Molecular and Antigenic Characterization of Triple‐Reassortant H3N2 Swine Influenza Viruses Isolated from Pigs,Turkey and Quail in Canada

In 2005, triple‐reassortant H3N2 (trH3N2) influenza A viruses were isolated from swine and turkeys in Canada. Subsequently, these viruses were isolated from humans and mink in 2006 and 2007, respectively. Following full genome sequencing, H3N2 viruses isolated from turkeys (2005), quail (2008) and swine (2009) in Canada, were characterized as trH3N2. The 2005 turkey isolate was found to be almost identical to other viruses isolated in that year, with quail and pig isolates related very closely to the 2005 trH3N2. Minimal antigenic evolution of the swine isolates relative to the reference 2005 virus was observed. These results suggest the establishment of a stable lineage of trH3N2 in Canadian pigs, with evidence for interspecies transmission to turkeys and quails.     

The Impact of Pandemic Influenza A H1N1 2009 on Australian Lung Transplant Recipients

Influenza A H1N1 2009 led to 189 deaths during the Australian pandemic. Community‐acquired respiratory viruses not only can cause prolonged allograft dysfunction in lung transplant recipients but have also been linked to bronchiolitis obliterans syndrome (BOS). We report the impact of the 2009 H1N1 pandemic on Australian lung transplant recipients. An observational study of confirmed H1N1 cases was conducted across five Australian lung transplant programs during the pandemic. An electronic database collected patient demographics, clinical presentation, management and outcomes up to a year follow‐up. Twenty‐four H1N1 cases (mean age 43 ± 14 years, eight females) were identified, incidence of 3%. Illness severity varied from upper respiratory tract symptoms only in 29% to lung allograft dysfunction (≥10% decline FEV1) in 75% to death in 5 (21%) cases (pre‐existing BOS grade 3, n = 4). Treatment with oseltamivir occurred in all but one case confirmed after death, reduced immunosuppression, n = 1, augmented corticosteroid therapy, n = 16, and mechanical/noninvasive ventilation, n = 4. There was BOS grade decline within a year in six cases (32%). In conclusion, Australian lung transplant recipients were variably affected by the H1N1 pandemic mirroring the broader community with significant morbidity and mortality. After initial recovery, a considerable proportion of survivors have demonstrated BOS progression.     

De Novo Anti‐HLA Antibody After Pandemic H1N1 and Seasonal Influenza Immunization in Kidney Transplant Recipients

In solid organ transplanted patients, annual influenza immunization is strongly recommended because of morbidity and mortality of influenza infections. In 2009, the rapid spread of a novel H1N1 influenza A virus led to the accelerated development of novel pandemic influenza vaccines. In Switzerland, the recipients received one dose of seasonal influenza and two doses of AS03‐adjuvanted H1N1 vaccines. This situation provided a unique opportunity to analyze the influence of novel adjuvanted influenza vaccines on the production of de novo anti‐HLA antibodies. We prospectively followed two independent cohorts including 92 and 59 kidney‐transplanted patients, assessing their anti‐HLA antibodies before, 6 weeks and 6 months after vaccination. Sixteen of 92 (17.3%) and 7 of 59 (11.9%) patients developed anti‐HLA antibodies. These antibodies, detected using the single antigen beads technology, were mostly at low levels and included both donor‐specific and non‐donor‐specific antibodies. In 2 of the 20 patients who were followed at 6 months, clinical events possibly related to de novo anti‐HLA antibodies were observed. In conclusion, multiple doses of influenza vaccine may lead to the production of anti‐HLA antibodies in a significant proportion of kidney transplant recipients. The long‐term clinical significance of these results remains to be addressed.     

Therapy With m‐TOR Inhibitors Decreases the Response to the Pandemic Influenza A H1N1 Vaccine in Solid Organ Transplant Recipients

