Clinical, cytogenetic and molecular genetic characterization of a tandem fusion translocation in a male Holstein cattle with congenital hypospadias and a ventricular septal defect

Autoři: Alessandra Iannuzzi aff001;  Marina Braun aff002;  Viviana Genualdo aff001;  Angela Perucatti aff001;  Sina Reinartz aff002;  Ioannis Proios aff003;  Maike Heppelmann aff003;  Jürgen Rehage aff003;  Kirsten Hülskötter aff004;  Andreas Beineke aff004;  Julia Metzger aff002;  Ottmar Distl aff002
Působiště autorů: Institute for the Animal Production System in Mediterranean Environment (ISPAAM), National Research Council (CNR), Naples, Italy aff001;  Institute for Animal Breeding and Genetics, University of Veterinary Medicine Hannover, Hannover, Germany aff002;  Clinic for Cattle, University of Veterinary Medicine Hannover, Hannover, Germany aff003;  Department of Pathology, University of Veterinary Medicine Hannover, Hannover, Germany aff004
Vyšlo v časopise: PLoS ONE 15(1)
Kategorie: Research Article
doi: 10.1371/journal.pone.0227117


Hypospadias, disorder of sex development (DSD), is a sporadic congenital abnormality of the genital region in male ruminants, which is characterized by a non-fused urethra during fetal development. Detailed clinical examination classified the hypospadias phenotype of a male Holstein calf studied here as the perineal type. In combined use of cytogenetic analysis and whole genome sequencing, a non-mosaic, pseudo-monosomy 59, XY + tan(18;27) was detected. This chromosomal aberration had its origin in a tandem fusion translocation of the bovine autosomes (BTA) 18 and 27 with an accompanying loss of genomic sequences mainly in the distal end of BTA 18 and the proximal end of BTA 27. The resulting phenotype included hypospadias, growth retardation and ventricular septal defect.

Klíčová slova:

Cattle – Genome sequencing – Metaphase – Penis – Telomeres – Y chromosomes – Hypospadias – Chromosomal translocations


1. Nassar N, Bower C, Barker A. Increasing prevalence of hypospadias in Western Australia, 1980–2000. Archives of disease in childhood. 2007;92(7):580–4. doi: 10.1136/adc.2006.112862 17405856

2. Goldblatt J, Wallis C, Viljoen D. A New hypospadias-mental retardation syndrome in three brothers. American Journal of Diseases of Children. 1987;141(11):1168–9. 3673966

3. Fredell L, Iselius L, Collins A, Hansson E, Holmner S, Lundquist L, et al. Complex segregation analysis of hypospadias. Human Genetics. 2002;111(3):231–4. doi: 10.1007/s00439-002-0799-y 12215834

4. Fukami M, Wada Y, Miyabayashi K, Nishino I, Hasegawa T, Nordenskjold A, et al. CXorf6 is a causative gene for hypospadias. Nature Genetics. 2006;38(12):1369–71. doi: 10.1038/ng1900 17086185

5. Van der Zanden L, Van Rooij I, Feitz W, Franke B, Knoers N, Roeleveld N. Aetiology of hypospadias: a systematic review of genes and environment. Human reproduction update. 2012;18(3):260–83. doi: 10.1093/humupd/dms002 22371315

6. King G, Johnson E. Hypospadias in a Himalayan cat. Journal of Small Animal Practice. 2000;41(11):508–10. doi: 10.1111/j.1748-5827.2000.tb03973.x 11105790

7. Rahal SC, da Mota LS, Mamprim MJ, Ciani RB. Perineal hypospadias in a dog. In: [internet]. [cited 15 July 2019]

8. Brink P, Schumacher J. Hypospadias in a Friesian gelding. Veterinary Surgery. 2011;40(1):120–3. doi: 10.1111/j.1532-950X.2010.00762.x 21198681

9. Bleul U, Theiss F, Rütten M, Kähn W. Clinical, cytogenetic and hormonal findings in a stallion with hypospadias–a case report. The Veterinary Journal. 2007;173(3):679–82. doi: 10.1016/j.tvjl.2006.03.005 16820309

10. De Lorenzi L, Genualdo V, Iannuzzi A, Di Meo GP, Perucatti A, Mancuso R, et al. Cytogenetic and genetic studies in a hypospadic horse (Equus caballus, 2n = 64). Sexual development: genetics, molecular biology, evolution, endocrinology, embryology, and pathology of sex determination and differentiation. 2010;4(6):352–7.

