Analyze Diet

Topic:DNA

DNA in horses refers to the genetic material that carries the hereditary information necessary for the growth, development, functioning, and reproduction of equine species. It consists of sequences of nucleotides that encode the genetic instructions used in the development and functioning of horses. DNA analysis in horses can provide insights into genetic diversity, lineage, and breed characteristics. It is also utilized in identifying genetic disorders, understanding hereditary traits, and assisting in selective breeding programs. This page compiles peer-reviewed research studies and scholarly articles that explore the structure, function, and applications of DNA analysis in equine genetics and breeding.
Characterization of a full-length endogenous beta-retrovirus, EqERV-beta1, in the genome of the horse (Equus caballus).
Viruses    June 1, 2011   Volume 3, Issue 6 620-628 doi: 10.3390/v3060620
van der Kuyl AC.Information on endogenous retroviruses fixed in the horse (Equus caballus) genome is scarce. The recent availability of a draft sequence of the horse genome enables the detection of such integrated viruses by similarity search. Using translated nucleotide fragments from gamma-, beta-, and delta-retroviral genera for initial searches, a full-length beta-retrovirus genome was retrieved from a horse chromosome 5 contig. The provirus, tentatively named EqERV-beta1 (for the first equine endogenous beta-retrovirus), was 10434 nucleotide (nt) in length with the usual retroviral genome structure of 5'...
Association of sequence variants in CKM (creatine kinase, muscle) and COX4I2 (cytochrome c oxidase, subunit 4, isoform 2) genes with racing performance in Thoroughbred horses.
Equine veterinary journal. Supplement    May 27, 2011   Issue 38 569-575 doi: 10.1111/j.2042-3306.2010.00181.x
Gu J, MacHugh DE, McGivney BA, Park SD, Katz LM, Hill EW.The wild progenitors of the domestic horse were subject to natural selection for speed and stamina for millennia. Uniquely, this process has been augmented in Thoroughbreds, which have undergone at least 3 centuries of intense artificial selection for athletic phenotypes. While the phenotypic adaptations to exercise are well described, only a small number of the underlying genetic variants contributing to these phenotypes have been reported. Objective: A panel of candidate performance-related genes was examined for DNA sequence variation in Thoroughbreds and the association with racecourse per...
Evaluation of ACE, SP17, and FSHB as candidates for stallion fertility in Hanoverian warmblood horses.
Animal reproduction science    May 24, 2011   Volume 126, Issue 3-4 200-206 doi: 10.1016/j.anireprosci.2011.05.007
Giesecke K, Hamann H, Stock KF, Klewitz J, Martinsson G, Distl O, Sieme H.The research of fertility in humans and other mammals has strongly advanced in the recent years. The examination of molecular mechanisms influencing horse fertility is relatively recent. We chose the angiotensin converting enzyme (ACE), the sperm autoantigenic protein 17 (SP17) and the follicle stimulating hormone (FSHB) as candidates for determining stallion fertility and to analyze associations of intragenic single nucleotide polymorphisms (SNPs), flanking microsatellites and candidate-gene linked haplotypes with the pregnancy rate per oestrus (PRO) in 179 Hanoverian stallions. Fertility tra...
Gametic phase disequilibrium between the syntenic multiallelic HTG4 and HMS3 markers widely used for parentage testing in Thoroughbred horses.
Molecular biology reports    May 24, 2011   Volume 39, Issue 2 1447-1452 doi: 10.1007/s11033-011-0881-4
Machado FB, de Vasconcellos Machado L, Bydlowski CR, Bydlowski SP, Medina-Acosta E.Validation of parentage and horse breed registries through DNA typing relies on estimates of random match probabilities with DNA profiles generated from multiple polymorphic loci. Of the twenty-seven microsatellite loci recommended by the International Society for Animal Genetics for parentage testing in Thoroughbred horses, eleven are located on five chromosomes. An important aspect in determining combined exclusion probabilities is the ascertainment of the genetic linkage status of syntenic markers, which may affect reliable use of the product rule in estimating random match probabilities. I...
Consistent detection of bovine papillomavirus in lesions, intact skin and peripheral blood mononuclear cells of horses affected by hoof canker.