Concern has been raised regarding the response to vaccination in solid organ transplant recipients (SOTR) undergoing immunosuppressant regimens and the possibility of rejection related to the immune response associated with pandemic influenza H1N1–2009 vaccination. The goal of this study was to assess the immunogenicity, efficacy and safety of the pandemic vaccine in SOTR. We performed a multicenter prospective study in SOTR receiving the pandemic vaccine. Immunological response was determined in serum 5 weeks after vaccination by microneutralization assays, and immunoglobulins were measured by ELISA. Three hundred and forty‐six SOTR were included. Preexisting seroprotection was detected in 13.6% of cases and rates of seroconversion and seroprotection after vaccination were 73.1% and 82.9%, respectively. Patients with baseline antibody titers had better geometric mean titers (GMT)‐post after pandemic vaccination (339.4 vs. 121.4, p < 0.001). Younger age, liver disease and m‐TOR inhibitor therapy were independently associated with lower seroprotection and GMT‐post. There were no major adverse effects or rejection episodes. Pandemic vaccine was safe in SOTR and elicited an adequate response, although lower than in healthy individuals. This is the first study describing a decreased response after vaccination in patients receiving mTOR inhibitors who presented lower seroprotection rates and lower GMT‐post.     

Factors Associated With H1N1 Influenza Vaccine Receipt in a High-Risk Population During the 2009-2010 H1N1 Influenza Pandemic

Background:

Persons with spinal cord injuries and disorders (SCI/D) are at high risk for respiratory complications from influenza. During pandemic situations, where resources may be scarce, uncertainties may arise in veterans with SCI/D.

Objective:

To describe concerns, knowledge, and perceptions of information received during the 2009-2010 H1N1 influenza pandemic and to examine variables associated with H1N1 vaccine receipt.

Methods:

In August 2010, a cross-sectional survey was mailed to a national sample of veterans with traumatic and nontraumatic SCI/D.

Results:

During the pandemic, 58% of veterans with SCI/D received the H1N1 vaccine. Less than two-thirds of non-H1N1 vaccine recipients indicated intentions to get the next season’s influenza vaccine. Being ≥50 years of age and depressed were significantly associated with higher odds of H1N1 vaccination. Being worried about vaccine side effects was associated with lower odds of H1N1 receipt. Compared to individuals who reported receiving an adequate amount of information about the pandemic, those who received too little information had significantly lower odds of receiving the H1N1 vaccine. Those who received accurate/clear information (vs confusing/conflicting) had 2 times greater odds of H1N1 vaccine receipt.

Conclusions:

H1N1 influenza vaccination was low in veterans with SCI/D. Of H1N1 vaccine nonrecipients, only 63% intend to get a seasonal vaccine next season. Providing an adequate amount of accurate and clear information is vital during uncertain times, as was demonstrated by the positive associations with H1N1 vaccination. Information-sharing efforts are needed, so that carry-over effects from the pandemic do not avert future healthy infection prevention behaviors.     

Evidence Obtained from anova to Reason Cross‐species Infection and Cross‐subtype Mutation in Neuraminidases of Influenza A Viruses

The current pandemic of A/H1N1 influenza raises a serious question on cross‐species infection and cross‐subtype mutation because our previous focus on possible influenza pandemic laid on H5N1 subtype and the cross‐species infection between avian and human. In this study, we analyse 3874 neuraminidases from influenza A viruses using anova to answer the question of if there is barrier between species and between subtypes. The results show that there is no cross‐species barrier in some species, and the intra‐species variation is larger than the inter‐species variation in some species hosting the viruses, and the cross‐subtype mutation is possible because there is no cross‐subtype barrier in some subtypes and the intra‐subtype variation is larger than the inter‐subtype variation in some subtypes. These results highlight the state of barrier of influenza A virus, which can help us understand the current pandemic and manufacture more effective vaccines.     

Co‐circulation and characterization of HPAI‐H5N1 and LPAI‐H9N2 recovered from a duck farm,Yogyakarta, Indonesia

In July 2016, an avian influenza outbreak in duck farms in Yogyakarta province was reported to Disease Investigation Center (DIC), Wates, Indonesia, with approximately 1,000 ducks died or culled. In this study, two avian influenza (AI) virus subtypes, A/duck/Bantul/04161291‐OR/2016 (H5N1) and A/duck/Bantul/04161291‐OP/2016 (H9N2) isolated from ducks in the same farm during an AI outbreak in Bantul district, Yogyakarta province, were sequenced and characterized. Our results showed that H5N1 virus was closely related to the highly pathogenic AI (HPAI) H5N1 of clade 2.3.2.1c, while the H9N2 virus was clustered with LPAI viruses from China, Vietnam and Indonesia H9N2 (CVI lineage). Genetic analysis revealed virulence characteristics for both in avian and in mammalian species. In summary, co‐circulation of HPAI‐H5N1 of clade 2.3.2.1c and LPAI‐H9N2 was identified in a duck farm during an AI outbreak in Yogyakarta province, Indonesia. Our findings raise a concern of the potential risk of the viruses, which could increase viral transmission and/or threat to human health. Routine surveillance of avian influenza viruses should be continuously conducted to understand the dynamic and diversity of the viruses for influenza prevention and control in Indonesia and SEA region.     