11. Röder P. Hypospadie beim Pferd. Berliner und Münchener Tierärztliche Wochenschrift. 1929;47:797–8.

12. Smith K, Brown P, Parkinson T. Hypospadias in rams. Veterinary Record: Journal of the British Veterinary Association. 2006;158(23).

13. Sakhaee E, Azari O. Hypospadias in goats. Iranian Journal of Veterinary Research. 2009;10(3):298–301.

14. Misk NA, Misk TN, Semieka MA. Diagnosis and treatment of affections of the urethra in male ruminants: A review of 403 cases. International Journal of Veterinary Medicine: Research and Reports. 2013;Article ID 715907.

15. Kumi-Diaka J, Osori D. Perineal hypospadias in two related bull calves, a case report. Theriogenology. 1979;11(2):163–4. doi: 10.1016/0093-691x(79)90068-2 16725401

16. Cassata R, Iannuzzi A, Parma P, De Lorenzi L, Peretti V, Perucatti A, et al. Clinical, cytogenetic and molecular evaluation in a dog with bilateral cryptorchidism and hypospadias. Cytogenetic and Genome Research. 2008;120(1–2):140–3. doi: 10.1159/000118753 18467838

17. Switonski M, Payan-Carreira R, Bartz M, Nowacka-Woszuk J, Szczerbal I, Colaco B, et al. Hypospadias in a male (78,XY; SRY-positive) dog and sex reversal female (78,XX; SRY-negative) dogs: clinical, histological and genetic studies. Sexual development: genetics, molecular biology, evolution, endocrinology, embryology, and pathology of sex determination and differentiation. 2012;6(1–3):128–34.

18. Switonski M, Dzimira S, Aleksiewicz R, Szczerbal I, Nowacka-Woszuk J, Krzeminska P, et al. Hypospadias Is Not Rare in Dogs: Five new cases, a retrospective study, and a review of the literature. Sexual development: genetics, molecular biology, evolution, endocrinology, embryology, and pathology of sex determination and differentiation. 2018;12(5):244–50.

19. Hynes P, Fraher J. The development of the male genitourinary system. I. The origin of the urorectal septum and the formation of the perineum. British Journal of Plastic Surgery. 2004;57(1):27–36. doi: 10.1016/j.bjps.2003.08.019 14672675

20. Kluth D, Lambrecht W, Reich P. Pathogenesis of hypospadias—more questions than answers. Journal of Pediatric Surgery. 1988;23(12):1095–101. doi: 10.1016/s0022-3468(88)80322-1 3236174

21. Yamada G, Satoh Y, Baskin LS, Cunha GR. Cellular and molecular mechanisms of development of the external genitalia. Differentiation. 2003;71(8):445–60. doi: 10.1046/j.1432-0436.2003.7108001.x 14641326

22. Switonski M, Payan-Carreira R, Bartz M, Nowacka-Woszuk J, Szczerbal I, Colaço B, et al. Hypospadias in a male (78, XY; SRY-positive) dog and sex reversal female (78, XX; SRY-negative) dogs: clinical, histological and genetic studies. Sexual development: genetics, molecular biology, evolution, endocrinology, embryology, and pathology of sex determination and differentiation. 2011;6(1–3):128–34.

23. Alam M, Shin S, Lee H, Choi I, Kim N. Hypospadias in three calves: a case report. Veterinarni Medicina -Praha-. 2005;50(11):506.

24. Usta Z, Distl O. Atresia ani, hypospadia and rudimentary external genitalia in two German Holstein calves. Atatürk Üniversitesi Veteriner Bilimleri Dergisi. 2017;12(2):195–202.

