010). Virus isolation from dead wild birds at Lake Qinghai in 2005 revealed a new strain of HPAI H5N1, designated Clade 2.2 (World Health Organization/World Organisation for Animal Health/Food and Agriculture Organization, 2008), which has subsequently spread to Europe and Asia (Feare, 2007). Although there had been outbreaks in poultry in Lanzhou, approximately 300 km from Lake Qinghai, and in the vicinity of Lhasa, Tibet (OIE, 2010), the evolution of HPAI H5N1 viruses was not studied in northern China (Smith et al., 2006), and the origin of the Qinghai stain remains a mystery. After the 2005 HPAI H5N1 outbreak at Lake Qinghai, one of the authors (T.K.) discovered Chinese-language internet media bulletins that described captive rearing of Bar-headed Geese on China’s central high plateau. These reports lack detail but describe practices that promote contact between captive-bred geese and domesticated and wild birds. Our aim here is to summarize the salient information from these reports because they present a possible scenario for emergence of the Qinghai Clade 2.2 viruses and its occurrence in wild birds. Captive breeding of Bar-headed Geese began in 2003 at Yamzho Yumco Lake, about 100 km south of Lhasa, Tibet, by the ‘‘Lhasa Nida Natural Ecology Development Bar-headed Goose Artificial Breeding Company.’’ Several hundred abandoned Bar-headed Goose eggs were collected from a declining colony, claimed to comprise 30,000 birds, at the lake and were artificially incubated, with 99% reared to fledging. The enterprise was extended to Xaina County, Nagqu Prefecture (about 300 km NE of Lhasa), where 500 Bar-headed Geese were successfully raised ‘‘to meet market demand.’’ The company also contracted production to a villager in Gonggar (Meldrogonkar) County, south of Lhasa. In 2006 another breeding unit was established in the same town, at a wetland and reservoir suitable

Comparative analysis of six genome sequences of three novel picornaviruses, turdiviruses 1, 2 and 3, in dead wild birds, and proposal of two novel genera, Orthoturdivirus and Paraturdivirus, in the family Picornaviridae

In this territory-wide molecular epidemiology study of picornaviruses, involving 6765 dead wild birds from 201 species in 50 families over a 12 month period, three novel picornaviruses, turdiviruses 1, 2 and 3 (TV1, TV2 and TV3), were identified from birds of different genera in the family Turdidae. In contrast to many other viruses in birds of the family Turdidae or viruses of the family Picornaviridae, TV1, TV2 and TV3 were found exclusively in the autumn and winter months. Two genomes each of TV1, TV2 and TV3 were sequenced. Regions P1, P2 and P3 of the three turdiviruses possessed, respectively, <40, <40 and <50 % amino acid identities with those of other picornaviruses. Moreover, P1, P2 and P3 of TV1 also possessed, respectively, <40, <40 and <50 % amino acid identities with those of TV2 and TV3. Phylogenetic analysis revealed that TV1, TV2 and TV3 were distantly related to members of the genus Kobuvirus. Among the three turdiviruses, TV2 and TV3 were always clustered together, with high bootstrap supports of 1000. The genomic features of TV2 and TV3 were also distinct from TV1, including lower G+C contents, shorter leader protein and a preference for codon sequence NNT rather than NNC for amino acids that can use either NNT or NNC as codons (P<0.001 by χ(2)-test). Based on our results we propose two novel genera, Orthoturdivirus for TV1, and Paraturdivirus for TV2 and TV3, in the family Picornaviridae. The type of internal ribosomal entry site for TV1, TV2 and TV3 remains to be determined.

Geographical spread of highly pathogenic avian influenza virus H5N1 during the 2006 outbreak in Austria.

