CA2994352A1 - Methods and compositions using bifidobacterium longum to optimize breastfeeding - Google Patents

Abstract

B. longum ATCC BAA-999, or a composition comprising B. longum ATCC BAA-999, for use in an individual to; support breastfeeding initiation and/or to optimise breastfeeding duration and/or to optimize the quality and/or quantity ofbreast milk and/or to prevent or treat peripartum emotional distress and/or to regulate amygdala activity and/or to regulate the release of CRH, and to prevent or treat the negative effects of peripartum emotional distress in the offspring of individuals.

Description

Title: METHODS AND COMPOSITIONS USING BIFIDOBACTERIUM LONGUM TO OPTIMIZE
BREASTFEEDING
BACKGROUND
Breastfeeding is known to have beneficial health effects for both mother and baby. Because of its benefits, the World Health Organisation (WHO) recommends exclusive breastfeeding of infants for the first 6mths following birth and continued breastfeeding (along with giving appropriate complementary foods) up until the infant or child is 2 years of age or beyond.
However, despite this recommendation from the WHO, and the known health benefits of breastfeeding, many women, especially in developed countries, are not exclusively breastfeeding or are not breastfeeding for the recommended time frames.
Whilst there are a variety of factors that influence a woman's decision to initiate breastfeeding and/or to continue breastfeeding either exclusively or non-exclusively, research has shown that after adjusting for confounding socio-demographic factors there is as strong and robust association between the initiation and duration of breastfeeding, and peripartum emotional distress.
Peripartum emotional distress in some form or another is a condition though to effect more than 50% of pregnant women in one form or another.
Medications such as antidepressants, anti-anxiety drugs, or beta-blockers may be used as treatments for peripartum emotional distress. However, these may have side effects and many may not be safe for use or desirable to use perinatally.
Accordingly, there is a need to prevent or treat peripartum emotional distress and to support breastfeeding initiation and optimise breastfeeding duration. In particular, there is a need for new ingredients that do not suffer from side effects of known ingredients.
Surprisingly it has now been found that administration of Bifidobacterium longum NCC3001 (ATCC BAA-999) to an individual may improve anxiety and depressive symptoms, may down regulate activity in brain centers involved in the control of emotions and mood (limbic regions such as the amygdala), and may down regulate the expression of Corticotropin Releasing Hormone (hereinafter CRH) in the hypothalamus.

SUMMARY OF THE INVENTION
The invention is set out in the claims and in further detail in the detailed description included herein.
The present invention encompasses B. longum ATCC BAA-999, or a composition comprising B.
longum ATCC BAA-999 for use in an individual to; support breastfeeding initiation and/or to optimise breastfeeding duration and/or to optimize the quality and/or quantity of breast milk and/or to prevent or treat peripartum emotional distress and/or to regulate amygdala activity and/or to regulate the HPA axis. Said individual may be a mammal and in particular may be a human trying to get pregnant, pregnant or lactating.
The HPA axis may be regulated for example by regulating it's activity which may be regulated for example by regulating the release of CRH.
Peripartum emotional distress may be a condition selected from the group consisting of: peripartum low mood, peripartum anxiety, peripartum depression and peripartum traumatic stress disorder.
The B. longum ATCC BAA-999, or composition comprising B. longum ATCC BAA-999 may be particularly effective for use as specified above if administered to the individual for at least 3 weeks or at least 6 weeks. Said administration may be daily.
The B. longum ATCC BAA-999, or composition comprising B. longum ATCC BAA-999 may be administered at any time during the perinatal period e.g. pre-pregnancy to an individual trying to get pregnant, to a pregnant individual or to an individual that has had a baby e.g. a lactating individual.
At least a portion of the B. longum ATCC BAA-999 or B. longum ATCC BAA-999 comprises in the composition may be alive.
At least a portion of the B. longum ATCC BAA-999 or B. longum ATCC BAA-999 comprises in the composition may be non-replicating.

2 The B. longum ATCC BAA-999 or composition comprising B. longum ATCC BAA-999 may be administered to an individual in any therapeutically effective dose.
Particularly useful doses may be daily doses comprising between 104 and 1012 cfu of the B. longum ATCC BAA-999, more particularly between 109 and 1012 for example 1010 .
The composition comprising B. longum ATCC BAA-999 may further comprises an ingredient selected from the group consisting of a fat, a protein, a prebiotic, a carbohydrate, a vitamin, myo-inositol, another probiotic, and combinations thereof.
The prebiotic may be selected from the group consisting of an oligosaccharide, a dietary fiber, and a combination thereof None limiting examples dietary fiber include but are not restricted to pectins, mucilages, gums, galacto-oligosaccharides, oligofructan, inulin, polyfructoses, arabinoglactans, hemicelullose, oligosaccharides or mixtures of thereof.
The fat may be selected from the group consisting of DHA, EPA, DPA, and a combination thereof The composition comprising B. longum ATCC BAA-999 may be any type of composition suitable for perinatal consumption and in particular may be a maternal supplement.
The B. longum ATCC BAA-999 or composition comprising B. longum ATCC BAA-999 may be simultaneously, sequentially or separately administered to another ingredient e.g. a pharmaceutical, that may; support breastfeeding initiation and/or optimise breastfeeding duration and/or optimize the quality and/or quantity of breast milk and/or prevent or treat peripartum emotional distress and/or regulate amygdala activity and/or to regulate HPA axis e.g. the release of CRH, in an individual.
Said B. longum ATCC BAA-999 or composition comprising B. longum ATCC BAA-999, optionally simultaneously sequentially or separately administered to said another ingredient, may

3 also be administered in combination with a psychosocial or psychological intervention e.g.
cognitive behavioural therapy, interpersonal therapy, parent training programs, and/or alternative therapy and/or complementary therapy e.g. yoga therapy, music therapy, art therapy.
B. longum ATCC BAA-999 or a composition comprising B. longum ATCC BAA-999 may also be used in a method of supporting breastfeeding initiation and/or optimising breastfeeding duration and/or optimising the quality and/or quantity of breast milk and/or preventing or treating peripartum emotional distress and/or regulating amygdala activity and/or regulating the HPA axis e.g. the release of CRH, in an individual, said method comprising administering B. longum ATCC
BAA-999 or a composition comprising B. longum ATCC BAA-999 to said individual.
Said method may be a natural therapy.
B. longum ATCC BAA-999 or a composition comprising B. longum ATCC BAA-999 may also be used in the manufacture of a composition for use in an individual to;
support breastfeeding initiation and/or to optimise breastfeeding duration and/or to optimize the quality and/or quantity of breast milk and/or to prevent or treat peripartum emotional distress and/or to regulate amygdala activity and/or to regulate the HPA axis e.g. the release of CRH.
B. longum ATCC BAA-999 or a composition comprising B. longum ATCC BAA-999 may also be used in an individual to; to support breastfeeding initiation and/or to optimise breastfeeding duration and/or to optimize the quality and/or quantity of breast milk and/or to prevent or treat peripartum emotional distress and/or to regulate amygdala activity and/or to regulate the HPA axis e.g. the release of CRH. Said use may be non-therapeutic.
B. longum ATCC BAA-999, or a composition comprising B. longum ATCC BAA-999, may also be used to prevent or treat the negative effects of peripartum emotional distress in the offspring of individuals.
An advantage of the present invention is that B. longum ATCC BAA-999 is effective, readily available, low-priced, and safe to administer perinatally without unwanted side effects. Another