25. Jurka P, Galanty M, Zielinska P, Max A, Sysa P. Hypospadias in six dogs. The Veterinary Record. 2009;164(11):331–3. doi: 10.1136/vr.164.11.331 19287029

26. McFarland L. Perineal hypospadias in male lambs. Journal of the American Veterinary Medical Association. 1958;133(1):81–2. 13549337

27. Phiri AM, Sakala M, Soko J. Management of atresia ani with concurrent hypospadias and penile aplasia in a 3-day-old calf. International Journal of Livestock Research. 2016;6(1):98–103.

28. Abd-El-Hady A, El-Din MMM. Hypospadia and urethral diverticulum in a female pseudohermaphrodite calf. Scholars Journal of Agriculture and Veterinary Sciences. 2014;1(4B):288–292.

29. Maiti S, Raghuvanshi P, Divya M, Sangeetha P, Deepesh G, Naveen K. Surgical correction of urethral diverticulum in a female pseudo-hermaphrodite crossbred calf. Iranian Journal of Veterinary Research. 2018;19(1):57. 29805465

30. Misk N, Rilat J. Surgical management of congenital anomalies of ruminants. Agri-Practice. 1984;5:38–47.

31. Murakami T. Anatomical examination of hypospadias in cattle. Journal of the Japan Veterinary Medical Association (Japan). 2008;61:931–935.

32. Jeong W-S, Seong Y-S, Kim I-S, Jang H-S, Oh T-H, Lee K-W, et al. Radiographic diagnosis of hypospadias in three Korean native calves. Journal of Veterinary Clinics. 2008;25(6):557–9.

33. Pamuk K, Korkmaz M, Saritaş ZK, Orhan İÖ. Simental Irkı Bir Buzağıda Hypospadiasis ve Urogenital Anomali. Kocatepe Veterinary Journal. 2010;3(2).

34. Vidal G, Traslavina RP, Cabrera C, Gaffney PM, Lane VM. Theriogenology question of the month. Journal of the American Veterinary Medical Association. 2011;239(10):1295–7. doi: 10.2460/javma.239.10.1295 22044324

35. Park Y-S, Yang H-S, Ko M-H, Ko J-S, Cho S-R, Kim N-Y, et al. A case of urinary tract infection in a calf with hypospadias. Journal of Veterinary Clinics 2012;29(4):352–5.

36. Mihsler L, Hussein H, Wehrend A. Perineal hypospadia in a red Holstein-calf. Tierärztliche Praxis Großtiere. 2015;43(2):105–8.

37. Potliya S, Chaudhary R, Pal M, Kumar S, Kumar S. Hypospadias with atresia ani in a cow calf. Haryana Veterinarian 2016;55(1):118–9.

38. Torres AAA, Lhamas CL, da Graça Macoris D, de Oliveira Vasconcelos R. Macroscopic and microscopic findings in a set of congenital anomalies in two calves produced through in vitro production. Brazilian Journal of Veterinary Pathology. 2017;6(2):65–8.

39. Volpato R, Araujo MS, Vexenate SC, Oba E. Hypospadias in buffalo (Bubalus Bubalis) mediterranean breed. Journal of Buffalo Science. 2013;2:85–7.

40. Smith K, Brown P, Barr F. A survey of congenital reproductive abnormalities in rams in abattoirs in south west England. Reproduction in Domestic Animals. 2012;47(5):740–5. doi: 10.1111/j.1439-0531.2011.01952.x 22136299

41. Veena P, Sankar P, S RV, Kokila KS. Congenital absence of tail with atresia ani and hypospadias in a kid-A case report. Journal of Advanced Veterinary Research. 2011;1(2):76–9.

42. Omidi A, Monjezi S, Nabipour A. Hypospadias, diverticulum, and agenesis in the penile shaft of a goat kid (Capra hircus). Comparative Clinical Pathology. 2011;20(4):417–9.

43. Bokhari S. Hypospadias and urethral diverticulum in two goat kids: a case report. Journal of Animal and Plant Science. 2013;23:675–7.

44. Almubarak AM, Abdelghafar RM, Gameel AA, Osman NM. Penile urethral hypospadias with two fistulae and diverticulum in a Saanen kid. Hindawi Publishing Corporation. 2016;Article ID 6534062.

