Experimental infection of house sparrows (Passer domesticus) with West Nile virus isolates of Euro-Mediterranean and North American origins

West Nile virus (WNV) is a zoonotic arboviral pathogen transmitted by mosquitoes in a cycle involving wild birds as reservoir hosts. The virus has recently emerged in North America and re-emerged in Europe. North American WNV outbreaks are often accompanied by high mortality in wild birds, a feature that is uncommon in Europe. The reason for this difference is unknown, but the intrinsic virulence of the viruses circulating in each continent and/or the susceptibility to the disease of Palearctic as opposed to Nearctic wild bird species could play a role. To assess this question, experimental inoculations with four lineage 1 WNV strains, three from southern Europe (Italy/2008, Italy/2009 and Spain/2007) and one from North America (NY99) were performed on house sparrows (Passer domesticus), a wild passerine common in both continents. Non-significant differences which ranged from 0% to 25% were observed in mortality for the different WNV strains. Viremias lasted from 1 to 5–6 days post-inoculation (dpi) in all cases; individuals inoculated with NY99 had significantly higher titres than those inoculated with any of the Euro-Mediterranean strains. Remarkably, host competence was found to be higher for NY99 than for the other strains. Consequently, albeit being pathogenic for house sparrows, some Euro-Mediterranean strains had reduced capacity for replication in -and transmission from- this host, as compared to the NY99 strain. If applicable also to other wild bird host species, this relatively reduced transmission capacity of the Euro-Mediterranean strains could explain the lower incidence of this disease in wild birds in the Euro-Mediterranean area.     

CHLAMYDIA AVIUM DETECTION FROM A RING-NECKED PARAKEET (PSITTACULA KRAMERI) IN FRANCE

The crossing of host species barriers, through the spreading populations of introduced pet animals that become established in the wild, sets the stage for zoonotic pathogen (re)emergence. A literature review on pathogens that are hosted by the ring-necked parakeet (Psittacula krameri), a worldwide introduced pet, highlighted local infections of captive birds by chlamydial agents with high sanitary risk for human health in its introduced range. We searched for these pathogens through cloacal swabs collected from 85 individuals in an invasive population established in the suburban areas of Paris (Ile-de-France) from 5 localities during the winter seasons between 2011 and 2014. Based on quantitative PCR analysis, Chlamydiaceae shedding was detected at too low levels for species identification in 5 birds, but 1 parakeet (found dead) was positive for Chlamydiaceae typed as Chlamydia avium. The only known hosts recorded for C. avium in Europe are feral pigeons (Columba livia) and captive psittacines. This result raises the question of the sanitary risks associated with new pathogen transmission from exotic pets released in the wild, which could locally affect birds and potentially people who feed birds.     

Ecological Surveillance for West Nile in Catalonia (Spain), Learning from a Five‐Year Period of Follow‐up

To enhance early detection of West Nile virus (WNV) transmission, an integrated ecological surveillance system was implemented in Catalonia (north‐eastern Spain) from 2007 to 2011. This system incorporated passive and active equine surveillance, periodical testing of chicken sentinels in wetland areas, serosurveillance wild birds and testing of adult mosquitoes. Samples from 298 equines, 100 sentinel chickens, 1086 wild birds and 39 599 mosquitoes were analysed. During these 5 years, no acute WNV infection was detected in humans or domestic animal populations in Catalonia. WNV was not detected in mosquitoes either. Nevertheless, several seroconversions in resident and migrant wild birds indicate that local WNV or other closely related flaviviruses transmission was occurring among bird populations. These data indicate that bird and mosquito surveillance can detect otherwise silent transmission of flaviviruses and give some insights regarding possible avian hosts and vectors in a European setting.     

