Milk in Life Conditions (MiLC): Bacterial Composition of Human Milk Pumped and Stored in "Real-Life" Conditions

• Sponsor: Cornell University (Other)
• Overall Status: Completed
• CT.gov ID: NCT03123874
• Collaborator: National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) (NIH), University of Idaho (Other), United States Department of Agriculture (USDA) (U.S. Fed)
• Enrollment: 52
• Location: 1
• Arms: 2
• Actual Duration (Months): 4.4
• Patients Per Site Per Month: 11.9

Study Details

Study Description

Brief Summary

The MiLC trial is a randomized control trial of two different breast pump set-ups: mother's own and sterile. The objective of this trial is to investigate the bacterial composition of human milk pumped and stored in "real-life" conditions. To meet this objective, lactating mothers will fully express breast milk from one breast on two consecutive pumping sessions at home, once with the participant's own pumps and collection kits (own pump set-up) and once with a hospital-grade pump and disposable, sterile collection kits (sterile pump set-up). Randomization will be used to determine which pump participants use first. From the total volume of milk pumped during each pumping session, the researchers will collect 1 oz. Milk from both pumps will be stored at home and sampled on days 0, 2, 4, and 30 after expression for analysis of its bacterial composition.

Condition or Disease	Intervention/Treatment	Phase
Human Milk Microbiome; Bacterial Growth	Device: Sterile pump set-up; Device: Mother's Own pump set-up	N/A

Detailed Description

Participants will donate 1 oz of human milk during each of two consecutive pumping sessions, for a total volume of 2 oz donated on one day. To collect this human milk, participants will be asked to fully express one breast during each pumping session. Participants will pump once with the participant's own pump set-up and once with the sterile pump set-up (provided by the research team). Women will be randomized to which pump is used first.

Randomization will be done using stratified randomization as follows: participants were stratified by how their infants were fed, namely whether infants were fed human milk only vs. human milk and complementary foods). Then, randomization was conducted within each strata. The researchers aim to have a minimum of 25 participants in each stratum.

All human milk collections will occur at participants' homes between 0700 and 1100 hours. The second pumping session must begin 3 hours (+/-30 minutes) after the beginning of the first pumping session (e.g. the first pumping session at 7:30 am and the second at 10:30 am). Participants will elect from which breast to donate human milk and that breast will be used for both pumping sessions. Participants will be asked not to nurse from or pump that breast during the 2 hours before the first pumping session and not until after the second pumping session (a total of ~5.5 hours).

From the milk produced during each pumping session (which could be up to ~6 oz), researchers will collect 1 oz using a sterile, plastic syringe. Participants will keep the remaining volume of milk. Each ounce of milk collected will be separated into 5 sterile containers (provided). Participants will store donated milk at home until it is picked up by a researcher 2, 4, and 30 days after pumping.

Study Design

• Study Type: Interventional
• Actual Enrollment: 52 participants
• Allocation: Randomized
• Intervention Model: Crossover Assignment
• Intervention Model Description: All participants will pump their milk with two pump set-ups: own and sterile supplies. The pump set-up each woman used first was randomized according to infant feeding status (human milk only vs. human milk and complementary foods).All participants will pump their milk with two pump set-ups: own and sterile supplies. The pump set-up each woman used first was randomized according to infant feeding status (human milk only vs. human milk and complementary foods).
• Masking: None (Open Label)
• Primary Purpose: Basic Science
• Official Title: Milk in Life Conditions (MiLC): Characterizing the Bacterial Composition of Human Milk Pumped and Stored in "Real-life" Conditions
• Actual Study Start Date: May 22, 2017
• Actual Primary Completion Date: Oct 2, 2017
• Actual Study Completion Date: Oct 2, 2017

Arms and Interventions

Arm	Intervention/Treatment
Experimental: Sterile pump set-up first Participants will pump with sterile pump set-ups first. Approximately 3 hours later, participants will pump with their own pump set-ups.	Device: Sterile pump set-up; Medela symphony breast pump (model number 0240108) and disposable, sterile collection kits (model number 67399S).; Device: Mother's Own pump set-up; Mother's own electric breast pump and own collection kit (previously used and cleaned at home using her usual practices).
Experimental: Mother's Own pump set-up first Participants will pump with their own pump set-ups first. Approximately 3 hours later, participants will pump with sterile pump set-ups.	Device: Sterile pump set-up; Medela symphony breast pump (model number 0240108) and disposable, sterile collection kits (model number 67399S).; Device: Mother's Own pump set-up; Mother's own electric breast pump and own collection kit (previously used and cleaned at home using her usual practices).