4

5 advantage of one or more embodiments provided by the present disclosure is to provide a better safety profile relative to known mood-regulating compounds.
A further advantage of one or more embodiments provided by the present disclosure is to minimize or avoid completely the side effects from known mood-regulating compounds.
An additional advantage of one or more embodiments provided by the present disclosure is to improve the effect of and/or reduce the dose of one or more known mood-regulating compounds which are co-administered with the composition disclosed herein i.e. exert an adjuvant effect.
Yet another advantage of one or more embodiments provided by the present disclosure is to minimize or avoid completely unnecessary costs related to healthcare assistance.
A further advantage of one or more embodiments provided by the present disclosure is to use a bacterial strain that provides other health benefits as well.
Additional features and advantages of the invention are described herein and will be apparent from the following Detailed Description and the Figures.
BRIEF DESCRIPTION OF THE FIGURES
FIG. 1 shows the design of the clinical trial disclosed herein.
FIG. 2 shows graphs demonstrating the primary outcome from administration of B. longum ATCC
BAA-999, improvement in depression and anxiety dichomotous scores.
FIG. 3 shows graphs demonstrating the secondary outcome from administration of B. longum ATCC BAA-999, improvement in depression and anxiety continuous scores.
FIGS. 4 and 5 respectively show graphs and a table demonstrating that administration of B. longum ATCC BAA-999 significantly improved the physical global domain as well as general physical health (physical functioning) and problems with work of other daily activities (role physical) and resulted in an improvement trend in the mental subdomains of vitality and role emotional.
FIG. 6 shows fMRI images demonstrating greater engagement of the visual association and parietal cortices in the group administered B. longum ATCC BAA-999 relative to the placebo group and lesser engagement of brain centers involved in emotion and mood (amygdala and fronto-limbic region) in the group administered B. longum ATCC BAA-999 relative to the placebo group.
FIG. 7 shows a graph demonstrating the effect of the administration of B.
longum ATCC BAA-999 on CRH in the hypothalamus.

DETAILED DESCRIPTION
As used in this disclosure and the appended claims, the singular forms "a,"
"an" and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a bacterial strain" or "the bacterial strain" includes two or more bacterial strains.
The words "comprise," "comprises" and "comprising" are to be interpreted inclusively rather than exclusively. Likewise, the terms "include," "including" and "or" should all be construed to be inclusive, unless such a construction is clearly prohibited from the context.
Nevertheless, the compositions disclosed herein may lack any element that is not specifically disclosed. Thus, a disclosure of an embodiment using the term "comprising"
includes a disclosure of embodiments "consisting essentially of' and "consisting of' the components identified.
Similarly, the methods disclosed herein may lack any step that is not specifically disclosed herein.
Thus, a disclosure of an embodiment using the term "comprising" includes a disclosure of embodiments "consisting essentially of' and "consisting of' the steps identified.
The term "and/or" used in the context of "X and/or Y" should be interpreted as "X," or "Y," or "X
and Y." Where used herein, the terms "example" and "such as," particularly when followed by a listing of terms, are merely exemplary and illustrative and should not be deemed to be exclusive or comprehensive. Any embodiment disclosed herein can be combined with any other embodiment disclosed herein unless explicitly stated otherwise.
As used herein, "about" and "approximately" are understood to refer to numbers in a range of numerals, for example the range of -10% to +10% of the referenced number, preferably within -5% to +5% of the referenced number, more preferably within -1% to +1% of the referenced number, most preferably within -0.1% to +0.1% of the referenced number.
Furthermore, all numerical ranges herein should be understood to include all integers, whole or fractions, within the range. The term "between" includes the end points of the identified range.
Moreover, these numerical ranges should be construed as providing support for a claim directed to any number or subset of numbers in that range. For example, a disclosure of from 1 to 10 should

6 be construed as supporting a range of from 1 to 8, from 3 to 7, from 1 to 9, from 3.6 to 4.6, from 3.5 to 9.9, and so forth.
"Animal" includes, but is not limited to, mammals, which includes but is not limited to, rodents, aquatic mammals, domestic animals such as dogs and cats, farm animals such as sheep, pigs, cows and horses, and humans. Where "animal," "mammal" or a plural thereof is used, these terms also apply to any animal that is capable of the effect exhibited or intended to be exhibited by the context of the passage.
As used herein, the terms "individual" and "patient" are understood to include an animal, especially a mammal trying to get pregnant, pregnant or lactating, and more especially a human trying to get pregnant, a pregnant human or a human that has given birth e.g. a lactating human. Even more especially the terms will include a human at risk of peripartum emotional distress.
A person at risk of peripartum emotional distress may be a person that has previously suffered from peripartum emotional distress, has a family history of peripartum emotional stress or has previously suffered from emotional distress e.g. anxiety or depression.
However, while the terms "individual" and "patient" are often used herein to refer to a human, the present disclosure is not so limited. Accordingly, the terms "individual" and "patient" refer to any animal, mammal that can benefit from the treatment.
The terms "treatment" and "treating" include any effect that results in the improvement of the condition or disorder, for example lessening/ameliorating, reducing, modulating, or eliminating the condition or disorder. The term does not necessarily imply that a subject is treated until total recovery. Non-limiting examples of "treating" or "treatment of' a condition or disorder include:
(1) inhibiting the condition or disorder, i.e. arresting the development of the condition or disorder or its clinical symptoms and (2) relieving the condition or disorder, i.e.
causing the temporary or permanent regression of the condition or disorder or its clinical symptoms. A
treatment can be patient- or doctor-related.

7 The terms "prevention" or "preventing" mean causing the clinical symptoms of the referenced condition or disorder to not develop or reducing the risk of their development in an individual. The individual may be exposed or predisposed to the condition or disorder but does not yet experience or display symptoms of the condition or disorder. The terms "condition" and "disorder" mean any disease, condition, symptom, or indication but more particularly mean any peripartum emotional distress condition or disorder.
The relative term "optimize or optimise" as used herein mean to improve, increase, or enhance.
With respect to the optimization of breastfeeding duration the term can mean to increase the chance that an individual will breastfeed for the period they desire without needing to stop breastfeeding or excusive breastfeeding e.g. because of emotional distress e.g. to minimise the risk of early cessation of breastfeeding because of peripartum emotional distress.
With respect to the optimization of the quantity of breast milk, the term optimise can mean to increase the amount of breastmilk produced by an individual.
With respect to the optimization of the quality of breast milk, the term optimise can mean to improve the composition of breastmilk and for example to bring it in line or closer to the composition of breastmilk produced by an individual that is not or has not suffered from peripartum emotional distress. It may for example mean to reduce or modulate the concentration of stress hormones e.g. cortisol, in breastmilk and in particular to bring the concentration of these ingredients in line or closer to the concentration found in breastmilk produced by an individual that is not or has not suffered from peripartum emotional distress and more particularly the medium or mean of the concentration of a stress hormone e.g. cortisol, found in the breastmilk of individuals not suffering from perinatal emotional distress.
Stress hormones e.g. cortisol, are believed to be present in higher concentrations in breastmilk produced by of individuals that have suffered or are suffering from peripartum emotional distress.
Accordingly, this may be particularly beneficial in individuals that have or are suffering from peripartum emotional distress or at are risk of suffering from peripartum emotional distress.

