key: cord-0773588-w3918erb
authors: Rivero, Ricardo; Garay, Evelin; Botero, Yesica; Serrano-Coll, Héctor; Gastelbondo, Bertha; Muñoz, Marina; Ballesteros, Nathalia; Castañeda, Sergio; Patiño, Luz Helena; Ramirez, Juan David; Calderon, Alfonso; Guzmán, Camilo; Martinez-Bravo, Caty; Aleman, Ader; Arrieta, Germán; Mattar, Salim
title: Human-to-dog transmission of SARS-CoV-2, Colombia
date: 2022-05-12
journal: Sci Rep
DOI: 10.1038/s41598-022-11847-9
sha: 4081364ed42fe3cae660ceb8527165fba2c460a9
doc_id: 773588
cord_uid: w3918erb

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the causative agent of the current COVID-19 pandemic, has evolved to have a wide range of hosts, including non-human primates, wild and domestic animals. The ACE2 protein has a high level of conservation and is the common receptor invertebrate species for a viral infection to occur; this receptor could give rise to anthroponotic events. This article describes the first event of symptomatic transmission in Latin America from a human to a dog by the B.1.625 lineage of SARS-CoV-2. We found 21 shared mutations in the complete genomes of viral sequences from owners and dogs. Further phylogenetic and molecular analysis showed that 100% co-localization of the clade helps to understand human-animal transmission. Prediction of the Spike protein structure of the sequenced virus and docking analyzes showed that the E484K mutation in the receptor-binding domain (RBD) could contribute to the viral affinity of dACE2. Therefore, close contact between SARS-CoV-2-infected humans and pets should be avoided to prevent the emergence of novel mutations of public health importance from anthroponotic events.

Receptor Binding Domain and S2 of the B.1625 S protein (Fig. 3 ). Docking analyses for B.1.625 RBD-dACE2 and wtRBD-dAC2 showed a standard free binding energy (ΔG°) of − 295.58 and − 264.11 kJ mol −1 , respectively, which suggests a higher affinity of B.1.625's RBD towards dACE2 compared to wild-type SARS-CoV-2 RBD. Also, electrostatic potential mapping evidenced that aminoacid substitutions found in B.1.625's RBD predicted structure accounted for more positively charged residues when compared to wild-type protein, which could play a role in augmenting viral protein's affinity towards the receptor (Fig. 4) . Molecular docking analysis between B.1.625 RBD and dACE2, identified 27 interface residues within 5 Å distance including N501, K484 and K417 (Fig. 5 ).

Our results report the first case of human-to-dog transmission of SARS-CoV-2 in Latin America with successful isolation and sequencing of both viral full-genomes, and show concordance with previously reported studies on SARS-CoV-2 infection in dogs that had prolonged close contact with their owners 3, 17 . A seroprevalence study carried out in Croatia found a 7.56% seropositivity in 172 randomly selected dogs living with healthcare, laboratory and veterinary personnel 17 . On the other hand, studies done in Spain and Italy could not obtain positive RT-qPCR from dog samples exhibiting pulmonary complications, but found a seroprevalence of 25% from dogs living in households, this indicates their susceptibility to SARS-CoV-2 infection and its relationship to close-contact with humans rather than dog-to-dog 7, 18 .

An experimental infection carried by Bosco-Lauth et al. 8 in cats and dogs that were inoculated with 3.0e5 and 1.45e5 PFU intranasally evidenced viral shedding occurring up to 5 days post-infection (DPI) in cats and moderate ulcerative, suppurative lympho-plasmocytic rhinitis in the nasal turbinates along with mild lymphoplasmacytic tracheitis but showed no signs of clinical disease, no weight-loss and body temperature < 39.5 °C, viral isolation was accomplished from trachea, nasal turbinates and esophagus of necropsied cats on day 5 postinfection. Contrastingly, no viral shedding nor clinical signs of disease was found in the experimental group of infected dogs with a seroconversion against RBD at 14 DPI 8 . In our study, although we did not perform viral isolation from samples, the upper respiratory tract clinical signs of disease observed in the infected dog could suggest active viral replication in lung tissue as well as in intestinal tract evidenced by the appearance of gastrointestinal signs such as vomiting, diarrhea, adynamia and lack of appetite.

The molecular and phylogenetic analyses of the obtained sequences support our hypothesis of a transmission event between the dog and its human owner, with both genomes being located within the same clade with a branch support of 100 and a total of 21 mutations shared between both sequences across isolated viral genomes. It is remarkable that these infections were due a Colombian SARS-CoV-2 lineage (B.1.625) which carries several mutations in the spike protein that confers augmented transmissibility and resistance to neutralization by naturally acquired and vaccine-elicited antibodies such as E484K mutation, which has been related to attenuation of anti-RBD neutralization 19 and N440K has been observed to reduce viral susceptibility (up to 28-fold) to monoclonal antibody-based antiviral treatment 14 . This mutations reduces complementarity and electrostatic affinity between neutralizing antibodies and RBD, which could promote an enhanced viral immune evasion in both humans and animals, as well as an increase in reinfection cases and reduction the efficacy of anti-SARS-CoV-2 vaccines 12, 20, 21 .

Other spike mutations found in the obtained sequences such as N440K and D614G have been observed to reduce the efficacy of monoclonal antibody-based antiviral treatment up to 28-fold and augment the viral infectivity 14, 22 . Furthermore, it is essential to emphasize that mutation D164G confers increased transmissibility to SARS-CoV-2 by stabilizing Spike protein, this prevents the S1 and S2 segments to be cleaved before RBD interacts with ACE2 receptor 23 . This could be associated with the greater affinity observed by Zhang et al. 24 towards dog ACE2 (dACE2) of variants carrying D614G mutation when compared to Wu-1 SARS-CoV-2. Therefore, mutation D614G could play a key role for anthropozoonotic transmission of SARS-CoV-2 24 .

On the other hand, in vitro studies have reported that N501Y mutation found in several variants such as Alpha (B.1.1.7), Gamma (P.1) and Delta (B.1.617.2) enhances the affinity of RBD towards dACE2, accounting for a threefold increase in affinity (K D = 37.1). Hence, wild-type RBD (K D = 123 nM), augmented affinity and higher viral loads found in lineages carrying D614G mutation that was found in the present study, and it could cause an increase in human-to-animal transmission 22, 24 .

The virological landscape in Colombia during the time of the study was characterized by the co-circulation of several lineages including Alpha, Gamma and Delta VOCs as well as Mu VOI which accounted for a percentage of 2. 17, 12.8, 8.87 and 57.3 of all the reported sequenced between January and August of 2021, showing that the circulation of B.1.625 was very limited to the persistence and dominance of other lineages with enhanced transmissibility and immune escape 25-28 . Zooanthroponosis has occurred in a mink farm in Denmark. this led to the culling of more than 17 million animals, and the emergence of a novel SARS-CoV-2 lineage with reduced susceptibility to antibody-neutralization (Mink cluster V) 6, [29] [30] [31] . Due to the broad host range of SARS-CoV-2, self-isolation measures should be taken by humans to avoid the occurrence of zooanthroponosis events and public health risks which could lead to the emergence of novel lineages with new mutations of importance 32 .