Equine veterinary journal    May 20, 2011   Volume 43, Issue 2 202-209 doi: 10.1111/j.2042-3306.2010.00147.x
Brandt S, Schoster A, Tober R, Kainzbauer C, Burgstaller JP, Haralambus R, Steinborn R, Hinterhofer C, Stanek C.Equine hoof canker is a chronic proliferative pododermatitis of as yet unknown aetiology. Like equine sarcoid disease, canker is a therapy-resistant disorder characterised by hyperkeratosis, acanthosis and a marked tendency to recur. Objective: There is an association of sarcoid-inducing bovine papillomaviruses of types 1 and 2 (BPV-1, BPV-2) with hoof canker disease. Methods: Using PCR-based techniques, we assessed canker tissue, intact skin and/or peripheral blood mononuclear cells (PBMCs) of 25 canker-affected horses for the presence of sarcoid-associated BPV-1 and -2. Results: Conventional...
β-Tubulin genotypes in six species of cyathostomins from anthelmintic-naive Przewalski and benzimidazole-resistant brood horses in Ukraine.
Parasitology research    May 7, 2011   Volume 109, Issue 4 1199-1203 doi: 10.1007/s00436-011-2426-0
Blackhall WJ, Kuzmina T, von Samson-Himmelstjerna G.Resistance to benzimidazoles (BZ) in the gastrointestinal nematodes of livestock is characterised by the presence of specific polymorphisms in the β-tubulin isotype 1 protein, a component of microtubules. The most prevalent polymorphism associated with resistance in nematodes infecting cattle, sheep, and goats is found at codon 200, with minor occurrences of polymorphisms at codons 167 and 198. In the cyathostomins that infect horses, however, a polymorphism at codon 167 appears to be more common than the codon 200 polymorphism. In the present study, a focussed analysis of PCR-amplified β-tu...
Direct detection of equine herpesvirus type 1 DNA in nasal swabs by loop-mediated isothermal amplification (LAMP).
The Journal of veterinary medical science    May 6, 2011   Volume 73, Issue 9 1225-1227 doi: 10.1292/jvms.11-0065
Nemoto M, Ohta M, Tsujimura K, Bannai H, Yamanaka T, Kondo T, Matsumura T.We evaluated loop-mediated isothermal amplification (LAMP) as a means of detecting equine herpesvirus type 1 (EHV-1) DNA directly from nasal swabs. To increase the sensitivity, we added a step in which the samples were heat-treated to the original LAMP procedure. The detection limit of the LAMP assay with heat treatment was 10 times more sensitive than the original LAMP assay even when the DNA extraction step was omitted. In addition, the LAMP assay with heat treatment was more sensitive than the original LAMP assay and the polymerase chain reaction using clinical samples. The LAMP assay with ...
Age-related changes in genomic stability of horses.
Mechanisms of ageing and development    April 30, 2011   Volume 132, Issue 5 257-268 doi: 10.1016/j.mad.2011.04.009
Wnuk M, Bugno-Poniewierska M, Lewinska A, Oklejewicz B, Zabek T, Bartosz G, Słota E.Recently, the old horse has been proposed as a model to study telomere-dependent senescence, immunosenescence and inflamm-aging. In the present paper, we used 80 Hucul and Anglo-Arabian horses divided into 3 age groups (juvenile, adult, old) to evaluate age-dependent changes at the genomic and DNA level and in cell proliferative potential. The level of positive TUNEL cells (both apoptotic and with DNA fragmentation), oxidative DNA damage (8-oxoG immunostaining), sister chromatid exchange and bleomycin-induced chromatid breaks were significantly increased in the combined old group compared to t...
The association of the presence of seminal plasma and its components with sperm longevity in fractionated stallion ejaculates.
Reproduction in domestic animals = Zuchthygiene    April 28, 2011   Volume 46, Issue 6 1073-1081 doi: 10.1111/j.1439-0531.2011.01789.x
Kareskoski M, Sankari S, Johannisson A, Kindahl H, Andersson M, Katila T.Semen jets in the stallion's ejaculate differ in both the biochemical composition of seminal plasma (SP) and in sperm concentration, forming sperm-rich fractions (HIGH) and sperm-poor fractions (LOW). This study examined (i) the association of the levels of selected components of SP [alkaline phosphatase (AP), acid phosphatase (ACP), Na(+), Cl(-), K(+), Ca, Mg and prostaglandin E(2)] with semen quality, sperm longevity and fertility; (ii) the effects of the presence of SP on sperm survival during cooled storage; (iii) the differences in the composition of SP between sperm-rich and sperm-poor e...