8 The term "support" as used herein means to facilitate, enable, or to remove barriers. With respect to supporting the initiation of breastfeeding, the term support can mean to increase the chance that an individual will successfully initiate breastfeeding.
The terms "regulate" and "modulate" as used herein mean to bring back into balance or to maintain balance or homeostatis e.g. to normalize acitivity or the concentration of a compound and bring it in-line with that seen in a healthy or optimally functioning individual.
The term "maternal supplement" means a supplement that can be administered to a mammal in the perinatal period.
The terms "food," "food product" and "food composition" mean a product or composition that is intended for ingestion by an individual such as a human and provides at least one nutrient to the individual. The compositions o f the present disclosure, including the many embodiments described herein, can comprise, consist of, or consist essentially of the essential elements and limitations described herein, as well as any additional or optional ingredients, components, or limitations described herein or otherwise useful in a diet.
As used herein, "complete nutrition" contains sufficient types and levels of macronutrients (protein, fats and carbohydrates) and micronutrients to be sufficient to be a sole source of nutrition for the animal to which the composition is administered. Individuals can receive 100%
of their nutritional requirements from such complete nutritional compositions.
As used herein "perinatal or peripartum period" refers to the period before pregnancy when an individual is trying to get pregnant, to the pregnancy period, and to the post pregnancy period following birth including the lactation period.
In a first aspect of the present invention there is provided B. longum ATCC
BAA-999, or a composition comprising B. longum ATCC BAA-999 for use in an individual to;
support breastfeeding initiation and/or to optimise breastfeeding duration and/or to optimize the quality

9 and/or quantity of breast milk and/or to prevent or treat peripartum emotional distress and/or to regulate amygdala activity and/or to regulate the HPA axis e.g. the release of CRH.
In an embodiment the individual is a mammal trying to get pregnant, a pregnant mammal or a mammal that has given birth e.g. a lactating mammal. In a more specific embodiment the individual is a human trying to get pregnant, a pregnant human or a human that has given birth e.g. a lactating human.
The term peripartum emotional distress as used herein refers to any emotional distress occurring in the peripartum period. The emotional distress can be mild e.g. the baby blues to severe. Non limiting examples of conditions encompassed by the term peripartum emotional distress are, peripartum low mood, peripartum anxiety, peripartum depression and peripartum traumatic stress disorder.
In a particular embodiment the peripartum emotional distress is selected from the group consisting of peripartum low mood, peripartum anxiety, peripartum depression and peripartum traumatic stress disorder.
Without wishing to be bound by theory the inventors believe that B. longum ATCC BAA-999 may prevent or treat peripartum emotional distress, support breastfeeding initiation and/or, optimise breastfeeding duration, and/or to optimize the quality and/or quantity of breast milk in an individual by modulating activity in the in brain centers involved in the control of emotions and mood (limbic regions such as the amygdala), and/or by regulating hypothalamic-pituitary adrenal axis (hereinafter HPA) activity by modulating the release of CRH e.g. in the hypothalamus, and thereby regulating cortisol levels.
It is thought that dysregulation of the HPA in pregnancy, and the resulting raised cortisol levels, possibly brought about, at least in part, by an increased or an accelerated increase in CRH, may be linked to peripartum emotional distress. It is also thought that high cortisol concentrations in an individual are linked with decreased milk production and higher cortisol concentration in breastmilk.

B. longum ATCC BAA-999 is also known as BL999 and NCC3001 and may be obtained commercially from specialist suppliers, for example from Morinaga Milk Industry Co. Ltd. of Japan under the trademark BB536. The term "B. longum ATCC BAA-999" includes the bacterium, parts of the bacterium, and/or a growth medium fermented by the bacterium.
B. longum ATCC BAA-999 was deposited by the Assignee of the present application as NCC 3001 on January 29, 2001 at the Institut Pasteur, 28 rue du Docteur Roux, F-75024 Paris Cedex 15, France. All restrictions upon public access to the deposits will be irrevocably removed upon grant of a patent on this application, and the deposits will be replaced if viable samples cannot be dispensed by the depository.
The B. longum ATCC BAA-999 may be cultured according to any suitable method.
B. longum ATCC BAA-999 may be added to a composition in any technically feasible form e.g. a freeze-dried or spray-dried form.
The B. longum ATCC BAA-999 or composition comprising B. longum ATCC BAA-999 may be administered for any time period. However, administration daily for a time period of at least 3, 4, 5 or 6 weeks may be particularly effective. Accordingly, in an embodiment, the B. longum ATCC
BAA-999 or composition comprising B. longum ATCC BAA-999 can be administered to the individual daily for a time period that is at least three weeks, at least four weeks in some embodiments, at least five weeks in other embodiments, and at least six weeks in yet other embodiments.
The B. longum ATCC BAA-999, or composition comprising B. longum ATCC BAA-999 may be administered at any time during the perinatal period e.g. pre-pregnancy to an individual trying to get pregnant, to a pregnant individual or to an individual post birth (postnatally) e.g. during the lactating period in a lactating individual.
Administration to an individual desiring to get pregnant may be at any time during which an individual is trying to become pregnant however, ordinarily administration will be during at least the 12 months preceeding the pregnancy or desired pregnancy and more particularly 1, 2, 3 or 4 months preceding the pregnancy or desired pregnancy.
Administration during pregnancy may be at any point e.g. during the 1st, 2nd and/or 3rd trimester.
Administration after an individual has given birth may be at any point in particular may be for 2 years, lyear or 6months following birth (postpartum) and in particular for the time period when an individual is lactating.
Research indicates that dysregulation of the HPA axis and in particular accelerated increases in CRH during mid pregnancy are linked to an increased likelihood of peripartum anxiety/depression. Accordingly, treatment in early pregnancy and mid pregnancy may be particularly beneficial as it may prevent or minimise said accelerated increase in CRH.
Early pregnancy could be any time in the first trimester, mid pregnancy would be any time in the second trimester.
It may be particularly beneficial if individuals at risk of suffering from peripartum emotional distress are proactively given B. longum ATCC BAA-999, or composition comprising B. longum ATCC BAA-999 as a prophylactic treatment.
In an embodiment the B. longum ATCC BAA-999, or composition comprising B.
longum ATCC
BAA-999 is administered during pregnancy and post pregnancy e.g. during the lactation period. In a more specific embodiment B. longum ATCC BAA-999, or composition comprising B. longum ATCC BAA-999, is administered for the 2nd and/or 3rd trimester of a pregnancy and during the lactation period of an individual, in particular an individual at risk of peripartum emotional distress.
As stated above, the B. longum ATCC BAA-999 or composition can be administered to treat or prevent peripartum emotional distress. One way B. longum ATCC BAA-999 or the composition comprising B. longum ATCC BAA-999 can do this is by stabilising and/or improving mood of an individual in the peripartum period.