Our study shows in silico that these mutations could account for an increase in the risk of zooanthroponosis events. However, in vitro experiments should be carried out for determining the affinity of VOC and VOI RBD 'hCoV-19/mink/Netherlands/NB-EMC-57-2/2020| EPI ISL 632457|2020-09-28' 'hCoV-19/mink/Netherlands/NB-EMC-60-5/2020|EP I ISL 632474|2020' 'hCoV-19/mink/Netherlands/NB-EMC-58-1/2020|EPI ISL 632462|2020'

'hCoV-19/mink/Netherlands/NB-EMC-60-2/2020|EPI ISL 632471|2020'

'hCoV-19/mink/Netherlands/NB-EMC-60-3/2020|EPI ISL 632472|2020'

'hCoV-19/mink/Netherlands/NB-EMC-57-5/2020|EPI ISL 632459|2020-09-28'

'hCoV-19/mink/Netherlands/NB-EMC-65-1/2020|EPI ISL 632489|2020-10-14'

'hCoV-19/mink/Netherlands/NB-EMC-65-10/2020|EPI ISL 632491|2020-10-14'

'hCoV-19/mink/Netherlands/NB-EMC-65-3/2020|EPI ISL 632490|2020-10-14' 'hCoV-19/mink/Netherlands/NB-EMC-65-6/2020|EPI ISL 632353|2020-10-14' 'hCoV-19/mink/Netherlands/NB-EMC-65-2/2020|EPI ISL 632324|2020-10-14' 'hCoV-19/mink/Netherlands/NB-EMC-58-4/2020|EPI ISL 632351|2020' 'hCoV-19/mink/Netherlands/NB-EMC-60-1/2020|EPI ISL 632470|2020'

'hCoV-19/mink/Netherlands/NB-EMC/2020|EPI ISL 632492|2020-10-12' 'hCoV-19/mink/Netherlands/NB-EMC-57-1/2020|EPI ISL 632456|2020-09-28' 'hCoV-19/mink/Netherlands/NB-EMC-60-4/2020|EPI ISL 632473|2020' 'hCoV-19/mink/Netherlands/NB-EMC-58-5/2020|EPI ISL 632464|2020' 'hCoV-19/mink/Netherlands/NB-EMC-58-3/2020|EPI ISL 632463|2020'

'hCoV-19/mink/Netherlands/NB-EMC-40-3/2020|EPI ISL 577787|2020-08-25'

'hCoV-19/mink/Netherlands/NB-EMC-62-2/2020|EPI ISL 632434|2020-10-05'

'hCoV-19/mink/Netherlands/NB-EMC-62-4/2020|EPI ISL 632436|2020-10-05'

'hCoV-19/mink/Netherlands/NB-EMC-62-6/2020|EPI ISL 632438|2020-10-05'

'hCoV-19/mink/Netherlands/NB-EMC-62-5/2020|EPI ISL 632437|2020-10-05' 'hCoV-19/mink/Netherlands/NB-EMC-62-3/2020|EPI ISL 632435|2020-10-05' 'hCoV-19/mink/Netherlands/NB-EMC-67-19/2020|EPI ISL 722416|2020-10-21' 'hCoV-19/mink/Netherlands/NB-EMC-67-1/2020|EPI ISL 722414|2020- [10] [11] [12] [13] [14] [15] [16] [17] [18] [19] [20] [21] 'hCoV-19/mink/Denmark/mDK-277/2020|EPI ISL 683127|2020-11-10' 'hCoV-19/mink/Denmark/mDK-278/2020|EPI ISL 683128|2020-11-10' 'hCoV-19/mink/Denmark/mDK-180/2020|EPI ISL 683031|2020-11-04' 'hCoV-19/mink/Denmark/mDK-178/2020|EPI ISL 683029|2020-11-04' 'hCoV-19/mink/Denmark/mDK-179/2020|EPI ISL 683030|2020-11-04' 'hCoV-19/mink/Denmark/mDK-222/2020|EPI ISL 683073|2020-11-02' 'hCoV-19/mink/Denmark/mDK-224/2020|EPI ISL 683075|2020-11-02' 'hCoV-19/mink/Denmark/mDK-223/2020|EPI ISL 683074|2020-11-02' 'hCoV-19/mink/Denmark/mDK-76/2020|EPI ISL 641468|2020-09-24' 'hCoV-19/mink/Denmark/mDK-126/2020|EPI ISL 641505|2020-10-08' 'hCoV-19/mink/Denmark/mDK-124/2020|EPI ISL 641503|2020-10-08' 'hCoV- 19 'hCoV-19/Colo mbia/G UR-045 33/202 0|EPI ISL 941975 |2020-0 4-24' 'hCoV-19/Colo mbia/G UR-064 05/202 0|EPI ISL 941979 |2020-0 4-30' 'hCoV-1 9/Colom bia/GU R-0649 4/2020 |EPI ISL 942000 |2020-0 5-03' 'hCoV-1 9/Colom bia/GU R-0647 4/2020 |EPI ISL 941980 |2020-0 5-02' 'hCoV-1 9/Colom bia/GU R-0640 6/2020 |EPI ISL 941964 |2020-0 4-30' 'hCoV- 1 www.nature.com/scientificreports/ towards animal hosts to assess the impact of the emergence of these novel and more transmissible lineages in non-human hosts.

In conclusion, the present study was the first reported case of a human-to-dog transmission event in Latin America supported by the sequencing and molecular characterization of both viral genomes. The identification of the B.1.625 lineage as the causative agent of this infection adds to the discussion on the importance of selfisolation measures between infected humans and pets. Considering the emergence of novel and more transmissible variants of SARS-CoV-2 could have an enhanced capability of infecting non-human hosts. Epidemiological and molecular surveillance of zooanthroponosis in pets co-habiting with SARS-CoV-2 patients is mandatory since it could represent an animal mutations risk for public health. www.nature.com/scientificreports/

Sample collection and detection of SARS-CoV-2. Two hundred ninety samples were collected from domestic animals in 8 municipalities of the Cordoba department located in the north-western Colombian Caribbean, informed consent was obtained from human participants for sample collection in this study. Oropharyngeal and rectal swabs were collected for felines and canines as well as nasopharyngeal samples for humans as a part of the genomic and epidemiologic surveillance of pets living in COVID-19(+) patients' households. Samples were conserved in a Viral Transport medium for SARS-CoV-2 detection by RT-qPCR. Briefly, RNA extraction was carried out using Thermofisher Genejet viral RNA/DNA extraction kit by following the manufacturer's instructions for swab samples. Then, purified RNA was tested by RT-qPCR detection of SARS-CoV-2 E Gene following Charité Berlin protocol 33 . Those animal samples with a Ct value ≤ 32 was then subjected to nextgeneration sequencing of the virus' complete genome. This study was approved by Universidad de Cordoba's Veterinary Medicine faculty and its Ethics Committee number 005 (May 26th, 2021). All methods were carried out in accordance with national and international guidelines, including Law 84 of 1989 of the Congress of the Republic of Colombia, national guidelines of animal protection, resolution 8430 of Colombia's Health Ministry and articles 87 and 88 of the 1989's Universal declaration of Animal Rights. Additionally, animals and humans were sampled in compliance of CDC's Guidelines for Safe Work Practices in Human and Animal Medical Diagnostic Laboratories and WHO Laboratory Biosafety Manual, ensuring human and animal wellbeing 34, 35 . All methods were reported in accordance with the ARRIVE guidelines for the reporting of animal experiments 36 .