Pathogenicity of two recent Western Mediterranean West Nile virus isolates in a wild bird species indigenous to Southern Europe: the red-legged partridge

ABSTRACT: West Nile virus (WNV) is an emerging zoonotic pathogen whose geographic spread and incidence in humans, horses and birds has increased significantly in recent years. WNV has long been considered a mild pathogen causing self-limiting outbreaks. This notion has changed as WNV is causing large epidemics with a high impact on human and animal health. This has been particularly noteworthy since its introduction into North America in 1999. There, native bird species have been shown to be highly susceptible to WNV infection and disease with high mortalities. For this reason, the effect of WNV infection in North American bird species has been thoroughly studied by means of experimental inoculations in controlled trials. To a lesser extent, European wild birds have been shown to be affected clinically by WNV infection. Yet experimental studies on European wild bird species are lacking. The red-legged partridge (Alectoris rufa) is a gallinaceous bird indigenous to the Iberian Peninsula, widely distributed in South Western Europe. It plays a key role in the Mediterranean ecosystem and constitutes an economically important game species. As such it is raised intensively in outdoor facilities. In this work, red-legged partridges were experimentally infected with two recent WNV isolates from the Western Mediterranean area: Morocco/2003 and Spain/2007. All inoculated birds became viremic and showed clinical disease, with mortality rates of 70% and 30%, respectively. These results show that Western Mediterranean WNV variants can be pathogenic for some European bird species, such as the red-legged partridge.     

The devil is in the details—Host disease and co‐infections are associated with zoonotic pathogen carriage in Norway rats (Rattus norvegicus)

Traditionally, zoonotic pathogen ecology studies in wildlife have focused on the interplay among hosts, their demographic characteristics and their pathogens. But pathogen ecology is also influenced by factors that traverse the hierarchical scale of biological organization, ranging from within‐host factors at the molecular, cellular and organ levels, all the way to the host population within a larger environment. The influence of host disease and co‐infections on zoonotic pathogen carriage in hosts is important because these factors may be key to a more holistic understanding of pathogen ecology in wildlife hosts, which are a major source of emerging infectious diseases in humans. Using wild Norway rats (Rattus norvegicus) as a model species, the purpose of this study was to investigate how host disease and co‐infections impact the carriage of zoonotic pathogens. Following a systematic trap and removal study, we tested the rats for the presence of two potentially zoonotic bacterial pathogens (Bartonella tribocorum and Leptospira interrogans) and assessed them for host disease not attributable to these bacteria (i.e., nematode parasites, and macroscopic and microscopic lesions). We fitted multilevel multivariable logistic regression models with pathogen status as the outcome, lesions and parasites as predictor variables and city block as a random effect. Rats had significantly increased odds of being infected with B. tribocorum if they had a concurrent nematode infection in one or more organ systems. Rats with bite wounds, any macroscopic lesion, cardiomyopathy or tracheitis had significantly increased odds of being infected with L. interrogans. These results suggest that host disease may have an important role in the ecology and epidemiology of rat‐associated zoonotic pathogens. Our multiscale approach to assessing complex intrahost factors in relation to zoonotic pathogen carriage may be applicable to future studies in rats and other wildlife hosts.     

Evidence of West Nile virus seropositivity in wild birds on the island of Cyprus

West Nile Virus (WNV)¹ is an emerging pathogen in Cyprus, with the first human case of infection reported in 2016, and another documented in 2018. A cluster of cases in humans was then reported in 2019. However, little is known regarding which avian species might bring WNV to Cyprus. Here, we investigated seroprevalence of WNV antibodies in migratory and resident birds, captured across Cyprus to assess to what extent human populations might be exposed to WNV. We used Enzyme-Linked Immunosorbent Assay (ELISA)² to test for the presence of WNV antibodies in 836 avian blood samples of 44 species captured between 2015 and 2020. A seropositivity rate of 1.3 % was found. The majority of seropositive wild birds belonged to the migratory species Sylvia atricapilla, a common and widespread migrant, implying a high risk of WNV being introduced throughout Cyprus.     