The term "mood" refers to a state or quality of feeling (an emotional state) at a particular time.
Moods differ from simple emotions in that they are less specific, less intense, and less likely to be triggered by a particular stimulus or event. Moods generally have either a positive or negative valence. An improved mood may comprise one or more of a decreased depressive level, a decreased anxiety level, a decreased stress level, an increased perceived energy level ("vitality"), a more positive emotional state, an increased self-esteem, a reduced amount and/or a reduced intensity of negative thoughts and/or negative tensions, a reduced risk of mood swings, or retention of a positive mood.
The B. longum ATCC BAA-999 or composition comprising B. longum ATCC BAA-999 may be administered orally and/or enterally At least a portion of the B. longum ATCC BAA-999 may be living bacterium.
Additionally or alternatively, at least a portion of the B. longum ATCC BAA-999 may be inactivated non-replicating bacterium.
"Non-replicating" means that no viable cells and/or colony forming units can be detected by classical plating methods. Such classical plating methods are summarized in the microbiology book: James Monroe Jay, Martin J. Loessner, David A. Golden. 2005. Modern food microbiology.
7th edition, Springer Science, New York, N.Y. 790 p. Typically, the absence of viable cells can be shown as follows: no visible colony on agar plates or no turbidity in liquid growth medium after inoculation with different concentrations of bacterial preparations ("non-replicating" samples) and incubation under appropriate conditions (aerobic and/or anaerobic atmosphere for at least 24 h).
In some embodiments, such as special sterile food products or medicaments, a non-replicating form of the B. longum ATCC BAA-999 may be preferable. For example, at least 80%, preferably at least 90%, more preferably at least 95% of the B. longum ATCC BAA-999 can be non-replicating in the composition.
In an embodiment, at least a part of the B. longum ATCC BAA-999 are alive and preferably arrive alive in the intestine. For example, at least 5%, preferably at least 10%, more preferably at least 15% of the B. longum ATCC BAA-999 can be viable in the composition. As a result, the alive B.
longum ATCC BAA-999 can persist in the intestine and may increase their effectiveness by multiplication. The alive B. longum ATCC BAA-999 may also be effective by interacting with the commensal bacteria and/or the host.
The B. longum ATCC BAA-999 or composition comprising B. longum ATCC BAA-999 may be administered in any amount that is effective in achieving the objective of the present invention.
An amount adequate to accomplish this purpose is defined as "a therapeutically effective dose".
Amounts effective for this purpose will depend on a number of factors known to those of skill in the art, such as the severity of the condition and the weight and general state of the patient.
In prophylactic applications, the composition can be administered to a patient susceptible to or otherwise at risk of peripartum emotional distress in an amount that is sufficient to at least partially reduce the risk of developing said condition. Such an amount is "a prophylactically effective dose." Again, the precise amounts depend on a number of patient-specific factors, such as the patient's state of health and weight.
An individual at risk of peripartum emotional distress may be a person that has previously suffered from peripartum emotional distress, has a family history of peripartum emotional stress or has previously suffered from emotional distress e.g. anxiety or depression.
The B. longum ATCC BAA-999 or composition comprising B. longum ATCC BAA-999 is preferably administered in an amount that provides a therapeutically effective dose and/or in a prophylactic effective dose of the B. longum ATCC BAA-999. If at least a portion of the B.
longum ATCC BAA-999 is present in a viable form, the B. longum ATCC BAA-999 is theoretically effective in any concentration because the B. longum ATCC BAA-999 can colonize the gut and multiply therein.

Nevertheless, a daily dose of the B. longum ATCC BAA-999 or composition comprising B.
longum ATCC BAA-999 preferably provides between 104 and 1012 cfu (colony forming units) of the B. longum ATCC BAA-999, more preferably from 104 to 1011 cfu, most preferably from 104 to 1010 cfu. The composition may comprise between 102 and 1010 cfu, preferably 102 to 109 cfu, more preferably 102 to 108 cfu of the B. longum ATCC BAA-999 per gram dry weight of the composition.
In the case of inactivated and/or non-replicating B. longum ATCC BAA-999, the B. longum ATCC BAA-999 or composition comprising B. longum ATCC BAA-999 can be administered in a dose comprising between 102 and 1010 non-replicating cells of the B. longum per gram of dry weight of the composition, preferably 103 to 1010 non-replicating cells per gram of dry weight of the composition, more preferably 105to 1010 non-replicating cells per gram of dry weight of the composition.
Non-replicating micro-organisms do not form colonies, so the term "cells"
indicates the amount of non-replicating micro-organisms obtained from the specified amount of replicating bacterial cells. This amount includes micro-organisms that are inactivated, non-viable or dead, or present as fragments such as DNA or cell wall materials.
The B. longum ATCC BAA-999 or composition comprising B. longum ATCC BAA-999 may be administered or employed in any form suitable for ingestion by an individual.
In particular the composition will be a maternal supplement in the form of a powdered nutritional composition to be consumed directly or to be reconstituted in for example milk, juice or water, or sprinkled on food before consumption, a food product, a drink, a tablet or pill for example a soft gel capsule.
If the composition comprising B. longum ATCC BAA-999 is a powder it may be a powder having a water activity less than 0.2, preferably less than 0.15. The composition may be a shelf-stable powder. The low water activity can provide this shelf stability and can ensure that the B.

longum ATCC BAA-999 and any additional probiotic micro-organism will remain viable even after long storage times. Water activity (aw) is a measurement of the energy status of the water in a system and is defined as the vapor pressure of water divided by that of pure water at the same temperature; therefore, pure distilled water has a water activity of exactly one.
The composition comprising B. longum ATCC BAA-999 may comprise a protein. Non-limiting examples of suitable proteins include animal proteins (such as milk protein, meat protein or egg protein), a vegetable protein (such as soy protein, wheat protein, rice protein, or pea protein);
mixtures of free amino acids; or combinations thereof Milk proteins such as casein and whey, and soy proteins are particularly preferred.
The proteins may be intact, hydrolyzed, or a mixture of intact and hydrolyzed proteins. Partially hydrolyzed proteins (degree of hydrolysis between 2 and 20%) may be advantageous for human subjects and/or animals at risk of developing cows' milk allergy. Furthermore, pre-hydrolyzed protein sources are generally easier digested and absorbed by an impaired gastro-intestinal tract.
If hydrolyzed proteins are used, the hydrolysis process may be carried out as desired and as known in the art. For example, a whey protein hydrolysate may be prepared by enzymatically hydrolyzing the whey fraction in one or more steps. If the whey fraction used as the starting material is substantially lactose-free, the protein can suffer much less lysine blockage during the hydrolysis process. This enables the extent of lysine blockage to be reduced from about 15% by weight of total lysine to less than about 10% by weight of lysine; for example about 7% by weight of lysine which greatly improves the nutritional quality of the protein source.
The composition comprising B. longum ATCC BAA-999 may comprise a carbohydrate and/or a fat.