Sequencing of RT-qPCR positive samples. Samples were subjected to whole-genome sequencing.

Sequence libraries were prepared from RNA extracted from each nasopharyngeal/oropharyngeal swab per individual using the ARTIC Network protocol (https:// artic. netwo rk/ ncov-2019). Long-read Oxford Nanopore MinION sequencing was performed with the MinKNOW application (v1.5.5). Initially, raw Fast5 files were base called, and demultiplexed using Guppy; subsequently, reads were filtered by quality and length, eliminating possible chimeric and low-quality reads. Finally, genome assemblies were obtained following the MinION pipeline described in the ARTIC bioinformatics pipeline (https:// artic. netwo rk/ ncov-2019/ ncov2 019-bioin forma tics-sop. html). Each assembly was typed based on the PANGOLIN nomenclature lineage allocator and SNPs identification was performed using the clade assignment, mutation calling and sequence quality checking tool, NextClade, which identifies differences between sequences and a reference sequence used by Nextstrain.

Phylogenetic analysis of the sequences. Briefly, two datasets were generated from virus sequences reported in GISAID. For humans, 1855 viruses have been reported from Colombia in GISAID, incomplete sequences (< 29,000 bp) and sequences with low coverage (> 5% Ns) were excluded from the analysis resulting in a final dataset of 1002 sequences including the dog and owner. On the other hand, another dataset was created with sequences of SARS-CoV-2 detected in Canis lupus familiaris (dog), Felis catus (cat), Panthera leo (lion), Panthera tigris (tiger) and Neovison vison (mink) reported in GISAID globally by following the selection criteria as mentioned above to evaluate phylogenetic between globally reported sequences from cohabiting animals. The two datasets composed of a total of 1002 and 1057 sequences were subjected to multiple sequence alignment (MSA) against reference sequence NC_04551 in MAFFT v 7.48 37 using "-addfragments" 37 functionality within the webserver. Then, alignments were manually edited in Bioedit v.7.2.5 in order to add sample collection date to FASTA accession IDs as defined by REGEX (\d\d\d\d-\d\d-\d\d). Maximum-likelihood phylogenetic trees were reconstructed for each dataset in IQtree v.2.1.3 38 , ModelFinder 39 was run for the datasets to determine the bestfit substitution model, this resulted in GTR + F + I being selected for tree reconstruction, branch support was calculated using UFBoot2 40 within IQTree2. The consensus tree was visualized and tailored in FigTree v.1.4.4 41 Figure 5 . B.1.625 SARS-CoV-2-dACE2 interface residues. Mutation E484K is presented in red; residues 417 and 501 of importance in transmissibility enhancement are highlighted in orange.

hCoV-19/Colombia /ANT-CWOHC-VG -SEC00255U/2021 |EPI ISL 1960056|2021-04-05' 'hCoV-19/Colombia /ANT-CWOHC-VG-SEC00447E/2021| EPI ISL 2321624|2021-04-1 6' 'hCoV-19/Colombia /COR-INS-VG-4017 /2021|EPI ISL 2827774|2021-06-2 1

hCoV-19/Colombia/A NT-CWOHC-VG-SEC 00403M/2021|EPI ISL 2136326|2021-04-15' 'hCoV-19/Colombia/LA G-INS-VG-2789/2021| EPI ISL 2828046|2021-04-02' 'hCoV-19/Colombia/SA N-INS-VG-2946/2021|E PI ISL 2674349|2021-05-18' 'hCoV-19/Colombia/SAP -INS-VG-2989/2021|EP I ISL 2674362|2021-05-12' 'hCoV-19/Colombia/SAN -INS-VG-2181/2021|EPI ISL 1821071|2021-04-11' 'hCoV-19/Colombia/DC-IN S-VG-2117/2021|EPI ISL 1821067|2021-04-12' 'hCoV-19/Colombia/SAN-IN S-VG-2189/2021|EPI ISL 1821075|2021-04-05' 'hCoV-19/Colombia/ANT-CW OHC-VG-SEC00213E/2021 |EPI ISL 1960080|2021-04-09' 'hCoV-19/Colombia/ANT-CW OHC-VG-SEC00722T/2021| EPI ISL 2621177|2021-05-07' 'hCoV-19/Colombia/CES-INS-VG-2686/2021|EPI ISL 2674284|2021-04-26' 'hCoV-19/Colombia/BOY-INS-V G-2887/2021|EPI ISL

hCoV-19/Colombia/DC-A005/2021|EPI ISL 2339799|2021-04-13' 'hCoV-19/Colombia/DC-A056/2021|EPI ISL 2339847|2021-04-10' 'hCoV-19/Colombia/ATL-INS-VG-1633/2021|EPI ISL 1820927|2021-03-04' 'hCoV-19/Colombia/BOL-INS-VG-2023/2021|EPI ISL 2828029|2021-04-04' 'hCoV-19/Colombia/ATL-A021/2021|EPI ISL 2339814|2021-04-09' 'hCoV-19/Colombia/CHO-INS-VG-3119/2021|EPI ISL 2500941|2021-04-22' 'hCoV-19/Colombia/CHO-INS-VG-3303/2021|EPI ISL 2500966|2021-05-08' 'hCoV-19/Colombia/CHO-INS-VG-3265/2021|EPI ISL 2500963|2021-04-30' 'hCoV-19/Colombia/ANT-CWOHC-VG-SEC00422F/2021|EPI ISL 2321600|2021-04-14' 'hCoV-19/Colombia/ANT-INS-VG-2423/2021|EPI ISL 2158348|2021-04-16' 'hCoV-19/Colombia/ATL-A023/2021|EPI ISL 2339816|2021-04-09' 'hCoV-19/Colombia/ANT-CWOHC-VG-SEC00360V/2021|EPI ISL 2136287|2021-04-05' 'hCoV-19/Colombia/ANT-CWOHC-VG-SEC00818L/2021|EPI ISL

