Sustainable and Inclusive Use of Alternative Proteins in Mediterranean Supply Chains (CIPROMED) (CIPROMED)

Circular and Inclusive Use of Alternative Proteins in Mediterranean Supply Chains (CIPROMED)

This study investigates the effects of foods enriched with alternative protein sources, including edible insects, microalgae, hemp, and legumes, on appetite regulation, satiety, food preferences, and metabolic health in healthy adults and individuals with overweight. The study is part of the CIPROMED project, which aims to support sustainable and circular food systems in the Mediterranean area.

The study consists of two phases. In the acute phase, participants will consume different protein-enriched bread products in a controlled setting following a randomized crossover design. Each participant will be exposed to multiple test conditions, allowing within-subject comparisons of postprandial responses. Outcomes assessed during this phase include satiety, hunger, food preference, craving, and short-term energy intake, measured using validated scales and dietary assessment tools.

In the chronic phase, participants will follow structured dietary interventions over a longer period within a Mediterranean dietary framework. Participants will be assigned to different dietary patterns including alternative protein-based foods and control products. This phase aims to evaluate the effects of repeated consumption of alternative protein sources on metabolic parameters, gastrointestinal tolerance, nutritional status, and behavioral responses.

The study aims to assess the acceptability and physiological effects of alternative protein sources and to compare their impact with that of traditional protein sources commonly used in Mediterranean diets.

Study Overview

• Status: Recruiting
• Conditions: Food Preferences; Gut Microbiota; Appetite Regulation; Overweight (BMI > 25); Satiety and Food Intake
• Intervention / Treatment: Other: Mediterranean Diet with Control Products (chronic phase); Other: Mediterranean Diet with Insect Protein Bread (chronic phase); Other: Mediterranean Diet with Seaweed-Based Burger (chronic phase)

Detailed Description

The acute phase of the study is designed to evaluate the short-term effects of foods enriched with alternative protein sources on appetite regulation, satiety, and eating behavior.

Healthy volunteers and individuals with overweight will participate in a controlled, within-subject crossover study conducted at the Department of Agricultural and Food Sciences (DISTAL), University of Bologna. Participants will attend five test sessions over five consecutive weeks.

During each session, participants will consume one of five test products: bread enriched with alternative protein sources (edible insect protein [Acheta domesticus], microalgae [spirulina], hemp, or legumes) or a wheat-based control bread. All products will be standardized and comparable in terms of energy content.

The order of product administration will be randomized according to a Latin square crossover design, ensuring balanced exposure to all test conditions. Randomization will be stratified by gender and weight status (normal weight/overweight).

At each test session, participants will undergo assessments of postprandial satiety, hunger, food preferences, craving responses, and short-term energy intake using validated scales and dietary assessment tools. This design allows within-subject comparisons across different protein sources while minimizing potential confounding factors.

• Study Type: Interventional
• Enrollment (Estimated): 40
• Phase: Not Applicable

Contacts and Locations

• Study Contact: Name: Maria Letizia Petroni, Professor; Phone Number: +39 051/2143688; Email: marialetizia.petroni@unibo.it

Study Locations

• Italy
  • BO
    • Bologna, BO, Italy, 40138
      • Recruiting
      • University of Bologna, Department of Medical and Surgical Sciences
      • Contact: Maria Letizia Petroni, Professor; Phone Number: +39 051/2143688; Email: marialetizia.petroni@unibo.it
      • Principal Investigator: Maria Letizia Petroni, Professor

Participation Criteria

Eligibility Criteria

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

Description

Inclusion Criteria:

• Providing signed informed consent,
• Being between 18 and 65 years of age,
• Having a body mass index (BMI) within the normal range, defined as between 18.5 and 24.9 kg/m² or Having a body mass index (BMI) between 25.0 and 34.9 kg/m², inclusive, corresponding to the overweight category or, at most, class I obesity.