Appearance of reassortant European avian‐origin H1 influenza A viruses of swine in Vietnam

Three subtypes—H1N1, H1N2 and H3N2—of influenza A viruses of swine (IAV s‐S) are currently endemic in swine worldwide, but there is considerable genotypic diversity among each subtype and limited geographical distribution. Through IAV s‐S monitoring in Vietnam, two H1N2 influenza A viruses were isolated from healthy pigs in Ba Ria‐Vung Tau Province, Southern Vietnam, on 2 December 2016. BLAST and phylogenetic analyses revealed that their HA and NA genes were derived from those of European avian‐like H1N2 IAV s‐S that contained avian‐origin H1 and human‐like N2 genes, and were particularly closely related to those of IAV s‐S circulating in the Netherlands, Germany or Denmark. In addition, the internal genes of these Vietnamese isolates were derived from human A(H1N1)pdm09 viruses, suggesting that the Vietnamese H1N2 IAV s‐S are reassortants between European H1N2 IAV s‐S and human A(H1N1)pdm09v. The appearance of European avian‐like H1N2 IAV s‐S in Vietnam marks their first transmission outside Europe. Our results and statistical analyses of the number of live pigs imported into Vietnam suggest that the European avian‐like H1N2 IAV s‐S may have been introduced into Vietnam with their hosts through international trade. These findings highlight the importance of quarantining imported pigs to impede the introduction of new IAV s‐S.     

Evidence for Vertical Transmission of Novel Duck‐Origin Goose Parvovirus‐Related Parvovirus

In 2015, novel duck‐origin goose parvovirus‐related parvovirus (N‐GPV) infection progressively appeared in commercial Cherry Valley duck flocks in North China. Diseased ducks were observed to have beak atrophy and dwarfism syndrome (BADS). A previous study showed that a high seropositive rate for N‐GPV indicated a latent infection in most breeder duck flocks. To investigate this possibility in hatching eggs collected from N‐GPV‐infected breeder ducks, 120 eggs were collected at various stages of embryonic development for viral DNA detection and an N‐GPV‐specific antibody test. N‐GPV DNA was present in nine hatching eggs, eleven duck embryo and eight newly hatched ducklings. Of the newly hatched ducklings, 58.33% (21/36) were seropositive. Further, two isolates were obtained from a 12‐day‐old duck embryo and a newly hatched duckling. N‐GPV infection did not reduce the fertilization rate and hatchability. These results indicate possible vertical transmission of N‐GPV and suggest that it may be transmitted from breeder ducks to ducklings in ovo.     

A Meta‐Analysis of the Prevalence of Influenza A H5N1 and H7N9 Infection in Birds

Despite a much higher rate of human influenza A (H7N9) infection compared to influenza A (H5N1), and the assumption that birds are the source of human infection, detection rates of H7N9 in birds are lower than those of H5N1. This raises a question about the role of birds in the spread and transmission of H7N9 to humans. We conducted a meta‐analysis of overall prevalence of H5N1 and H7N9 in different bird populations (domestic poultry, wild birds) and different environments (live bird markets, commercial poultry farms, wild habitats). The electronic database, Scopus, was searched for published papers, and Google was searched for country surveillance reports. A random effect meta‐analysis model was used to produce pooled estimates of the prevalence of H5N1 and H7N9 for various subcategories. A random effects logistic regression model was used to compare prevalence rates between H5N1 and H7N9. Both viruses have low prevalence across all bird populations. Significant differences in prevalence rates were observed in domestic birds, farm settings, for pathogen and antibody testing, and during routine surveillance. Random effects logistic regression analyses show that among domestic birds, the prevalence of H5N1 is 47.48 (95% CI: 17.15–133.13, P < 0.001) times higher than H7N9. In routine surveillance (where surveillance was not conducted in response to human infections or bird outbreaks), the prevalence of H5N1 is still higher than H7N9 with an OR of 43.02 (95% CI: 16.60–111.53, P < 0.001). H7N9 in humans has occurred at a rate approximately four times higher than H5N1, and for both infections, birds are postulated to be the source. Much lower rates of H7N9 in birds compared to H5N1 raise doubts about birds as the sole source of high rates of human H7N9 infection. Other sources of transmission of H7N9 need to be considered and explored.     