The fat may for example be a long chain polyunsaturated fatty acids, such as arachidonic acid (ARA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) and/or docosapentaenoic acid (DPA).
Non-limiting examples of suitable carbohydrates include sucrose, lactose, glucose, fructose, corn syrup solids, maltodextrins, and mixtures thereof Additionally or alternatively, a dietary fiber may be added. Dietary fiber passes through the small intestine undigested by enzymes and functions as a natural bulking agent and laxative. Dietary fiber may be soluble or insoluble and generally a blend of the two types is preferred. Non-limiting examples of suitable dietary fibers include soy, pea, oat, pectin, guar gum, partially hydrolyzed guar gum, gum Arabic, fructo-oligosaccharides, acidic oligosaccharides, galacto-oligosaccharides, sialyl-lactose and oligosaccharides derived from animal milks. A preferred fiber blend is a mixture of inulin with shorter chain fructo-oligosaccharides. In an embodiment, the fiber content is between 2 and 40 g/L of the composition, for example between 4 and 10 g/L.
The composition comprising B. longum ATCC BAA-999 may comprise minerals and/or micronutrients such as trace elements and vitamins in accordance with the recommendations of Government bodies such as the USRDA. For example, the composition may comprise, per daily dose, one or more of the following micronutrients, preferably in the ranges given: 300 to 500 mg calcium, 50 to 100 mg magnesium, 150 to 250 mg phosphorus, 5 to 20 mg iron, 1 to 7 mg zinc, 0.1 to 0.3 mg copper, 50 to 200 iug iodine, 5 to 15 iug selenium, 1000 to 3000 iug beta carotene,

10 to 80 mg Vitamin C, 1 to 2 mg Vitamin Bl, 0.5 to 1.5 mg Vitamin B6, 0.5 to 2 mg Vitamin B2, 5 to 18 mg niacin, 0.5 to 2.0 iug Vitamin B12, 100 to 800 iug folic acid, 30 to 70 iug biotin, 1 to 5 iug Vitamin D, and/or 3 to 10 iug Vitamin E.
The composition comprising B. longum ATCC BAA-999 may also comprise myo-inositol.
The composition comprising B. longum ATCC BAA-999 may also comprise an additional food grade micro-organism (i.e., in addition to the B. longum ATCC BAA-999). "Food grade" micro-organisms are micro-organisms that are safe for use in food. The food grade micro-organisms can comprise food-grade yeast. The food grade bacteria may be selected from the group consisting of lactic acid bacteria, bifidobacteria, propionibacteria and mixtures thereof Non-limiting examples of suitable food grade yeast include Saccharomyces cerevisiae and/or Saccharomyces boulardii.
The food grade bacteria can comprise another/additional probiotic bacteria, although in some embodiments the B. longum ATCC BAA-999 is the only probiotic bacteria in the composition.
"Probiotic" means microbial cell preparations or components of microbial cells with a beneficial effect on the health or well-being of the host. (Salminen S., Ouwehand A., Benno Y. et al "Probiotics: how should they be defined" Trends Food Sci. Technol. 1999:10 107-10).
Probiotic bacteria are preferably selected from the group consisting of lactic acid bacteria, bifidobacteria, propionibacteria and mixtures thereof Probiotic bacteria may be any lactic acid bacteria or bifidobacteria with established probiotic characteristics. For example, probiotic bacteria may be capable of promoting the development of a bifidogenic intestinal microbiota.
Non-limiting examples of suitable probiotic bacteria include Bifidobacterium, Lactobacillus, Streptococcus, Saccharomyces and mixtures thereof, in particular selected from the group consisting of Bifidobacterium longum, Bifidobacterium lactis, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus paracasei, Lactobacillus johnsonii, Lactobacillus plantarum, Lactobacillus salivarius, Enterococcus faecium, Saccharomyces boulardii and and mixtures thereof, preferably selected from the group consisting of Lactobacillus johnsonii (NCC533; CNCM 1-1225), Bifidobacterium longum (NCC490; CNCM 1-2170), Bifidobacterium longum (NCC2705; CNCM 1-2618), Bifidobacterium lactis (2818;

3446), Lactobacillus paracasei (NCC2461; CNCM 1-2116), Lactobacillus rhamnosus GG
(ATCC53103), Lactobacillus rhamnosus (NCC4007; CGMCC 1.3724), Enterococcus faecium SF
68 (NCIMB10415), and mixtures thereof B. longum ATCC BAA-999 and any additional probiotic micro-organism may be provided or comprised in the composition comprising B. longum ATCC BAA-999 in an encapsulated form.
Encapsulation of the bacteria can have therapeutical and technical advantages.
For example, encapsulation can increase the survival of the bacteria and thus the number of live bacteria which arrive in the intestine. Furthermore, the bacteria can be gradually released, allowing a prolonged action of the bacteria on the health of the subject. For example, the bacteria may be freeze or spray dried and incorporated into a gel.
The composition comprising B. longum ATCC BAA-999 may comprise at least one prebiotic.
"Prebiotic" means a food substance intended to promote the growth of probiotic bacteria in the intestines. A prebiotic can promote the growth of certain food grade bacteria, in particular growth of probiotic bacteria, in the intestines and can thus enhance the effect of B. longum ATCC BAA-999 and any additional probiotic bacteria. Preferably the prebiotic is selected from the group consisting of oligosaccharides and optionally fructose, galactose, mannose, soy and/or inulin; dietary fibers; or mixtures thereof The composition may further contain one or more of the following: a protective hydrocolloid (such as a gum, a protein, a modified starch), a binder, a film-forming agent, an encapsulating agent, a wall/shell material, a matrix compound, a coating, an emulsifier, a surface active agent, a solubilizing agent (such as an oil, a fat, a wax, a lecithin), an adsorbent, a carrier, a filler, a co-compound, a dispersing agent, a wetting agent, a processing aid (such as a solvent), a flowing agent, a taste masking agent, a weighting agent, a jellifying agent, a gel forming agent, an antioxidant or an antimicrobial. The composition may also contain a conventional pharmaceutical additive, adjuvant, excipient or diluent, including, but not limited to, water, gelatin of any origin, vegetable gum, ligninsulfonate, talc, a sugar, a starch, gum arabic, a vegetable oil, polyalkylene glycol, a flavoring agent, a preservative, a stabilizer, a, emulsifying agent, a buffer, a lubricant, or a colorant. Such further components are preferably selected having regard to their suitability for the intended recipient.