0|EPI ISL 1629735|2020-12-29' 'hCoV-19/Colombia

PI ISL 1324135|2020-11-22' 'hCoV-19/Colombia/BOY-INS-VG-553/2021|EPI ISL 1220047|2021-01-21' 'hCoV-19/Colombia/DC-C88/2020|EPI ISL

00041O/2021|EPI ISL 1629736|2021-01-20

hCoV-19/Colombia/ANT-CWOHC-V G-RSEC00002B/2020|EPI ISL 1091785|2020-10-05' 'hCoV-19/Colombia/SAP-INS-SPS AN01/2020|EPI ISL 791087|2020-12-27' 'hCoV-19/Colombia/SAP-INS-SP SAN02/2020|EPI ISL 791088|2020-12-28' 'hCoV-19/Colombia/GVI-161641 /2020|EPI ISL 941994|2020-06-24' 'hCoV-19/Colombia/GVI-13954 2/2020|EPI ISL

1642/2020|EPI ISL 941951|2020-06-24' 'hCoV-19/Colombia/COR

U33/2020|EPI ISL 2603127|2020-07-05' 'hCoV-19/Colombia/COR -U31/2020|EPI ISL

hCoV-19/Colombia/NA R-INS-VG-2360/2021|E PI ISL 2009061|2021-04-10' 'hCoV-19/Colombia/BO Y-INS-VG-508/2021|EP I ISL 1220046|2021-01-18

hCoV-19/Co lombia/TOL -INS-0345-T 20/2020|EP I ISL 526963|202 0-04-14' 'hCoV-19/Co lombia/TOL-INS-0330-T 5/2020|EPI ISL 526965|202 0-04-14' 'hCoV-19/Co lombia/un-C IAT-01/2020 |EPI ISL 445085|202 0-04-02' 'hCoV-19/Co lombia/VAC-UdeV-01-2/2 020|EPI ISL 445219|2020 -04-02' 'hCoV-19/Co lombia/ANT-A140/2021|E PI ISL 2603172|202 1-04-05' 'hCoV-19/Co lombia/ANT-C WOHC-VG-S EC00408R/2 021|EPI ISL 2136331|202 1-04-14' 'hCoV-19/Co lombia/ANT-A 100/2021|EP I ISL 2603150|202 1-04-07' 'hCoV-19/Col ombia/ANT-C WOHC-VG-S EC00411U/20 21|EPI ISL 2136334|202 1-04-14' 'hCoV-19/Col ombia/ANT-A 077/2021|EPI ISL 2603136|202 1-04-09' 'hCoV-19/Colo mbia/ANT-A1 35/2021|EPI ISL 2603167|2021 -04-05' 'hCoV-19/Colo mbia/ANT-CW OHC-VG-SEC 00505K/2021| EPI ISL 2321649|2021 -04-28' 'hCoV-19/Colo mbia/ANT-A13 6/2021|EPI ISL 2603168|2021 -04-05' 'hCoV-19/Colo mbia/ANT-A08 0/2021|EPI ISL 2603138|2021 -04-09' 'hCoV-19/Colo mbia/ANT-A14 1/2021|EPI ISL

hCoV-19/Colomb ia/ANT-CWOHC-VG-SEC00311Y/2 021|EPI ISL 1960041|2021-04 -13' 'hCoV-19/Colomb ia/CHO-INS-VG-3 124/2021|EPI ISL 2500946|2021-04 -21

hCoV-19/Colombia /ANT-CWOHC-VG-SEC00442Z/2021|E PI ISL 2321619|2021-04-1 5' 'hCoV-19/Colombia /VAC-INS-VG-2852 /2021|EPI ISL 2674315|2021-04-2 6' 'hCoV-19/Colombia/ ANT-CWOHC-VG-S EC00448F/2021|EP I ISL 2321625|2021-04-16

hCoV-19/Colombia/C AQ-INS-VG-4007/202 1|EPI ISL 2827779|2021-06-10' 'hCoV-19/Colombia/CA Q-INS-VG-4008/2021| EPI ISL 2827783|2021-06-10' 'hCoV-19/Colombia/AN T-CWOHC-VG-SEC003 77M/2021|EPI ISL 2136300|2021-04-07' 'hCoV-19/Colombia/ANT -CWOHC-VG-SEC0042 8L/2021|EPI ISL 2321606|2021-04-15' 'hCoV-19/Colombia/ANT-A103/2021|EPI ISL

hCoV-19/Colombia/ANT-CW OHC-VG-SEC00379O/2021| EPI ISL 2136302|2021-04-07' 'hCoV-19/Colombia/ANT-CWO HC-VG-SEC00314B/2021|EP I ISL 1960053|2021-04-16

hCoV-19/Colombia/ANT-CWOHC -VG-SEC00405O/2021|EPI ISL 2136328|2021-04-14' 'hCoV-19/Colombia/ANT-CWOHC-V G-SEC00449G/2021|EPI ISL 2321626|2021-04-16' 'hCoV-19/Colombia/DC-G015891/202 1|EPI ISL 2157196|2021-04-08' 'hCoV-19/Colombia/ANT-CWOHC-VG-S EC00438V/2021|EPI ISL

hCoV-19/Colombia/ANT-CWOHC-VG-SEC00415Y /2021|EPI ISL 2321593|2021-04-14' 'hCoV-19/Colombia/ANT-CWOHC-VG-SEC00388X/20 21|EPI ISL 2136311|2021-04-08' 'hCoV-19/Colombia/ANT-CWOHC-VG-SEC00406P/2021|E PI ISL 2136329|2021-04-14' 'hCoV-19/Colombia/ANT-CWOHC-VG-SEC00260Z/2021|EPI ISL 1960077|2021-04-14' 'hCoV-19/Colombia/DC-379559/2021|EPI ISL 2157208|2021-04-12' 'hCoV-19/Colombia/QUI-COV-VG-3890/2021|EPI ISL 2894632|2021-06-13' 'hCoV-19/Colombia/DC-UANDES-339014/2021|EPI ISL 1626619|2021-04-03' 'hCoV-19/Colombia/ANT-A105/2021|EPI ISL