Exclusion Criteria:

• Positive and documented history of allergy to legumes, insects, microalgae, shellfish, mollusks, crustaceans, snails, insect venom, house dust mites, or any component (ingredient or additive) present in the food prototypes tested, or a confirmed or suspected diagnosis of Celiac Disease.
• History of severe allergic reactions to any type of allergen.
• Pregnancy or Breastfeeding

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Basic Science
• Allocation: Randomized
• Interventional Model: Crossover Assignment
• Masking: None (Open Label)

Number of Arms

4

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: Control (A) to Insect Bread (B) Sequence; Participants follow a Mediterranean diet including control products (legume bread and/or standard fish burger) during the first intervention period, followed by a Mediterranean diet including insect protein-enriched bread (Acheta domesticus) during the second period. Each period lasts approximately 2 months. Participants are assigned to one of four randomized sequences according to a Latin square crossover design.	Other: Mediterranean Diet with Control Products (chronic phase); Mediterranean diet including control foods such as legume-based bread and standard fish burger, consumed five times per week for a period of 2 months.; Other: Mediterranean Diet with Insect Protein Bread (chronic phase); Mediterranean diet including bread enriched with edible insect protein (Acheta domesticus), consumed five times per week over a 2-month intervention period.
Experimental: Insect Bread (B) to Control (A) Sequence; Participants follow a Mediterranean diet including insect protein-enriched bread (Acheta domesticus) during the first intervention period, followed by a Mediterranean diet including control products (legume bread and/or standard fish burger) during the second period. Each period lasts approximately 2 months. Participants are assigned to one of four randomized sequences according to a Latin square crossover design.	Other: Mediterranean Diet with Control Products (chronic phase); Mediterranean diet including control foods such as legume-based bread and standard fish burger, consumed five times per week for a period of 2 months.; Other: Mediterranean Diet with Insect Protein Bread (chronic phase); Mediterranean diet including bread enriched with edible insect protein (Acheta domesticus), consumed five times per week over a 2-month intervention period.
Experimental: Control (A) to Seaweed Burger (C) Sequence; Participants follow a Mediterranean diet including control products (legume bread and/or standard fish burger) during the first intervention period, followed by a Mediterranean diet including a seaweed-based burger during the second period. Each period lasts approximately 2 months. Participants are assigned to one of four randomized sequences according to a Latin square crossover design.	Other: Mediterranean Diet with Control Products (chronic phase); Mediterranean diet including control foods such as legume-based bread and standard fish burger, consumed five times per week for a period of 2 months.; Other: Mediterranean Diet with Seaweed-Based Burger (chronic phase); Mediterranean diet including a burger made from edible seaweed, consumed five times per week over a 2-month intervention period.
Experimental: Seaweed Burger (C) to Control (A) Sequence; Participants follow a Mediterranean diet including a seaweed-based burger during the first intervention period, followed by a Mediterranean diet including control products (legume bread and/or standard fish burger) during the second period. Each period lasts approximately 2 months. Participants are assigned to one of four randomized sequences according to a Latin square crossover design.	Other: Mediterranean Diet with Control Products (chronic phase); Mediterranean diet including control foods such as legume-based bread and standard fish burger, consumed five times per week for a period of 2 months.; Other: Mediterranean Diet with Seaweed-Based Burger (chronic phase); Mediterranean diet including a burger made from edible seaweed, consumed five times per week over a 2-month intervention period.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Postprandial Satiety Assessed by Visual Analogue Scales (VAS)	Subjective appetite sensations are assessed using a customized 100 mm Visual Analogue Scale (VAS). The scale evaluates four primary domains: hunger, fullness (satiety), desire to eat, and prospective food consumption. Participants mark a point on a 100 mm line anchored by "not at all" (0 mm) and "extremely" (100 mm). Total scores for each domain range from 0 to 100, where higher scores for satiety indicate a greater feeling of fullness, and higher scores for hunger indicate a greater urge to eat. Post-prandial ratings are measured at baseline and at specific intervals following the consumption of protein-enriched bread products.	At each test session following product consumption (5-week period)
Food Preference and Craving Responses Assessed by HTAS, LFP, and Hedonic Scale	Food preference, motivation, and attitudes were assessed following consumption of different protein-enriched bread products using three distinct validated instruments: Health and Taste Attitude Scales (HTAS): The validated Italian version of the HTAS was used to evaluate psychological attitudes toward food. The questionnaire comprises six subscales divided into two dimensions: Health-related attitudes (General health interest, Light product interest, Natural product interest) and Taste-related attitudes (Craving for sweet foods, Pleasure orientation, Using food as a reward). 9-Point Hedonic Scale: Sensory liking of the test foods was evaluated using a 9-point scale ranging from 1 ('Extremely unpleasant') to 9 ('Extremely pleasant'). Leeds Food Preference Questionnaire (LFP): This psychometric tool was used to assess food motivation via food images. It measures two distinct components: 'Liking' (perceived pleasantness) and 'Wanting' (motivation to consume the food at testing).	At each test session following product consumption (5-week period)
Fasting Blood Glucose	Fasting plasma glucose concentration, assessed following the dietary intervention with alternative protein sources. Unit of Measure: mg/dL	At baseline, 2 months, and 4 months
Fasting Serum Insulin	Fasting serum insulin concentration measured following dietary interventions with alternative protein sources. Unit of Measure: µIU/mL.	At baseline, 2 months, and 4 months
Serum Triglyceride Levels	Fasting serum triglyceride concentration measured following dietary interventions with alternative protein sources. Unit of Measure: mg/dL	At baseline, 2 months, and 4 months
Total Cholesterol Levels	Fasting serum high-density lipoprotein (HDL) cholesterol concentration measured following dietary interventions with alternative protein sources. Unit of Measure: mg/dL	At baseline, 2 months, and 4 months
LDL Cholesterol Levels	Fasting serum low-density lipoprotein (LDL) cholesterol concentration, calculated using the Friedewald formula, measured following dietary interventions with alternative protein sources. Unit of Measure: mg/dL	At baseline, 2 months, and 4 months
Gastrointestinal Symptoms Assessed by Nepean Dyspepsia Index (NDI)	Gastrointestinal symptoms were assessed following dietary interventions with different protein sources using the Nepean Dyspepsia Index (NDI). This validated instrument measures symptom severity, quality of life, and the impact of functional dyspepsia on daily living. The study utilizes the version validated through translation from Australian English to Italian.	At baseline, 2 months, and 4 months

