This study aimed to examine and compare the effectiveness of a novel spray-dried inoculant and a commercial freeze-dried additive on the fermentation quality, aerobic stability and bacterial population of vetch-oat silage. An entirely random design used a 3 × 4 factorial arrangement of treatments, with and without lactic acid bacteria (LAB) inoculants and four fermentation periods. Physicochemical parameters, microbiological counts and 16S rRNA gene sequencing analysis on Nanopore MinION were conducted to characterize the ensiling process. Both LAB inoculants increased dry matter, crude protein, lactic, acetic and propionic acid contents, while reducing pH, neutral detergent fibre, ammonia nitrogen/total nitrogen and ethanol concentrations compared to the control group. Overall, the native inoculant decreased the cell load of coliforms, yeasts and moulds. In addition, bio-inoculants enhanced the aerobic stability of vetch-oat intercrops. After ensiling, bacterial alpha diversity decreased noticeably; inoculation reduced the number of observed operational taxonomic units and the Shannon and Simpson indices. Notably, the relative abundance of Lactobacillus in the control group was lower than in treated silages, while the relative values of Staphylococcus increased sharply in the uninoculated group. In conclusion, the native strains showed promise for usage as a bio-inoculant in the ensiling of vetch-oat at a mixture rate of 1:1, producing an immediate impact as well as a favourable effect on the post-opening phase. This represents the first report on 16S rRNA gene-based nanopore metagenomics applied to the bacterial analysis of vetch-oat silage, providing a microbiological insight where native and commercial strains dominate the natural epiphytic community.

Animal husbandry is increasingly under pressure to meet world food demand. Thus, strategies are sought to ensure this productivity increment. The objective of this review was to gather advances in the use of bacterial probiotics in animal production. Lactobacilli correspond to the most used bacterial group, with several beneficial effects already reported and described, as well as the Enterococcus and Pediococcus genera – being the latter expressively used in aquaculture. Research on the Bifidobacterium genus is mostly focused on human health, which demonstrates great effects on blood biochemical parameters. Such results sustain the possibility of expanding its use in veterinary medicine. Other groups commonly assessed for human medicine but with prospective expansion to animal health are the genera Leuconostoc and Streptococcus, which have been demonstrating interesting effects on the prevention of viral diseases, and in dentistry, respectively. Although bacteria from the genera Bacillus and Lactococcus also have great potential for use in animal production, a complete characterization of the candidate strain must be previously made, due to the existence of pathogenic and/or spoilage variants. It is noteworthy that a growing number of studies have investigated the genus Propionibacterium, but still in very early stages. However, the hitherto excellent results endorse its application. In this way, in addition to the fact that bacterial probiotics represent a promising approach to promote productivity increase in animal production, the application of other strains than the traditionally employed genera may allow the exploitation of novel mechanisms and enlighten unexplored possibilities.

In the present research communication, we report on identification and quantification of four main lactic acid bacteria (LAB) genera (Lactococcus, Lactobacillus, Streptococcus and Leuconostoc), most common in Greek cheeses, by a novel culture-independent method. More specifically, new primers were designed to be used in both multiplex PCR for simultaneous identification and in real-time PCR for quantification of the LAB. The method was validated by applying it in parallel to culture-dependent method in a variety of cheeses from different Greek geographical locations, of different animal milk origins and of different production methods. While the standard plate culture method showed absence of Leuconostoc sp. in all cheeses, the culture-independent methods detected all four LAB genera studied. Furthermore, the relative presence of the four genera detected by the culture-independent method showed a pattern present in almost all cheese samples tested, indicating Lactococcus genus as the dominant one.

The present study aimed to evaluate the effects of chitosan instead of microbial inoculants on fermentation profile, losses, chemical composition, in vitro degradation, and aerobic stability of sugarcane silage (SS). Forty experimental silos (PVC tubes with 28 cm i.d., 25 cm height) were used in a randomized block design to evaluate the following treatments: (I) Control (CON): SS with no additive; (II) LB: SS ensiled with 5.0 × 105 colony forming units (CFU) of Lactobacillus buchneri (NCIM 40788)/g as-fed; (III) LPPA: SS ensiled with 1.6 × 105 CFU of L. plantarum and 1.6 × 105 CFU of Pediococcus acidilactici/g as-fed; and (IV) Chitosan (CHI): SS ensiled with 6 g/kg dry matter (DM) of chitosan. Microbial inoculation of SS reduced (P ≤ 0.05) silage pH relative to CON and CHI treatment. The LPPA decreased ammonia-nitrogen and LB decreased (P ≤ 0.05) ethanol content and increased acetic acid content relative to other treatments. The LPPA-silos had higher (P ≤ 0.05) gas losses and lower (P ≤ 0.05) DM recovery than other treatment silos. Consequently, LPPA reduced (P ≤ 0.05) DM and non-fibre carbohydrates and increased (P ≤ 0.05) neutral detergent fibre (NDF) silage content compared to other treatments. Treatments did not affect (P ≥ 0.212) DM and NDF in vitro degradation and silage pH after aerobic exposure. However, LB reduced silage temperature after aerobic exposure. Thus, LB reduces alcoholic fermentation and improves SS aerobic stability. Inoculation of LPPA reduces DM recovery and negatively affects SS chemical composition. Although CHI positively affects SS conservation relative to CON, it shows higher gas losses and decreased SS temperature after aerobic exposure compared to LB.

