Printer Friendly

Serologic evidence of influenza a (H14) virus introduction into North America.

To the Editor: Although a diverse population of influenza A viruses (IAVs) is maintained among ducks, geese, shorebirds, and gulls, not all of the 16 avian hemagglutinin (HA) subtypes are equally represented (1). The 14th HA subtype, commonly known as the H14 subtype, was historically limited to isolates from the former Soviet Union in the 1980s (2) and was not subsequently detected until 2010, when isolated in Wisconsin, USA from long-tailed ducks and a white-winged scoter (3-5). In the United States, the H14 subtype has since been isolated in California (6), Mississippi, and Texas (7); and has been reported in waterfowl in Guatemala (7). In this study, we examined whether there was serologic evidence of H14 spread among ducks in North America before (2006-2010) and after (2011-2014) the initial detection of the H14 subtype virus on this continent.

This report was reviewed and approved by United States Geological Survey under the Fundamental Science Practices policy ( Serum samples from blue-winged teal, American green-winged teal, and mallard ducks were screened by using blocking ELISA (FlockCheck AI MultiS-Screen antibody test kit; IDEXX Laboratories, Westbrook, ME, USA) to detect antibodies against the influenza virus nucleoprotein. Positive samples were tested by microneutralization assays as described (7) against viruses representing H14 and H3 subtypes. H3 is commonly detected in ducks found in North America (8) (online Technical Appendix Table 1, 21/12/15-0413-Techapp1.pdf).

Antibodies against H3 were detected during 20062014 in Michigan, Minnesota, New Jersey, Texas, and Louisiana (Table); titers ranged from 20 to 320. Antibodies against H14 were detected in 1 duck in 2007 and in 24 ducks sampled in 2012 after August. H14 antibodies were detected in all years and most locations studied after 2012; antibody titers ranged from 20 to 160. Thus, antibody prevalence was consistent with the relative prevalence of H3 reported among ducks in North America (1,4,8) and the timing of initial detection of H14 viruses.

To address the possibility of cross-neutralizations between HA subtypes, we tested the 2007 H14-positive serum samples and 22 of the H14-positive serum samples from 2012-2014 against HA subtypes 1-12 (online Technical Appendix Table 1) by virus neutralization (online Technical Appendix Table 2). Among humans, broadly neutralizing antibodies within HA groups targeting conserved regions in the HA stalk have been described (9), and if present in samples from mallards, these could contribute to cross-neutralizations. The H14-positive serum samples from 2007 reacted to subtypes H3, H4, H7, and H11, and high titers were identified for H3 and H4, which are within the same clade. Samples from 17 of these birds tested antibody-positive for additional HA subtypes and 5 tested positive only to H14. An H14 virus was recovered by virus isolation from the same blue-winged teal population sampled in March 2013, from which serum samples were obtained (7); however, although H14 antibodies have been detected in Minnesota, an H14 virus has not yet been isolated in that state.

Our serologic results are temporally consistent with H14 isolation reports and suggest that H14 subtype viruses were not circulating among ducks in North America before initial virus isolation. However, there are potential challenges with serologic-based investigations. For example, the overall prevalence of H14 antibodies after the initial detection of H14 viruses (2011-2014) was low (3.5% of blocking ELISA positive samples), thus requiring a large sample size (n = 670) for H14 antibody detection. However, an even lower prevalence was observed by using virus isolation; we isolated only 1 H14 IAV during parallel sampling of these sites (n = 8,875) during 2011-2014.

Differences in pre- and post-H14 detection also varied between species, location, and season. Differences in H14 antibody prevalence were observed in all ducks sampled pre- and post- (0.3%-3.5%, p = 0.0103) H14 detection, but not in the mallard-only subset (0.3%-2.1%, p = 0.0963). A significant difference in seroprevalence also was detected between species (mallard [2%] vs teal [6%]) in the 2011-2014 samples (p = 0.0104). IAV show strong seasonal patterns in prevalence, and the observed differences in antibodies may be associated with the probability of IAV infection before sampling and the persistence of antibody responses in these species. Mallards (primarily hatch-year birds) were sampled at the beginning of fall migration ([approximately equal to] 3-4 months of potential IAV exposure for hatch-year birds), whereas teal were sampled later, during spring migration ([approximately equal to] 9-10 months of potential IAV exposure for birds hatched the previous spring or summer). It is apparent that the sampling approach used can affect results.

