Skip to content
Licensed Unlicensed Requires Authentication Published by De Gruyter October 30, 2013

Inactivation of high-risk human papillomaviruses by Holder pasteurization: implications for donor human milk banking

  • Manuela Donalisio , Valeria Cagno , Marta Vallino , Guido E. Moro EMAIL logo , Sertac Arslanoglu , Paola Tonetto , Enrico Bertino and David Lembo


Aims: Several studies have recently reported the detection of oncogenic human papillomaviruses (HPV) in human milk of a minority of lactating mothers. These findings raised safety concerns in the context of human donor milk banking given the potential risk of HPV transmission to recipient infants. The aim of this study was to investigate whether the Holder pasteurization, a procedure currently in use in human donor milk banks for milk pasteurization, completely inactivates high-risk and low-risk HPV.

Methods: HPV pseudoviruses (PsV) were generated, spiked into cell culture medium or donor human milk and subjected to thermal inactivation. HPV PsV infectivity and morphological integrity was analyzed by cell-based assay and by electron microscopy, respectively.

Results: The Holder pasteurization completely inactivated the infectivity of high-risk (types 16 and 18) and low-risk (type 6) HPV both in cell culture medium and in human milk causing PsV particle disassembly.

Conclusions: The results presented here indicate that the Holder pasteurization is an efficient procedure to inactivate high-risk and low-risk HPV thus preventing the potential risk of their transmission through human donor milk.

Corresponding author: Guido E. Moro, Italian Association of Human Milk Banks (AIBLUD), c/o Biomedia, Via Libero Temolo No 4, 20126, Milan, Italy; and World Association of Perinatal Medicine, Working Group on Nutrition, Tel.: +39 3485659614, E-mail:

This research has been partly supported by the Iolanda Minoli Foundation, San Giuseppe University Hospital, Milan, Italy. A special thank you to the Board of Directors of the Italian Association of Human Milk Banks (Associazione Italiana Banche del Latte Umano Donato, AIBLUD: the continuous support to promote new studies with the aim to improve the quality of donor human milk.


[1] Arslanoglu S, Bertino E, Tonetto P, De Nisi G, Ambruzzi AM, Biasini A, et al. Guidelines for the establishment and operation of a donor human milk bank. J Matern Fetal Neonatal Med. 2010;23(S 2):1–20.10.3109/14767058.2010.512414Search in Google Scholar

[2] Arslanoglu S, Moro GE, Bellù R, Turoli D, De Nisi G, Tonetto P, et al. Presence of human milk bank is associated with elevated rate of exclusive breastfeeding in VLBW infants. J Perinat Med. 2012;41:129–31.Search in Google Scholar

[3] Balmer SE, Williams AF. Guidelines for the establishment and operation of human milk banks in the UK. J Hosp Infect. 1995;31:155–6.10.1016/0195-6701(95)90173-6Search in Google Scholar

[4] Bidawid S, Farber JM, Sattar SA, Hayward S. Heat inactivation of hepatitis A virus in dairy foods. J Food Prot. 2002;63:522–28.10.4315/0362-028X-63.4.522Search in Google Scholar

[5] Bosch FX, de Sanjose S. Chapter 1, Human papillomavirus and cervical cancer--burden and assessment of causality. J Natl Cancer Inst Monogr. 2003;31:3–13.Search in Google Scholar

[6] Buck CB, Pastrana DV, Lowy DR, Schiller JT. Efficient intracellular assembly of papillomaviral vectors. J Virol. 2004;78:751–7.10.1128/JVI.78.2.751-757.2004Search in Google Scholar

[7] Buck CB, Pastrana DV, Lowy DR, Schiller JT. Generation of HPV pseudovirions using transfection and their use in neutralization assays. Methods Mol Med. 2005;119:445–62.10.1385/1-59259-982-6:445Search in Google Scholar

[8] Buck CB, Thompson CD, Roberts JN, Müller M, Lowy DR, Schiller JT. Carrageenan is a potent inhibitor of papillomavirus infection. PLoS Pathog. 2006;2:e69.10.1371/journal.ppat.0020069Search in Google Scholar

[9] Cazzaniga M, Gheit T, Casadio C, Khan N, Macis D, Valenti F, et al. Analysis of the presence of cutaneous and mucosal papillomavirus types in ductal lavage fluid, milk and colostrum to evaluate its role in breast carcinogenesis. Breast Cancer Res Treat. 2009;114:599–605.10.1007/s10549-008-0040-3Search in Google Scholar

[10] Donalisio M, Rusnati M, Civra A, Bugatti A, Allemand D, Pirri G, et al. Identification of a dendrimeric heparan sulfate-binding peptide that inhibits infectivity of genital types of human papillomaviruses. Antimicrob Agents Chemother. 2010;54:4290–9.10.1128/AAC.00471-10Search in Google Scholar

[11] Dworsky M, Stagno S, Pass RF, Cassady G, Alford C. Persistence of cytomegalovirus in human milk after storage. J Pediatr. 1982;101:440–3.10.1016/S0022-3476(82)80081-4Search in Google Scholar

[12] Eglin RP, Wilkinson AR. HIV infection and pasteurization of breast milk. Lancet. 1987;1:1093.Search in Google Scholar

[13] Glenn WK, Whitaker NJ, Lawson JS. High risk human papillomavirus and Epstein Barr virus in human breast milk. BMC Res Notes. 2012;5:477.10.1186/1756-0500-5-477Search in Google Scholar PubMed PubMed Central

[14] Hamprecht K, Maschamann J, Muller D, Dietz K, Besenthal I, Goelz R, et al. Cytomegalovirus (CMV) inactivation in breastmilk: reassessment of pasteurization and freeze-thawing. Pediatr Res. 2004;56:529–35.10.1203/01.PDR.0000139483.35087.BESearch in Google Scholar PubMed