[Molecular basis and applicability in equine color genetics].
Yi chuan = Hereditas    April 26, 2011   Volume 32, Issue 11 1133-1140 
Li B, He XL, Zhao YP, Wang XJ, Manglai D, Zhang YR.Coat color can be used not only in identifying individuals and species, but also in diagnosing certain diseases. The coat color of horse mainly depends on the distribution and proportion of eumelanin and phaeomelanin, generated by melanocytes, which are regulated by the genes such as MC1R, ASIP, KIT, TYRP, and EDNRB. In addition, STX17, MATP, and PMEL17 also play a role in the formation of coat color of horse. In this review, the action mechanism of candidate genes for coat color and association of DNA sequence polymorphism of these candidate genes with color pattern phenotypes and melanoma we...
A cohort study of racing performance in Japanese Thoroughbred racehorses using genome information on ECA18.
Animal genetics    April 19, 2011   Volume 43, Issue 1 42-52 doi: 10.1111/j.1365-2052.2011.02201.x
Tozaki T, Hill EW, Hirota K, Kakoi H, Gawahara H, Miyake T, Sugita S, Hasegawa T, Ishida N, Nakano Y, Kurosawa M.Using 1710 Thoroughbred racehorses in Japan, a cohort study was performed to evaluate the influence of genotypes at four single nucleotide polymorphisms (SNPs) on equine chromosome 18 (ECA18), which were associated in a previous genome-wide association study for racing performance with lifetime earnings and performance rank. In males, both g.65809482T>C and g.65868604G>T were related to performance rank (P= 0.005). In females, g.65809482T>C (P = 1.76E-6), g.65868604G>T (P=6.81E-6) and g.66493737C>T (P=4.42E-5) were strongly related to performance rank and also to lifetime ...
Congenital stationary night blindness is associated with the leopard complex in the Miniature Horse.
Veterinary ophthalmology    April 19, 2011   Volume 15, Issue 1 18-22 doi: 10.1111/j.1463-5224.2011.00903.x
Sandmeyer LS, Bellone RR, Archer S, Bauer BS, Nelson J, Forsyth G, Grahn BH.  To determine if congenital stationary night blindness (CSNB) exists in the Miniature Horse in association with leopard complex spotting patterns (LP), and to investigate if CSNB in the Miniature Horse is associated with three single nucleotide polymorphisms (SNPs) in the region of TRPM1 that are highly associated with CSNB and LP in Appaloosas. Methods:   Three groups of Miniature Horses were studied based on coat patterns suggestive of LP/LP (n=3), LP/lp (n=4), and lp/lp genotype (n=4). Methods:   Horses were categorized based on phenotype as well as pedigree analysis as LP/LP, LP/lp,...
Population studies of 17 equine STR for forensic and phylogenetic analysis.
Animal genetics    April 14, 2011   Volume 42, Issue 6 627-633 doi: 10.1111/j.1365-2052.2011.02194.x
van de Goor LH, van Haeringen WA, Lenstra JA.As a consequence of the close integration of horses into human society, equine DNA analysis has become relevant for forensic purposes. However, the information content of the equine Short Tandem Repeat (STR) loci commonly used for the identification or paternity testing has so far not been fully characterized. Population studies were performed for 17 polymorphic STR loci (AHT4, AHT5, ASB2, ASB17, ASB23, CA425, HMS1, HMS2, HMS3, HMS6, HMS7, HTG4, HTG6, HTG7, HTG10, LEX3 and VHL20) including 8641 horses representing 35 populations. The power of parental exclusion, polymorphic information content...
Genomic structure and expression analyses of the PYGM gene in the thoroughbred horse.
Zoological science    April 7, 2011   Volume 28, Issue 4 276-280 doi: 10.2108/zsj.28.276
Nam GH, Ahn K, Bae JH, Han K, Lee CE, Park KD, Lee SH, Cho BW, Kim HS.Muscle glycogen Phosphorylase (PYGM) has been shown to catalyze the degradation of glycogen to glucose-1-phosphate. The PYGM gene can contribute to providing energy to the body by disassembling the glycogen in muscle. Here, we analyzed the genomic structure and expression of the PYGM gene in the thoroughbred horse. The PYGM gene, containing several transposable elements (MIRs, LINEs, and MERs), was highly conserved in mammalian genomes. In order to understand the expression of the horse PYGM gene, we performed quantitative RT-PCR using 11 thoroughbred horse tissue samples. The horse PYGM gene ...