The current study aimed to evaluate the effects of cactus pear as a moistening additive on fermentative and microbiological characteristics, aerobic stability (AS), chemical composition and in situ rumen degradability of corn grain silage at different opening times. A completely randomized experimental design was adopted in a 4 × 3 factorial scheme with four levels of dry matter (DM) (50; 60; 70 and 80% of DM) and three opening times (30; 60 and 120 days after ensiling), with four replications. There was an effect of interaction (P < 0.05) between the DM levels and opening times on silage yeast population, effluent losses, gas losses, dry matter recovery (DMR), AS of the silage and on lactic acid bacteria, mould and yeast populations after AS trial. The 60% DM level presented DMR values above 930 g/kg of DM. However, the lowest AS time (96.52 h) was observed in silages with 60% DM at 60 days after ensiling, although all silages have shown high AS. The DM in situ degradability of the ensiled mass increased after the ensiling process at all DM levels and opening times, with the 60% DM content showing the best result. When using cactus pear as a corn grain moistening additive, the 60% DM level is recommended when the opening time is up 120 days.

Our objective was to analyze the diversity of the microbiota over 180 d of ripening of eight batches of artisanal goatskin Tulum cheeses by culture-dependent and culture-independent (PCR-DGGE) methods. V3 region of the bacterial 16S rRNA gene was amplified with the PCR after direct DNA isolation from the cheese samples. Nine different species and five genera were determined by culturing, while 11 species were identified in the PCR-DGGE technique. This diversity revealed the uniqueness of artisanal cheese varieties. The dominant genera in all the cheese samples were composed of Enterococcus species. The culture-dependent method revealed five genera (Enterococcus,Bacillus,Lactococcus,Lactobacillus, Sphingomonas) while three genera (Enterococcus, Streptococcus, Lactococcus) were detected in the culture-independent method. It was concluded that combining the two methods is important for characterizing the whole microbiota of the Tulum cheese varieties produced in the Anamur region.

This paper reveals the technological properties of lactic acid bacteria isolated from raw milk (colostrum and mature milk) of Wagyu cattle raised in Okayama Prefecture, Japan. Isolates were identified based on their physiological and biochemical characteristics as well as 16S rDNA sequence analysis. Streptococcus lutetiensis and Lactobacillus plantarum showed high acid and diacetyl-acetoin production in milk after 24 h of incubation at 40 and 30°C, respectively. These strains are thought to have potential for use as starter cultures and adjunct cultures for fermented dairy products.

This study aimed to compare the effect of different ratios of Streptococcus thermophilus to Lactobacillus bulgaricus (3 : 1, 1 : 1, and 1 : 3) under the various stressful temperatures (37 and 45°C) on the fatty acid profiles quality of Kermanshahi roghan (yogurt by-product) and sour cream to obtain a formula for producing a kind of animal fat healthier than milk and cream. Stresses such as fermentation play an important role in bacterial behavior and consequently in food quality. Our findings presented a significant difference between roghan and sour cream fatty acid levels only at 37°C. Furthermore, starter culture 3 : 1 was the best starter for producing products with a higher quality of fatty acid profile at 37°C, and a 1 : 1 S. thermophilus to L. bulgaricus ratio was optimal at 45°C. It seems that bacteria adapt to harsh growth conditions by changing the fatty acid profiles, and these changes warrant consideration in the production of a kind of animal fat with the best fatty acid profiles. In conclusion, the roghan fatty acid profile is more suitable than sour cream only at 37°C.

