Evaluation of human milk microbiota by 16S rRNA gene next-generation sequencing (NGS) and cultivation/MALDI-TOF mass spectrometry identification

Treven, Primož (author), Mahnič, Aleksander (author), Rupnik, Maja (author), Golob, Majda (author), Pirš, Tina (author), Bogovič Matijašić, Bojana (author), Mohar Lorbeg, Petra (author)

The aim of the present study was to characterize human milk microbiota (HMM) with 16S rRNA gene amplicon next-generation sequencing and cultivation/matrix-assisted laser desorption/ionization (MALDI)-time of flight (TOF) mass spectrometry (MS) identification approaches. We analyzed 31 human milk samples from healthy Slovenian mothers. To check the accuracy of MALDI-TOF MS identification, several colonies representing most abundant genera and those, which could not be reliably identified by MALDITOF, were subjected to Sanger sequencing of their 16S rRNA gene. We showed that cultivation/MALDI-TOF MS was a suitable tool for culture-dependent determination of HMM. With both approaches, Staphylococcus and Streptococcus were found as predominant genera in HMM and the abundance of Staphylococcus was associated with decreased microbial diversity. In addition, we characterized factors that might influence HMM. The use of a breast pump was significantly associated with composition of HMM, higher microbial load, and lower abundance of cultivable staphylococci. Moreover, our study suggests that administration of probiotics to the suckling infant might influence HMM by increased abundance of lactobacilli and the presence of viable probiotic bacteria in human milk. However, since our study was observational with relatively small sample size, more targeted studies are needed to study possible transfer of probiotics to the mammary gland via an external route and the physiological relevance of these events.

• Language: English
• Keywords: milk microbiota, microbiota assembly, breastfeeding, breast pump, probiotic drops
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: BF - Biotechnical Faculty; VF - Veterinary Faculty
• Publication status: Published
• Publication version: Version of Record
• Year: 2019
• Number of pages: 12 str.
• Numbering: Vol. 10, November, art. 2612
• PID: 20.500.12556/RUL-113787
• UDC: 579
• ISSN on article: 1664-302X
• DOI: 10.3389/fmicb.2019.02612
• COBISS.SI-ID: 4326792
• Publication date in RUL: 04.02.2020

Record is a part of a journal

• Title: Frontiers in microbiology
• Shortened title: Front. microbiol.
• Publisher: Frontiers Research Foundation
• ISSN: 1664-302X
• COBISS.SI-ID: 4146296

Licences

• License: CC BY 4.0, Creative Commons Attribution 4.0 International
• Link: http://creativecommons.org/licenses/by/4.0/
• Description: This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.
• Licensing start date: 15.11.2019

Secondary language

• Language: Slovenian
• Keywords: mikrobiologija, humano mleko, mikrobiota, genetika, masna spektrometrija

Projects

• Funder: ARRS - Slovenian Research Agency
• Project number: Z3-8198
• Name: Karakterizacija in vitro modela za študij mikrobiote mlečne žleze (akronim - MAGIC)

• Funder: ARRS - Slovenian Research Agency
• Project number: P4-0097
• Name: Prehrana in mikrobna ekologija prebavil