Major microbiota of lactic acid bacteria from Matsoni, a traditional Georgian fermented milk

A total of 26 samples of Matsoni were collected in Georgia. From these samples 80 strains of lactic acid cocci and 173 strains of lactobacilli were isolated. The number of lactic acid bacteria varied between 10⁵ and 10¹⁰ colony forming unit (CFU)/mL. All the isolated lactic acid bacteria were thermophilic bacteria that could grow at 45°C. The predominant lactic acid bacteria were Streptococcus thermophilus and Lactobacillus delbrueckii spp. bulgaricus in 25 samples of Matsoni, while Lactobacillus helveticus was also a predominant species, together with the two previous species in one sample of Matsoni. We showed there was diversity in both S. thermophilus and L. delbrueckii spp. bulgaricus at the strain level by randomly amplified polymorphic DNA analysis.     

Probiotics and broiler growth performance: a meta-analysis of randomised controlled trials

1. The aim of this meta-analysis was to investigate the effects of probiotics on the growth performance of broilers. PubMed, Scopus and Scholar Google databases were searched in all languages from 1980 to 2012. The studies in the meta-analysis were only selected if they were randomised and controlled experiments using broilers without apparent disease and the results were published in peer-reviewed journals.

2. A total of 48 and 46 studies were included to assess probiotic effects on body weight gain (BWG) and feed efficiency (FE), respectively. Probiotics increased BWG compared to controls (SMD = 0.661, 95% CI 0.499 to 0.822) and improved FE (SMD = ? 0.281, 95% CI ?0.404 to ?0.157) in the pooled standardised mean difference random effect model, considering the source of heterogeneity and publication biases. However, there are evidences of publication bias and heterogeneity, so the results of this meta-analysis should be considered with caution. Applying the Duval and Tweedie’s trim-and-fill methods, the adjusted value for BWG was 0.0594 (95% CI ?0.122 to 0.242), and the adjusted value for FE did not show any modifications.

3. The meta-analysis showed that application of probiotics via water resulted in greater BGW and FE than administration through the feed. The effect was not related to the use of mono-strain or multi-strain probiotics, although it may depend on the strain used. The number of broilers and the duration of the experiments had an impact on the outcomes.

4. Additional studies should be conducted with the aim to identify the covariates which can explain the differences in the estimated effect sizes.     

Selection of useful probiotic lactic acid bacteria from the Lactobacillus acidophilus group and their applications to functional foods

In the present review, a new mass screening system for selecting probiotic strains from Lactobacillus (L) acidophilus group lactic acid bacteria (LAB) with strong adhesion to the human intestinal tract is described. Characteristics of antimicrobial peptides (bacteriocin), lactose‐hydrolyzing enzymes and immunostimulative oligo DNA motifs in L. gasseri strains are also described. Finally, the use of L. acidophilus LAB, selected by our screening method, that have strong adhesion to the human colonic mucosa in functional yogurt products is described. Adhesiveness to the human intestine is one of the most important characteristics of probiotic LAB. A new screening system that involves a combination of three methods is proposed: rat colonic mucin (RCM)‐micro plate assay, Carnoy's histochemical staining method and carbohydrate probe binding assay. By using an RCM‐coated poly‐vinylidene‐diflouride membrane that mimics the human colonic mucous layer, a new lectin was isolated and its structure was clarified by gene cloning. Furthermore, the structures and functions of a new cyclic bacteriocin (gassericin A), new lactose‐hydrolyzing enzymes, and new immunostimulating oligo DNA motifs from Lactobacillus gasseri (B₁ subgroup) were clarified. A new functional yogurt ‘Fit down’ is proposed, that is fermented by an adhesive strain of L. acidophilus LA67 selected by our screening and contains antihypertensive peptides derived from whey proteins by protease digestion. In the future, superior functional foods containing more effective probiotic LAB are expected to be developed by the use of the proposed mass screening system.     

猪源乳酸菌的分离、筛选、鉴定及产酸性能的研究

试验旨在分离、筛选出具有强抗逆性、产酸能力强的同源菌株。从28日龄断奶健康仔猪粪便分离得到30株乳酸菌,应用体外筛选法,筛选出在pH值2.5处理2h后的菌株13株;将此13株菌株通过浓度为0.2%胆盐后剩7株。采用牛津杯法进行抑菌试验,从7株菌中筛选出对大肠杆菌、沙门氏菌有较强抑菌作用的4株,分别为R11、R15、R18、R22,对此4株菌株进行产酸试验验证,都表现出强的产酸能力。对这4株菌进行鉴定性试验,结果R11为屎肠球菌,R15为鸡肠球菌,R18为嗜酸乳杆菌,R22为口乳杆菌。     