H1N1 infection in emergency surgery: A cautionary tale

Pandemic 2009 influenza A H1N1 has spread rapidly since its first report in Mexico in March 2009. This is the first influenza pandemic in over 40 years and it atypically affects previously healthy young adults, with higher rates of morbidity and mortality. The medical literature has been inundated with reports of H1N1 infection, the majority found in critical care and internal medicine journals with a relative paucity in the surgical literature. Despite this, it remains an important entity that can impact greatly on acute surgical emergencies. We present a case of previously healthy 31-year-old male who underwent open appendectomy. His post-operative recovery was complicated by acute respiratory distress syndrome secondary to H1N1 infection. This case report highlights the impact that H1N1 virus can have on acute surgical emergencies and how it can complicate the post-operative course.     

Cover Image

Influenza A (H1N1) viruses are distributed worldwide and pose a threat to public health. Swine, as a natural host and mixing vessel of influenza A (H1N1) virus, play a critical role in the transmission of this virus to humans. Furthermore, swine influenza A (H1N1) viruses have provided all eight genes or some genes to the genomes of influenza strains that historically have caused human pandemics. Hence, persistent surveillance of influenza A (H1N1) virus in swine herds could contribute to the prevention and control of this virus. Here, we report a novel reassortant influenza A (H1N1) virus generated by reassortment between 2009 pandemic H1N1 viruses and swine viruses. We also found that this virus is prevalent in swine herds in Shandong Province, eastern China. Our findings suggest that surveillance of the emergence of the novel reassortant influenza A (H1N1) virus in swine is imperative.     

Infection Dynamics of Pandemic 2009 H1N1 Influenza Virus in a Two‐Site Swine Herd

Influenza A viruses are common causes of respiratory disease in pigs and can be transmitted among multiple host species, including humans. The current lack of published information on infection dynamics of influenza viruses within swine herds hinders the ability to make informed animal health, biosecurity and surveillance programme decisions. The objectives of this serial cross‐sectional study were to describe the infection dynamics of influenza virus in a two‐site swine system by estimating the prevalence of influenza virus in animal subpopulations at the swine breeding herd and describing the temporal pattern of infection in a selected cohort of growing pigs weaned from the breeding herd. Nasal swab and blood samples were collected at approximately 30‐day intervals from the swine breeding herd (Site 1) known to be infected with pandemic 2009 H1N1 influenza virus. Sows, gilts and neonatal pigs were sampled at each sampling event, and samples were tested for influenza virus genome using matrix gene RRT‐PCR. Influenza virus was detected in neonatal pigs, but was not detected in sow or gilt populations via RRT‐PCR. A virus genetically similar to that detected in the neonatal pig population at Site 1 was also detected at the wean‐to‐finish site (Site 2), presumably following transportation of infected weaned pigs. Longitudinal sampling of nasal swabs and oral fluids revealed that influenza virus persisted in the growing pigs at Site 2 for at least 69 days. The occurrence of influenza virus in neonatal pigs, but not breeding females, at Site 1 emphasizes the potential for virus maintenance in this dynamic subpopulation, the importance of including this subpopulation in surveillance programmes and the potential transport of influenza virus between sites via the movement of weaned pigs.     

Swine H1N1 infection in a renal transplant recipient

Influenza pandemics have been observed in several periods throughout history. The first influenza pandemic of the 21st century began in Mexico in 2009 and has spread rapidly all over the world. Swine H1N1 has been officially declared a pandemic by the World Health Organization in June 2009. As has been observed in previous pandemics, pregnant women, adolescents, and immunosuppressed individuals are affected more severely in this pandemic. Despite several reports about the pandemic, there have not been any reports of swine H1N1 infection in individuals who underwent renal transplant. The aim of the current study was to present oseltamivir therapy in a swine H1N1-infected patient who underwent renal transplant 10 months earlier, and was thus under immunosuppressive treatment. To the best of our knowledge, this is the first case report of a swine H1N1 infection in a renal transplant recipient.     