Exploration of stress-induced immunosuppression in chickens reveals both stress-resistant and stress-susceptible antigen responses

In the present study, depriving chickens of foraging material was shown to induce stress. The impact of this type of stress on the immune response was compared with feeding of corticosterone (1.5 mg per bird per day), a hormone known to be immunosuppressive and to be the major stress hormone of chickens. Corticosterone feeding induced stress as revealed by higher heterophil/lymphocyte (H/L) ratios, longer tonic immobility (TI) reaction, reduced body weight gain and reduced egg production. Blood corticosterone levels were increased. Corticosterone feeding decreased the antibody response to tetanus toxoid and SRBC, DTH to PPD from Mycobacterium tuberculosis and the inflammatory response to PHA. Housing chickens on slats also induced chronic stress, as evidenced by increased H/L ratios, prolonged TI duration and decreased egg production. Corticosterone levels were slightly but not significantly enhanced. This novel form of chronic stress strongly suppressed humoral and cellular immune responses as evidenced by lower antibody titers to sheep red blood cells (SRBC) and tetanus toxoid (TT) decreased DTH reaction to PPD and inflammatory reaction to PHA in the skin. In contrast, the antibody response to human serum albumin (HSA) was neither influenced by corticosterone feeding nor by keeping the birds on slats. Even the combination of corticosterone feeding and housing the birds on slats did not significantly impair antibody responses to HSA. In conclusion, the present study showed that chronic stress induced by depriving the birds of foraging material led to a similar impairment of humoral and cell-mediated immunity as did feeding with corticosterone. More importantly, it showed for the first time that depending on the antigen tested, there are stress-resistant and stress-susceptible antigen responses.     

Pathogenesis of West Nile virus lineage 1 and 2 in experimentally infected large falcons

West Nile virus (WNV) is a zoonotic flavivirus that is transmitted by blood-suckling mosquitoes with birds serving as the primary vertebrate reservoir hosts (enzootic cycle). Some bird species like ravens, raptors and jays are highly susceptible and develop deadly encephalitis while others are infected subclinically only. Birds of prey are highly susceptible and show substantial mortality rates following infection. To investigate the WNV pathogenesis in falcons we inoculated twelve large falcons, 6 birds per group, subcutaneously with viruses belonging to two different lineages (lineage 1 strain NY 99 and lineage 2 strain Austria). Three different infection doses were utilized: low (approx. 500 TCID50), intermediate (approx. 4 log10 TCID50) and high (approx. 6 log10 TCID50). Clinical signs were monitored during the course of the experiments lasting 14 and 21 days. All falcons developed viremia for two weeks and shed virus for almost the same period of time. Using quantitative real-time RT-PCR WNV was detected in blood, in cloacal and oropharyngeal swabs and following euthanasia and necropsy of the animals in a variety of neuronal and extraneuronal organs. Antibodies to WNV were first time detected by ELISA and neutralization assay after 6 days post infection (dpi). Pathological findings consistently included splenomegaly, non-suppurative myocarditis, meningoencephalitis and vasculitis. By immunohistochemistry WNV-antigens were demonstrated intralesionally. These results impressively illustrate the devastating and possibly deadly effects of WNV infection in falcons, independent of the genetic lineage and dose of the challenge virus used. Due to the relatively high virus load and long duration of viremia falcons may also be considered competent WNV amplifying hosts, and thus may play a role in the transmission cycle of this zoonotic virus.     

Pathogenicity of West Nile virus for turkeys

In the fall of 1999, West Nile virus (WNV) was isolated during an outbreak of neurologic disease in humans, horses, and wild and zoological birds in New York, Connecticut, and New Jersey. Turkeys could potentially be a large reservoir for WNV because of the high-density turkey farming and the presence of large wild turkey populations in the eastern seaboard of the United States. Little is known about the pathogenicity of WNV in domestic or wild turkeys. Specific-pathogen-free 3-wk-old turkeys were inoculated subcutaneously with 10(3.3) mean tissue culture infective doses of a WNV strain isolated fromthe index case in a New York crow. No clinical signs were observed in the turkeys over the 21 days of the experiment. One turkey died abruptly at 8 days postinoculation (DPI). Many turkeys developed viremia between 2 and 10 DPI, but the average level of virus was very low, less than needed to efficiently infect mosquitos. Low levels of WNV were detected in feces on 4 and 7 DPI, but no virus was isolated from oropharyngeal swabs. WNV wasnot transmitted from WNV-inoculated to contact-exposed turkeys. All WNV-inoculated poults seroconverted on 7 DPI. In the turkey that died, WNV was not isolated from intestine, myocardium, brain, kidney, or cloacal and oropharyngeal swabs, but sparse viral antigen was demonstrated by immunohistochemistry in the heart and spleen. Turkeys in contact with WNV-inoculated turkeys and sham-inoculated controls lacked WNV specific antibodies,and WNV was not isolated from plasma and cloacal and oropharyngeal swabs. These data suggest that WNV lacks the potential to be a major new disease of turkeys and that turkeys will not be a significant amplifying host for infecting mosquitos.     