Potential effect of two Bacillus probiotic strains on performance and fecal microbiota of breeding sows and their piglets

The effect of long-term administration of two Bacillus strains was tested on 98 breeding sows and their litters allotted into three treatments: a control group (CON); supplemented with 5 × 10⁸ cfu/kg B. subtilis − 541 (BSU); or with 5 × 10⁸ cfu/kg B. amyloliquefaciens − 516 (BAM). Reproductive and performance variables were recorded over three cycles with 56 dams remaining through the third lactation. Blood and fecal samples were taken longitudinally from 12 sows per treatment on days 8 and 21 of the third lactation and milk samples were taken on day 21. Feces from one piglet per litter was sampled on days 21 and 33 and jejunal gene expression was assessed in two piglets on day 21. Changes in fecal microbiota were assessed by 16S rRNA gene sequencing (Illumina MiSeq) and gene expression by Open-Array technology. Metabolomic responses were analyzed in milk by NMR and Ig-G and Ig-A specific antibodies were determined by ELISA. No significant differences were observed on feed intake, body weight, or fat mobilization of the sows. However, a significant increase in the total number of piglets born was observed in supplemented sows. Although the increase was seen from the first cycle with BAM, improvements were not seen with BSU until the third cycle. BAM also increased the number of born-alive and weaned piglets. NMR analysis showed an impact of BAM on milk composition. No differences were found in milk or blood immunoglobulins. A different structure of the fecal microbiota was found in supplemented sows, with changes across phylum, family, and genus. These changes were greater at day 8, suggesting a relevant role of probiotics establishing a new intestinal balance after labor. Shifts in the microbiota were also seen in the piglets, with a clearer impact post-weaning than in suckling. In this regard, correlations between microbial groups of sows and piglets showed a higher link with weaned (d33) than with suckling pigs (d21), reinforcing the idea of an early maternal carry-over. No changes due to treatment in jejunal gene expression were detected; however, piglet size had a clear impact on different genes. In summary, the addition of both probiotics, and particularly Bacillus amyloliquefaciens, demonstrated potential benefits on the prolificacy of sows. Daily feeding of Bacillus amyloliquefaciens resulted in an increase in the number of weaned piglets. The high correlations between the compositions of the microbiota of sows and their piglets are evidence of maternal imprinting, with effects lasting beyond weaning.     

Effect of trehalose supplementation in milk replacer on the incidence of diarrhea and fecal microbiota in preweaned calves

Trehalose, a nonreducing disaccharide consisting of d-glucose with α,α-1,1 linkage, was evaluated as a functional material to improve the gut environment in preweaned calves. In experiment 1, 173 calves were divided into two groups; the trehalose group was fed trehalose at 30 g/animal/d with milk replacer during the suckling period, and the control group was fed nonsupplemented milk replacer. Medication frequency was lower in the trehalose group (P < 0.05). In experiment 2, calves (n = 20) were divided into two groups (control group [n = 10] and trehalose group [n = 10]) based on their body weight and reared under the same feeding regimens as in experiment 1. Fresh feces were collected from individual animals at the beginning of the trial (average age 11 d), 3 wk after trehalose feeding (experimental day 22), and 1 d before weaning, and the fecal score was recorded daily. Fecal samples were analyzed for fermentation parameters and microbiota. The fecal score was significantly lower in the trehalose group than in the control group in the early stage (at an age of 14 to 18 d; P < 0.05) of the suckling period. Calves fed trehalose tended to have a higher proportion of fecal butyrate on day 22 than calves in the control group (P = 0.08). Population sizes of Clostridium spp. were significantly lower (P = 0.036), whereas those of Dialister spp. and Eubacterium spp. tended to be higher in the feces of calves in the trehalose group on day 22 (P = 0.060 and P = 0.083). These observations indicate that trehalose feeding modulated the gut environment and partially contributed to the reduction in medication frequency observed in experiment 1.     

Vaginal lactic acid bacteria in healthy and ill bitches and evaluation of in vitro probiotic activity of selected isolates

The concentrations of lactic acid bacteria (LAB) in 42 vaginal samples from healthy and ill bitches were determined. Eight isolates belonging to the genera Lactobacillus and Enterococcus were selected and identified and their in vitro antimicrobial activity against canine pathogens and their ability to adhere to canine vaginal epithelial cells were determined. There was no correlation between the concentrations of vaginal LAB and clinical status, body temperature, vaginal pH, or age. Although the animals were either well or suffering from various illnesses, LAB were found in almost every sample, and the selected isolates showed promising probiotic-related features. These findings are significant for the design of strategies for the modulation of vaginal microbiota by vaginal LAB isolates.     