[15] Howley PM, Lowy DR. Papillomaviruses and their replication. In: Fields BN, Knipe DM, Howley PM, editors. Fields virology. Philadelphia: Lippincott-Raven; 2001:2197–2229.Search in Google Scholar

[16] Human Milk Banking Association of North America (HMBANA). Guidelines for the establishment and operation of a donor human milk bank. Sandwich, MA: HMBANA; 2013.Search in Google Scholar

[17] Jones F. History of North American donor milk banking: one hundred years of progress. J Hum Lact. 2003;19:313–8.10.1177/0890334403255857Search in Google Scholar PubMed

[18] Kjaer SK, Chackerian B, van den Brule AJ, Svare EI, Paull G, Walbomers JM, et al. High-risk human papillomavirus is sexually transmitted; evidence from a follow-up study of virgins starting sexual activity (intercourse). Cancer Epidemiol Biomarkers Prev. 2001;10:101–16.Search in Google Scholar

[19] Lacey CJ, Lowndes CM, Shah KV. Chapter 4: Burden and management of non-cancerous HPV-related conditions: HPV-6/11 disease. Vaccine. 2006;24(S3):S35–41.10.1016/j.vaccine.2006.06.015Search in Google Scholar PubMed

[20] Lembo D, Donalisio M, Rusnati M, Bugatti A, Cornaglia M, Cappello P, et al. Sulfated K5 Escherichia coli polysaccharide derivatives as wide-range inhibitors of genital types of human papillomavirus. Antimicrob Agents Chemother. 2008;52:1374–81.10.1128/AAC.01467-07Search in Google Scholar PubMed PubMed Central

[21] Lowy DR, Howley PM. Papillomaviruses. In: Fields BN, Knipe DM, Howley PM, editors. Fields virology. Philadelphia: Lippincott-Raven; 2001:2231–2264.Search in Google Scholar

[22] Mammas IN, Zaravinos A, Sourvinos G, Myriokefalitakis N, Theodoridou M, Spandidos DA. Can ‘high-risk’ human papillomaviruses (HPVs) be detected in human breast milk? Acta Paediatr. 2011;100:705–7.10.1111/j.1651-2227.2010.02129.xSearch in Google Scholar PubMed

[23] Marchini A, Liu H, Zhu H. Human cytomegalovirus with IE-2 (UL122) deleted fails to express early lytic genes. J Virol. 2001;75:1870–8.10.1128/JVI.75.4.1870-1878.2001Search in Google Scholar PubMed PubMed Central

[24] National Institute for Health and Clinical Excellence (NICE). Donor breast milk banks: the operation of donor milk bank services. NICE clinical guidelines 93; 2010.Search in Google Scholar

[25] Orloff SL, Wallingford JC, McDougal JS. Inactivation of human immunodeficiency virus type I in human milk: effects of intrinsic factors in human milk and of pasteurization. J Hum Lact. 1993;9:13–7.10.1177/089033449300900125Search in Google Scholar PubMed

[26] Paaso AE, Louvanto K, Syrjänen KJ, Waterboer T, Grénman SE, Pawlita M, et al. Lack of type-specific concordance between human papillomavirus (HPV) serology and HPV DNA detection in the uterine cervix and oral mucosa. J Gen Virol. 2011;92:2034–46.10.1099/vir.0.032011-0Search in Google Scholar PubMed

[27] Parry JV, Mortimer PP. The heat sensitivity of hepatitis A virus determined by a simple tissue culture method. J Med Virol. 1984;14:277–83.10.1002/jmv.1890140312Search in Google Scholar PubMed

[28] Sarkola M, Funtala M, Grenman S, Syganen S. Human papillomavirus DNA detected in breast milk. Ped Inf Dis J. 2008;27:557–8.10.1097/INF.0b013e318169ef47Search in Google Scholar PubMed

[29] Schlegel A, Immelmann A, Kempf C. Virus inactivation of plasma-derived proteins by pasteurization in the presence of guanidine hydrochloride. Transfusion. 2001;41:382–9.10.1046/j.1537-2995.2001.41030382.xSearch in Google Scholar

[30] Strazynski M, Kramer J, Becker B. Thermal inactivation of poliovirus type 1 in water, milk and yoghurt. Int J Food Microbiol. 2002;74:73–8.10.1016/S0168-1605(01)00708-5Search in Google Scholar

[31] Suligoi B, Galli C, Ciuta S, Decker R. Low seroprevalence of HTLV-I and HTLV-II in patients with a sexually transmitted disease. Study Group for HTLV and STDs. Eur J Epidemiol. 1999;15:225–9.10.1023/A:1007534600034Search in Google Scholar

[32] Tomasula PM, Kozempel MF, Konstance RP, Gregg D, Boettcher S, Baxt B, et al. Thermal inactivation of foot-and-mouth disease virus in milk using high-temperature, short-time pasteurization. J Dairy Sci. 2007;90:3202–11.10.3168/jds.2006-525Search in Google Scholar PubMed

[33] Yoshida K, Furumoto H, Abe A, Kato T, Nishimura M, Kuwahara A, et al. The possibility of vertical transmission of human papillomavirus through maternal milk. J Obstet Gynaecol. 2011;31:503–6.10.3109/01443615.2011.570814Search in Google Scholar PubMed

The authors stated that there are no conflicts of interest regarding the publication of this article.

Received: 2013-8-2
Accepted: 2013-9-8
Published Online: 2013-10-30
Published in Print: 2014-01-01

©2014 by Walter de Gruyter Berlin Boston

Downloaded on 31.3.2023 from
Scroll to top button