For this research communication, 90 samples of a Brazilian dairy were combined into four groups (raw material, final product, food-contact and non-food contact surfaces) and analyzed by metataxonomics based on 16S rRNA gene sequencing. The results showed high alpha-diversity indexes for final product and non-food contact surfaces but, overall, beta-diversity indexes were low. The samples were separated in two main clusters, and the core microbiota was composed by Macrococcus, Alkaliphilus, Vagococcus, Lactobacillus, Marinilactibacillus, Streptococcus, Lysinibacillus, Staphylococcus, Clostridium, Halomonas, Lactococcus, Enterococcus, Bacillus and Psychrobacter. These results highlight that rare taxa occur in dairies, and this may aid the development of strategies for food protection.

The aim of the work presented in this Research Communication was to evaluate the effect of fermented whey (FW) with Lactobacillus rhamnosus RC007 in a mice model. BALB/c mice were divided into three groups: control group: animals received orally 0.1 ml of phosphate buffered saline (PBS); FW group: animals received orally 0.1 ml of FW; whey (W) group: animals received orally 0.1 ml of W without fermentation with probiotic bacterium. After 10 d mice were sacrificed. Small intestines were collected for determination of IL-10; IL-6, TNFα, goblet cells and intraepithelial lymphocytes. Increases of all the cytokines assayed were observed in mice that received FW compared to control and W group. The ratio between the anti and pro-inflammatory cytokines (IL-10/TNFα) increased in the group of mice that received FW. The number of goblet cells and intraepithelial lymphocytes were also increased in animals that received FW. The results showed that FW with L. rhamnosus RC007 was able to stimulate and to modulate mouse immune system. Whey fermented by this probiotic bacterium is an interesting alternative for development of a new food additive for pig production, taking advantage of the beneficial properties of probiotic bacterium and the nutritional properties of whey.

The experiments reported in this research paper aimed to determine the technological properties of indigenous Lactococcus lactis strains isolated from Lait caillé, a spontaneous fermented milk, from the perspective of starter culture development. Fermentations were conducted to determine the acidification patterns. The ropy character, growth in 0.04 g/ml NaCl and citrate metabolism were additionally tested. Furthermore, the rheological properties of samples from selected strains and the impact of cold storage were evaluated. Based on the rate of acidification, the indigenous strains were divided into 2 groups depending on their fermentation time, i.e. 10–13 h (fast acidifier), and up to 72 h (slow acidifier), respectively. The physiological tests suggested that most of these strains produced exopolysaccharides but none could ferment citrate. The flow properties of the samples inoculated by the fast acidifier strains showed a time-dependent shear thinning behaviour, while their viscoelastic properties corresponded structurally to those of weak gels. Cold storage decreased the viscosity and CFU counts for most of the indigenous strains tested. This study is a step towards the definition of starter cultures for African spontaneous fermented milks such as Lait caillé.

This review aims to familiarize the reader with research efforts on the cultivation media of lactic acid bacteria (LAB). We have also included a brief discussion on standard ingredients used in LAB media and chemically defined media as related to bacterial growth requirements. Recent research has focused on modifying standard media for the enumeration, differentiation, isolation, and identification of starter cultures and probiotics. Even though large numbers of these media have been developed to serve dairy microbial control, they have failed to provide consistent results. The research consequently points to the need to develop a reliable lactobacilli growth medium for the dairy industry.

Scientific studies demonstrate the importance of intestinal microbiota to human health and how probiotic microorganisms can positively affect health when administered regularly and in adequate amounts. Probiotic bacteria can be part of fermented products and their functional importance is associated mainly with their metabolism. They are thought to benefit individuals to maintain their health and also to strengthen resistance against various types of diseases. The acceptance of probiotic cultures and products by consumers increased when these bacteria were marketed as natural cultures that help in digestion and health. Considering this, the food industry has an increasing demand for new candidates as probiotic cultures, and the dairy industry has a particular interest for fermented milks and other dairy products, since these are the most common food vehicles for probiotic cultures. Therefore, the dairy industries are increasingly seeking to improve their products with these beneficial bacteria. However, the legal peculiarities and excess of control agencies in Brazil makes the registration of these products and the collection of data very complex. Prospective analysis suggests that probiotic foods have the potential to effect a considerable expansion of the dairy industry, allowing the dairy sector to grow and for these products to be increasingly sought by consumers globally. For this, not only actions on research and innovation are necessary, but also official clarifications on the claims for considerations of microbiological security and functionality of these products. This review aims to elucidate important probiotic research regarding the isolation and characterization of beneficial cultures in Brazil, and to demonstrate the relevance of the dairy chain as a potential source of novel cultures for the development of new probiotic products to expand the Brazilian dairy industry.