ISL 1960052|2021-04-14' 'hCoV-19/Colombia/DC-G016158/2021|EPI ISL

hCoV-19/Colombia/ANT-CWOHC-VG-SEC00273M/2021|EPI ISL 1960078|2021-03-18' 'hCoV-19/Colombia/ANT-CWOHC-VG-SEC00391A/2021|EPI ISL 2136314|2021-04-12' 'hCoV-19/Colombia/DC-G015834/2021|EPI ISL

hCoV-19/Colombia/ANT-CWOHC-VG-SEC00472D/2021|EPI ISL 2321630|2021-04-28' 'hCoV-19/Colombia/DC-A130/2021|EPI ISL 2603164|2021-04-05' 'hCoV-19/Colombia/VAC-A035/2021|EPI ISL 2339828|2021-04-09' 'hCoV-19/Colombia/VAC-INS-VG-3552/2021|EPI ISL 2657878|2021-04-25' 'hCoV-19/Colombia/ANT-CWOHC-VG-SEC00432P/2021|EPI ISL 2321610|2021-04-15' 'hCoV-19/Colombia/VAC-A039/2021|EPI ISL 2339831|2021-04-09' 'hCoV-19/Colombia/CUN-S2/2021|EPI ISL 1824180|2021-04-10' 'hCoV-19/Colombia/VAC-INS-VG-101/2020|EPI ISL 794654|2020-12-30' 'hCoV-19/Colombia/GUR-06537/2020|EPI ISL 941981|2020-04-30' 'hCoV-19/Colombia/GUR-06539/2020|EPI ISL 941945|2020-04-30' 'hCoV-19/Colombia/NAR-GUR-0627/2020|EPI ISL 447749|2020-03-29' 'hCoV-19/Colombia

hCoV-19/Colombia/VAU-INS-VG-3103/20 21|EPI ISL 2500930|2021-04-09' 'hCoV-19/Colombia/VAU-INS-VG-3201/2 021|EPI ISL 2500961|2021-04-29

hCoV-19/Colombia/SAN-INS-VG-1415/2021|EPI ISL 1494948|2021-02-03' 'hCoV-19/Colombia/BOY-INS-VG -129/2020|EPI ISL 1017701|2020-12-26' 'hCoV-19/Colombia/GVI-133304 /2020|EPI ISL 941986|2020-05-29' 'hCoV-19/Colombia

hCoV-19/Colombia/SAN -INS-80123/2020|EPI ISL 456149|2020-03-15' 'hCoV-19/Colombia/SAN -INS-94514/2020|EPI ISL 456156|2020-04-06' 'hCoV-19/Colombia/DC -324777/2021|EPI ISL 1738784|2021-03-29' 'hCoV-19/Colombia/DC -INS-78390/2020|EPI ISL 418262|2020-03-06' 'hCoV-19/Colombia/VA C-GVI-93537/2020|EP I ISL

20|EPI ISL

20|EPI ISL

EPI ISL 941983|2020-04-2 3

3-26' 'hCoV-1 9/Colom bia/CUN -INS-VG -2268/2 021|EP I ISL 215504 3|2021-04-12' 'hCoV-1 9/Colom bia/DC-A120/2 021|EP I ISL 260315 8|2021-04-06' 'hCoV-1 9/Colom bia/DC-308450 /2021|E PI ISL 173878 3|2021-03-24' 'hCoV-1 9/Colom bia/CAU -INS-VG -3043/2 021|EP I ISL 267433 1|2021-05-09' 'hCoV-1 9/Colom bia/NAR -INS-VG -2354/2 021|EP I ISL 215504 7|2021-04-13' 'hCoV-1 9/Colom bia/VAC -A010/2 021|EP I ISL /CUN -INS-T1 /2020|EP I ISL 498168| 2020-03 -11' 'hCoV-1 9/Colom bia/CUN -INS-T8 /2020|EP I ISL 498169| 2020-03 -11' 'hCoV-1 9/Colom bia/CUN -INS-T12 /2020|EP I ISL 498170| 2020-03 -11' 'hCoV-19 /Colomb ia/TOL-I NS-0344 /2020|EP I ISL 653749| 2020-04 -14' 'hCoV-19 /Colomb ia/CAL-I NS-7948 9

Colom bia/CUN -INS-VG -2268/2 021|EP I ISL 215504 3|2021-04-12

hCoV-19/Colo mbia/A TL-INS -79743 /2020|E PI ISL 456145 |2020-0 3-14' 'hCoV-19/Colo mbia/D C-INS-84861/ 2020|E PI ISL 456116 |2020-0 3-22' 'hCoV-19/Colo mbia/R IS-INS -80245 /2020|E PI ISL 456155 |2020-0 3-16' 'hCoV-19/Col ombia/ VAC-G VI-972 23/202 0|EPI ISL 447814 |2020-0 4-06

EPI ISL 941983|2020-04-2 3' 'hCoV-19/Colombia

20|EPI ISL

20|EPI ISL

EPI ISL 447804|2020-04-04

hCoV-19/Colombia/DC -INS-78390/2020|EPI ISL 418262|2020-03-06' 'hCoV-19/Colombia/DC -324777/2021|EPI ISL 1738784|2021-03-29' 'hCoV-19/Colombia/SAN -INS-94514/2020|EPI ISL 456156|2020-04-06

hCoV-19/Colombia/SAN-UW -651275441501/2020|EPI ISL 1324144|2020-11-30' 'hCoV-19/Colombia/SAN-5769 00-010-RNA/2020|EPI ISL 1582589|2020-12-04' 'hCoV-19/Colombia/GVI-133304 /2020|EPI ISL 941986|2020-05-29' 'hCoV-19/Colombia/BOY-INS-VG -129/2020|EPI ISL 1017701|2020-12-26' 'hCoV-19/Colombia/SAN-INS-VG-1415/2021|EPI ISL

hCoV-19/Colombia/VAU-INS-VG-3201/2 021|EPI ISL 2500961|2021-04-29' 'hCoV-19/Colombia/VAU-INS-VG-3103/20 21|EPI ISL

hCoV-19/Colombia/GUR-06538/2020|EPI ISL 941944|2020-04-30' 'hCoV-19/Colombia/NAR-GUR-0627/2020|EPI ISL 447749|2020-03-29' 'hCoV-19/Colombia/GUR-06539/2020|EPI ISL 941945|2020-04-30' 'hCoV-19/Colombia/GUR-06537/2020|EPI ISL 941981|2020-04-30' 'hCoV-19/Colombia/VAC-INS-VG-101/2020|EPI ISL 794654|2020-12-30' 'hCoV-19/Colombia/CUN-S2/2021|EPI ISL 1824180|2021-04-10' 'hCoV-19/Colombia/VAC-A039/2021|EPI ISL 2339831|2021-04-09' 'hCoV-19/Colombia/ANT-CWOHC-VG-SEC00432P/2021|EPI ISL 2321610|2021-04-15' ' hCoV-19/Colombia/VAC-INS-VG-3552/2021|EPI ISL 2657878|2021-04-25' 'hCoV-19/Colombia/VAC-A035/2021|EPI ISL 2339828|2021-04-09' 'hCoV-19/Colombia/DC-A130/2021|EPI ISL 2603164|2021-04-05' 'hCoV-19/Colombia/ANT-CWOHC-VG-SEC00472D/2021|EPI ISL 2321630|2021-04-28' 'hCoV-19/Colombia/ANT-A030/2021|EPI ISL 2339823|2021-04-09' 'hCoV-19/Colombia/VAC-A034/2021|EPI ISL 2339827|2021-04-09' 'hCoV-19/Colombia/VAC-A011/2021|EPI ISL 2339805|2021-04-12' 'hCoV-19/Colombia/VAC-INS-VG-2075/2021|EPI ISL 2155040|2021-04-12' 'hCoV-19/Colombia/VAC-A157/2021|EPI ISL 2603182|2021-04-05' 'hCoV-19/Colombia/VAC-A033/2021|EPI ISL 2339826|2021-04-09' 'hCoV-19/Colombia/VAC-A015/2021|EPI ISL 2339809|2021-04-12' 'hCoV-19/Colombia/AMA-INS-VG-632/2021|EPI ISL 956288|2021-01-13' 'hCoV-19/Colombia