Outcome Measures

Primary Outcome Measures

1. Bacterial Community Richness [0 days after pumping]

   Richness is the total number of different bacterial taxa detected in the sample. This metric will be assessed on data collected via high-throughput sequencing of the bacterial 16S rRNA gene present in milk.

2. Bacterial Community Diversity [0 days after pumping]

   Bacterial community diversity will be assessed using the the Shannon diversity index. The Shannon diversity index is a type of entropy measure and is a function of the distribution of the total number of organisms across all of the species. If S is the total number of species in the sample and p_i is the number of organisms in the i-th species divided by the total number of organisms, then Diversity = -Σ p_i log(p_i). This metric will be assessed on data collected via high-throughput sequencing of the bacterial 16S rRNA gene present in milk.

3. Total Live Aerobic Bacterial Counts [0 days after pumping]

   Number of live total aerobic bacteria in milk assessed by aerobic culturing of milk on plate count agar. Reported as colony-forming units (CFU)/mL.

Eligibility Criteria

Criteria

• Ages Eligible for Study: 18 Years and Older
• Sexes Eligible for Study: Female
• Accepts Healthy Volunteers: Yes

Inclusion Criteria:

• Lactating women over the age of 18 years

• Self-reported as healthy women and infants

• Use an electric breast pump

• Confident of ability to donate 1 oz of milk from one breast during each of two consecutive pumping sessions where pumping sessions are 3 hours (+/- 30 minutes) apart and between 0700-1100 hours.

• Able to store donated milk at home for 30 days

• Have infants who do not consume formula or only consume formula episodically as long as the most recent formula-feeding occurred > 2 weeks before the day milk is pumped for this study.

Exclusion Criteria:

• Not confident of ability to donate 1 oz of milk from one breast during each of two consecutive pumping sessions, 3 hours (+/- 30 minutes) apart and between the hours of 0700-1100 hours.

• Infant consumption of formula in the past 2 weeks

• Current indication of breast infection (e.g., breast pain, discomfort, lumps, mastitis with fever, red streaks, or hard red portions of the breast)

• Breast pain that the woman does not consider "normal" for lactation/breastfeeding

• Signs/symptoms of acute illness in woman or infant in past 7 days including fever (rectal or temporal temperature ≥99.5 F), dark green nasal discharge, diarrhea (abrupt onset of 3 or more excessively "loose" stools in one day), vomiting (where infant vomiting is not associated with feeding), or severe cough.

Contacts and Locations

Locations

	Site	City	State	Country	Postal Code
1	Hay Laboratory, B75C Wing Hall, Cornell University	Ithaca	New York	United States	14853

Sponsors and Collaborators

• Cornell University
• National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
• University of Idaho
• United States Department of Agriculture (USDA)

Investigators

• Principal Investigator: Kathleen M. Rasmussen, ScD, Cornell University
• Principal Investigator: Anthony Hay, PhD, Cornell University
• Study Director: Sarah Reyes, MS, Cornell University

Study Documents (Full-Text)

• Study Protocol and Statistical Analysis Plan - May 2, 2017

More Information

Publications

None provided.

• Responsible Party: Cornell University
• ClinicalTrials.gov Identifier: NCT03123874
• Other Study ID Numbers: IRB #: 1608006566; 2T32DK007158-42; NYC-399436
• First Posted: Apr 21, 2017
• Last Update Posted: May 21, 2019
• Last Verified: Apr 1, 2019

• Individual Participant Data (IPD) Sharing Statement: Yes
• Plan to Share IPD: Yes
• Studies a U.S. FDA-regulated Drug Product: No
• Studies a U.S. FDA-regulated Device Product: Yes
• Keywords provided by Cornell University: human milk; breast milk; expressed human milk; bacterial communities; microbiome; breast pump; bacterial growth
• Additional relevant MeSH terms: Disease

Study Results

Participant Flow

Arm/Group Title	Sterile Pump Set-up First	Mother's Own Pump Set-up First
Arm/Group Description	Participants will pump with sterile pump set-ups first. Approximately 3 hours later, participants will pump with their own pump set-ups.	Participants will pump with their own pump set-ups first. Approximately 3 hours later, participants will pump with sterile pump set-ups.
Period Title: Overall Study
STARTED	27	25
COMPLETED	27	25
NOT COMPLETED	0	0