45. Ducos A, Revay T, Kovacs A, Hidas A, Pinton A, Bonnet-Garnier A, et al. Cytogenetic screening of livestock populations in Europe: an overview. Cytogenetic and Genome Research. 2008;120(1–2):26–41. doi: 10.1159/000118738 18467823

46. Pinheiro LEL, Carvalho TB, Oliveira DAA, Popescu CP, Basrur PK. A 4/21 tandem fusion in cattle. Hereditas. 1995;122:99:102. doi: 10.1111/j.1601-5223.1995.00099.x 7558885

47. Popescu CP. Les anomalies chromosomiques des bovins (Bos taurus L.). Etat actuel des connaissances. Annales de Génétique et de Sélection Animale. 1977;9(4):463:70. doi: 10.1186/1297-9686-9-4-463 22896549

48. Bruere A, McDonald M, Marshall R. Cytogenetical analysis of an ovine male pseudohermaphrodite and the possible role of the Y chromosome in cryptorchidism of sheep. Cytogenetic and Genome Research. 1969;8(2):148–57.

49. Thauvin‐Robinet C, Faivre L, Cusin V, Khau Van Kien P, Callier P, Parker KL, et al. Cloacal exstrophy in an infant with 9q34. 1‐qter deletion resulting from a de novo unbalanced translocation between chromosome 9q and Yq. American Journal of Medical Genetics Part A. 2004;126(3):303–7.

50. Horiike S, Taniwaki M, Misawa S, Nishigaki H, Okuda T, Yokota S, et al. The unbalanced 1; 7 translocation in de novo myelodysplastic syndrome and its clinical implication. Cancer. 1990;65(6):1350–4. doi: 10.1002/1097-0142(19900315)65:6<1350::aid-cncr2820650617>;2-j 2306683

51. Riegel M, Hargreaves P, Baumer A, Guc-Scekic M, Ignjatovic M, Schinzel A. Unbalanced 18q/21q translocation in a patient previously reported as monosomy 21. European Journal of Medical Genetics. 2005;48(2):167–74. doi: 10.1016/j.ejmg.2005.01.026 16053908

52. Zneimer SM, Cotter PD, Stewart SD. Telomere‐telomere (end to end) fusion of chromosomes 7 and 22 with an interstitial deletion of chromosome 7p11. 2→ p15. 1: phenotypic consequences and possible mechanisms. Clinical Genetics. 2000;58(2):129–33. doi: 10.1034/j.1399-0004.2000.580207.x 11005146

53. Van der Zanden LF, Van Rooij IA, Feitz WF, Knight J, Donders ART, Renkema KY, et al. Common variants in DGKK are strongly associated with risk of hypospadias. Nature Genetics. 2011;43(1):48. doi: 10.1038/ng.721 21113153

54. Richard MA, Sok P, Canon S, Brown AL, Peckham‐Gregory EC, Nembhard WN, et al. The role of genetic variation in DGKK on moderate and severe hypospadias. Birth Defects Research. 2019; 111:932–937. doi: 10.1002/bdr2.1522 31102501

55. Kalfa N, Philibert P, Sultan C. Is hypospadias a genetic, endocrine or environmental disease, or still an unexplained malformation? International Journal of Andrology. 2009;32(3):187–97. doi: 10.1111/j.1365-2605.2008.00899.x 18637150

56. Stratmann N, Wehrend A. Perineale Hypospadie bei einem Chihuahua-Mischling. Ein Fallbericht. Tierärztliche Praxis Kleintiere. 2004;32(5):294–7.

57. Saunders P, Ladds P. Congenital and developmental anomalies of the genitalia of slaughtered bulls. Australian Veterinary Journal. 1978;54(1):10–3. doi: 10.1111/j.1751-0813.1978.tb00261.x 26328

58. Herzog A. Pareys Lexikon der Syndrome: Erb-und Zuchtkrankheiten der Haus-und Nutztiere. Parey; 2001.

59. Hynes P, Fraher J. The development of the male genitourinary system: II. The origin and formation of the urethral plate. British Journal of Plastic Surgery. 2004;57(2):112–21. doi: 10.1016/j.bjps.2003.08.018 15037165

60. Buczinski S, Fecteau G, DiFruscia R. Ventricular septal defects in cattle: A retrospective study of 25 cases. The Canadian Veterinary Journal. 2006;47(3):246. 16604981