In spring 2006, highly pathogenic avian influenza virus (HPAIV) of subtype H5N1 was detected in Austria in 119 dead wild birds. The hemagglutinin cleavage site showed that the amino acid sequence motif was identical to that of the Qinghai lineage. For detailed analysis, the hemagglutinin (HA) and neuraminidase (NA) genes of 27 selected Austrian H5N1 viruses originating from different regions and wild bird species were analyzed phylogenetically, which revealed two clearly separated Austrian subclusters, both belonging to European cluster EMA-1. Subcluster South (SCS) contains virus isolates from the south of Austria as well as from Slovenia, Turkey, Egypt, and Nigeria. The second subcluster, Northwest (SCN), covered a larger group of viruses originating from different locations and wild bird species in the northern and very western parts of Austria, as well as from Bavaria and Switzerland. Surprisingly, virus isolates originating from two mute swans and one wild duck found on the north side of the Alps did not cluster with SCN but with SCS. Together with isolates from Bavarian, the Czech Republic, Italy, and Slovakia, they form a genuine subgroup, named subgroup Bavaria (SGB). This subgroup forms a link to SCN, indicating a spread of the virus from south to north. There has been a general assumption that the generic HPAI introduction route into Europe was from Russia to north Germany, introducing cluster EMA-2 into Europe. Interestingly, our findings support the assumption of an alternative introduction of the HPAI H5N1 virus from Turkey to central Europe, where it spread as cluster EMA-1 during the outbreak of 2006.

Visualization and Analysis of the Danish 2006 Highly Pathogenic Avian Influenza Virus H5N1 Wild Bird Surveillance Data by a Prototype Avian Influenza BioPortal

Infection with highly pathogenic avian influenza virus H5N1 occurred for the first time in Denmark in 2006 during the last part of the European epidemic that mainly affected migrating wild birds. The total number of Danish wild bird cases was 45, of which only one was found through active surveillance using fecal sampling from resting areas for migrating species, whereas passive surveillance of dead wild birds provided 44 cases. One backyard, mixed poultry flock also became infected late in the epidemic. This study describes the spatial and temporal distribution of cases, initially characterized by a spatial-temporal cluster of affected tufted ducks that led to further spread to other wild bird species in the vicinity. The surveillance data also indicate an apparent die-off of the regional epidemic. As a tool in visualizing the spatial and temporal development of the epidemic, a prototype avian influenza (AI) BioPortal was used to provide online web-based access to the data. The AI BioPortal tools include mapping, graphing, phylogenetic tree construction, playback scenarios, and visualization of results of temporal-spatial analyses. Several of the features of this surveillance system compare favorably to the design of existing national and international surveillance information systems, and the AI BioPortal may become a useful tool for disease surveillance and for decision support in the event of future AI epidemics, both at national and international levels.

Virological Monitoring of White Storks (Ciconia ciconia) for Avian Influenza

Between 2003 and 2008, more than 600 white stork (Ciconia ciconia) nestlings in the German federal state of Brandenburg were ringed and examined for influenza A viruses. With the spread of highly pathogenic avian influenza virus (HPAIV) of subtype H5N1 among wild birds in Germany in spring 2006, dead wild birds, including 88 white storks, were tested for infection with HPAIV. Furthermore, fresh fecal samples were examined by RT-PCR to monitor the occurrence of HPAIV in adult storks. While the monitoring of nestlings and adult white storks failed to yield evidence of influenza A virus infections in these birds, two storks found dead in April 2006 in the same location tested positive for HPAIV H5N1. Sequence analysis revealed that the virus isolated from one of the storks belonged to clade 2.2, which was commonly found in wild birds in the north of Germany and other European countries during the epidemic in 2006. Despite these two cases, white storks seemed to serve as neither a vector nor as a reservoir for HPAIV in Germany. The risk of white storks transmitting HPAIV to domestic poultry and humans is low.