Interpretation of subtype-specific serologic data can be complex, especially in birds that are normally infected with several IAV subtypes during their lives. Nevertheless, this study demonstrates the value of a subtype-specific serologic approach to detect even relatively minor changes in subtype diversity and clearly shows that new viruses can establish in duck populations in North America. Serologic techniques also can be optimized to detect incursions of novel viruses such as the highly pathogenic Eurasian H5 viruses (10) among wild birds.



We thank B. Wilcox, D. Carter, J. Slusher, N. Davis-Fields, C. Kienzle, G. Knutsen, R. Poulson, P. Link, P. Walther, and G. Newsome for assistance acquiring the serum samples used in this study. We also thank M. Wille and 2 anonymous persons for providing reviews. Positive control antisera were provided by the National Veterinary Services Laboratories, Veterinary Services, United States Department of Agriculture.

The trapping, handling, and sampling of birds was approved through the University of Georgia Institutional Animal Care and Use Committee Animal Use Proposal A2013 05-021.

The work was funded through the National Institute of Allergy and Infectious Diseases, Centers of Excellence for Influenza Research and Surveillance (contract number:

HHSN272201400006C) and by the United States Geological Survey through the Wildlife Program of the Ecosystem Mission Area. N.L.-M. was supported by postdoc grants from the Wenner-Gren Foundations, Stockholm, Sweden and the Swedish Research Council.


(1.) Olsen B, Munster VJ, Wallensten A, Waldenstrom J, Osterhaus ADME, Fouchier RAM. Global patterns of influenza A virus in wild birds. Science. 2006; 312:384-8 10.1126/science.1122438.

(2.) Kawaoka Y, Yamnikova S, Chambers TM, Lvov DK, Webster RG. Molecular characterization of a new hemagglutinin, subtype-H14, of influenza-A virus. Virology. 1990; 179:759-67 10.1016/0042-6822(90)90143-F.

(3.) Fries AC, Nolting JM, Bowman AS, Killian ML, Wentworth DE, Slemons RD. Genomic analyses detect Eurasian-lineage H10 and additional H14 influenza A viruses recovered from waterfowl in the Central United States. Influenza Other Respir Viruses. 2014; 8:493-8

(4.) Fries AC, Nolting JM, Danner A, Webster RG, Bowman AS, Krauss S, et al. Evidence for the circulation and inter-hemispheric movement of the H14 subtype influenza A virus. PLoS ONE. 2013; 8 [cited 2015 Jun 22]. journal.pone.0059216

(5.) Nolting JFA, Slemons RD, Courtney C, Hines N, Pedersen J. Recovery of H14 influenza A virus isolates from sea ducks in the Western Hemisphere. PLOS Currents Influenza 2012 (Edition 1).

(6.) Boyce WM, Schobel S, Dugan VG, Halpin R, Lin X, Wentworth DE, et al. Complete genome sequence of a reassortant H14N2 avian influenza virus from California. Genome Announc. 2013; 1:e00543-13. genomeA.00543-13

(7.) Ramey AM, Poulson RL, Gonzalez-Reiche AS, Perez DR, Stallknecht DE, Brown JD. Genomic characterization of H14 subtype influenza A viruses in new world waterfowl and experimental infectivity in mallards (Anas platyrhynchos). PLoS ONE. 2014; 9 [cited 2015 Jun 22]. journal.pone.0095620

(8.) Wilcox BR, Knutsen GA, Berdeen J, Goekjian V, Poulson R, Goyal S, et al. Influenza-A viruses in ducks in northwestern Minnesota: fine scale spatial and temporal variation in prevalence and subtype diversity. PLoS ONE. 2011; 6:e24010 10.1371/journal.pone.0024010.