European domestic horses originated in two holocene refugia.
PloS one    March 30, 2011   Volume 6, Issue 3 e18194 doi: 10.1371/journal.pone.0018194
Warmuth V, Eriksson A, Bower MA, Cañon J, Cothran G, Distl O, Glowatzki-Mullis ML, Hunt H, Luís C, do Mar Oom M, Yupanqui IT, Ząbek T, Manica A.The role of European wild horses in horse domestication is poorly understood. While the fossil record for wild horses in Europe prior to horse domestication is scarce, there have been suggestions that wild populations from various European regions might have contributed to the gene pool of domestic horses. To distinguish between regions where domestic populations are mainly descended from local wild stock and those where horses were largely imported, we investigated patterns of genetic diversity in 24 European horse breeds typed at 12 microsatellite loci. The distribution of high levels of gen...
Equine sperm nuclei with different ploidy levels: relationship between the nuclear DNA content and the nuclear area.
Andrologia    March 23, 2011   Volume 43, Issue 4 248-253 doi: 10.1111/j.1439-0272.2010.01059.x
Spirito S, Campi S, Boquet M, Fernández H, Ferrari M.The aims of this study were to estimate the ability of the Feulgen reaction to identify equine sperm nuclei with different ploidy levels, to determine the frequency of haploid, diploid and polyploid sperm nuclei in the semen of fertile equines and to evaluate the relationship between the nuclear DNA content and the nuclear area. Determination of the ploidy level of Feulgen-stained spermatozoa using a scanning microspectrophotometer was very similar to the subjective estimations made with a light microscope. This indicates that the Feulgen reaction is a simple, inexpensive and reliable techniqu...
Comparative evolutionary analyses of beta globin gene in eutherian, dinosaurian and neopterygii taxa.
Journal of vector borne diseases    March 17, 2011   Volume 48, Issue 1 27-36 
Awasthi G, Srivastava G, Das A.Comparative genomics and evolutionary analyses of conserved genes have enabled us to understand the complexity of genomes of closely related species. For example: β-globin gene present in human hemoglobin is one such gene that has experienced many genetic changes in many related taxa and produced more than 600 variants. One of the variant, HBS causes sickle-cell anemia in humans but offers protection against severe malaria due to Plasmodium falciparum. In the present study, we characterized and performed evolutionary comparative analyses of the ?-globin gene in different related and unrelated...
A suspect horse excluded by analysis of species- and male-specific DNA and mtDNA.
Forensic science international    March 3, 2011   Volume 207, Issue 1-3 e66-e68 doi: 10.1016/j.forsciint.2011.02.006
Xu H, Han W, Zhao C, Manglai D.No abstract available
Immunomodulation during and after castration under inhalation anaesthetic without genotoxic effects on equine lymphocytes.
Research in veterinary science    February 26, 2011   Volume 92, Issue 2 306-310 doi: 10.1016/j.rvsc.2011.01.021
Strasser A, Kühnel H, Velde K, Dadak A.Genotoxic DNA damage due to inhalation anaesthesia has been demonstrated in human lymphocytes. In order to evaluate anaesthesia-associated changes in cell-mediated immunity on the basis of a potential DNA damage as a health risk in horses, single cell gel electrophoresis and lymphocyte proliferation assay were performed on equine lymphocytes which were obtained before, during and after regular castration under inhalation anaesthetic. No significant lymphocytic DNA damage due to isoflurane anaesthesia was observed, whereas lymphocyte proliferative reactivity and lymphocyte counts decreased sign...
Five novel KIT mutations in horses with white coat colour phenotypes.
Animal genetics    February 23, 2011   Volume 42, Issue 3 337-339 doi: 10.1111/j.1365-2052.2011.02173.x
Haase B, Rieder S, Tozaki T, Hasegawa T, Penedo MC, Jude R, Leeb T.No abstract available
Host blood meal identification by multiplex polymerase chain reaction for dispersal evidence of stable flies (Diptera:Muscidae) between livestock facilities.