Evolutionary genetics of highly pathogenic H5N1 avian influenza viruses isolated from whooper swans in northern Japan in 2008

In April and May 2008, highly pathogenic avian influenza viruses subtype H5N1 were isolated from dead or moribund whooper swans in Aomori, Akita and Hokkaido prefectures in northern Japan. To trace the genetic lineage of the isolates, the nucleotide sequences of all eight genes were determined and phylogenetically analyzed. The Japanese strains were nearly identical to chicken viruses isolated in Russia in April 2008 and closely related to viruses isolated from dead wild birds in Hong Kong in 2007-2008. Their HA genes clustered in clade 2.3.2. On the other hand, NA and the other internal genes were closely related to those of clade 2.3.4 viruses (genotype V) whose NP genes originated from an HA clade 2.3.2 virus. In conclusion, the H5N1 viruses isolated in Japan, Russia and Hong Kong were derived from a common ancestor virus belonging to genotype V that was generated from genetic reassortment events between viruses of HA clades 2.3.2 and 2.3.4.

Analysis of H5N1 avian influenza infections from wild bird surveillance in Hong Kong from January 2006 to October 2007

Intensive surveillance of dead wild birds for H5N1 avian influenza infection is conducted in Hong Kong. Between January 2006 and October 2007 pooled cloacal and tracheal swabs from 17692 dead wild birds (from 16 different orders including 82 genera) were tested and 33 H5N1 highly pathogenic avian influenza viruses were isolated. No H5N1 infection has occurred in poultry farms since January 2003, or in live poultry markets in Hong Kong since November 2003 until a recent detection of H5N1 virus by surveillance of live poultry markets in June 2008. The gross and histopathology in the various H5N1-infected avian species is described, along with the performance of the virus isolation and polymerase chain reaction (PCR) tests used. This evaluation also included determination of virus titres and detection limits for the H5 haemagglutinin gene (H5)and matrix gene real-time reverse-transcription PCR tests in cloacal and tracheal swabs from 12 wild birds. The viruses isolated belonged to Clades 2.3.2 and 2.3.4, and within Clade 2.3.4 some clustering was evident based on H5 haemagglutinin gene haemagglutinating sequencing. There were no differences in the pathology findings between these subgroupings and the various diagnostic tests gave similar results for these viruses, except for a loss in sensitivity of the H5 real-time reverse-transcription PCR for several viruses in one cluster from birds submitted in February 2007. The use of multiple test methods was important in maintaining the diagnostic sensitivity for detecting avian influenza viruses with high genetic variability.

Epidemiological and Ornithological Aspects of Outbreaks of Highly Pathogenic Avian Influenza Virus H5N1 of Asian Lineage in Wild Birds in Germany, 2006 and 2007

In Germany, two distinct episodes of outbreaks of highly pathogenic avian influenza virus of subtype H5N1 (HPAIV H5N1) in wild birds occurred at the beginning of 2006, and in summer 2007. High local densities of wild bird populations apparently sparked clinically detectable outbreaks. However, these remained restricted in (i) number of birds, (ii) species found to be affected, (iii) time, and (iv) location despite the presence of several hundred thousands of susceptible wild birds and further stressors (food shortage, harsh weather conditions and moulting). Northern and southern subpopulations of several migratory anseriform species can be distinguished with respect to their preference for wintering grounds in Germany. This corroborates viral genetic data by Starick et al. (2008) demonstrating the introduction of two geographically restricted virus subpopulations of Qinghai-like lineage (cluster 2.2.A and 2.2.B) into northern and southern Germany, respectively, in 2006. The incursion of virus emerging in 2007, found to be distinct from the clusters detected in 2006 (Starick et al., 2008), may have been associated with moulting movements. Intensive past-outbreak investigations with negative results of live and dead wild birds and of terrestrial scavengers excluded continued circulation of virus on a larger scale. However, persistence of virus in small pockets of local wild bird populations could not be ruled out resiliently. 1.5% of investigated sera originating from cats sampled at the epicentres of the Ruegen 2006-outbreak contained H5-antibodies. Passive monitoring was found to be highly superior to live bird surveillance when aiming at the detection of HPAIV H5N1 in wild birds (P < 0.0001).

