Spanish company Puleva has shown that human breast milk contains valuable probiotics. Arjan Geerlings, nutrition director of Puleva Biotech, explains how the company has now isolated three new Lactobacillus strains for commercial health applications
Human breast milk is known to be the best food for rapidly growing infants because it fulfils all the nutritional requirements. Additionally, several studies have shown that breast-feeding protects the newborn against infectious diseases. This effect may be due to the combined action of several breast milk components, such as immunoglobulins, immunocompetent cells and different antimicrobial compounds. Human breast milk also contains prebiotic substances, which selectively stimulate the growth of intestinal bacteria that exert positive effects in the newborn gut.
The key influence of human breast milk in the development of intestinal microflora, immune-modulating activity and prevention of infections is well documented. However, the presence of bacteria in human breast milk has never been investigated because it has always been considered a sterile fluid.
The loss of antimicrobial activity exhibited by fresh human milk after pasteurisation together with the discovery of the presence of bacteria in the umbilical cord blood and the meconium of newborns, led Puleva Biotech Exxentia to initiate a research project to clarify the bacterial composition present in human breast milk.
Initially, the company researched the presence and characterisation of human breast milk microbiota in samples of eight healthy breast-feeding mothers. General and specific culture media were used to isolate lactic acid bacteria from all the breast milk samples.1 In order to discard the presence of bacteria in human breast milk that was due to cross-contamination during sample collection, samples of breast skin were also collected. The absence of similarity among bacteria isolated from breast milk and from breast skin ruled out the contamination of human breast milk with breast skin and environmental bacteria.
Puleva Biotech Exxentia has demonstrated the presence of commensal bacteria in human breast milk and the existence of a natural transfer of commensal bacteria from the mother to her newborn infant. Human breast milk is the major factor in the initiation and development of the gut microbiota of breast-fed infants since it represents a continuous source of lactic acid bacteria to the infant gut.
It is estimated that an infant consuming approximately 800ml per day of breast milk ingests about 1 x 105 to 1 x 107 commensal bacteria. The commensal bacteria transfer by human breast milk is another means by which mothers protect the newborn against infections and also initiate a healthy microbiota that leads to the correct development of gut microbiota and immune system of the infant.
Puleva Biotech Exxentia obtained a high number of strains from the breast milk of healthy women. The company used an "in house" screening method to select the best strains based on their probiotic potential. The selection process included more than 40 different criteria that considered safety, functionality and technological aspects. The result was the selection of three strains naturally found in human breast milk that possessed the best probiotic characteristics,2,3 Lactobacillus gasseri (CECT5714), Lactobacillus fermentum (CECT5716) and Lactobacillus salivarius (CECT5713). The three probiotic strains were registered under the trade mark Hereditum, and patented (WO/2004/003235).
beneficial flora
Puleva has demonstrated, for the first time, that human breast milk is not sterile but in fact is an important source of beneficial bacteria for the infant. Three new Lactobacillus strains were isolated and are now available commercially for different applications. One of Puleva's applications is infant formulas; however, other novel applications are under development.
The fact that these specific strains can be found in human breast milk opened up a line of research into infections of the mammarian gland. From the company's research it is known that some Lactobacillus strains, hence those naturally present in human breast milk, are able to be transported from the gut to the mammarian gland in lactating women. This led to the hypothesis that Lactobacillus strains might compete, after oral administration, with pathogens in the female breast.
Indeed, women suffering from mastitis, an infection caused by Staphylococcus species, recovered significantly from an oral treatment with these probiotics strains. 4
Breast mastitis is an infection that commonly affects women who are breast-feeding (especially during the first two months after childbirth) but can occur in all women at any time. Mastitis is a benign (non-cancerous) condition that can usually be treated successfully with antibiotics. Signs of mastitis include red, hot, painful or inflamed breasts and other flu-like symptoms such as headache, nausea, high temperature, or chills.
It is estimated that around 15% of women who are breast-feeding suffer from this disease, and it is one of the main reasons why women stop giving breast milk during the first months after giving birth. The only treatment currently available is an antibiotic treatment.
Twenty women with staphylococcal mastitis were randomly divided in two groups. Those in the probiotic group daily ingested 10 log(10) CFU of Lactobacillus salivarius (CECT5713) and the same quantity of Lactobacillus gasseri (CECT5714) for four weeks, while those in the control group only ingested the excipient. Both Lactobacillus strains were originally isolated from breast milk.
On day 0, the mean staphylococcal counts in the probiotic and control groups were similar (4.74 and 4.81 log(10) CFU/ml, respectively), but lactobacilli could not be detected. On day 30, the mean staphylococcal count in the probiotic group (2.96 log (10) CFU/ml) was lower than that of the control group (4.79 log(10) CFU/ml). L. salivarius (CECT5713) and L. gasseri (CECT5714) were isolated from the milk samples of six of the 10 women of the probiotic group. At day 14, no clinical signs of mastitis were observed in the women assigned to the probiotic group, but mastitis persisted throughout the study period in the control group.
In conclusion, L. salivarius (CECT5713) and L. gasseri (CECT5714) appear to be an efficient alternative for the treatment of lactational infectious mastitis during lactation.
Before the publication of these results,5 a PCT patent was filed to protect the use of the Hereditum probiotics and compositions containing them for their use in the treatment of mastitis.
New studies are underway to support further the Hereditum probiotics for the treatment of mastitis, one of the principal reasons why women stop breastfeeding.
CPhI Innovation Award Winners
The CPhI Innovation Awards recognise innovation in drug discovery, development, production, packaging or in business models that benefit pharma manufacturing.
Arjan Geerlings collected the 2009 Gold Innovation Award at the CPhI Exhibition in Madrid on behalf of Puleva Biotech Exxentia for the company's novel probiotic products.
The Silver and Bronze winning innovations will be featured in forthcoming issues of Manufacturing Chemist.