Plasma concentrations of corticosterone and thyroid hormones in laying fowls from different housing systems

Plasma concentrations of corticosterone, thyroxine (T4) and triiodothyronine (T3) were measured in representative mature female domestic fowls from three housing systems: cages, covered strawyard and range. There were no between-system differences for T4 but T3 was lower in birds from range than in those from cages. Corticosterone concentrations were lower in birds from strawyards than in those from range and cages. In the strawyards there were no differences in corticosterone concentrations between normal and low-ranking hens. The latter were selected on the basis that they remained apart from the flock, were under-weight, poorly feathered and unable to feed at will. Low-ranking hens had significantly higher plasma concentrations of T3 and T4 than normal birds. This was attributed mainly to the poor feather covering of most low-ranking birds. It was concluded that, in the assessment of different housing systems, plasma corticosterone and thyroid hormones were not useful measures of long-term stress or welfare. Different factors relevant to welfare may have contrary effects on the plasma concentrations of these hormones.     

Wild birds as sentinels for multiple zoonotic pathogens along an urban to rural gradient in greater Chicago, Illinois

Wild birds are important in the maintenance and transmission of many zoonotic pathogens. With increasing urbanization and the resulting emergence of zoonotic diseases, it is critical to understand the relationships among birds, vectors, zoonotic pathogens, and the urban landscape. Here, we use wild birds as sentinels across a gradient of urbanization to understand the relative risk of diseases caused by three types of zoonotic pathogens: Salmonella pathogens, mosquito-borne West Nile virus (WNV) and tick-borne pathogens, including the agents of Lyme disease and human anaplasmosis. Wild birds were captured using mist nets at five sites throughout greater Chicago, Illinois, and blood, faecal and ectoparasite samples were collected for diagnostic testing. A total of 289 birds were captured across all sites. A total of 2.8% of birds harboured Ixodes scapularis--the blacklegged tick--of which 54.5% were infected with the agent of Lyme disease, and none were infected with the agent of human anaplasmosis. All infested birds were from a single site that was relatively less urban. A single bird, captured at the only field site in which supplemental bird feeding was practised within the mist netting zone, was infected with Salmonella enterica subspecies enterica. While no birds harboured WNV in their blood, 3.5% of birds were seropositive, and birds from more urban sites had higher exposure to the virus than those from less urban sites. Our results demonstrate the presence of multiple bird-borne zoonotic pathogens across a gradient of urbanization and provide an assessment of potential public health risks to the high-density human populations within the area.     

Pathology and tissue tropism of natural West Nile virus infection in birds: a review

West Nile virus (WNV) is a globally distributed arthropod-borne flavivirus capable of infecting a wide variety of vertebrates, with birds as its natural reservoir. Although it had been considered a pathogen of little importance for birds, from the 1990’s, and especially after its introduction in the North American continent in 1999, thousands of birds have succumbed to West Nile infection. This review summarizes the pathogenesis and pathology of WNV infection in birds highlighting differences in lesion and antigen distribution and severity among bird orders and families. Despite significant species differences in susceptibility to infection, WNV associated lesions and viral antigen are present in the majority of organs of infected birds. The non-progressive, acute or more prolonged course of the disease accounts for part of the differences in lesion and viral antigen distribution and lesion severity. Most likely a combination of host variables and environmental factors in addition to the intrinsic virulence and pathogenicity of the infecting WNV strain influence the pathogenesis of the infection.     