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Postprandial Energy Intake and Energy Compensation Assessed by 24-Hour Dietary Recall	Postprandial energy intake and energy compensation measured using 24-hour dietary recall following consumption of different protein-enriched bread products.	Within 24 hours after each test session (5-week period)
Liver Fat Content and Hepatic Stiffness Assessed by FibroScan and Controlled Attenuation Parameter (CAP)	Liver fat content and hepatic stiffness measured using transient elastography (FibroScan) and Controlled Attenuation Parameter (CAP) to evaluate changes following dietary interventions with different protein sources.	At baseline, 2 months, and 4 months
Gut Microbiota Composition	Gut microbiota composition assessed through analysis of fecal samples to evaluate changes following dietary interventions with different protein sources.	At baseline and 4 months
Appetite Sensations Assessed by Visual Analogue Scales (VAS)	Subjective appetite sensations are assessed using a customized 100 mm Visual Analogue Scale (VAS). The scale evaluates four primary domains: hunger, fullness (satiety), desire to eat, and prospective food consumption. Participants mark a point on a 100 mm line anchored by "not at all" (0 mm) and "extremely" (100 mm). Total scores for each domain range from 0 to 100, where higher scores for satiety indicate a greater feeling of fullness, and higher scores for hunger indicate a greater urge to eat. Post-prandial ratings are measured at baseline and at specific intervals following the consumption of protein-enriched bread products.	At baseline, 2 months, and 4 months
Alanine Aminotransferase (ALT) Levels	Serum alanine aminotransferase (ALT) concentration measured following dietary interventions with different protein sources. Unit of Measure: U/L	At baseline, 2 months, and 4 months
Aspartate Aminotransferase (AST) Levels	Serum aspartate aminotransferase (AST) concentration measured following dietary interventions with different protein sources. Unit of Measure: U/L	At baseline, 2 months, and 4 months
Gamma-glutamyl Transferase (GGT) Levels	Serum gamma-glutamyl transferase (GGT) concentration measured following dietary interventions with different protein sources. Unit of Measure: U/L	At baseline, 2 months, and 4 months
Fatty Liver Index (FLI)	Fatty Liver Index (FLI) score calculated following dietary interventions with different protein sources. Unit of Measure: Score (o "Unitless")	At baseline, 2 months, and 4 months
Complete Blood Count (CBC)	Hematological parameters (including red blood cells, white blood cells, and platelets) assessed following dietary interventions with different protein sources. Unit of Measure: cells/µL	At baseline, 2 months, and 4 months
Serum Creatinine and Urea	Serum creatinine and urea levels assessed following dietary interventions with different protein sources. Unit of Measure: mg/dL	At baseline, 2 months, and 4 months
Urinary Renal Markers	Microalbuminuria and urinary nitrogen levels assessed following dietary interventions with different protein sources. Unit of Measure: mg/24h	At baseline, 2 months, and 4 months
C-Reactive Protein (CRP)	Serum C-reactive protein (CRP) levels assessed following dietary interventions with different protein sources. Unit of Measure: mg/L	At baseline, 2 months, and 4 months
Blood Pressure	Systolic and diastolic blood pressure assessed following dietary interventions with different protein sources. Unit of Measure: mmHg	At baseline, 2 months, and 4 months
Serum Immunoglobulin Levels	Serum immunoglobulin levels (including total IgE) assessed following dietary interventions with different protein sources. Unit of Measure: IU/mL	At baseline, 2 months, and 4 months
Body Weight	Body weight measured following dietary interventions with different protein sources. Unit of Measure: kg	At baseline, 2 months, and 4 months
Body Circumferences	Body circumferences (including waist, hip, and abdominal) measured following dietary interventions with different protein sources. Unit of Measure: cm	At baseline, 2 months, and 4 months
Mood Assessed by the Beck Depression Inventory-II (BDI-II)	Assesses the severity of depressive symptoms following dietary interventions with different protein sources. The BDI-II is a 21-item questionnaire. Total scores range from 0 to 63, where higher scores indicate greater severity of depressive symptoms (worse outcome). Unit of Measure: Score on a scale (0-63)	At baseline, 2 months, and 4 months
Food Neophobia Assessed by the Food Neophobia Scale (FNS)	Assesses the reluctance to taste new foods following dietary interventions with different protein sources. The FNS consists of 10 items measured on a 7-point Likert scale, with total scores ranging from 8 to 40. Higher scores indicate an increasing neophobic attitude (worse outcome/reluctance to try new foods). Unit of Measure: Score on a scale (8-40)	At baseline, 2 months, and 4 months
Food-Related Attentional Bias Assessed by the Pictorial Dot-Probe Task	Evaluates selective attention towards food stimuli (healthy vs. unhealthy) following dietary interventions with different protein sources. Attentional bias is measured as the reaction time difference (in milliseconds) to probes appearing in the location of emotionally salient food stimuli. Faster reaction times to probes appearing in the location of the food stimulus suggest greater attentional bias. Unit of Measure: milliseconds (ms)	At baseline and 4 months
Incidence of Adverse Events	Participants will complete a 72-hour recall questionnaire to document any adverse reactions they may have experienced following product consumption.	Within 72 hours after each test session (5-week period)
Changes in Concomitant Medication Use	Participants will be asked to provide a detailed account of any medications they are currently taking.	At baseline and 4 months

