Influenza Virus in a Natural Host, the Mallard: Experimental Infection Data

Wild waterfowl, particularly dabbling ducks such as mallards (Anas platyrhynchos), are considered the main reservoir of low-pathogenic avian influenza viruses (LPAIVs). They carry viruses that may evolve and become highly pathogenic for poultry or zoonotic. Understanding the ecology of LPAIVs in these natural hosts is therefore essential. We assessed the clinical response, viral shedding and antibody production of juvenile mallards after intra-esophageal inoculation of two LPAIV subtypes previously isolated from wild congeners. Six ducks, equipped with data loggers that continually monitored body temperature, heart rate and activity, were successively inoculated with an H7N7 LPAI isolate (day 0), the same H7N7 isolate again (day 21) and an H5N2 LPAI isolate (day 35). After the first H7N7 inoculation, the ducks remained alert with no modification of heart rate or activity. However, body temperature transiently increased in four individuals, suggesting that LPAIV strains may have minor clinical effects on their natural hosts. The excretion patterns observed after both re-inoculations differed strongly from those observed after the primary H7N7 inoculation, suggesting that not only homosubtypic but also heterosubtypic immunity exist. Our study suggests that LPAI infection has minor clinically measurable effects on mallards and that mallard ducks are able to mount immunological responses protective against heterologous infections. Because the transmission dynamics of LPAIVs in wild populations is greatly influenced by individual susceptibility and herd immunity, these findings are of high importance. Our study also shows the relevance of using telemetry to monitor disease in animals

Тепловой расходомер

Полезная модель относится к области измерительной техники, а именно к тепловым устройствам для измерения расхода жидких или газообразных сред, и может быть использована в химической, электронной, энергетической и других отраслях промышленности

Chewing lice Trichodectes pinguis pinguis in Scandinavian brown bears ( Ursus arctos )

In April 2014 and 2015, we noted localized alopecia (neck, forelimbs, and chest) and hyperpigmentation on two adult brown bears (Ursus arctos) captured in central-south Sweden for ecological studies under the Scandinavian Brown Bear Research Project. In spring 2015, a brown bear was shot because of human- wildlife conflict in the same region. This bear also had extensive alopecia and hyperpigmentation. Ec- toparasites were collected from the affected skin areas in all three individuals and preserved in ethanol for identification. Based on morphological characteristics, the lice were identified as Trichodectes spp. and Trichodectes pinguis pinguis. To our knowledge, these are the first reported cases of chewing lice in free- ranging brown bears in Scandinavia

Tissue distribution of angiotensin-converting enzyme 2 (ACE2) receptor in wild animals with a focus on artiodactyls, mustelids and phocids

Natural cases of zooanthroponotic transmission of SARS-CoV-2 to animals have been reported during the COVID-19 pandemic, including to free-ranging white-tailed deer (Odocoileus virginianus) in North America and farmed American mink (Neovison vison) on multiple continents. To understand the potential for angiotensin-converting enzyme 2 (ACE2)-mediated viral tropism we characterised the distribution of ACE2 receptors in the respiratory and intestinal tissues of a selection of wild and semi-domesticated mammals including artiodactyls (cervids, bovids, camelids, suids and hippopotamus), mustelid and phocid species using immunohistochemistry. Expression of the ACE2 receptor was detected in the bronchial or bronchiolar epithelium of several European and Asiatic deer species, Bactrian camel (Camelus bactrianus), European badger (Meles meles), stoat (Mustela erminea), hippopotamus (Hippopotamus amphibious), harbor seal (Phoca vitulina), and hooded seal (Cystophora cristata). Further receptor mapping in the nasal turbinates and trachea revealed sparse ACE2 receptor expression in the mucosal epithelial cells and occasional occurrence in the submucosal glandular epithelium of Western roe deer (Capreolus capreolus), moose (Alces alces alces), and alpaca (Vicunga pacos). Only the European badger and stoat expressed high levels of ACE2 receptor in the nasal mucosal epithelium, which could suggest high susceptibility to ACE2-mediated respiratory infection. Expression of ACE2 receptor in the intestinal cells was ubiquitous across multiple taxa examined. Our results demonstrate the potential for ACE2-mediated viral infection in a selection of wild mammals and highlight the intra-taxon variability of ACE2 receptor expression, which might influence host susceptibility and infection

Environmental Levels of the Antiviral Oseltamivir Induce Development of Resistance Mutation H274Y in Influenza A/H1N1 Virus in Mallards