Baseline Characteristics

Arm/Group Title	Sterile Pump Set-up First	Mother's Own Pump Set-up First	Total
Arm/Group Description	Participants will pump with sterile pump set-ups first. Approximately 3 hours later, participants will pump with their own pump set-ups.	Participants will pump with their own pump set-ups first. Approximately 3 hours later, participants will pump with sterile pump set-ups.	Total of all reporting groups
Overall Participants	27	25	52
Age (Count of Participants)
<=18 years	0 0%	0 0%	0 0%
Between 18 and 65 years	27 100%	25 100%	52 100%
>=65 years	0 0%	0 0%	0 0%
Age (years) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [years]	33.6 (4.1)	33.6 (4.0)	33.6 (4.0)
Sex: Female, Male (Count of Participants)
Female	27 100%	25 100%	52 100%
Male	0 0%	0 0%	0 0%
Race/Ethnicity, Customized (Count of Participants)
Caucasian/white	23 85.2%	22 88%	45 86.5%
African American/black	0 0%	1 4%	1 1.9%
Asian or Pacific Islander	2 7.4%	1 4%	3 5.8%
Native American	1 3.7%	0 0%	1 1.9%
Other	0 0%	2 8%	2 3.8%
Region of Enrollment (participants) [Number]
United States	27 100%	25 100%	52 100%
Infant feeding status, number fed human milk only (Count of Participants)
Count of Participants [Participants]	13 48.1%	14 56%	27 51.9%

Outcome Measures

1. Primary Outcome

Title	Bacterial Community Richness
Description	Richness is the total number of different bacterial taxa detected in the sample. This metric will be assessed on data collected via high-throughput sequencing of the bacterial 16S rRNA gene present in milk.
Time Frame	0 days after pumping

Outcome Measure Data

Analysis Population Description

Data analyzed for all participants

Arm/Group Title	Sterile Pump Set-up First	Mother's Own Pump Set-up First
Arm/Group Description	Participants will pump with sterile pump set-ups first. Approximately 3 hours later, participants will pump with their own pump set-ups.	Participants will pump with their own pump set-ups first. Approximately 3 hours later, participants will pump with sterile pump set-ups.
Measure Participants	27	25
In Milk Collected with Sterile Pump Set-Up	64.2 (38.3)	58.9 (26.9)
In Milk Collected with Mother's Own Pump Set-Up	64.0 (30.7)	60.8 (32.8)

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	Sterile Pump Set-up First, Mother's Own Pump Set-up First
	Comments
	Type of Statistical Test	Other
	Comments	Linear mixed effects models testing the effect of pump set-up on the number of live aerobic bacterial counts. The fixed effect of interest will be pump set-up. Model will also be adjusted for infant feeding status (human milk only vs. human milk and complementary foods) and randomization schedule (which pump set-up went first).
Statistical Test of Hypothesis	p-Value	0.9
	Comments	Results were considered statistically significant at p<0.05. No adjustments were made to p-values.
	Method	Mixed Models Analysis
	Comments
Method of Estimation	Estimation Parameter	Mean Difference (Final Values)
	Estimated Value	0.827
	Confidence Interval	(2-Sided) %; to
	Parameter Dispersion	Type: Value:
	Estimation Comments	Ratio of mean bacterial taxa in milk collected with own / sterile pump set-ups.

2. Primary Outcome

Title	Bacterial Community Diversity
Description	Bacterial community diversity will be assessed using the the Shannon diversity index. The Shannon diversity index is a type of entropy measure and is a function of the distribution of the total number of organisms across all of the species. If S is the total number of species in the sample and p_i is the number of organisms in the i-th species divided by the total number of organisms, then Diversity = -Σ p_i log(p_i). This metric will be assessed on data collected via high-throughput sequencing of the bacterial 16S rRNA gene present in milk.
Time Frame	0 days after pumping

Outcome Measure Data

Analysis Population Description

Data analyzed from all participants.

Arm/Group Title	Sterile Pump Set-up First	Mother's Own Pump Set-up First
Arm/Group Description	Participants will pump with sterile pump set-ups first. Approximately 3 hours later, participants will pump with their own pump set-ups. Sterile pump set-up: Medela symphony breast pump (model number 0240108) and disposable, sterile collection kits (model number 67399S). Mother's Own pump set-up: Mother's own electric breast pump and own collection kit (previously used and cleaned at home using her usual practices).	Participants will pump with their own pump set-ups first. Approximately 3 hours later, participants will pump with sterile pump set-ups. Sterile pump set-up: Medela symphony breast pump (model number 0240108) and disposable, sterile collection kits (model number 67399S). Mother's Own pump set-up: Mother's own electric breast pump and own collection kit (previously used and cleaned at home using her usual practices).
Measure Participants	27	25
In Milk Collected with Sterile Pump Set-Up	3.89 (1.18)	3.56 (1.13)
In Milk Collected with Mother's Own Pump Set-Up	3.84 (1.08)	3.93 (1.26)

