Shaping Microbiome in the First 1,000 Days of Life (AMAMI)

Prenatal and Postnatal Determinants in Shaping Offspring's Microbiota in the First Year of Life: A Study Protocol

Events occurring in the early stages of life play an important role in the development of chronic diseases; diet and lifestyle during pregnancy are recognized to be crucial determinants in modulating offspring microbiota, throughout a vertical transfer of dysbiotic maternal ambient. Moreover, the intestinal colonization is maximized in the first two years of life through newborn's type and time of feeding. This study will provide the starting point for a future prospective observational study to assessing the association between maternal lifestyle with infant microbiota and their influence future health.

Study Overview

• Status: Completed
• Conditions: Microbial Colonization

Detailed Description

Evidence from the Developmental Origins of Health and Disease (DOHaD) theory has pointed out biological mechanisms supporting the intergenerational nature of non communicable diseases (NCDs), which are an increasing public health problem in many countries. Furthermore, scientific evidence confirms that the events occurring in the early stages of life play a critical role in fostering the development of chronic diseases (e.g. overweight and obesity, allergies, cardiovascular and metabolic diseases, neurocognitive impairment and cancers), throughout the life-course, indicating the high relevance of "the maternal environment" impact on the life of the future child. This crucial window goes from the conception to the two years of life, and it is known as 'first thousand days'.

Nutritional status during pregnancy is one of the crucial determinants in modulating the characteristics of the "maternal environment" in which the foetus originates and develops. Therefore, adequate nutrition just before conception, during pregnancy, infancy and early childhood is essential to ensure growth, health, and development of children to their full potential.

Epigenetic links have been confirmed in mice between the prenatal diet and the increased incidence of obesity, type 2 diabetes and other metabolic associated conditions in their offspring. However, specific epigenetic regulation mechanisms are still unclear. Recently, an increasing number of animal and human studies have proposed the hypothesis that the gut microbiota may be considered an important mediator of vital microbiome-host interaction(s) under health or disease conditions. Moreover, microbiota has been shown to pass features from one generation to the next via maternal contact; in fact, recent observation of microbial presence in placenta, meconium and amniotic fluid has suggested a significant interplay between the environmental microbes and the developing gastrointestinal tract of the fetus even before delivery. Thus, the alteration of microbiota composition, driving to increased long-term susceptibility to diseases, can begin early in life.

Examination of the neonatal gut microbiome immediately after delivery revealed that it varied by virtue of maternal gestational diet suggesting that the colonization of the infant gut is likely to occur prior to parturition.

During the first months after birth, the bacterial flora in the gut is known to be affected by many factors, including mode of delivery, use of antibiotics or probiotics, geographical location, type and time of feeding.

Moreover, maternal exposures during pregnancy to several environmental chemicals are known to induce perturbations in the composition of the gastrointestinal microbiome. Among these substances, endocrine disruptor chemicals (EDCs), such as phthalates, bisphenol and hydroxypyrene, which can be found in oil and water resistant textile coatings, non-stick cookware, food container coatings, floor polish, fire-fighting foam, and industrial surfactants, are a class of suspected obesogenic contaminants persisting in the environment and humans. EDCs have been also detected in cord blood samples, suggesting that exposure starts prenatally and early life EDC exposures may perturb neuroendocrine systems involved in growth, energy metabolism, appetite, adipogenesis, and glucose-insulin homeostasis promoting childhood obesity development.

After this first period, the gut microbial composition continues to evolve until the age of 2-3 years; these first years represent as "a window of opportunity" for microbial modulation, influenced by complementary feeding period, gradual transition from milk-based infant feeding to weaning and solid foods, usually occurring between 6 and 24 months.

A.MA.MI is a longitudinal, prospective, observational study that includes a group of mother-infant pairs (n=60) attending the Neonatal Unit, Fondazione IRCCS Policlinico San Matteo, Pavia (Italy). The study was planned to provide data collected before discharge (T0) and at 1,6,12 months after birth (T1,T2,T3). Maternal and infant anthropometric measurements are assessed at each time. Other variables evaluated are pre-pregnancy/gestational weight status (T0), maternal dietary habits/physical activity (T1-T3); infant medical history, type of feeding, antibiotics/probiotics/supplements use, environment exposures (e.g cigarette smoking, pets, environmental temperature) (T1-T3). A child stool sample was planned to be collected at each time and analyzed using metagenomics 16S ribosomal RNA gene sequence-based methods. Maternal urine samples were planned to be collected at T3 to investigate pollutants exposure (Phthalates, Bisphenol A and Hydroxypyrene).