Seroprevalence of highly pathogenic avian influenza (H5N1) virus infection among humans in mainland China: A systematic review and meta‐analysis

Although the effective transmission of the H5N1 virus from humans to humans has yet to be further observed, humans are at increased risk of a pandemic caused by H5N1. In order to fully evaluate the seroprevalence and risk factor of highly pathogenic avian influenza A (H5N1) virus infection among in mainland China, we performed a systematic review and meta‐analysis. In this review, we searched literature on the seroprevalence of H5N1 infection among humans in mainland China from 1 January 1997 to 20 October 2018 in English and Chinese databases, including PubMed, Google scholar, Cochrane library, Clinical Trial, VIP, CNKI and WanFang database. We made a selection according to the title and the abstract of paper, and then, we excluded duplicated literature, and data incomplete literature according to the exclusion criteria we formulated. Finally, we extracted how many humans have H5N1 infection from the obtained studies to establish the seroprevalence of H5N1 infection among humans in mainland China. A total of 56 studies (including data of 35,159 humans) were compliant with our criteria. In China, the overall seroprevalence of H5N1 infection among humans was 2.45% (862/35,159), while the seroprevalence of H5N1 infection among humans from central China was 7.32% (213/2,911), higher than those in other regions of China. The seroprevalence of H5N1 infection was associated with test method, sampling time and demographic characteristics of humans. However, there was no significant difference in the effect of gender on the seroprevalence of H5N1 among humans in China. The purpose of this review was to better understand the real infection rate of H5N1 virus among humans and evaluate the potential risk factors for the zoonotic spread of H5N1 virus to humans. Sufficient epidemiological data are important to explore and understand the prevalent status of AIVs throughout the country and to disease control.     

2009年杭州市甲型H1N1流感监测现状分析

目的探讨2009年杭州市甲型H1N1流感的流行规律、临床表现和病原学特征。方法对2009年杭州市13家流感监测哨点医院的每日门诊流感样病例和甲型H1N1流感确诊病例的流行病学进行分析。采用RT—PCR方法对咽拭子中血凝素（HA）基因进行扩增和测序,并用DNASTAR软件进行序列分析。全文数据采用SPSS12．0软件进行统计分析,率的比较采用χ^2检验。结果流感样病例监测显示,第28周之前,疫情处于平稳状态,之后全市流感样病例逐渐增多。第35周时达到高峰,全市流感样病例就诊比例为7．47％（5442／72859）。之后疫情回落,但到第41周时又再次回升,至第46周出现第二次高峰,全市流感样病例就诊比例达到11．32％（11034／97436）。第38周以前,杭州市人群中主要流行株是季节性H1N1型,其次有少量的乙型和季节性流感H1N1型,到第44周,甲型H1N1流感成为唯一的优势流行株,主要易感人群是11～25岁的青少年,以学生居多。序列分析显示,2009年杭州市甲型H1N1流感病毒株与北美株、中国其他地区的毒株以及疫苗株同源性达到99％,关键位点高度保守,但与季节性流感病毒株同源性只有70％。结论2009年杭州市流感疫情发展迅速,人群中流行的优势毒株为甲型流感H1N1型,易感人群为青少年,目前未发现变异株及高致病株,但仍需进一步监测。     

Evaluation of a Point‐of‐Care Influenza Antigen Test for the Detection of Highly Pathogenic Avian Influenza H5N1 Virus in Cats

Quick diagnosis of H5N1 infection in cats is important because of the zoonotic and pandemic potential of this virus. Human rapid influenza antigen tests are also sold commercially to veterinarians for use in cats. The point‐of‐care test actim^™ Influenza A&B (Medix Biochemica, Kauniainen, Finland) was evaluated for the diagnosis of H5N1 infection in cats. The test showed a very low sensitivity and did not detect virus in samples of experimentally infected cats, so that its application cannot be recommended for the diagnosis of H5N1 infection in cats.     