0|EPI ISL 944635|2020-04-17' 'hCoV-19/Colombia

020|EPI ISL 944637|2020-04-16' 'hCoV-19/Colombia/MET

456154|2020-04-26' 'hCoV-19/Colombia/NAR-INS-103 303-2/2020|EPI ISL 526955|2020-04-28' 'hCoV-19/Colombia

2020|EPI ISL 941982|2020-04-30' 'hCoV-19/Colombia

ISL 1017708|2020-11-26' 'hCoV-19/Colombia/NAR-IN

VG-2176/2021|EPI ISL

hCoV-19/Colombia/ANT-A028/2021|EPI ISL 2339821|2021-04-12' 'hCoV-19/Colombia/DC-G015834/2021|EPI ISL 2157195|2021-04-07' 'hCoV-19/Colombia/ANT-CWOHC-VG-SEC00391A/2021|EPI ISL 2136314|2021-04-12' 'hCoV-19/Colombia/ANT-CWOHC-VG-SEC00273M/2021|EPI ISL

hCoV-19/Colombia/ANT-A105/2021|EPI ISL 2603153|2021-04-05' 'hCoV-19/Colombia/DC-UANDES-339014/2021|EPI ISL 1626619|2021-04-03' 'hCoV-19/Colombia/QUI-COV-VG-3890/2021|EPI ISL 2894632|2021-06-13' 'hCoV-19/Colombia/DC-379559/2021|EPI ISL 2157208|2021-04-12' 'hCoV-19/Colombia/ANT-CWOHC-VG-SEC00260Z/2021|EPI ISL 1960077|2021-04-14' 'hCoV-19/Colombia/ANT-CWOHC-VG-SEC00406P/2021|E PI ISL 2136329|2021-04-14' 'hCoV-19/Colombia/ANT-CWOHC-VG-SEC0 0490V/2021|EPI ISL 2321643|2021-04-17' 'hCoV-19/Colombia/ANT-CWOHC-VG-SE C00439W/2021|EPI ISL 2321616|2021-04-15' 'hCoV-19/Colombia/ANT-CWOHC-VG-S EC00438V/2021|EPI ISL 2321615|2021-04-15' 'hCoV-19/Colombia/ANT-CWOHC-VG-SEC00388X/20 21|EPI ISL 2136311|2021-04-08' 'hCoV-19/Colombia/ANT-CWOHC-VG-SEC00415Y /2021|EPI ISL 2321593|2021-04-14' 'hCoV-19/Colombia/ANT-CWOHC-VG-SEC003 81Q/2021|EPI ISL 2136304|2021-04-07' 'hCoV-19/Colombia/DC-G015891/202 1|EPI ISL 2157196|2021-04-08' 'hCoV-19/Colombia/ANT-CWOHC-V G-SEC00449G/2021|EPI ISL

hCoV-19/Colombia/ANT-CWO HC-VG-SEC00314B/2021|EP I ISL 1960053|2021-04-16' 'hCoV-19/Colombia/ANT-CW OHC-VG-SEC00379O/2021| EPI ISL

hCoV-19/Colombia/ARA-INS-VG-3462/2021|EPI ISL 2657861|2021-05-19' 'hCoV-19/Colombia/ANT-A103/2021|EPI ISL 2603152|2021-04-07' 'hCoV-19/Colombia/ANT -CWOHC-VG-SEC0042 8L/2021|EPI ISL 2321606|2021-04-15' 'hCoV-19/Colombia/AN T-CWOHC-VG-SEC003 77M/2021|EPI ISL

hCoV-19/Colombia/C AQ-INS-VG-3079/20 21|EPI ISL 2674338|2021-04-28 ' 'hCoV-19/Colombia/ ANT-CWOHC-VG-S EC00448F/2021|EP I ISL 2321625|2021-04-16 ' 'hCoV-19/Colombia /VAC-INS-VG-2852 /2021|EPI ISL 2674315|2021-04-2 6' 'hCoV-19/Colombia /ANT-CWOHC-VG-SEC00442Z/2021|E PI ISL 2321619|2021-04-1 5' 'hCoV-19/Colombia /ANT-CWOHC-VG -SEC00402L/2021 |EPI ISL 2136325|2021-04-15' 'hCoV-19/Colombi a/ANT-CWOHC-V G-SEC00252R/20 21|EPI ISL 1960044|2021-04-13

hCoV-19/Colom bia/ANT-CWOH C-VG-SEC0089 3I/2021|EPI ISL 2651229|2021-0 5-21' 'hCoV-19/Colom bia/ANT-CWOH C-VG-SEC0024 1G/2021|EPI ISL 1960047|2021-04-19' 'hCoV-19/Colom bia/ANT-A070/ 2021|EPI ISL 2603133|2021-04-09' 'hCoV-19/Colo mbia/ANT-A14 1/2021|EPI ISL 2603173|2021 -04-05' 'hCoV-19/Colo mbia/ANT-A08 0/2021|EPI ISL 2603138|2021 -04-09' 'hCoV-19/Colo mbia/ANT-A13 6/2021|EPI ISL 2603168|2021 -04-05' 'hCoV-19/Colo mbia/ANT-CW OHC-VG-SEC 00505K/2021| EPI ISL 2321649|2021 -04-28' 'hCoV-19/Colo mbia/ANT-A1 35/2021|EPI ISL 2603167|2021 -04-05' 'hCoV-19/Col ombia/ANT-A 077/2021|EPI ISL 2603136|202 1-04-09' 'hCoV-19/Co lombia/ANT-C WOHC-VG-S EC00408R/2 021|EPI ISL 2136331|202 1-04-14' 'hCoV-19/Co lombia/ANT-A140/2021|E PI ISL 2603172|202 1-04-05' 'hCoV-19/Col ombia/ANT-C WOHC-VG-S EC00411U/20 21|EPI ISL 2136334|202 1-04-14' 'hCoV-19/Co lombia/ANT-A 100/2021|EP I ISL