61. Iannuzzi L, Di Meo GP, Leifsson PS, Eggen A, Christensen K. A case of trisomy 28 in cattle revealed by both banding and FISH-mapping techniques. Hereditas. 2001;134(2):147–51. doi: 10.1111/j.1601-5223.2001.00147.x 11732851

62. Berry D, Wolfe A, O’Donovan J, Byrne N, Sayers R, Dodds K, et al. Characterization of an X-chromosomal non-mosaic monosomy (59, X0) dairy heifer detected using routinely available single nucleotide polymorphism genotype data. Journal of Animal Science. 2017;95(3):1042–9. doi: 10.2527/jas.2016.1279 28380529

63. Fiorentino F, Bono S, Biricik A, Nuccitelli A, Cotroneo E, Cottone G, et al. Application of next-generation sequencing technology for comprehensive aneuploidy screening of blastocysts in clinical preimplantation genetic screening cycles. Human Reproduction. 2014;29(12):2802–13. doi: 10.1093/humrep/deu277 25336713

64. Metzger J, Rau J, Naccache F, Conn LB, Lindgren G, Distl O. Genome data uncover four synergistic key regulators for extremely small body size in horses. BMC Genomics. 2018;19(1):492. doi: 10.1186/s12864-018-4877-5 29940849

65. Aslan H, Karaca N, Basaran S, Ermis H, Ceylan Y. Prenatal diagnosis of Wolf-Hirschhorn syndrome (4p-) in association with congenital hypospadias and foot deformity. BMC Pregnancy and Childbirth. 2003;3(1):1. doi: 10.1186/1471-2393-3-1 12546710

66. Garcia NM, Allgood J, Santos LJ, Lonergan D, Batanian JR, Henkemeyer M, et al. Deletion mapping of critical region for hypospadias, penoscrotal transposition and imperforate anus on human chromosome 13. Journal of Pediatric Urology. 2006;2(4):233–42. doi: 10.1016/j.jpurol.2006.03.006 17476316

67. Ravnan JB, Tepperberg JH, Papenhausen P, Lamb AN, Hedrick J, Eash D, et al. Subtelomere FISH analysis of 11 688 cases: an evaluation of the frequency and pattern of subtelomere rearrangements in individuals with developmental disabilities. Journal of Medical Genetics. 2006;43(6):478–89. doi: 10.1136/jmg.2005.036350 16199540

68. Almind GJ, Brondum-Nielsen K, Bangsgaard R, Baekgaard P, Gronskov K. 11p Microdeletion including WT1 but not PAX6, presenting with cataract, mental retardation, genital abnormalities and seizures: a case report. Molecular Cytogenetics. 2009;2:6. doi: 10.1186/1755-8166-2-6 19222835

69. Belligni EF, Biamino E, Molinatto C, Messa J, Pierluigi M, Faravelli F, et al. Subtelomeric FISH analysis in 76 patients with syndromic developmental delay/intellectual disability. Italian Journal of Pediatrics. 2009;35(1):9. doi: 10.1186/1824-7288-35-9 19490664

70. Iannuzzi A, Perucatti A, Genualdo V, Pauciullo A, Incarnato D, Musilova P, et al. The utility of chromosome microdissection in clinical cytogenetics: a new reciprocal translocation in sheep. Cytogenetic and Genome Research. 2014;142(3):174–8. doi: 10.1159/000361000 24733039

71. Genualdo V, Perucatti A, Marletta D, Castiglioni B, Bordonaro S, Iannaccone M, et al. Cytogenetic investigation in two endangered pig breeds raised in Southern-Italy: Clinical and environmental aspects. Livestock Science. 2018;216:36–43.