Characterization of Avian Influenza Viruses A (H5N1) from Wild Birds, Hong Kong, 2004–2008

From January 2004 through June 2008, surveillance of dead wild birds in Hong Kong, People's Republic of China, periodically detected highly pathogenic avian influenza (HPAI) viruses (H5N1) in individual birds from different species. During this period, no viruses of subtype H5N1 were detected in poultry on farms and in markets in Hong Kong despite intensive surveillance. Thus, these findings in wild birds demonstrate the potential for wild birds to disseminate HPAI viruses (H5N1) to areas otherwise free from the viruses. Genetic and antigenic characterization of 47 HPAI (H5N1) viruses isolated from dead wild birds in Hong Kong showed that these isolates belonged to 2 antigenically distinct virus groups: clades 2.3.4 and 2.3.2. Although research has shown that clade 2.3.4 viruses are established in poultry in Asia, the emergence of clade 2.3.2 viruses in nonpasserine birds from Hong Kong, Japan, and Russia raises the possibility that this virus lineage may have become established in wild birds.

Comparative Analysis of Complete Genome Sequences of Three Avian Coronaviruses Reveals a Novel Group 3c Coronavirus

In this territory-wide molecular epidemiology study of coronaviruses (CoVs) in Hong Kong involving 1,541 dead wild birds, three novel CoVs were identified in three different bird families (bulbul CoV HKU11 [BuCoV HKU11], thrush CoV HKU12 [ThCoV HKU12], and munia CoV HKU13 [MuCoV HKU13]). Four complete genomes of the three novel CoVs were sequenced. Their genomes (26,396 to 26,552 bases) represent the smallest known CoV genomes. In phylogenetic trees constructed using chymotrypsin-like protease (3CL(pro)), RNA-dependent RNA polymerase (Pol), helicase, spike, and nucleocapsid proteins, BuCoV HKU11, ThCoV HKU12, and MuCoV HKU13 formed a cluster distantly related to infectious bronchitis virus and turkey CoV (group 3a CoVs). For helicase, spike, and nucleocapsid, they were also clustered with a CoV recently discovered in Asian leopard cats, for which the complete genome sequence was not available. The 3CL(pro), Pol, helicase, and nucleocapsid of the three CoVs possessed higher amino acid identities to those of group 3a CoVs than to those of group 1 and group 2 CoVs. Unique genomic features distinguishing them from other group 3 CoVs include a distinct transcription regulatory sequence and coding potential for small open reading frames. Based on these results, we propose a novel CoV subgroup, group 3c, to describe this distinct subgroup of CoVs under the group 3 CoVs. Avian CoVs are genetically more diverse than previously thought and may be closely related to some newly identified mammalian CoVs. Further studies would be important to delineate whether the Asian leopard cat CoV was a result of interspecies jumping from birds, a situation analogous to that of bat and civet severe acute respiratory syndrome CoVs.

Surveillance of H5 Avian Influenza Virus in Wild Birds Found Dead

Wild birds are suspected to play a role in the spread of avian influenza H5N1; however, much remains unknown about the ecology and epidemiology of H5N1 in wild birds. Lake Constance is an important wetland area and was a focus for surveillance of dead wild birds between February and June 2006. Dead wild birds collected from the lake and surrounding regions were tested for avian influenza H5. This article provides a descriptive and spatial analysis of the data collected during this period and includes discussion of the strengths and limitations of this type of surveillance. The sampling of dead birds may provide a rapid and cost-effective means of detecting the presence of H5N1; however, such sampling is prone to certain biases and lacks sensitivity in detecting asymptomatic infections. The benefit of such surveillance will be enhanced by detailed ornithologic information, greater accuracy of spatially referenced data, and quantification of surveillance effort.