1|EPI ISL

021|EPI ISL 1303371|2021-01-19' 'hCoV-19/Colombia/ANT-CWOHC-VG-SEC0021

7I/2021|EPI ISL

ISL 2674295|2021-04-30

ISL 2674294|2021-04-30

hCoV-19/Colombia/ARA-INS-VG-2149/2021|EPI ISL 1805637|2021-04-09' 'hCoV-19/Co lombia/VAC-UdeV-01-2/2 020|EPI ISL 445219|2020 -04-02' 'hCoV-19/Co lombia/un

hCoV-19 /Colombia /MET-INS -VG-1463 /2021|EP I ISL 1582983| 2021-02-19' 'hCoV-19 /Colombi a/SAN-IN S-VG-272 5/2021|E PI ISL 2828056| 2021-04-24' 'hCoV-19 /Colombi a/DC-A10 9/2021|E PI ISL 2603157| 2021-04-07' 'hCoV-19 /Colombi a/VAC-IN S-VG-35 38/2021| EPI ISL 2657870 |2021-05 -07

hCoV-19/Colo mbia/G UR-045 33/202 0|EPI ISL 941975 |2020-0 4-24' 'hCoV-19/Colo mbia/C OR-GV I-97527 /2020|E PI ISL 447817 |2020-0 4-09' 'hCoV-19/Colo mbia/V AC-GV I-97181 /2020|E PI ISL 447792 |2020-0 4-03' 'hCoV-19/Col ombia/ CAU

I ISL 941943 |2020-0 6-17' 'hCoV-19/Col ombia/ GVI-15 0146/2 020|EP I ISL

0|EPI ISL 233987 7|2020 -08-08 ' 'hCoV-19/Col ombia

0|EPI ISL 941984 |2020-0 5-06

|EPI ISL 941978

bia/ME T-406/2 020|EP I ISL 215107

PI ISL 942005

I ISL 942004 |2020-0 6-30'

PI ISL 941993 |2020-0 6-23' 'hCoV-1

Colomb ia/AMA-G UR-0680 9/2020|E PI ISL 941151|2 020-05-0 4

PI ISL

0|EPI ISL

|EPI ISL

PI ISL

|EPI ISL

20|EPI ISL 653761|202 0-06-08' 'hCoV-19/C olombia/GU R-40200/20 20|EPI ISL 941996|202 0-06-26' 'hCoV-19/Co lombia/BOY

020|EPI ISL 447743|2020 -03-28' 'hCoV-19/Co lombia/ANT-GVI-92161/2 020|EPI ISL 447763|2020 -03-31' 'hCoV-19/Co lombia/AMA

2020|EPI ISL 941157|2020 -05-04' 'hCoV-19/Co lombia/AMA

2020|EPI ISL 941156|2020 -05-04' 'hCoV-19/Co lombia/AMA

2603126|2020 -07-13' 'hCoV-19/Colo mbia/BOY-GV I-97795/2020|

EPI ISL 447805|2020-04-04' 'hCoV-19/Colo mbia

PI ISL 447739|2020-03-27

|EPI ISL 941141|2020-0 4-25

hCoV-19/Colo mbia/BOY-GUR -0209/2020|EP I ISL 447740|2020-0 3-28' 'hCoV-19/Colom bia/AMA-GUR-06733/2020|EP I ISL 941112|2020-0 5-03' 'hCoV-19/Colom bia/BOY

hCoV-19/Colom bia/AMA-INS-10 6856/2020|EPI ISL

hCoV-19/Colomb ia/AMA-GUR-067 32/2020|EPI ISL 941111|2020-05-0 3' 'hCoV-19/Colombi a/ANT-CWOHC-V G-SEC00067O/20 20|EPI ISL 1629757|2020-12-16' 'hCoV-19/Colombia /DC-C110/2020|EP I ISL

|EPI ISL 794653|2020-12-30'

hCoV-19/Colombia/BO Y-INS-VG-508/2021|EP I ISL 1220046|2021-01-18' 'hCoV-19/Colombia/NA R-INS-VG-2360/2021|E PI ISL 2009061|2021-04-10

I ISL 2834730|2021-04-21' 'hCoV-19/Colombia/COR -U31/2020|EPI ISL

U33/2020|EPI ISL

hCoV-19/Colombia/GVI-13954 2/2020|EPI ISL 941942|2020-06-02' 'hCoV-19/Colombia/GVI-161641 /2020|EPI ISL 941994|2020-06-24

hCoV-19/Colombia/ANT-CWOHC-V G-RSEC00002B/2020|EPI ISL 1091785|2020-10-05' 'hCoV-19/Colombia/ANT-CWOHC-VG -RSEC00001A/2020|EPI ISL 1091784|2020-10-04' 'hCoV-19/Colombia/ANT-CWOHC-VG-S EC00050X/2020|EPI ISL 1629745|2020-12-19' 'hCoV-19/Colombia/ANT-CWOHC-VG-SEC 00041O/2021|EPI ISL

PI ISL 1324135|2020-11-22

hCoV-19/Colombia/ANT-CWOHC-VG-SEC00013M/2021|EPI ISL 1091781|2021-01-14' 'hCoV-19/Colombia/DC-290716/2021|EPI ISL 1738785|2021-03-18' 'hCoV-19/Colombia/HUI-INS-VG-1378/2021|EPI ISL 1494944|2021-02-26' 'hCoV-19/Colombia/SAN-INS-VG-1428/2021|EPI ISL 1494950|2021-02-25' 'hCoV-19/Colombia/DC-336078/2021|EPI ISL 1738787|2021-04-01' 'hCoV-19/Colombia/VAC-A093/2021|EPI ISL 2603146|2021-04-08' 'hCoV-19/Colombia/DC-336093/2021|EPI ISL 1738789|2021-04-02' 'hCoV-19/Colombia/DC-INS-VG-68/2020|EPI ISL 794659|2020-12-30' 'hCoV-19/Colombia/TOL-INS-VG-1906/2021|EPI ISL 1805632|2021-02-25' 'hCoV-19/Colombia/TOL-INS-VG-847/2021|EPI ISL

ISL 1424061|2020-07-05' 'hCoV-19/Colombia/ANT-INS-5/2020|EPI ISL

ISL 794666|2021-01-05' 'hCoV-19/Colombia/ANT-C72/2020|EPI ISL

63K/2020|EPI ISL 1629753|2020-12-28

0056D/2020|EPI ISL 1629751|2020-12-17

C00065M/2020|EPI ISL 1629755|2020-12-19

EC00052Z/2021|EPI ISL 1629747|2021-01-06' 'hCoV-19/Colombia/ANT-CWOHC-VG -SEC00049W/2020|EPI ISL

PI ISL 2339890|2020-07-07' 'hCoV-19/Colombia/ANT-INS-CO

EPI ISL 2674353|2021-05-20

PI ISL 1821073|2021-04-08' 'hCoV-19/Colombia/DC -A054/2021|EPI ISL 2339845|2021-04-10' 'hCoV-19/Colombia/CAQ -INS-VG-2179/2021|EPI ISL