Journal of medical entomology    February 23, 2011   Volume 48, Issue 1 53-60 doi: 10.1603/me10123
Pitzer JB, Kaufman PE, Tenbroeck SH, Maruniak JE.A species-specific multiplex polymerase chain reaction targeting the cytochrome b gene of cattle, horses, humans, and dogs was developed to determine the blood meal sources of stable flies, Stomoxys calcitrans (L.), collected from Florida equine facilities. Of 595 presumptive blood-fed stable flies analyzed, successful host amplification was obtained in 350, for a field host-detection efficiency of 58.8%. The majority of analyzed stable flies had fed on cattle (64.6%), followed by horses (24.3%), humans (9.5%), and dogs (1.6%). A survey of animal-enclosed pastures occurring within 3 km of stab...
Autologous chondrocyte implantation drives early chondrogenesis and organized repair in extensive full- and partial-thickness cartilage defects in an equine model.
Journal of orthopaedic research : official publication of the Orthopaedic Research Society    February 11, 2011   Volume 29, Issue 7 1121-1130 doi: 10.1002/jor.21366
Nixon AJ, Begum L, Mohammed HO, Huibregtse B, O'Callaghan MM, Matthews GL.Autologous chondrocyte implantation (ACI) has been used clinically for over 15 years and yet definitive evidence of chondrocyte persistence and direct impact on cartilage repair in full-thickness lesions is scant and no data are available on ACI in partial-thickness defects in any animal model. This study assessed the effect of chondrocytes secured using periosteal overlay in partial- and full-thickness cartilage defects in the equine model. Paired cartilage defects 15 mm in diameter were made in the patellofemoral joint of 16 horse and repaired with ACI or periosteal flap alone. Response wa...
Successful treatment of hyperkalaemic periodic paralysis in a horse during isoflurane anaesthesia.
Veterinary anaesthesia and analgesia    February 10, 2011   Volume 38, Issue 2 113-120 doi: 10.1111/j.1467-2995.2010.00592.x
Pang DS, Panizzi L, Paterson JM.A 3-year-old, 400 kg, gelding Quarter Horse was presented for investigation of epistaxis. Methods: The horse was bright, alert and responsive with rectal temperature, heart rate and respiration rate within normal limits. Results: During a second general anaesthetic for surgical treatment of guttural pouch mycosis by balloon-tipped catheter occlusion of the right major palatine artery and ligation of the right external carotid artery, signs consistent with hyperkalaemic periodic paralysis (HYPP) were exhibited. These included concurrent hyperkalaemia, hypercapnoea, sinus tachycardia, and muscle...
Identification of Anaplasma spp. rickettsia isolated from horses from clinical disease cases in Poland.
Zoonoses and public health    February 1, 2011   Volume 58, Issue 7 514-518 doi: 10.1111/j.1863-2378.2011.01394.x
Adaszek Ł, Winiarczyk S.This study was aimed at determining the cause of the diseases in five horses exhibiting symptoms of fever, joint oedema and ataxia and thrombocytopenia. The PCR technique revealed the presence in the blood of 16S RNA Anaplasma/Ehrlichia spp. genetic material. DNA amplification with primers EHR 521 and EHR 747 gave a product with a size of 247 bp. The sequence of the PCR product obtained showed a 97.6-99.6% similarity with a sequence of a fragment of 16S RNA Anaplasma phagocytophilum, gene number EU 090186 from GenBank. Intravenous administration of oxytetracycline at a dose of 8 mg/kg of body ...
Lyme neuroborreliosis in 2 horses.
Veterinary pathology    February 1, 2011   Volume 48, Issue 6 1151-1157 doi: 10.1177/0300985811398246
Imai DM, Barr BC, Daft B, Bertone JJ, Feng S, Hodzic E, Johnston JM, Olsen KJ, Barthold SW.Lyme neuroborreliosis--characterized as chronic, necrosuppurative to nonsuppurative, perivascular to diffuse meningoradiculoneuritis--was diagnosed in 2 horses with progressive neurologic disease. In 1 horse, Borrelia burgdorferi sensu stricto was identified by polymerase chain reaction amplification of B burgdorferi sensu stricto-specific gene targets (ospA, ospC, flaB, dbpA, arp). Highest spirochetal burdens were in tissues with inflammation, including spinal cord, muscle, and joint capsule. Sequence analysis of ospA, ospC, and flaB revealed 99.9% sequence identity to the respective genes in...