Review of clinical symptoms and spectrum in humans with influenza A/H5N1 infection

Influenza A/H5N1 infection has become the major emerging infectious disease of global concern again since late 2003. A history of exposure to dead or sick poultry or wild birds occurs in over 60% of cases of human H5N1 infection. The incubation period of avian-to-human transmission is generally between 2 and 5 days and the median duration of symptoms before hospitalization is about 4.5 days. The clinical spectrum has ranged from asymptomatic infection or mild influenza-like illness to severe pneumonia and multi-organ failure. Fever > 38 degrees C, cough and dyspnoea are the major symptoms on presentation, whereas gastrointestinal symptoms such as watery diarrhoea, vomiting and abdominal pain are common early in the course of the disease. In contrast, upper respiratory tract symptoms are less prominent in human H5N1 infection when compared to seasonal influenza. Laboratory features of human H5N1 infection include leucopoenia, especially lymphopenia, elevated amino-transaminases, thrombocytopenia, prolonged prothrombin time and activated partial thromboplastin time, increased D-Dimer, increased serum lactate dehydrogenase and creatinine phospho-kinase, and hypoalbuminemia. A low absolute lymphocyte count on admission is associated with more severe disease and death. Radiographic abnormalities include multi-focal airspace consolidation, interstitial infiltrates, patchy or lobar involvement, with rapid progression to bilateral and diffuse ground-glass opacities consistent with ARDS. However, none of the clinical, laboratory and radiographic features are specific to H5N1 infection. A detailed exposure history needs to be elicited, including any close contact with sick or dead poultry, wild birds, other severely ill persons, travel to an area with A/H5N1 activity or work in laboratory handling samples possibly containing A/H5N1 virus.

Aviäre Influenza: Wildvogelmonitoring in der Schweiz zwischen 2003–2006

During various surveillance programs more than 3500 cloacal swabs and organ samples from songbirds, waterbirds and poultry have been tested for avian influenza using real time RT-PCR. Switzerland carried out the first wildbird monitoring between autumn 2003 and spring 2005. 1053 samples, mostly from songbirds, were tested. LPAI-strains were found in two cases. A second intensified surveillance program started in October 2005 along with the first ban on free range poultry farming. Until the end of April 2006 2455 cloacal swabs from dead wildbirds have been analysed. By the end of february H5N1 was for the first time detected in Switzerland and by the end of march 32 waterbirds have been found positive for H5N1. 146 poultry flocks with a special permission for free range management proved to be AI negative.

Phylogenetic analyses of highly pathogenic avian influenza virus isolates from Germany in 2006 and 2007 suggest at least three separate introductions of H5N1 virus

In spring 2006, highly pathogenic avian influenza virus (HPAIV) of subtype H5N1 was detected in Germany in 343 dead wild birds, as well as in a black swan ( Cygnus atratus) kept in a zoo, three stray cats, one stone marten ( Martes foina), and in a single turkey farm. In 2007 (June–July) the virus reoccurred in 96 wild birds at six geographically separate locations in the Southeast of Germany. In addition, a backyard mixed duck and goose holding was affected. Real-time RT-PCR [Hoffmann, B., Harder, T., Starick, E., Depner, K., Werner, O., Beer, M., 2007. Rapid and highly sensitive pathotyping of avian influenza A H5N1 virus by using real-time reverse transcription-PCR. J. Clin. Microbiol. 45, 600–603] and nucleotide sequencing confirmed that these H5-viruses belonged to the Qinghai lineage of HPAIV H5N1 (clade 2.2). For a more detailed analysis, the hemagglutinin and neuraminidase genes of 27 selected German H5N1 viruses isolated 2006 or 2007 and originating from different regions and animal species were sequenced and analysed phylogenetically. As a result, three closely related but distinguishable H5N1 subclades could be defined: In 2006 a ‘Northern type’ (subclade 2.2.2), representing virus isolates from the German federal states Mecklenburg-Western Pomerania, Schleswig-Holstein, Brandenburg, and Lower Saxony, and a ‘Southern type’ (subclade 2.2.1) from Baden-Württemberg and Bavaria were detected. Interestingly, representatives of both types were present in Central Germany and caused the outbreak in turkeys (subclade 2.2.2) and in a case in a tufted duck ( Aythya fuligula) (subclade 2.2.1) in Saxony. Furthermore, one isolate from the South of Germany was identified as 2.2.2 and vice versa a 2.2.1-like isolate was found in Northern Germany. H5N1 viruses isolated in 2007 belonged to a third type (subclade 2.2.3) which was not detected in 2006. Our data suggest the introduction of three distinct H5N1 variants into the wild bird population of Germany. The source of these viruses and the exact time of introduction remain obscure. Based on the identification of closely related H5N1 viruses from Southern and Central Russia, a recent introduction via wild birds on winter escape from these regions, early in 2006 constitutes the most likely scenario for the 2006 outbreaks. The viruses detected in 2007 most likely represent another new incursion from an as yet unknown source.