ISL 2339802|2021-04-12' 'hCoV-19/Colombia/ATL-IN

hCoV-19/Colombia/BOL-INS -VG-2019/2021|EPI ISL

ISL 2657858|2021-05-19

ISL 2158349|2021-04-13' 'hCoV-19/Colombia/ANT-CWOHC

VG-SEC00397G/2021|EPI ISL

021|EPI ISL 2674339|2021-04-30

1|EPI ISL

ISL 2674313|2021-04-26' 'hCoV-19/Colombia/ATL-A019/2021|EPI

hCoV-19/Colombia/ANT-CWOHC-VG-SEC00366B/2021|EPI ISL 2136291|2021-04-05' 'hCoV-19/Colombia/ANT-CWOHC-VG-SEC00818L/2021|EPI ISL 2621189|2021-05-18' 'hCoV-19/Colombia/ANT-CWOHC-VG-SEC00360V/2021|EPI ISL

hCoV-19/Colombia/CHO-INS-VG-3303/2021|EPI ISL 2500966|2021-05-08' 'hCoV-19/Colombia/CHO-INS-VG-3119/2021|EPI ISL 2500941|2021-04-22' 'hCoV-19/Colombia/ATL-A023/2021|EPI ISL

hCoV-19/Colombia/BOL-INS-VG-2023/2021|EPI ISL 2828029|2021-04-04' 'hCoV-19/Colombia/ATL-INS-VG-1633/2021|EPI ISL 1820927|2021-03-04' 'hCoV-19/Colombia/DC-A056/2021|EPI ISL 2339847|2021-04-10' 'hCoV-19/Colombia/DC-A005/2021|EPI ISL 2339799|2021-04-13' 'hCoV-19/Colombia/ANT-CWOHC-VG-SEC00386 V/2021|EPI ISL 2136309|2021-04-08' 'hCoV-19/Colombia/MAG-INS-VG-1624/2021|E PI ISL

hCoV-19/Colombia/BOY-INS-V G-2887/2021|EPI ISL 2674322|2021-05-06' 'hCoV-19/Colombia/CES-INS-VG-2686/2021|EPI ISL 2674284|2021-04-26' 'hCoV-19/Colombia/ANT-CW OHC-VG-SEC00722T/2021| EPI ISL 2621177|2021-05-07' 'hCoV-19/Colombia/ANT-CW OHC-VG-SEC00213E/2021 |EPI ISL 1960080|2021-04-09' 'hCoV-19/Colombia/SAN-IN S-VG-2189/2021|EPI ISL 1821075|2021-04-05' 'hCoV-19/Colombia/DC-IN S-VG-2117/2021|EPI ISL 1821067|2021-04-12' 'hCoV-19/Colombia/SAN -INS-VG-2181/2021|EPI ISL 1821071|2021-04-11' 'hCoV-19/Colombia/SAP -INS-VG-2989/2021|EP I ISL 2674362|2021-05-12' 'hCoV-19/Colombia/SA N-INS-VG-2946/2021|E PI ISL 2674349|2021-05-18' 'hCoV-19/Colombia/LA G-INS-VG-2789/2021| EPI ISL 2828046|2021-04-02' 'hCoV-19/Colombia/A NT-CWOHC-VG-SEC 00403M/2021|EPI ISL

hCoV-19/Colomb ia/ANT-CWOHC-V G-SEC00467Y/20 21|EPI ISL 2321627|2021-04 -15' 'hCoV-19/Colomb ia/ANT-CWOHC-VG-SEC00211C/ 2021|EPI ISL 1960055|2021-04 -08' 'hCoV-19/Colomb ia/CUN-INS-VG-2815/2021|EPI ISL 2674306|2021-05 -08' 'hCoV-19/Colom bia/ANT-CWOHC -VG-SEC00276P /2021|EPI ISL 1960082|2021-0 4-22' 'hCoV-19/Colom bia/ANT-CWOH C-VG-SEC0020 6X/2021|EPI ISL 1960042|2021-0 4-10' 'hCoV-19/Colom bia/ANT-CWOH C-VG-SEC0026 2B/2021|EPI ISL 1960081|2021-0 4-10' 'hCoV-19/Colom bia/ANT-CWOH C-VG-SEC0026 4D/2021|EPI ISL 1960057|2021-0 4-08' 'hCoV-19/Colom bia/ANT-CWOH C-VG-SEC0072 4V/2021|EPI ISL 2621179|2021-05-08' 'hCoV-19/Colom bia/ANT-CWOH C-VG-SEC004 12V/2021|EPI ISL 2136335|2021-04-14' 'hCoV-19/Colo mbia/ANT-CW OHC-VG-SEC 00723U/2021|E PI ISL 2621178|2021-05-08' 'hCoV-19/Colo mbia/ANT-CW OHC-VG-SEC 00446D/2021| EPI ISL 2321623|2021 -04-15' 'hCoV-19/Colo mbia/BOY-INS -VG-2884/202 1|EPI ISL 2674320|2021 -05-07' 'hCoV-19/Colo mbia/ANT-CW OHC-VG-SEC 00843K/2021| EPI ISL 2621202|2021 -05-15' 'hCoV-19/Colo mbia/COR

1 (Gamma) B.1.1.7 (Alpha) C.37 (Lambda)

Phylogenetic trees of sequences available in GISAID from minks, lions, tigers, cats, and dogs, U117 is highlighted in red. (B) Phylogenetic tree comprising Colombian sequences, VOC, and lineages of interest are annotated within the tree

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The authors acknowledge the work done by laboratory staff of Instituto de Investigaciones Biologicas del Tropico in SARS-CoV2 detection of the samples. We appreciate the support of dog owner which allowed samples to be collected and this research to be published. Authors greatly acknowledge the originating and submitting laboratories for sharing genetic sequences and associated data through the GISAID initiative for SARS-CoV-2 genome sequences. This research was funded by Colombia's Science Ministry (Minciencias), BPIN 20200000100090.

R.R. contributed to the design of the study, protein structure prediction and writing of the manuscript; E.G. and R.R. contributed by doing phylogenetic analysis and writing; M.M., N.B., S.C., L.H.P. and J.D.R. contributed to genome isolation, sequencing, assembly, analysis and quality control. Y.B., A.A. and C.M.B. contributed to sample collection, viral detection and clinical case characterization. R.R., H.S.C., B.G., J.D.R., A.C., C.G., G.A. and S.M. contributed to data analysis, result discussion, and reviewing of the manuscript. All the authors reviewed the manuscript.

The authors declare no competing interests.

The online version contains supplementary material available at https:// doi. org/ 10. 1038/ s41598-022-11847-9.Correspondence and requests for materials should be addressed to S.M.Reprints and permissions information is available at www.nature.com/reprints.Publisher's note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

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