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Human milk provides complete nutrition for infants and at the same time promotes the growth of specific bacteria in the infant gastrointestinal tract. Breastfeeding can often be discontinued due to mastitis which is an inflammation of the breast tissue. We isolated 18 Staphylococcus aureus strains from milk donated by healthy (n = 6), subclinical (n = 6), and mastitic (n = 6) mothers, two strains of which were VISA (Vancomycin Intermediate S. aureus). All tested strains (n = 12) were able to form biofilms. We then examined the impact of nisin A and vancomycin alone and in combination on biofilm formation and eradication of selected strains (n = 8). We observed strain-specific responses, with the combinatorial treatment at 1/4X MIC (for both singularly) significantly inhibiting biofilm formation for seven out of eight strains when compared with nisin A or vancomycin alone. None of the selected treatments were able to eradicate pre-formed biofilms. Finally, we selected two strains, namely a VISA (APC3814H) and a strong biofilm former (APC3912CM) and used confocal microscopy to evaluate the effects of the antimicrobial agents at 1X MIC on biofilm inhibition and eradication. All treatments inhibited biofilm formation of APC3814H but were ineffective in eradicating a pre-formed biofilm. Single treatments at 1X MIC against APC3912CM cells did not prevent biofilm formation whereas combination treatment caused increased death of APC3912CM cells. Finally, the combination treatment reduced the thickness of the pre-formed APC3912CM biofilm as compared with the single treatments.

Bovine mastitis is a disease with a multi-etiological nature, defined as an inflammation of the udder. The main treatment for mastitis is the administration of antibiotics - usually directly to the udder. There is an urgent need for novel therapies to treat and prevent the disease, given the widespread emergence of antibiotic resistance and concomitant problems in the treatment of human and animal infections. We provide an overview of treatments for bovine mastitis, with emphasis on probiotics, bacteriocins, bacteriophages (phages), and phage endolysins. Probiotics have in recent years proved to be particularly efficacious in bovine mastitis treatment and prevention. In this case, the mode of action is most likely to be due to stimulation of the host immune response which clears the mastitis pathogen. Bacteriocins have the potential to be incorporated into teat washes and wipes, thus preventing pathogen spread on the farm. Phage therapy is limited by the inability of some phages to replicate in raw milk, as reported for some staphylococcal phages, and by their narrow host specificity. The use of phage endolysins is more promising, by enabling the development of broad host range potent antimicrobials, but additional research is required in terms of efficacy, safety and production.

peer-reviewed Bovine mastitis, an inflammation of the udder, is associated with increases in milk somatic cell count usually resulting from bacterial infection. We analysed 50 mastitic milk samples via cultivation, 16S rRNA sequencing and a combination of the two (culturomics) to define the complete microbial content of the milk. Most samples contained over 10,000 cfu mL-1 total bacterial counts including isolates that were haemolysin positive (n = 36). Among colonies isolated from blood agar plates, Streptococcus uberis was dominant (11/50) followed by Streptococcus dysgalactiae (6/50), Pseudomonas (6/50), Enterococcus faecalis (6/50), Escherichia coli (6/50), Staphylococcus argenteus (4/50), Bacillus (4/50) and Staphylococcus aureus (2/50). 16S rRNA profiling revealed that amplicons were dominated by Rhodococcus, Staphylococcus, Streptococcus and Pseudomonas. A higher inter-sample diversity was noted in the 16S rRNA readouts, which was not always reflected in the plating results. The combination of the two methods highlights the polymicrobial complexity of bovine mastitis.