In an embodiment, the composition is a nutritionally complete formula.
In another aspect of the present invention B. longum ATCC BAA-999 or a composition comprising B. longum ATCC BAA-999 may be simultaneously, sequentially or separately administered to another ingredient that may; support breastfeeding initiation and/or optimise breastfeeding duration and/or optimize the quality and/or quantity of breast milk and/or prevent or treat peripartum emotional distress and/or regulate amygdala activity and/or to regulate the HPA axis e.g. the release of CRH, in an individual.
This may be beneficial as it will enable a lesser amount of said other ingredient to be used e.g a lower dose or less frequent administration.
Non limiting examples of such other ingredients include: anti-depressants, anti-anxiety drugs, beta and herbal remedies for emotional distress such as St. John's wart, extracts of passion flower, valerian, kali phos.
The B. longum ATCC BAA-999 or composition comprising B. longum ATCC BAA-999, optionally simultaneously sequentially or separately administered to said another ingredient, may also be administered in combination with a form of psychosocial or psychological intervention e.g.
cognitive behavioural therapy, interpersonal therapy, parent training programs, and/or alternative therapy and/or complementary therapy e.g. yoga therapy, music therapy, art therapy.
This may be beneficial as it may increase the effectiveness of the psychosocial or psychological intervention or alternative and/or complementary therapy.
In another aspect of the invention there is provided a method of supporting breastfeeding initiation and/or optimising breastfeeding duration and/or optimising the quality and/or quantity of breast milk and/or preventing or treating peripartum emotional distress and/or regulating amygdala activity and/or regulating the HPA axis e.g. the release of CRH, in an individual; said method comprising administering B. longum ATCC BAA-999 or a composition comprising B.
longum ATCC BAA-999 to said individual. Said method may be a natural therapy.

In another aspect of the present invention there is provided B. longum ATCC
BAA-999 or a composition comprising B. longum ATCC BAA-999 for use in the manufacture of a composition for use in an individual to; support breastfeeding initiation and/or to optimise breastfeeding duration and/or to optimize the quality and/or quantity of breast milk and/or to prevent or treat peripartum emotional distress and/or to regulate amygdala activity and/or to regulate the HPA
axis e.g. the release of CRH.
In another apect of the present invention there is provided use of B. longum ATCC BAA-999, or a composition comprising B. longum ATCC BAA-999 to; support breastfeeding initiation and/or to optimise breastfeeding duration and/or to optimize the quality and/or quantity of breast milk and/or to prevent or treat peripartum emotional distress and/or to regulate amygdala activity and/or to regulate the HPA axis e.g. the release of CRH, in an individual.
Said use may be non-therapeutic.
In another aspect of the invention there is provided B. longum ATCC BAA-999, or a composition comprising B. longum ATCC BAA-999, for use to prevent or treat the negative effects of peripartum emotional distress in the offspring of individuals.
Said negative effects may for example be caused by over exposure to stress hormones e.g. cortisol, in-utero or in breastmilk The term stress hormones as used herein refers to one or more hormones secreted in response to stress.
Non limiting examples of stress hormones are: cortisol, noradrenaline, CRH, adrenocorticotrophine (ACTH), and glucocorticoids.
Over exposure to stress hormones e.g. cortisol, refers to an exposure e.g.
intake of stress hormones such as cortisol in breastmilk, that is above an optimum amount. An optimum amount may be an amount falling within a range found in the breastmilk of individuals not suffering from perinatal emotional distress and more particularly the medium or mean of the concentration of a stress hormone e.g. cortisol, found in the breastmilk of individuals not suffering from perinatal emotional distress.
Over exposure to stress hormones, such as cortisol, in-utero and/or through breastmilk has been linked to neurobehavioural changes and to an increased risk of the development of one or more of the following conditions: attachment insecurity, emotional dysregulation, poor self-control, internalizing and externalizing problems, difficulties in cognitive functioning and in social interactions with parents and peers, impaired adaptive functioning and psychopathology including conduct disorders, affective disorders and anxiety and depressive disorders including a reduced to tolerance to stressful situations and increased sensitivity to anxiety . It has also been linked to an increased risk of the development of ADHD and learning disabilities.
By reducing and/or regulating stress in an individual the overexposure of offspring to stress hormones such as cortisol e.g. concentrations in breastmilk, may be prevented and/or minimized and one or more of these conditions may be prevented or treated e.g. The risk of them occurring may be reduced or their severity may be mitigated.
It is also thought that overexposure to stress hormones such as cortisol, in-utero or through breastmilk may increase crying time in infants and negatively affect infant sleeping. Accordingly, it is thought that by reducing and/or regulating over exposure to stress hormones such as cortisol, may minimize or regulate crying time and/or regulate sleeping.
Without wishing to be bound by theory it is thought that these positive effects may, at least in part, be the result of the regulation of HPA activity in the offspring. HPA activity plays a critical part not only in stress regulation but also in sleep, feeding, emotions, emotion-regulation, and in the equilibrium of the gut microbiota.
Offspring of individuals suffering from peripartum emotional distress are at an increased risk of overexposure to stress and stress hormines such as cortisol in-utero and through breastmilk.