West Nile virus outbreak in captive and wild raptors,Czech Republic, 2018

West Nile virus lineage 2 (WNV‐2) was detected in the brain of 17 goshawks (Accipiter gentilis) that succumbed to neuroinvasive disease in the Czech Republic during 2018: twelve birds were captive and five wild. Furthermore, two wild sparrowhawks (Accipiter nisus) and three other captive birds of prey (golden eagle Aquila chrysaetos, hybrid saker falcon Falco cherrug × F. rusticolus and Harris's hawk Parabuteo unicinctus) also died due to WNV encephalitis. The 2018 outbreak in Czech raptors clearly reflects a new epidemiological situation and indicates an increasing risk of both raptor and human infection with WNV‐2 in the country.     

Experimental West Nile virus infection in Eastern Screech Owls (Megascops asio)

Eastern Screech Owls (EASOs) were experimentally infected with the pathogenic New York 1999 strain of West Nile virus (WNV) by subcutaneous injection or per os. Two of nine subcutaneously inoculated birds died or were euthanatized on 8 or 9 days postinfection (DPI) after <24 hr of lethargy and recumbency. All subcutaneously inoculated birds developed levels of viremia that are likely infectious to mosquitoes, with peak viremia levels ranging from 10(5.0) to 10(9.6) plaque-forming units/ml. Despite the viremia, the remaining seven birds did not display signs of illness. All birds alive beyond 5 DPI seroconverted, although the morbid birds demonstrated significantly lower antibody titers than the clinically normal birds. Cagemates of infected birds did not become infected. One of five orally exposed EASOs became viremic and seroconverted, whereas WNV infection in the remaining four birds was not evident. All infected birds shed virus via the oral and cloacal route. Early during infection, WNV targeted skin, spleen, esophagus, and skeletal muscle. The two morbid owls had myocardial and skeletal muscle necrosis and mild encephalitis and nephritis, whereas some of the clinically healthy birds that were sacrificed on 14 DPI had myocardial arteritis and renal phlebitis. WNV is a significant pathogen of EASOs, causing pathologic lesions with varying clinical outcomes.     

West Nile virus: lessons from the 21st century

Introduction: West Nile virus (WNV) first appeared in the United States in 1999, causing illness and death in birds, horses, and humans. While the initial outbreak of this sometimes deadly viral disease was limited to the northeastern United States, the virus had an inexorable migration across the continental United States over the next 3 years, causing huge losses among the affected species. The purpose of this review is to present currently available information regarding the epi‐demiology, diagnosis, treatment, and prevention of WNV infection. Veterinarians, particularly those in an emergency practice, serve as an important source of reliable information regarding this disease for animal owners and the public in general. Data sources: Data sources used for the preparation of this review include computer‐based searches of PubMed and Commonwealth Agricultural Bureaux (CAB) abstracts. A search in PubMed using ‘West Nile’ retrieved 1468 ‘hits’ or references, while a similar search in CAB abstracts produced 815 references. Additional information was obtained from various meeting proceedings, particularly data presented in abstract form, and from the Centers for Disease Control (CDC) website dedicated to WNV. Human data synthesis: Prior to the mid‐1990s, reported large‐scale epidemics of WNV infection in humans predominantly presented as acute, mild, febrile disease, sometimes associated with lymphadenopathy and skin rash. The recent large epidemic in the United States, in contrast, has prominently featured encephalitis, particularly among the elderly. Additionally, polio‐encephalomyelitis‐like complications resulting in long‐term neurologic sequelae have been reported. There are many WNV‐permissive native avian and mosquito hosts in the Unites States and there appear to be few limitations to the spread of the disease in the United States. It is expected that the virus will be identified in all 48 continental states, Mexico, and Canada by the end of 2003. Veterinary data synthesis: The horse is the animal species most affected by the recent WNV epidemic in the United States, and losses to the equine industry have been large and unprecedented. A United States Department of Agriculture (USDA)‐approved vaccine against WNV has been in use in horses since 2001 and appears to be effective in limiting the incidence of disease in well‐vaccinated populations. WNV infection has been documented in other species of mammals, including camelids (alpaca/llamas) and dogs, and veterinarians should include WNV as a differential diagnosis for animals presenting with clinical signs consistent with central nervous system infection. A large concern exists for endangered bird populations, particularly birds of prey, whether in zoos or in the wild.     