Oseltamivir (Tamiflu®) is the most widely used drug against influenza infections and is extensively stockpiled worldwide as part of pandemic preparedness plans. However, resistance is a growing problem and in 2008–2009, seasonal human influenza A/H1N1 virus strains in most parts of the world carried the mutation H274Y in the neuraminidase gene which causes resistance to the drug. The active metabolite of oseltamivir, oseltamivir carboxylate (OC), is poorly degraded in sewage treatment plants and surface water and has been detected in aquatic environments where the natural influenza reservoir, dabbling ducks, can be exposed to the substance. To assess if resistance can develop under these circumstances, we infected mallards with influenza A/H1N1 virus and exposed the birds to 80 ng/L, 1 µg/L and 80 µg/L of OC through their sole water source. By sequencing the neuraminidase gene from fecal samples, we found that H274Y occurred at 1 µg/L of OC and rapidly dominated the viral population at 80 µg/L. IC50 for OC was increased from 2–4 nM in wild-type viruses to 400–700 nM in H274Y mutants as measured by a neuraminidase inhibition assay. This is consistent with the decrease in sensitivity to OC that has been noted among human clinical isolates carrying H274Y. Environmental OC levels have been measured to 58–293 ng/L during seasonal outbreaks and are expected to reach µg/L-levels during pandemics. Thus, resistance could be induced in influenza viruses circulating among wild ducks. As influenza viruses can cross species barriers, oseltamivir resistance could spread to human-adapted strains with pandemic potential disabling oseltamivir, a cornerstone in pandemic preparedness planning. We propose surveillance in wild birds as a measure to understand the resistance situation in nature and to monitor it over time. Strategies to lower environmental levels of OC include improved sewage treatment and, more importantly, a prudent use of antivirals

Pathobiology of avian influenza in wild bird species

Avian influenza viruses, especially highly pathogenic avian influenza viruses (HPAIV), affect a wide range of species, including humans and have thus become a major concern for veterinary medicine and public health. A HPAIV-H5N1 belonging to clade 2.2, originally from South East Asia, spread across Eurasia and reached Sweden in 2006. Currently the most commonly isolated HPAIV-H5N1 from wild birds belong to clade 2.3.2. There is a growing concern that the H5N1 virus has evolved in such a way that it can be maintained in the wild bird population without causing severe disease. At the same time the role of natural hosts, such as mallards (Anas platyrhynchos), in the epidemiology of avian influenza is an ongoing concern. In order to characterize the natural disease in free ranging birds in Sweden and to assess the pathogenicity of clade 2.3.2 viruses, histopathology, polymerase chain reaction, virus isolation and immunohistochemistry (IHC) were used to investigate lesions and viral tissue targeting of HPAIV-H5N1 in naturally infected tufted ducks (Aythya fuligula) and in tufted ducks experimentally infected with a clade 2.3.2 virus. Since neurotropism is a key feature of HPAIV-H5N1 infection, the encephalitis in 9 wild bird species from the Swedish outbreak was characterized in more detail. Results were compared to mallards infected with a low pathogenic avian influenza virus H1N1. The studies highlight the range and variation of the presentation of the natural disease in wild birds. Experimentally infected ducks were highly susceptible to the current HPAIV-H5N1 clade and showed similar lesions and viral antigen distribution as the naturally infected ducks. The studies suggest that there are several routes of infection and dissemination of the virus including, respiratory, hematogenous and olfactory routes. The respiratory tract is probably the main route of excretion of HPAIV-H5N1 since no viral antigen was found in the intestine. This was in contrast to the experimentally infected mallards which had primarily intestinal replication with minimal lesions. The results highlight the importance of continued investigation of the pathobiology of both low- and HPAIV infections in wild birds which is essential in the understanding of their epidemiology and, in turn, can contribute to the design and implementation of preventive and control measures to protect the health of humans and animals

Alternate routes of influenza A virus infection in Mallard (Anas platyrhynchos)