Gut symbionts can augment resistance to pathogens by stimulating host-immune responses, competing for space and nutrients, or producing antimicrobial metabolites. Gut microbiota of social bees, which pollinate many crops and wildflowers, protect hosts against diverse infections and might counteract pathogen-related bee declines. Bumble bee gut microbiota, and specifically abundance of Lactobacillus ‘Firm-5’ bacteria, can enhance resistance to the trypanosomatid parasite Crithidia bombi. However, the mechanism underlying this effect remains unknown. We hypothesized that the Firm-5 bacterium Lactobacillus bombicola, which produces lactic acid, inhibits C. bombi via pH-mediated effects. Consistent with our hypothesis, L. bombicola spent medium inhibited C. bombi growth via reduction in pH that was both necessary and sufficient for inhibition. Inhibition of all parasite strains occurred within the pH range documented in honey bees, though sensitivity to acidity varied among strains. Spent medium was slightly more potent than HCl, d- and l-lactic acids for a given pH, suggesting that other metabolites also contribute to inhibition. Results implicate symbiont-mediated reduction in gut pH as a key determinant of trypanosomatid infection in bees. Future investigation into in vivo effects of gut microbiota on pH and infection intensity would test the relevance of these findings for bees threatened by trypanosomatids.

The objective of the study reported in this Research Communication was to investigate the effect of fermentation temperature (37 and 45 °C) and different ratios of Streptococcus thermophilus to Lactobacillus bulgaricus (3 : 1, 1 : 1 and 1 : 3) on Kermanshahi roghan and yoghurt fatty acid profiles (FAP) in order to obtain a product with optimized fatty acid profiles. Kermanshahi roghan is a yoghurt by-product in western Iran (Kermanshah). The results revealed that incubation temperature at 37 °C as compared to 45 °C had a better effect on fatty acid profiles of roghan and yoghurt. Furthermore, the results showed that fatty acid profile of roghan is better than yoghurt at two experimental temperatures. On the other hand, the roghan products made by equal ratio of S. thermophilus and L. bulgaricus (1 : 1) had the best quality of fatty acid profiles. Although a lower incubation temperature increases incubation time, our finding suggests that inoculation ratio 1 : 1 at 37 °C as compared to 45 °C can affect the quality of roghan and yoghurt fatty acid profiles.

Cronobacter sakazakii is a life-threatening foodborne pathogen found in powdered infant formula and dairy products. Kefir is a dairy probiotic product and its antimicrobial activity against C. sakazakii was reported in our previous study. To identify key microorganisms that mediate growth suppression, we tested the antimicrobial activity of culture supernatants derived from lactic acid bacteria found in kefir. Lactobacillus kefiri DH5, L. kefiranofaciens DH101, and Bifidobacterium longum 720 (a commercial probiotic strain that served as a positive control) all significantly inhibited the growth of C. sakazakii ATCC 29544, delaying the initiation of exponential growth from 3 to 9 h in the nutrient broth. Among them, L. kefiri DH5 exerted the strongest antimicrobial effects against C. sakazakii, showing bactericidal effect at the addition of 300 µl of supernatant in 1 ml of nutrient broth. Interestingly, the supernatant of L. kefiri DH5 has higher pH and lower titrable acidity than that of L. kefiranofaciens DH101, suggesting metabolites produced by heterofermentation of L. kefiri acted more effectively to antagonise the growth of C. sakazakii. In addition, the supernatant of L. kefiri DH5 induced the leakage of cytoplasmic materials including nucleic acid and proteins, suggesting L. kefiri DH5 disrupted the cellular membrane integrity of C. sakazakii. Considering that pH neutralisation reduced the L. kefiri-dependent growth suppression, it is inferred that this activity is mainly due to organic acids produced during the fermentation process.

Infectious diarrhoea is a worldwide problem in newborns. Optimal bacterial colonisation may enhance gut maturation and protect against pathogenic bacteria after birth. We hypothesised that lactic acid bacteria (LAB) administration prevents pathogen-induced diarrhoea in formula-fed newborns. Newborn caesarean-delivered, colostrum-deprived term piglets on parenteral nutrition for the first 15 h, were used as models for sensitive newborn infants. A commercially available probiotic strain, Lactobacillus paracasei F19 (LAP, 2·6×108 colony-forming units (CFU)/kg per d) and a novel LAB isolate, Pediococcus pentosaceus (PEP, 1·3×1010 CFU/kg per d), were administered for 5 d with or without inoculation of the porcine pathogen, Escherichia coli F18 (F18, 1010 CFU/d). This resulted in six treatment groups: Controls (n 9), LAP (n 10), PEP (n 10), F18 (n 10), F18–LAP (n 10) and F18–PEP (n 10). The pathogen challenge increased diarrhoea and density of F18 in the intestinal mucosa (P<0·05). LAB supplementation further increased the diarrhoea score, relative to F18 alone (P<0·01). Intestinal structure and permeability were similar among groups, whereas brush border enzymes were affected in variable intestinal regions with decreased activities in most cases after F18 and LAB inoculation. Bacterial density in colon mucosa increased after F18 inoculation (P<0·05) but was unaffected by LAB supplementation. In colon contents, acetic and butyric acids were increased by PEP (P<0·05). The LAB used in this study failed to reduce E. coli-induced diarrhoea in sensitive newborn pigs. In vulnerable newborns there may be a delicate balance among bacterial composition and load, diet and the host. Caution may be required when administering LAB to compromised newborns suffering from enteric infections.