Avian influenza virus H 5 N 1 subtype A diagnosed in sick and dead wild and domestic birds in Pavlodar oblast, Republic of Kazakhstan

This work describes laboratory tests conducted on biological material taken from sick and dead birds from unknown infection in the Pavlodar oblast, Republic of Kazakhstan. In the course of pathological material laboratory tests electron microscopy revealed virion of type A avian influenza virus. Type A AI virus common protein is recognized with the help of “Directigen Flu A” test system. The results of HAIT showed hemagglutinin agent H5, and NAIT results showed neuraminidase agent N1. On the basis of laboratory tests results it was determined that the cause of sickness and death of birds in the territory of Pavlodar oblast Irtysh region farm “NAN” was H5N1 type A AI virus. Key words: HAT, HAIT, NAIT, H5N1 type A avian influenza.

Possible Avian Influenza (H5N1) from Migratory Bird, Egypt

Possible Avian Influenza (H5N1) from Migratory Bird, Egypt

Validated H5 Eurasian Real-Time Reverse Transcriptase–Polymerase Chain Reaction and Its Application in H5N1 Outbreaks in 2005–2006

Real time reverse transcriptase (RRT)-polymerase chain reaction (PCR) for the detection of Eurasian H5 avian influenza virus (AIV) isolates was adapted from an existing protocol, optimized, and validated using a number of genetically diverse H5 isolates (n = 51). These included 34 "Asian lineage" H5N1 highly pathogenic avian influenza (HPAI) viruses (2004-2006), plus 12 other H5 isolates from poultry outbreaks and wild birds in the Eastern Hemisphere (1996-2005). All 51 were positive by H5 Eurasian RRT-PCR. Specificity was assessed by testing representative isolates from all other AL virus subtypes (n = 52), non-AI avian pathogens (n = 8), plus a negative population of clinical specimens derived from AI-uninfected wild birds and poultry (n = 604); all were negative by H5 Eurasian RRT-PCR. RNA was directly extracted from suspect HPAI H5N1 clinical specimens (Africa, Asia, and Europe; 2005-2006; n = 58) from dead poultry and wild birds, and 55 recorded as positive by H5 Eurasian RRT-PCR: Fifty-one of these 55 were in agreement with positive AIV isolation in embryonated chickens' eggs. H5 Eurasian RRT-PCR was invaluable in H5 outbreak diagnosis and management by virtue of its rapidity and high degree of sensitivity and specificity. This method provides a platform for automation that can be applied for large-scale intensive investigations, including surveillance.