Probiotic Feta cheese was produced using Propionibacterium freudenreichii subsp. shermanii as an adjunct since it has been shown to induce apoptosis of colon cancer cells in vitro and in vivo through the production of propionate and acetate. Microbiological and physicochemical analysis of the cheese was performed through the 60 d period of ripening. Counts of propionibacteria increased until day 7 and then remained constant until the end of ripening (approximately 9 log cfu g−1). Moreover, throughout ripening the presence of P. freudenreichii subsp. shermanii LMG 16424T was confirmed by 16S rRNA gene sequence analysis, while propionic acid was first detected on day 7 and reached a concentration of 52.1 m m on day 60. The ripened cheese containing P. freudenreichii subsp. shermanii was very well accepted by the sensory evaluation panellists. This is the first time that P. freudenreichii subsp. shermanii was studied as a probiotic adjunct in a white-brined cheese.

Raloxifene agonism of estrogen receptor (ER) in post-menopausal endometrium is not negligible. Based on a rational drug design workflow, we synthesized 14 analogues of raloxifene bearing a polar group in the aromatic ring of the basic side chain (BSC) and/or changes in the bulkiness of the BSC amino group. Analogues with a polar BSC aromatic ring and amino group substituents of increasing volume displayed increasing ER antagonism in Ishikawa cells. Analogues with cyclohexylaminoethoxy (13a) or adamantylaminoethoxy BSC (13b) lacking a polar aromatic ring displayed high ER-binding affinity and ER antagonism in Ishikawa cells higher than raloxifene and similar to fulvestrant (ICI182,780). The endometrial surface epithelium of immature female CD1 mice injected with 13b was comparable to that of vehicle-treated mice, while that of mice treated with estradiol, raloxifene or 13b in combination with estradiol was hyperplastic. These findings indicate that raloxifene analogues with a bulky BSC amino group could provide for higher endometrial safety treatment of the menopausal syndrome.

The ban of antibiotics at subtherapeutic levels as growth promoters in animals, along with the emerging challenges and prospects deriving from the use of probiotics and prebiotics in humans, opened the era of similar applications in the field of animal nutrition. Probiotics and prebiotics as novel additives in animal feeding aiming at host-protecting functions, which can maintain animal health and welfare, create fascinating options in favor of both animals and consumers of products of animal origin. In recent years, a considerable amount of knowledge regarding the potential of probiotics and prebiotics as supplements at prophylactic and therapeutic level has been accumulated. However, several bottlenecks are still observed, not only at the science, technology, and application level but also at the regulatory level. In this chapter, we summarize the current knowledge, along with the future trends on the applications of probiotics and prebiotics in animals and more specifically in polygastric, monogastric, and aquatic animals, pets, and bees. Moreover, we discuss the respective regulatory frame and its limitations.

Mastitis, which is generally described as an inflammation of breast tissue, is a common and debilitating disease which frequently results in the cessation of exclusive breastfeeding and affects up to 33% of lactating women. The condition is a primary cause of decreased milk production and results in organoleptic and nutritional alterations in milk quality. Recent studies employing culture-independent techniques, including metagenomic sequencing, have revealed a loss of bacterial diversity in the microbiome of mastitic milk samples compared to healthy milk samples. In those infected, the pathogens Staphylococcus aureus, Staphylococcus epidermidis and members of corynebacteria have been identified as the predominant etiological agents in acute, subacute and granulomatous mastitis, respectively. The increased incidence of antibiotic resistance in the causative species is also a key cause of concern for treatment of the disease, thus leading to the need to develop novel therapies. In this respect, probiotics and bacteriocins have revealed potential as alternative treatments.

Milk, a basic component of the human diet, is rich in all kinds of nutrients, such as proteins, carbohydrates, lipids, vitamins, and minerals. These compounds comprise an ideal growth substrate for both harmful and beneficial microorganisms, making milk a perishable raw material. As a result, milk contains numerous microorganisms that originate either from the animal itself, even when the animal is clinically healthy, or from the environment during the collection and storage of milk. The increasing social, economic, and research on non-cow types of milk and its products illustrates the renewed interest in the microbiota associated with these matrices. The present work is an overview of existing knowledge on the microbiota of sheep, goat, buffalo, camel, equine, yak, and human milk and their products. Relevant studies concerning both classical microbiological approaches as well as advanced molecular methods are included.