A metapopulation model to simulate West Nile virus circulation in Western Africa, Southern Europe and the Mediterranean basin

In Europe, virological and epidemiological data collected in wild birds and horses suggest that a recurrent circulation of West Nile virus (WNV) could exist in some areas. Whether this circulation is permanent (due to overwintering mechanisms) or not remains unknown. The current conception of WNV epidemiology suggests that it is not: this conception combines an enzootic WNV circulation in tropical Africa with seasonal introductions of the virus in Europe by migratory birds. The objectives of this work were to (i) model this conception of WNV global circulation; and (ii) evaluate whether the model could reproduce data and patterns observed in Europe and Africa in vectors, horses, and birds. The model was calibrated using published seroprevalence data obtained from African (Senegal) and European (Spain) wild birds, and validated using independent, published data: seroprevalence rates in migratory and resident wild birds, minimal infection rates in vectors, as well as seroprevalence and incidence rates in horses. According to this model, overwintering mechanisms are not needed to reproduce the observed data. However, the existence of such mechanisms cannot be ruled out.     

An epidemic of salmonellosis caused by Salmonella Typhimurium DT160 in wild birds and humans in New Zealand

AIM: This study reports an outbreak of salmonellosis due to S. Typhimurium DT160 which caused extensive mortality in wild birds and enteric disease in humans in New Zealand during the winter and spring months of the year 2000. METHODS: Necropsies were performed and microbiological examinations undertaken on wild birds from populations in which mass mortality was reported, and on captive indigenous birds which died suddenly during the winter and spring of 2000. Affected tissues were examined histologically and isolates of S. Typhimurium were phage typed and examined using pulsedfield gel electrophoresis (PFGE). Isolates of S. Typhimurium obtained from cases of human enteric disease which occurred during these months were phage typed, examined using PFGE and compared with the bird isolates. RESULTS: Central and northern areas of the South Island and the southern North Island were worst affected with die-offs of several hundreds of sparrows and other birds reported in rural areas. Mortalities reached a peak in winter (July-August) 2000 and decreased to small numbers during the spring and early summer. The birds usually died of an acute septicaemia with multifocal necrotising lesions in the liver and spleen. Human cases throughout the country increased gradually over the same period. Isolates from birds, livestock and humans examined using PFGE were indistinguishable from one another. CONCLUSION: This strain of Salmonellahas emerged as a major cause of septicaemia in wild birds in New Zealand. Because of the close association between house sparrows (Passer domesticus) and humans, the organism also poses a serious zoonotic risk. The possibility that the infection may spread to involve indigenous species needs investigation.     

Evidence of the role of European wild boar as a reservoir of Mycobacterium tuberculosis complex

Bovine tuberculosis (bTB) is caused by Mycobacterium bovis and closely related mycobacteria of the Mycobacterium tuberculosis complex. They have an extensive host range and may cause zoonotic TB. A major obstacle to bTB eradication in livestock is the implication of wildlife in the natural cycle of the pathogen. The identification of wildlife reservoir hosts is crucial for the implementation of effective control measures. The European wild boar (Sus scrofa) is frequently considered a spillover or dead end host rather than a true reservoir, and scientific evidence is conflicting outside Mediterranean Spain. The aim of this review is to update current scientific evidence of the wild boar as a TB reservoir and to underline those aspects that need further research. Evidences supporting that wild boar is a TB reservoir host include: (i) presence of common M. tuberculosis complex genotypes in wild boar, domestic and wild animals and humans, (ii) high prevalence of M. bovis among wild boar in estates fenced for decades in complete absence of contact with domestic livestock, and other wild ungulates (iii) TB lesions are frequently seen in thoracic lymph nodes and lungs, suggesting that respiratory infection and excretion may occur, and (iv) extensive tuberculous lesions in more than one anatomical region occur in a high proportion of juvenile wild boar that probably represents the main source of mycobacterial excretion. Hence, epidemiological, pathological and microbiological evidence strongly suggests that, at least in Spanish Mediterranean ecosystems, wild boar are able to maintain TB infection in the wild and are most probably able to transmit the disease to other species, acting as a true wildlife reservoir. These results expand the list of wildlife species that act as natural reservoirs of TB in different parts of the world and suggest the need to control the infection in wild boar populations for the complete eradication of the disease in Spain.