In an embodiment said individual is an individual suffering from peripartum emotional distress. In a more specific embodiment the individual is a lactating human, and even more particularly a lactating human suffering from peripartum emotional distress.
Those skilled in the art will understand that they can freely combine all features of the present invention disclosed herein. In particular, features described for different embodiments of the present invention may be combined. Where known equivalents exist to specific features, such equivalents are incorporated as if specifically referred to in this specification. Further advantages and features of the present invention are apparent from the figure and non-limiting example.
The present invention will now be described in further details by the way of the following examples EXAMPLES
Example 1 The following non-limiting example is a randomized, double blind, placebo-controlled trial illustrative of B. longum ATCC BAA-999 reducing brain emotional reactivity.
Introduction: Specific probiotic bacteria can improve gut symptoms of IBS, however, their efficacy in treating co-morbid anxiety or a depressive symptom in this population is unknown.
B. longum ATCC BAA-999 was previously shown to normalize anxiety-like behavior and hippocampal neurotrophin levels in murine models of low-grade gut inflammation. The present inventors also have unpublished data in depression models in mice and also showing improved sleeping patterns in mice.
Aims & Methods: To evaluate the effects of B. longum ATCC BAA-999 on anxiety and depressive symptoms in patients with IBS and to study the underlying mechanisms, the present inventors conducted a randomized, double-blind, placebo-controlled, single center study in adult patients with IBS with diarrhea or mixed stool pattern (Rome III criteria) and mild to moderate anxiety and/or a depressive symptom. There were no differences in demographics and baseline data between the two groups, except for HAD-D scores, which were higher in B.
longum group (p=0.046).
B. longum ATCC BAA-999 (1.0E+10 CFU daily) or placebo (maltodextrin) was administered daily for six weeks. Validated questionnaires were used to assess anxiety and a depressive symptom (HAD score (Hospital Anxiety and Depression) and STAI (State-Trait Anxiety Inventory) score), IBS symptoms (adequate relief question, IBS Birmingham and Bristol scale), quality of life (SF-36) and somatization (PHQ-15) before administration, at the end of administration, and one month after the treatment (follow-up). This experimental design is shown in FIG. 1.
The present inventors assessed brain activation patterns using the backward masked fear paradigm (fMRI), cognitive function (memory and concentration), serum BDNF and inflammatory markers, and gut microbiota profiles (16S rRNA Illumina). The fMRI paradigm utilized Blood Oxygenation Level Dependent (BOLD) activation in response to the presentation of emotional stimuli (fear and happy faces) that were masked by a neutral face, measured over four consecutive fMRI scan acquisitions in the scanner. The amygdala was selected as a priori region of interest. This analysis was performed on all subjects.
Results: The present inventors randomized 44 patients, and 38 of them (B.
longum ATCC BAA-999=18, placebo=20) completed the study. The results are shown in FIGS. 2-6.
At six weeks, depression scores improved in patients treated with B. longum ATCC BAA-999 compared with placebo (RR 2.94, 95% CI 1.05-8.23, p=0.01), and this beneficial effect was maintained at follow-up. More patients treated with B. longum ATCC BAA-999 than placebo reported adequate relief of overall IBS symptoms (RR 2.1, 95% CI 1.15-3.83, p=0.02) but no statistically significant changes were found in the IBS Birmingham scores. The physical subdomain of quality of life improved in the group treated with B. longum ATCC BAA-999 compared with placebo (p=0.03, Mann¨Whitney U=228.5), with trends for improvement in the mental subdomains of vitality and emotional role functioning.
The beneficial effect of B. longum ATCC BAA-999 on a depressive symptom was maintained at one month post-treatment, while IBS symptoms and quality of life returned to baseline.

Specifically, FIG. 2 shows that treatment with B. longum ATCC BAA-999 improved depression scores both by intention-to-treat analysis (ITT) and per protocol analysis (PP). The beneficial effect of the B. longum ATCC BAA-999 was maintained at one month post-treatment (follow-up visit, with both ITT and PP analysis).
FIG. 3 shows that, adjusting for baseline, depression improvement as a continuous variable was achieved in the B. longum ATCC BAA-999 group (ANCOVA, p=0.049). This beneficial effect was not maintained at one month post-treatment. Treatment with B. longum ATCC

did not improve anxiety scores when analyzed as continuous variables.
FIGS. 4 and 5 show that there was a statistically significant improvement in SF-36 physical global domain, as well as in general physical health (Physical functioning) and problems with work or other daily activities (Role physical), in the B. longum ATCC BAA-999 compared to placebo. Non-significant differences between treatment groups were observed in SF-36 mental global domain. However, when analyzing the mental subdomains, non-statistically significant trends for improvement in Vitality and Role emotional were observed in the B.
longum ATCC
BAA-999 treated group.
FIG. 6 shows that functional MRI revealed significant reductions from baseline in response to negative emotional stimuli in multiple brain areas involved in emotion processing, including amygdala, frontal and temporal brain regions (p<0.001), in patients treated with B. longum ATCC BAA-999 compared with placebo. Specifically, before treatment, there was no major difference in response to fear stimuli vs fixation between placebo and B.
longum groups, except for greater engagement of the visual association and parietal cortices in B.
longum group.
However, at the end of the treatment, there was greater engagement of the amygdala, frontal, and temporal cortices and reduced engagement of occipital regions in placebo group.
No statistically significant differences were observed in anxiety, cognitive function, inflammatory markers, serum BDNF levels or gut microbiota profiles in patients treated with B.
longum ATCC BAA-999 compared to placebo.
Conclusion: The results demonstrate that a six-week treatment with B. longum improves co-morbid depressive symptoms, overall gastrointestinal symptoms and quality of life in patients with IBS. This effect is associated with changes in the brain activation patterns in the amygdala and fronto¨limbic regions, suggesting that reduction in limbic reactivity may underlie the beneficial effect of B. longum ATCC BAA-999.
Example 2 Mice chronically infected with the parasite Trichuris muris showed an increase in the content of CRH mRNA in the paraventricular nucleus of the hypothalamus in comparison to uninfected mice (Fig. 7). Treatment of infected mice with B. longum ATCC BAA-999 resulted in a reduction in the content of CRH (Fig. 7).
Materials and Methods:
Bacterial culture conditions Probiotics were grown under anaerobic conditions in Man-Rogosa-Sharpe (MRS, BioMerieux) broth (bifidobacteria with 0.5% cysteine). After 24 h at 37 C, the number of bacteria was estimated by measuring the optical density at 600 nm (1 0D600 = 108 bacteria/mL).
Bacterial cells were pelleted by centrifugation at 5000 X g for 15 min at 4 C and further resuspended at a concentration of 101 /mL in their spent culture medium. Aliquots of 1 mL were kept frozen until use.
Animals Male BALB/c or AKR mice (Harlan, Canada) were purchases at age of 6-8 wks and housed in a conventional specific pathogen free unit at McMaster University Central Animal Facility. All experiments were conducted with approval from the McMaster University Animal Care Committee.
Design Chronic Trichuris muris infection:
Male AKR mice were gavaged with T. muris (300 eggs /mouse) (n=26) or with placebo (n=9).
Infected mice were then gavaged daily with B. longum ATCC BAA-999 or fresh MRS
from day for 10 days. Uninfected mice were gavaged with fresh MRS on a daily basis from day 30 to day 40. At the end of probiotic or placebo administration the mice were sacrificed thereafter and tissue samples were obtained. Brains were snap frozen in liquid nitrogen and stored for in situ hybridization.
Level of CRH
Levels of CRH in hypothalamus (paraventricular nucleus) were assessed by in situ hybridizations using 35S-labeled RNA probes on frozen brain sections (Whitfield et al., 1990;
Foster et al., 2002). Briefly, brains were removed and rapidly frozen by immersion in 2-methylbutane at -60 C, and stored at -70 C. Cryostat-cut 12-mm-thick coronal sections were thaw-mounted onto gelatin-coated slides, dried, and stored at -35 C. Tissue sections were fixed with 4%
formaldehyde, acetylated with 0.25% acetic anhydride in 0.1 M triethanolamine¨HC1, pH 8.0, dehydrated, and delipidated with chloroform. Anti-sense CRH ribonucleotide probe (gift of Dr.
James Herman, University of Cincinnati) was transcribed from linearized plasmid using the Riboprobe System (Promega Biotech, Burlington, ON) with a-35S-UTP (specific activity > 1000 Ci/
mmol; Perkin-Elmer, Boston, MA) and T3 and T7 polymerases respectively. Radiolabeled probes were diluted in a hybridization buffer (0.6 M NaC1, 10 mM Tris pH 8.0, 1 mM EDTA pH 8.0, 10% Dextran sulfate, 0.01% sheared salmon sperm DNA, 0.05% total yeast RNA, type XI, 0.01%
yeast tRNA, lx Denhardt's solution) and applied to brain sections (approximately 500,000 CPM/section).
Slides were incubated overnight at 55 C in a humidified chamber. To reduce nonspecific binding of the probe, slides were washed in 20 mg/ml RNase solution for 30 min at room temperature, followed by 1 h each in 2XSSC at 50 C, 0.2XSSC at 55 and 60 C. Slides were dehydrated and air-dried for autoadiography. Slides and 14C plastic standards were placed in x-ray cassettes, apposed to film (BioMax MR; Eastman Kodak, Rochester, NY) for 5 days and developed (Kodak Medical X-Ray Processor). Autoradiographic film images of brain sections and standards were digitized with a solid-state camera with a 60 mm Nikon lens using QCapture software (Qicam;
Quorum Technologies Inc., Guelph, ON) and a Macintosh computer-based image analysis system with Image software (http://rsb.info.nih.gov/nih-image). Light transmittance through the film was measured by outlining the structure on the monitor. For CRH mRNA, the density slice feature was utilized to measure both the light transmittance and the area of mRNA signal.
The calculated DPM
were then multiplied by area to produce a measurement of integrated density.