A Systematic Review of the Comparative Epidemiology of Avian and Human Influenza A H5N1 and H7N9 – Lessons and Unanswered Questions

The aim of this work was to explore the comparative epidemiology of influenza viruses, H5N1 and H7N9, in both bird and human populations. Specifically, the article examines similarities and differences between the two viruses in their genetic characteristics, distribution patterns in human and bird populations and postulated mechanisms of global spread. In summary, H5N1 is pathogenic in birds, while H7N9 is not. Yet both have caused sporadic human cases, without evidence of sustained, human‐to‐human spread. The number of H7N9 human cases in the first year following its emergence far exceeded that of H5N1 over the same time frame. Despite the higher incidence of H7N9, the spatial distribution of H5N1 within a comparable time frame is considerably greater than that of H7N9, both within China and globally. The pattern of spread of H5N1 in humans and birds around the world is consistent with spread through wild bird migration and poultry trade activities. In contrast, human cases of H7N9 and isolations of H7N9 in birds and the environment have largely occurred in a number of contiguous provinces in south‐eastern China. Although rates of contact with birds appear to be similar in H5N1 and H7N9 cases, there is a predominance of incidental contact reported for H7N9 as opposed to close, high‐risk contact for H5N1. Despite the high number of human cases of H7N9 and the assumed transmission being from birds, the corresponding level of H7N9 virus in birds in surveillance studies has been low, particularly in poultry farms. H7N9 viruses are also diversifying at a much greater rate than H5N1 viruses. Analyses of certain H7N9 strains demonstrate similarities with engineered transmissible H5N1 viruses which make it more adaptable to the human respiratory tract. These differences in the human and bird epidemiology of H5N1 and H7N9 raise unanswered questions as to how H7N9 has spread, which should be investigated further.     

A molecular epidemiological investigation of PEDV in China: Characterization of co‐infection and genetic diversity of S1‐based genes

Porcine epidemic diarrhoea virus (PEDV) is an emerging and re‐emerging epizootic virus of swine that causes substantial economic losses to the pig industry in China and other countries. The variations in the virus, and its co‐infections with other enteric viruses, have contributed to the poor control of PEDV infection. In the current study, a broad epidemiological investigation of PEDV was carried out in 22 provinces or municipalities of China during 2015–2018. The enteric viruses causing co‐infection with PEDV and the genetic diversity of the PEDV S1 gene were also analysed. The results indicated that, of the 543 diarrhoea samples, 66.85% (363/543) were positive for PEDV, and co‐infection rates of PEDV with 13 enteric viruses ranged from 3.58% (13/363) to 81.55% (296/363). Among these enteric viruses, the signs of diarrhoea induced by PEDV were potentially associated with co‐infections with porcine enterovirus 9/10 (PEV) and torque teno sus virus 2 (TTSuV‐2) (p < .05). The 147 PEDV strains identified in our study belong to Chinese pandemic strains and exhibited genetic diversity. The virulence‐determining S1 proteins of PEDV pandemic strains were undergoing amino acid mutations, in which S58_S58insQGVN–N135dup–D158_I159del‐like mutations were common patterns (97.28%, 143/147). When compared with 2011–2014 PEDV strains, the amino acid mutations of PEDV pandemic strains were mainly located in the N‐terminal domain of S1 (S1‐NTD), and 21 novel mutations occurred in 2017 and 2018. Furthermore, protein homology modelling showed that the mutations in pattern of insertion and deletion mutations of the S1 protein of PEDV pandemic strains may have caused structural changes on the surface of the S1 protein. These data provide a better understanding of the co‐infection and genetic evolution of PEDV in China.     

Experimental infection of clade 1.1.2 (H5N1), clade 2.3.2.1c (H5N1) and clade 2.3.4.4 (H5N6) highly pathogenic avian influenza viruses in dogs

Since the emergence of highly pathogenic avian influenza (HPAI) H5N1 in Asia, the haemagglutinin (HA) gene of this virus lineage has continued to evolve in avian populations, and H5N1 lineage viruses now circulate concurrently worldwide. Dogs may act as an intermediate host, increasing the potential for zoonotic transmission of influenza viruses. Virus transmission and pathologic changes in HPAI clade 1.1.2 (H5N1)‐, 2.3.2.1c (H5N1)‐ and 2.3.4.4 (H5N6)‐infected dogs were investigated. Mild respiratory signs and antibody response were shown in dogs intranasally infected with the viruses. Lung histopathology showed lesions that were associated with moderate interstitial pneumonia in the infected dogs. In this study, HPAI H5N6 virus replication in dogs was demonstrated for the first time. Dogs have been suspected as a “mixing vessel” for reassortments between avian and human influenza viruses to occur. The replication of these three subtypes of the H5 lineage of HPAI viruses in dogs suggests that dogs could serve as intermediate hosts for avian–human influenza virus reassortment if they are also co‐infected with human influenza viruses.