Collaborators and Investigators

• Sponsor: University of Bologna
• Collaborators: IRCCS Azienda Ospedaliero-Universitaria di Bologna; Azienda Usl di Bologna; University of Turin, Italy
• Investigators: Principal Investigator: Maria Letizia Petroni, Professor, University of Bologna

Study record dates

Study Major Dates

• Study Start (Actual): March 18, 2026
• Primary Completion (Estimated): October 31, 2026
• Study Completion (Estimated): January 31, 2027

Study Registration Dates

• First Submitted: March 30, 2026
• First Submitted That Met QC Criteria: April 20, 2026
• First Posted (Actual): April 28, 2026

Study Record Updates

• Last Update Posted (Actual): May 4, 2026
• Last Update Submitted That Met QC Criteria: April 28, 2026
• Last Verified: April 1, 2026

More Information

Terms related to this study

• Keywords: Overweight; Satiety; Gut Microbiota; Appetite Regulation; Metabolic Health; Alternative Proteins; Edible Insects; Hemp Protein; Legume Protein; Novel Foods; Microalgae (Protein); Food Acceptability
• Additional Relevant MeSH Terms: Nutrition Disorders; Overnutrition; Body Weight; Pathological Conditions, Signs and Symptoms; Behavior; Nutritional and Metabolic Diseases; Signs and Symptoms; Feeding Behavior; Overweight; Food Preferences; Therapeutics; Diet, Food, and Nutrition; Physiological Phenomena; Nutritional Physiological Phenomena; Diet, Plant-Based; Diet Therapy; Nutrition Therapy; Diet; Diet, Mediterranean
• Other Study ID Numbers: 399/2025/Sper/AOUBo

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: YES
• IPD Plan Description: De-identified individual participant data (IPD) underlying the results reported in this study will be made available upon reasonable request to the corresponding investigator, following publication. Data sharing will be subject to institutional policies and applicable data protection regulations.

Drug and device information, study documents

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