The aim of this study was to characterise the gut microbiota composition of piglets fed bovine colostrum (BC), milk replacer (MR) or sow milk (SM) in the post-weaning period. Piglets (n 36), 23-d old, were randomly allocated to the three diets. Faecal samples were collected at 23, 25, 27 and 30 d of age. Digesta from the stomach, ileum, caecum and mid-colon was collected at 30 d of age. Bacterial DNA from all samples was subjected to amplicon sequencing of the 16S rRNA gene. Bacterial enumerations by culture and SCFA analysis were conducted as well. BC-piglets had the highest abundance of Lactococcus in the stomach (P<0·0001) and ileal (P<0·0001) digesta, whereas SM-piglets had the highest abundance of Lactobacillus in the stomach digesta (P<0·0001). MR-piglets had a high abundance of Enterobacteriaceae in the ileal digesta (P<0·0001) and a higher number of haemolytic bacteria in ileal (P=0·0002) and mid-colon (P=0·001) digesta than SM-piglets. BC-piglets showed the highest colonic concentration of iso-butyric and iso-valeric acid (P=0·02). Sequencing and culture showed that MR-piglets were colonised by a higher number of Enterobacteriaceae, whereas the gut microbiota of BC-piglets was characterised by a change in lactic acid bacteria genera when compared with SM-piglets. We conclude that especially the ileal microbiota of BC-piglets had a closer resemblance to that of SM-piglets in regard to the abundance of potential enteric pathogens than did MR-piglets. The results indicate that BC may be a useful substitute for regular milk replacers, and as a feeding supplement in the immediate post-weaning period.

Kefir is fermented milk produced from grains that comprise a specific and complex mixture of bacteria and yeasts that live in a symbiotic association. The nutritional composition of kefir varies according to the milk composition, the microbiological composition of the grains used, the time/temperature of fermentation and storage conditions. Kefir originates from the Caucasus and Tibet. Recently, kefir has raised interest in the scientific community due to its numerous beneficial effects on health. Currently, several scientific studies have supported the health benefits of kefir, as reported historically as a probiotic drink with great potential in health promotion, as well as being a safe and inexpensive food, easily produced at home. Regular consumption of kefir has been associated with improved digestion and tolerance to lactose, antibacterial effect, hypocholesterolaemic effect, control of plasma glucose, anti-hypertensive effect, anti-inflammatory effect, antioxidant activity, anti-carcinogenic activity, anti-allergenic activity and healing effects. A large proportion of the studies that support these findings were conducted in vitro or in animal models. However, there is a need for systematic clinical trials to better understand the effects of regular use of kefir as part of a diet, and for their effect on preventing diseases. Thus, the present review focuses on the nutritional and microbiological composition of kefir and presents relevant findings associated with the beneficial effects of kefir on human and animal health.

Fermented beverages hold a long tradition and contribution to the nutrition of many societies and cultures worldwide. Traditional fermentation has been empirically developed in ancient times as a process of raw food preservation and at the same time production of new foods with different sensorial characteristics, such as texture, flavour and aroma, as well as nutritional value. Low-alcoholic fermented beverages (LAFB) and non-alcoholic fermented beverages (NAFB) represent a subgroup of fermented beverages that have received rather little attention by consumers and scientists alike, especially with regard to their types and traditional uses in European societies. A literature review was undertaken and research articles, review papers and textbooks were searched in order to retrieve data regarding the dietary role, nutrient composition, health benefits and other relevant aspects of diverse ethnic LAFB and NAFB consumed by European populations. A variety of traditional LAFB and NAFB consumed in European regions, such as kefir, kvass, kombucha and hardaliye, are presented. Milk-based LAFB and NAFB are also available on the market, often characterised as ‘functional’ foods on the basis of their probiotic culture content. Future research should focus on elucidating the dietary role and nutritional value of traditional and ‘functional’ LAFB and NAFB, their potential health benefits and consumption trends in European countries. Such data will allow for LAFB and NAFB to be included in national food composition tables.