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	Sterile Pump Set-up First, Mother's Own Pump Set-up First
	Comments
	Type of Statistical Test	Other
	Comments	Linear mixed effects models testing the effect of pump set-up on the number of live aerobic bacterial counts. The fixed effect of interest will be pump set-up. Model will also be adjusted for infant feeding status (human milk only vs. human milk and complementary foods) and randomization schedule (which pump set-up went first).
Statistical Test of Hypothesis	p-Value	0.3
	Comments	Results were considered statistically significant at p<0.05. No adjustments were made to p-values.
	Method	Mixed Models Analysis
	Comments
Method of Estimation	Estimation Parameter	Mean Difference (Final Values)
	Estimated Value	0.163
	Confidence Interval	(2-Sided) %; to
	Parameter Dispersion	Type: Value:
	Estimation Comments	Ratio of mean Shannon Diversity Index in milk collected with own / sterile pump set-ups

3. Primary Outcome

Title	Total Live Aerobic Bacterial Counts
Description	Number of live total aerobic bacteria in milk assessed by aerobic culturing of milk on plate count agar. Reported as colony-forming units (CFU)/mL.
Time Frame	0 days after pumping

Outcome Measure Data

Analysis Population Description

Data for all participants analyzed.

Arm/Group Title	Sterile Pump Set-up First	Own Pump Set-up First
Arm/Group Description	Participants will pump with sterile pump set-ups first. Approximately 3 hours later, participants will pump with their own pump set-ups. Sterile pump set-up: Medela symphony breast pump (model number 0240108) and disposable, sterile collection kits (model number 67399S). Mom's own pump set-up: Mom's own electric breast pump and own collection kit (previously used and cleaned at home using her usual practices).	Participants will pump with their own pump set-ups first. Approximately 3 hours later, participants will pump with sterile pump set-ups. Sterile pump set-up: Medela symphony breast pump (model number 0240108) and disposable, sterile collection kits (model number 67399S). Mom's own pump set-up: Mom's own electric breast pump and own collection kit (previously used and cleaned at home using her usual practices).
Measure Participants	27	25
In Milk Collected with Sterile Pump Set-Up	1848 (734)	1233 (490)
In Milk Collected with Mother's Own Pump Set-Up	9157 (3638)	6109 (2427)

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	Sterile Pump Set-up First, Mother's Own Pump Set-up First
	Comments
	Type of Statistical Test	Other
	Comments	Linear mixed effects models testing the effect of pump set-up on the number of live aerobic bacterial counts. The fixed effect of interest will be pump set-up. Model will also be adjusted for infant feeding status (human milk only vs. human milk and complementary foods) and randomization schedule (which pump set-up went first). Random effect was participant ID to account for multiple samples for each participant.
Statistical Test of Hypothesis	p-Value	0.0003
	Comments	Results were considered statistically significant at p<0.05. No adjustments were made to p-values.
	Method	Mixed Models Analysis
	Comments
Method of Estimation	Estimation Parameter	Mean Difference (Final Values)
	Estimated Value	4.95
	Confidence Interval	(2-Sided) %; to
	Parameter Dispersion	Type: Value:
	Estimation Comments	Ratio of mean bacterial counts in milk collected with own / sterile pump set-ups.

Adverse Events

Time Frame	No adverse events data collected for this study
Adverse Event Reporting Description	No adverse events data collected for this study
Arm/Group Title	Sterile Pump Set-up First		Mother's Own Pump Set-up First
Arm/Group Description	Participants will pump with sterile pump set-ups first. Approximately 3 hours later, participants will pump with their own pump set-ups.		Participants will pump with their own pump set-ups first. Approximately 3 hours later, participants will pump with sterile pump set-ups.
All Cause Mortality
	Sterile Pump Set-up First		Mother's Own Pump Set-up First
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	0/0 (NaN)		0/0 (NaN)
Serious Adverse Events
	Sterile Pump Set-up First		Mother's Own Pump Set-up First
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	0/0 (NaN)		0/0 (NaN)
Other (Not Including Serious) Adverse Events
	Sterile Pump Set-up First		Mother's Own Pump Set-up First
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	0/0 (NaN)		0/0 (NaN)

Limitations/Caveats

[Not Specified]

More Information

Certain Agreements

All Principal Investigators ARE employed by the organization sponsoring the study.

There is NOT an agreement between Principal Investigators and the Sponsor (or its agents) that restricts the PI's rights to discuss or publish trial results after the trial is completed.