FIG. 7 shows the effect of the administration of B. longum ATCC BAA-999 on CRH
in the hypothalamus.
Statistical analysis Data are presented as mean standard deviation or medians with interquartile ranges as appropriate. Data was analysed using either two-way ANOVA, test or non-paired t-test as appropriate. A p value of <0.05 was considered as statistically significant.
Example 3 An example composition comprising Bifidobacterium lactis BB122) further combined within vitamins, minerals and myo-inositol is set out in table 1. The composition in table I is for a nutritional supplement in a powder form, intended to be sprinkled on food.
Table 1 : Composition of Example 1 Ingredient Amount per daily dose Myo-inositol 4g Vitamin D 10 iLig Vitamin B6 2.6 mg Vitamin B12 5.2 iLig Vitamin B2 1.8 mg Zinc 10 mg I3-carotene 720 iLig Folic acid 400 iLig Iron 12 iLig Calcium 150 iLig Iodine 150 iLig B. longum ATCC BAA-999. lx101 cfu

Claims (16)

1. B. longum ATCC BAA-999, or a composition comprising B. longum ATCC BAA-999, for use in an individual to; support breastfeeding initiation and/or to optimise breastfeeding duration and/or to optimize the quality and/or quantity of breast milk and/or to prevent or treat peripartum emotional distress and/or to regulate amygdala activity and/or to regulate the HPA axis and more preferably the release of CRH.

2. B. longum ATCC BAA-999 or a composition comprising B. longum ATCC BAA-999 for use to according to claim 1 wherein the individual is a mammal and preferably a human trying to get pregnant, a pregnant human or lactating human.

3. B. longum ATCC BAA-999 or a composition comprising B. longum ATCC BAA-999 for use according to claim 1 or 2 wherein peripartum emotional distress is a condition selected from the group consisting of peripartum low mood, peripartum anxiety, peripartum depression, and peripartum traumatic stress disorder.

4. B. longum ATCC BAA-999 or a composition comprising B. longum ATCC BAA-999 for use according to anyone of claims 1 to 3 wherein said B. longum ATCC BAA-999 or a composition comprising B. longum ATCC BAA-999 is administered to the individual for at least 3 weeks.

5. B. longum ATCC BAA-999 or a composition comprising B. longum ATCC BAA-999 for use according to anyone of claims 1 to 4 wherein said B. longum ATCC BAA-999 or a composition comprising B. longum ATCC BAA-999 is administered to the individual for at least 6 weeks.

6. B. longum ATCC BAA-999 or a composition comprising B. longum ATCC BAA-999 for use according to any preceding claim wherein at least a portion of the B.
longum ATCC
BAA-999 is alive.

7. B. longum ATCC BAA-999 or a composition comprising B. longum ATCC BAA-999 for use according to any preceding claim wherein at least a portion of the B.
longum ATCC
BAA-999 is non-replicating cells.

8. B. longum ATCC BAA-999 or a composition comprising B. longum ATCC BAA-999 for use according to any preceding claim wherein said B. longum ATCC BAA-999 or composition is administered to the individual in a daily dose comprising between 104 to 1012cfu of the B. longum ATCC BAA-999.

9. A composition comprising B. longum ATCC BAA-999 for use according to any preceding claim wherein said composition further comprises a vitamin, a mineral, a prebiotic, myo-inositol, another probiotic, and/or a fatty acid.

10. B. longum ATCC BAA-999 or a composition comprising B. longum ATCC BAA-999 for use according to any preceding claim wherein said B. longum ATCC BAA-999 or composition is a maternal supplement.

11. B. longum ATCC BAA-999 or a composition comprising B. longum ATCC BAA-999 for use according to any preceding claim wherein said B. longum ATCC BAA-999 or composition is simultaneously, sequentially or separately administered to another ingredient that may; support breastfeeding initiation and/or optimise breastfeeding duration and/or optimize the quality and/or quantity of breast milk and/or prevent or treat peripartum emotional distress and/or regulate amygdala activity and/or to regulate the HPA axis and more preferably the release of CRH , in an individual.

12. B. longum ATCC BAA-999 or a composition comprising B. longum ATCC BAA-999 for use according to any preceding claim wherein said B. longum ATCC BAA-999 or a composition comprising B. longum ATCC BAA-999 is administered in combination with a form of psychosocial or psychological intervention and/or alternative therapy and/or complementary therapy.

13. A method of supporting breastfeeding initiation and/or optimising breastfeeding duration and/or optimising the quality and/or quantity of breast milk and/or preventing or treating peripartum emotional distress and/or regulating amygdala activity and/or regulating the HPA axis and more preferably the release of CRH, in an individual; said method comprising administering B. longum ATCC BAA-999 or a composition comprising B. longum ATCC
BAA-999 to said individual.

14. Use of B. longum ATCC BAA-999 in the manufacture of a composition for use in an individual to; support breastfeeding initiation and/or to optimise breastfeeding duration and/or to optimize the quality and/or quantity of breast milk and/or to prevent or treat peripartum emotional distress and/or to regulate amygdala activity and/or to regulate the HPA axis and more preferably the release of CRH.

15. Non-therapeutic use of B. longum ATCC BAA-999, or a composition comprising B.
longum ATCC BAA-999 to; support breastfeeding initiation and/or to optimise breastfeeding duration and/or to optimize the quality and/or quantity of breast milk and/or to prevent or treat peripartum emotional distress and/or to regulate amygdala activity and/or to regulate the HPA axis and more preferably the release of CRH, in an individual..

16. B. longum ATCC BAA-999, or a composition comprising B. longum ATCC BAA-999, for use to prevent or treat the negative effects of peripartum emotional distress in the offspring of individuals.