The avian flu: some lessons learned from the 2003 SARS outbreak in Toronto

As the H5N1 avian influenza virus races across the globe, leaving in its wake dead wild and domestic birds in many parts of Asia and Europe in the autumn of 2005, we may recall the not so distant memory of Severe Acute Respiratory Syndrome. During the initial outbreak period in Southern China in the early months of 2003, the withholding of epidemiological data by health authorities due to political reasons has made it difficult determine an exact number (see Davis 2005, 69), but it is estimated that eventually 8069 people were confirmed as infected and at least 773 died in a global SARS epidemic that spread mostly between large, global ized metropolitan centres in East Asia and North America. In Toronto, 44 people succumbed to the previously unknown corona virus, 213 were con firmed infected cases, thousands were quarantined, while millions of dollars were lost in business. What can the experience of how this disease was iden tified, monitored, controlled and ultimately beaten tell us about what we can expect in a major H5N1 outbreak? Admittedly, there are differences between the two diseases. Local SARS transmission was mostly limited to the hospital setting, with community infection rather rare although the role of 'super-spreaders' was an important mechanism for disease spread at the global level. Although both diseases had their origin in animals, SARS spread readily from humans to other humans. H5N1 does not (yet). It needs the vector of migrating or transported birds and seldom infects humans. Mike Davis names a few more differences: although SARS produces similar symptoms, it is not nearly as 'subtle' as influenza . . . SARS needs about five days to incubate and does not usually become contagious until well after the onset of fever and dry coughing; infectiousness takes about ten days to peak, and research has found few asymptotic infections without sickness. The old-fashioned tactics of isolation and quarantine, if ruthlessly implemented, can work effectively against such a slow-developing virus whose symptoms consistently signal infectiousness. (2005, 78)

WILD BIRD MORTALITY AND WEST NILE VIRUS SURVEILLANCE: BIASES ASSOCIATED WITH DETECTION, REPORTING, AND CARCASS PERSISTENCE

Surveillance targeting dead wild birds, in particular American crows (Corvus brachyrhynchos), plays a critical role in West Nile virus (WNV) surveillance in the United States. Using crow decoy surrogates, detection and reporting of crow carcasses within urban and rural environments of DeKalb County, Georgia were assessed for potential biases that might occur in the county's WNV surveillance program. In each of two replicated trials, during July and September 2003, 400 decoys were labeled with reporting instructions and distributed along randomly chosen routes throughout designated urban and rural areas within DeKalb County. Information-theoretic methods were used to compare alternative models incorporating the effects of area and trial on probabilities of detection and reporting. The model with the best empirical support included the effects of area on both detection and reporting of decoys. The proportion of decoys detected in the urban area (0.605, SE=0.024) was approximately twice that of the rural area (0.293, SE=0.023), and the proportion of decoys reported in the urban area (0.273, SE=0.023) was approximately three times that of the rural area (0.103, SE=0.028). These results suggest that human density and associated factors can substantially influence dead crow detection and reporting and, thus, the perceived distribution of WNV. In a second and separate study, the persistence and fate of American crow and house sparrow (Passer domesticus) carcasses were assessed in urban and rural environments in Athens-Clarke, Madison, and Oconee counties, Georgia. Two replicated trials using 96 carcasses of each species were conducted during July and September 2004. For a portion of the carcasses, motion sensitive cameras were used to monitor scavenging species visits. Most carcasses (82%) disappeared or were decayed by the end of the 6-day study. Carcass persistence averaged 1.6 days in rural areas and 2.1 days in urban areas. We analyzed carcass persistence rates using a known-fate model framework in program MARK. Model selection based on Akaike's Information Criteria (AIC) indicated that the best model explaining carcass persistence rates included species and number of days of exposure; however, the model including area and number of days of exposure received approximately equal support. Model-averaged carcass persistence rates were higher for urban areas and for crow carcasses. Six mammalian and one avian species were documented scavenging upon carcasses. Dead wild birds could represent potential sources of oral WNV exposure to these scavenging species. Species composition of the scavenger assemblage was similar in urban and rural areas but "scavenging pressure" was greater in rural areas.

Enhancing West Nile virus surveillance, United States.

We provide a method for constructing a county-level West Nile virus risk map to serve as an early warning system for human cases. We also demonstrate that mosquito surveillance is a more accurate predictor of human risk than monitoring dead and infected wild birds.