72. Eggen A, Gautier M, Billaut A, Petit E, Hayes H, Laurent P, et al. Construction and characterization of a bovine BAC library with four genome-equivalent coverage. Genetics, Selection, Evolution. 2001;33(5):543–8. doi: 10.1186/1297-9686-33-5-543 11712974

73. Hayes H, Di Meo GP, Gautier M, Laurent P, Eggen A, Iannuzzi L. Localization by FISH of the 31 Texas nomenclature type I markers to both Q- and R-banded bovine chromosomes. Cytogenetics and Cell Genetics. 2000;90(3–4):315–20. doi: 10.1159/000056795 11124540

74. Iannuzzi A, Perucatti A, Genualdo V, De Lorenzi L, Di Berardino D, Parma P, et al. Cytogenetic Elaboration of a Novel Reciprocal Translocation in Sheep. Cytogenetic and Genome Research. 2013;139(2):97–101. doi: 10.1159/000346193 23328281

75. Iannuzzi A, Pereira J, Iannuzzi C, Fu BY, Ferguson-Smith M. Pooling strategy and chromosome painting characterize a living zebroid for the first time. Plos One. 2017;12(7).

76. Cribiu EP, Di Berardino D, Di Meo GP, Eggen A, Gallagher DS, Gustavsson I, et al. International System for Chromosome Nomenclature of Domestic Bovids (ISCNDB 2000). Cytogenetics and Cell Genetics. 2001;92(3–4):283–99. doi: 10.1159/000056917 11435702

77. Iannuzzi A, Genualdo V, Perucatti A, Pauciullo A, Varricchio G, Incarnato D, et al. Fatal Outcome in a Newborn Calf Associated with Partial Trisomy 25q and Partial Monosomy 11q, 60,XX,der(11)t(11;25)(q11;q14 similar to 21). Cytogenetic and Genome Research. 2015;146(3):222–9. doi: 10.1159/000438973 26337016

78. Meinecke B, Kuiper H, Drögemüller C, Leeb T, Meinecke-Tillmann S. A mola hydatidosa coexistent with a foetus in a bovine freemartin pregnancy. Placenta. 2003;24(1):107–12. doi: 10.1053/plac.2002.0872 12495666

79. Schwarzenbacher H, Wurmser C, Flisikowski K, Misurova L, Jung S, Langenmayer MC, et al. A frameshift mutation in GON4L is associated with dwarfism in Fleckvieh cattle. bioRxiv. 2016:036889.

80. Braun M, Reinartz S, Heppelmann M, Rehage J, Sürie C, Distl O, et al. Curly coat caused by a keratin 27 variant was transmitted from Fleckvieh into German Angus. Animal Genetics. 2018; 49(4):349–350. doi: 10.1111/age.12669 29932459

81. Li H, Durbin R. Fast and accurate short read alignment with Burrows-Wheeler transform. Bioinformatics. 2009;25(14):1754–60. doi: 10.1093/bioinformatics/btp324 19451168

82. Li H, Handsaker B, Wysoker A, Fennell T, Ruan J, Homer N, et al. The sequence alignment/map format and SAMtools. Bioinformatics. 2009;25(16):2078–9. doi: 10.1093/bioinformatics/btp352 19505943

83. McKenna A, Hanna M, Banks E, Sivachenko A, Cibulskis K, Kernytsky A, et al. The genome analysis toolkit: a MapReduce framework for analyzing next-generation DNA sequencing data. Genome Research. 2010;20(9):1297–303. doi: 10.1101/gr.107524.110 20644199

84. Layer RM, Chiang C, Quinlan AR, Hall IM. LUMPY: a probabilistic framework for structural variant discovery. Genome Biology. 2014;15(6):R84. doi: 10.1186/gb-2014-15-6-r84 24970577

85. Cingolani P, Platts A, Wang LL, Coon M, Nguyen T, Wang L, et al. A program for annotating and predicting the effects of single nucleotide polymorphisms, SnpEff: SNPs in the genome of Drosophila melanogaster strain w1118; iso-2; iso-3. Fly. 2012;6(2):80–92. doi: 10.4161/fly.19695 22728672

86. McLaren W, Pritchard B, Rios D, Chen Y, Flicek P, Cunningham F. Deriving the consequences of genomic variants with the Ensembl API and SNP effect predictor. Bioinformatics. 2010;26(16):2069–70. doi: 10.1093/bioinformatics/btq330 20562413

87. Kumar P, Henikoff S, Ng PC. Predicting the effects of coding non-synonymous variants on protein function using the SIFT algorithm. Nature Protocols. 2009;4(7):1073. doi: 10.1038/nprot.2009.86 19561590

Článek vyšel v časopise


2020 Číslo 1