Abstract The natural reservoir for all influenza A viruses (IAVs) is wild birds, particularly dabbling ducks. During the autumn, viral prevalence can be very high in dabbling ducks (> 30%) in the Northern Hemisphere, and individuals may be repeatedly infected. Transmission and infection is through the fecal–oral route, whereby birds shed viruses in feces and conspecifics are infected though feeding in virus-contaminated water. In this study we wanted to assess two alternative infection routes: cloacal drinking and preening. Using experimental infections, we assessed patterns of infection using a combination of virus shedding, as assessed by real-time PCR from cloacal swabs, and patterns of viral replication using virus-immunohistochemistry of gastrointestinal tissues. The cloacal drinking experiment consisted of two trials using cloacal inoculation at two different time points to account for age differences, as well as a trial whereby ducks were allowed to take up virus-laden water through the cloaca. All ducks became infected, and rather than the bursa of Fabricius being the main site of replication, the colon had the highest intensity of replication, as inferred through immunohistochemistry. In experiments assessing preening, feathers were contaminated with virus-laden water and all ducks became infected, regardless of whether they were kept individually or together. Further, naive contacts were infected by the individuals whose feathers were virus-contaminated. Overall, we reinforce that IAV transmission in dabbling ducks is multifactorial—if exposed to virus-contaminated water ducks may be infected through dabbling, preening of infected feathers, and cloacal drinking

Mortalities due to constipation and dystocia caused by intraperitoneal radio- transmitters in Eurasian lynx (Lynx lynx)

Three lynx (Lynx lynx) were found dead following complications after a surgically implanted free floating intraperitoneal radio-transmitter became lodged within the pelvic canal. Two yearling lynx died due to consequences following severe constipation as the transmitter compressed the colon. Both were emaciated, with no abdominal or intrapelvic fat, which allowed the transmitter implant to fit into the pelvic canal. An adult female lynx died of dystocia when the pelvic birth canal was blocked by the transmitter when parturition began, leading to uterine rupture and subsequent peritonitis. A total of 41 lynx was implanted with this type of intraperitoneal transmitter in Scandinavia 1997-2002. After the three transmitter-associated mortalities, the transmitter type used in lynx cubs was exchanged for another model, and further fatalities due to the implants have not been documented

Influenza A(H7N9) Virus Acquires Resistance-Related Neuraminidase I222T Substitution When Infected Mallards Are Exposed to Low Levels of Oseltamivir in Water

Influenza A virus (IAV) has its natural reservoir in wild waterfowl, and new human IAVs often contain gene segments originating from avian IAVs. Treatment options for severe human influenza are principally restricted to neuraminidase inhibitors (NAIs), among which oseltamivir is stockpiled in preparedness for influenza pandemics. There is evolutionary pressure in the environment for resistance development to oseltamivir in avian IAVs, as the active metabolite oseltamivir carboxylate (OC) passes largely undegraded through sewage treatment to river water where waterfowl reside. In an in vivo mallard (Anas platyrhynchos) model, we tested if low-pathogenic avian influenza A(H7N9) virus might become resistant if the host was exposed to low levels of OC. Ducks were experimentally infected, and OC was added to their water, after which infection and transmission were maintained by successive introductions of uninfected birds. Daily fecal samples were tested for IAV excretion, genotype, and phenotype. Following mallard exposure to 2.5 μg/liter OC, the resistance-related neuraminidase (NA) I222T substitution, was detected within 2 days during the first passage and was found in all viruses sequenced from subsequently introduced ducks. The substitution generated 8-fold and 2.4-fold increases in the 50% inhibitory concentration (IC50) for OC (P < 0.001) and zanamivir (P = 0.016), respectively. We conclude that OC exposure of IAV hosts, in the same concentration magnitude as found in the environment, may result in amino acid substitutions, leading to changed antiviral sensitivity in an IAV subtype that can be highly pathogenic to humans. Prudent use of oseltamivir and resistance surveillance of IAVs in wild birds are warranted

Serological survey of Toxoplasma gondii infection in free-ranging Eurasian lynx (Lynx lynx) from Sweden

To investigate the prevalence of Toxoplasma gondii infection in free-ranging Eurasian lynx (Lynx lynx) in Sweden, serosanguinous fluids and feces were collected from 207 carcasses of lynx killed or found dead from 1996 to 1998. Sera were tested for antibodies against T. gondii by the direct agglutination test, and 156 (75.4%) of the sera tested positive at antibody titers>or=40. Antibody prevalence was significantly lower in lynx originating from the northern parts of Sweden than in lynx from the more southern regions that are more densely populated by humans. Age-related differences also were found, with a significantly lower prevalence (55%) in juvenile (<1-yr-old) than in subadult and adult animals (82%). There was no significant difference in seroprevalence between males and females. Oocysts typical of T. gondii were not detected in any of the fecal samples