Citation: Ortiz, L.; Geiger, G.; Ferreri, L.; Moran, D.; Mendez, D.; Gonzalez-Reiche, A.S.; Alvarez, D.; Motta, M.; Escobar, F.; Rajao, D.; et al. Blue-Winged Teals in Guatemala and Their Potential Role in the Ecology of H14 Subtype Influenza a Viruses. Viruses 2023, 15, 483. https:// doi.org/10.3390/v15020483 Academic Editor: Jianjun Chen Received: 20 January 2023 Revised: 3 February 2023 Accepted: 6 February 2023 Published: 9 February 2023 Copyright: © 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/). viruses Article Blue-Winged Teals in Guatemala and Their Potential Role in the Ecology of H14 Subtype Influenza a Viruses Lucia Ortiz 1,2 , Ginger Geiger 1 , Lucas Ferreri 3 , David Moran 2 , Dione Mendez 2 , Ana Silvia Gonzalez-Reiche 4 , Danilo Alvarez 2 , Mayra Motta 5 , Francisco Escobar 5 , Daniela Rajao 1 , Celia Cordon-Rosales 2 , Martha I. Nelson 6 and Daniel R. Perez 1, * 1 Poultry Diagnostic and Research Center, Department of Population Health, College of Veterinary Medicine, University of Georgia, Athens, GA 30602, USA 2 Centro de Estudios en Salud, Universidad del Valle de Guatemala, Guatemala City 01015, Guatemala 3 Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, GA 30322, USA 4 Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA 5 Laboratorio de Referencia Regional de Salud Animal, Facultad de Medicina Veterinaria y Zootecnia, Universidad del San Carlos de Guatemala, Guatemala City 01012, Guatemala 6 Computational Biology Branch, National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, Bethesda, MD 20894, USA * Correspondence: dperez1@uga.edu; Tel.: +1-706-5838-919 Abstract: Wild aquatic birds are considered the natural hosts of 16 HA (H1–H16) and 9 NA (N1–N9) subtypes of influenza A viruses (FLUAV) found in different combinations. H14 FLUAVs are rarely detected in nature. Since 2011, H14 FLUAVs have been consistently detected in Guatemala, leading to the largest collection of this subtype from a single country. All H14 FLUAVs in Guatemala were detected from blue-winged teal samples. In this report, 17 new full-length H14 FLUAV genome sequences detected from 2014 until 2019 were analyzed and compared to all published H14 sequences, including Guatemala, North America, and Eurasia. The H14 FLUAVs identified in Guatemala were mostly associated with the N3 subtype (n = 25), whereas the rest were paired with either N4 (n = 7), N5 (n = 4), N6 (n = 1), and two mixed infections (N3/N5 n = 2, and N2/N3 n = 1). H14 FLUAVs in Guatemala belong to a distinct H14 lineage in the Americas that is evolving independently from the Eurasian H14 lineage. Of note, the ORF of the H14 HA segments showed three distinct motifs at the cleavage site, two of these containing arginine instead of lysine in the first and fourth positions, not previously described in other countries. The effects of these mutations on virus replication, virulence, and/or transmission remain unknown and warrant further studies. Keywords: avian influenza; blue-winged teals; H14 subtype; Guatemala; surveillance 1. Introduction Influenza A viruses (FLUAV) infect a wide range of bird species and mammals, in- cluding humans [1]. The virus genome contains 8 segments of negative single-stranded RNA encoding 6 internal (PB2, PB1, PA, NP, M, and NS) and 2 surface (HA and NA) gene segments. Subtype classification is based on the antigenic properties of the surface glycopro- teins. To date, 18 HA (H1–H18) and 11 NA (N1–N11) subtypes have been described [2–4]. Wild aquatic birds in the orders Anseriformes and Charadriiformes are considered the natural hosts for 16 HA (H1–H16) and 9 NA (N1–N9) subtypes, playing an important role in the perpetuation of FLUAVs in nature. H3, H4, and H6 subtypes are usually detected at high frequencies in the breeding grounds of North America, while others such as H8, H12, H14, and H15 are rarely found [5,6]. The H14 subtype was initially detected in 1982 in 4 virus isolates obtained from 3 mallard ducks and 1 herring gull in the former Soviet Union (Russia and Kazakhstan) [7]. Viruses 2023, 15, 483. https://doi.org/10.3390/v15020483 https://www.mdpi.com/journal/viruses