Equine penile squamous cell carcinomas are associated with the presence of equine papillomavirus type 2 DNA sequences.
Veterinary pathology    January 31, 2011   Volume 48, Issue 6 1190-1194 doi: 10.1177/0300985810396516
Knight CG, Munday JS, Peters J, Dunowska M.Forty cases of equine penile disease were screened with polymerase chain reaction for the presence of papillomaviral DNA. Cases consisted of 20 squamous cell carcinomas (average age of horse, 23.9 years) and 20 non-squamous cell carcinoma diseases (average age of horse, 13.3 years). All horses but one originated from the Northeastern United States. Breeds were not recorded. As based on MY09/MY11 consensus primers, DNA sequences from equine papillomavirus type 2 were amplified from 9 of 20 horses (45%) with penile squamous cell carcinoma and only 1 of 20 horses (5%) with non-squamous cell carci...
Identification of Arcanobacterium haemolyticum isolated from postcastrational complications of a horse.
Folia microbiologica    January 21, 2011   Volume 55, Issue 6 666-668 doi: 10.1007/s12223-010-0108-4
Hijazin M, Ulbegi-Mohyla H, Alber J, Lämmler C, Hassan AA, Prenger-Berninghoff E, Weiss R, Zschöck M.An Arcanobacterium haemolyticum strain isolated from a postcastrational lesion of a horse was identified phenotypically and genotypically. The latter was performed by sequencing the 16S-23S rDNA intergenic spacer region (ISR), by amplification of the gene encoding A. haemolyticum phospholipase D, by amplification of A. haemolyticum specific parts of ISR-23S rDNA and by amplification of the newly described CAMP factor family protein encoding gene of A. haemolyticum. This indicates (as described previously for seven additional A. haemolyticum strains; Hassan et al. 2009) that A. haemolyticum see...
Transcriptional profiling of equine conceptuses reveals new aspects of embryo-maternal communication in the horse.
Biology of reproduction    January 5, 2011   Volume 84, Issue 5 872-885 doi: 10.1095/biolreprod.110.088732
Klein C, Troedsson MH.Establishment and maintenance of pregnancy are critically dependent on embryo-maternal communication during the preimplantation period. The horse is one of the few domestic species in which the conceptus-derived pregnancy recognition signal has not been identified. To gain new insights into the factors released by the equine conceptus, transcriptional profiling analyses of conceptuses retrieved 8, 10, 12, and 14 days after ovulation were performed using a whole-genome microarray. Selected array data were confirmed using quantitative PCR, and the expression of proteins of interest was confirmed...
Epidemiology of equine sarcoids in horses in western Canada.
The Canadian veterinary journal = La revue veterinaire canadienne    January 5, 2011   Volume 51, Issue 10 1103-1108 
Wobeser BK, Davies JL, Hill JE, Jackson ML, Kidney BA, Mayer MN, Townsend HG, Allen AL.Sarcoids are the most common tumor of the equine skin but only 1 study describing the epidemiology of sarcoids in Canadian horses has been published. The records of 5 veterinary diagnostic laboratories in western Canada were searched to identify submissions of sarcoids from horses. The submission records and diagnostic reports of 802 separate submissions of equine sarcoids were reviewed for age, breed, and gender of the horse and the number, location, and clinical type of sarcoid. From these records, the 307 submissions to laboratories in Saskatchewan were compared to a reference group to test...
Phylogenetic analysis of bovine papillomavirus E5 detected in equine sarcoids in Poland.
Polish journal of veterinary sciences    January 1, 2011   Volume 14, Issue 4 653-654 doi: 10.2478/v10181-011-0096-6
Szczerba-Turek A, Siemionek J, Bancerz-Kisiel A, Raś A, Szweda W.The aim of the study was to analyse a part of the sequence of the E5 gene of bovine papillomaviruses (BPV) associated with equine sarcoids in Polish horses. Samples of 40 skin lesions obtained from 29 horses were collected for molecular examination. The PCR amplicons of BPV DNA were detected in 38 specimens. After phylogenetic analysis 37 specimens were recognized as BPV-1 and one as BPV-2. Phylogenetic analysis has allowed the classification of the amplicons into two phylogenetic groups (A1,) and four separate isolates (2, 10, 16, 17).
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