Since there are still limited knowledge about the child's microbiota, its changes over time and the host and environmental interactions in shaping its composition, further prospective studies are needed to determine how different factors related to diet-related factors, lifestyle, drugs and anthropometric measures can influence microbial colonization of the gut during early childhood and influence the future health status.

• Study Type: Observational
• Enrollment (Actual): 60

Contacts and Locations

Study Locations

• Italy
  • Pavia, Italy, 27100
    • IRCCSSMP

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Child; Adult; Older Adult
• Accepts Healthy Volunteers: Yes
• Genders Eligible for Study: All

• Sampling Method: Non-Probability Sample

Study Population

60 dyads (mother/new-born) attending the Neonatal Unit, Fondazione IRCCS Policlinico San Matteo, Pavia (Italy)

Description

Inclusion Criteria:

• Infants of both sexes born to natural or caesarean delivery;
• Gestational age between 35 and 42 completed weeks;
• Italian-speaking parents;
• Ability of the parent/guardian to give informed consent;
• Ability of the mother to respond to the structured interview/questionnaires;

Exclusion Criteria:

Children or mothers with genetic/congenital diseases;

• Children hospitalized immediately after birth;
• Children selected for other clinical studies;
• Presence of gestational diabetes;
• Presence of hyperthyroidism during pregnancy;
• Parents refusing to sign the informed consent.

Study Plan

How is the study designed?

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Change in intestinal microbiota composition from 0 to 12 months of life	Investigate the intestinal microbiota development of infants at 0, 1, 6, 12 months after birth by: i) Identifying the relative abundance of the dominant organism genera; ii) Assessing the microbial diversity (α-diversity); iii) Assessing inter-individual variation (β-diversity).	0 months (2-3 days after birth); 1 months after birth; 6 months after birth; 12 months after birth.

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
examine the association of maternal and infants factor and the intestinal microbiota development of infants.	exploring the possible correlation between maternal factors before pregnancy (weight status), gestational exposures (maternal diet, physical activity, weight status, EDCs), mode of delivery, lactation (formula or breastfeeding), family environment exposure, infant diet, sleeping habits and infant intestinal microbiota composition, during the first year of life at different follow-up: immediately after the delivery (not more than 2-3 days after the delivery, T0);; 1 month after birth (T1);; 6 months after birth (T2);; 12 months after birth (T3).	0 months (2-3 days after birth); 1 months after birth; 6 months after birth; 12 months after birth.

Collaborators and Investigators

• Sponsor: University of Pavia
• Collaborators: IRCCS Policlinico S. Matteo; Istituti Clinici Scientifici Maugeri SpA; Azienda Ospedaliera Specializzata in Gastroenterologia Saverio de Bellis
• Investigators: Principal Investigator: Hellas Cena, Università di Pavia

Publications and helpful links

General Publications

• Vacca M, Raspini B, Calabrese FM, Porri D, De Giuseppe R, Chieppa M, Liso M, Cerbo RM, Civardi E, Garofoli F, Cena H, De Angelis M. The establishment of the gut microbiota in 1-year-aged infants: from birth to family food. Eur J Nutr. 2022 Aug;61(5):2517-2530. doi: 10.1007/s00394-022-02822-1. Epub 2022 Feb 25.
• Raspini B, Porri D, De Giuseppe R, Chieppa M, Liso M, Cerbo RM, Civardi E, Garofoli F, Monti MC, Vacca M, De Angelis M, Cena H. Prenatal and postnatal determinants in shaping offspring's microbiome in the first 1000 days: study protocol and preliminary results at one month of life. Ital J Pediatr. 2020 Apr 15;46(1):45. doi: 10.1186/s13052-020-0794-8.

Study record dates

Study Major Dates

• Study Start (Actual): September 23, 2018
• Primary Completion (Actual): October 7, 2019
• Study Completion (Actual): October 7, 2019

Study Registration Dates

• First Submitted: October 7, 2019
• First Submitted That Met QC Criteria: October 9, 2019
• First Posted (Actual): October 10, 2019

Study Record Updates

• Last Update Posted (Actual): October 10, 2019
• Last Update Submitted That Met QC Criteria: October 9, 2019
• Last Verified: October 1, 2019

More Information

Terms related to this study

• Keywords: pregnancy; diet; microbiota; 1000 days; maternal lifestyle
• Additional Relevant MeSH Terms: Pathologic Processes; Disease Attributes; Infections; Communicable Diseases
• Other Study ID Numbers: 20191004

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No