(9.) Ekiert DC, Friesen RHE, Bhabha G, Kwaks T, Jongeneelen M, Yu WL, et al. A highly conserved neutralizing epitope on group 2 influenza A viruses. Science. 2011; 333:843-50

(10.) Gilbert M, Koel BF, Bestebroer TM, Lewis NS, Smith DJ, Fouchier RAM. Serological evidence for non-lethal exposures of Mongolian wild birds to highly pathogenic avian influenza H5N1 virus. PLoS ONE. 2014; 9 [cited 2015 Jun 22].

Neus Latorre-Margalef, Andrew M. Ramey, Alinde Fojtik, David E. Stallknecht

University of Georgia, Athens, Georgia, USA (N. Latorre-Margalef, A.M. Ramey, A. Fojtik, E. Stallknecht); Lund University, Lund, Sweden (N. Latorre-Margalef); US Geological Survey Alaska Science Center, Anchorage, Alaska, USA (A.M. Ramey)

Address for correspondence: Neus Latorre-Margalef, College of Veterinary Medicine, The University of Georgia, 589 D. W. Brooks Dr, Athens, GA 30602, USA; email:
Table. H3 and H14 microneutralization assay data from ducks sampled
during 2006-2014, North America *

Year          Month of        State                 Species

2006            Aug         Michigan                Mallard
              Aug/Sep       Minnesota               Mallard
2007          Aug/Sep       Minnesota               Mallard
2008          Aug/Sep       Minnesota               Mallard
2009          Aug/Sep       Minnesota               Mallard
                Aug        New Jersey      Domestic and wild mallard
2010          Aug/Sep       Minnesota               Mallard
                Aug        New Jersey      Domestic and wild mallard
2011          Aug/Sep       Minnesota               Mallard
2012          Feb/Mar         Texas             Blue-winged teal
              Aug/Sep       Minnesota               Mallard
2013          Feb/Mar    Texas/Louisiana        Blue-winged teal
              Feb/Mar    Texas/Louisiana   American green-winged teal
              Aug/Sep       Minnesota               Mallard
2014          Feb/Mar         Texas             Blue-winged teal
                Sep         Minnesota               Mallard
  2006-2010      NA            NA                  All ducks
                 NA            NA                Mallards only
  2011-2014      NA            NA                  All ducks
                 NA            NA                Mallards only
                 NA            NA             Blue-winged teal and
                                           American green-winged teal

Year          No.    H3N8,     H14N5,
                    no. (%)   no. (%)

2006          29    6 (21)       0
              39     3 (8)       0
2007          46    8 (17)     1 (2)
2008          44    8 (18)       0
2009          29    10 (34)      0
              36     1 (3)       0
2010          29    6 (21)       0
              20    5 (25)       0
2011          124   37(30)       0
2012          19    3 (16)       0
              188   11 (6)     2 (1)
2013          120   13 (11)   12 (10)
              91     5 (5)     2 (2)
              65    8 (12)     7 (11)
2014          22     1 (5)     1 (5)
              41    4 (10)       0
  2006-2010   272   47 (17)   1 (0.3)
              272   47 (17)   1 (0.3)
  2011-2014   670   82 (12)   24 (3.5)
              418   60 (14)    9(2.1)
              252   22 (9)     15 (6)

* NA, not applicable.
COPYRIGHT 2015 U.S. National Center for Infectious Diseases
No portion of this article can be reproduced without the express written permission from the copyright holder.
Copyright 2015 Gale, Cengage Learning. All rights reserved.

Article Details
Printer friendly Cite/link Email Feedback
Title Annotation:LETTERS
Author:Latorre-Margalef, Neus; Ramey, Andrew M.; Fojtik, Alinde; Stallknecht, David E.
Publication:Emerging Infectious Diseases
Article Type:Letter to the editor
Date:Dec 1, 2015
Previous Article:NADC30-like strain of porcine reproductive and respiratory syndrome virus, China.
Next Article:Disseminated infection caused by Francisella philomiragia, France, 2014.

Terms of use | Privacy policy | Copyright © 2019 Farlex, Inc. | Feedback | For webmasters