乳酸菌发酵液体饲料对生长猪生长性能和粪中微生物区系的影响

<正>发酵液体饲料(Fermented liquid feed,FLF)作为一种新型饲料,最早使用是在20世纪80年代末的荷兰,当时的发酵液体饲料实际上就是湿拌料(陈文斌,2004)。随着现代饲料工业技术的发展,发酵液体饲料正朝着可人工控制发酵过程的方向发展(王金全,2009)。     

Cultivable microbiota of Lithobates catesbeianus and advances in the selection of lactic acid bacteria as biological control agents in raniculture

The cultivable microbiota of skin and cloaca of captive Lithobates catesbeianus includes microorganisms generally accepted as beneficial and potentially pathogenic bacteria. In order to select a group of potentially probiotic bacteria, 136 isolates were evaluated for their surface properties and production of antagonistic metabolites. Then, 11 lactic acid bacteria (LAB) strains were selected and identified as Lactobacillus plantarum, Lb. brevis, Pediococcus pentosaceus, Lactococcus lactis, L. garvieae and Enterococcus gallinarum. Studies of compatibility indicate that all the strains could be included in a multi-strain probiotic, with the exception of Ent. gallinarum CRL 1826 which inhibited LAB species through a bacteriocin-like metabolite. These results contribute to the design of a probiotic product to improve the sanitary status of bullfrogs in intensive culture systems, to avoid the use of antibiotics and thus to reduce production costs. It could also be an alternative to prevent infectious diseases during the ex situ breeding of amphibian species under threat of extinction.     

Effects of the supplementation of a calcium soap containing medium-chain fatty acids on the fecal microbiota of pigs,lactating cows,and calves

Medium-chain fatty acids (MCFAs) have antialgal, antibacterial, antifungal, antiprotozoan, and antiviral activities. However, antibacterial activities of MCFAs in the hindgut of pigs and cattle are still unknown. We report the effects of the supplementation of MCFAs on fecal bacteria of pigs, lactating cows, and Japanese Black calves. Lactobacillus spp., Bifidobacterium spp., eaeA(+) Escherichia coli, Salmonella spp., Campylobacter jejuni, and Clostridium perfringens in the feces of animals were quantified by real-time PCR assay. There was no significant increase or decrease in Lactobacillus spp. and Bifidobacterium spp. in the three animals. In the pig feces, eaeA(+) E. coli was reduced to less than a third in the treatment group (P < 0.01). C. jejuni in the pig feces was also significantly less in the treatment group compared with the control (P < 0.01). In the lactating cow, eaeA(+) E. coli was reduced to one fifth of that in the control (P < 0.01). Salmonella spp. was halved in calf feces (P < 0.01). Thus, a reduction in Gram-negative pathogenic bacteria was observed. In conclusion, supplementation of a MCFA calcium soap in the diet would be beneficial to growing pigs, lactating cow, and calves by reducing pathogenic bacteria.     

乳酸菌在奶牛上应用的研究进展

乳酸菌作为一种新型的微生态添加剂,因其安全、无污染的特性受到广泛关注。乳酸菌在奶牛上的应用包括制备青贮、饲喂奶牛、防治疾病、增强免疫力和提高生产性能等方面,本文对其进行了论述。     

Test schemes for evaluation of the effect of a probiotic feed additive on the gut microflora of young calves and newly hatched turkeys

Probiotic feed additives have to undergo a strict assessment according to EU regula-tions, including the evaluation of the influence on the gut microflora of the host animal. A probiotic strain of the genus Enterococcus faecium and two different host animal species, young calves and newly hatched turkeys, were chosen as a test model for the establishment of schemes for feeding trials. Investigations included faeces samples and pooled swab samples of faeces as well as samples from gut sections derived from diagnostic slaughtering. The samples were tested for aerobic total count, for enterococci count and for Enterobacteriaceae and E. coli. The test schemes were appropriate for the evaluation of the influence of the probiotic feed additive on the physiological gut microflora of the host animals. Additional information could be gathered concerning safety aspects and effectiveness.     

Interventions for atopic dermatitis in dogs: a systematic review of randomized controlled trials

The objective of this systematic review, which was performed following the guidelines of the Cochrane collaboration, was to assess the effects of interventions for treatment of atopic dermatitis (AD) in dogs. Citations identified from three databases (MEDLINE, Thomson's Science Citation Index Expanded and CAB Abstracts) and trials published by December 2007 were selected. Proceedings books from the major veterinary dermatology international congresses were hand searched for relevant citations. The authors selected randomized controlled trials (RCTs), published from January 1980 to December 2007, which reported the efficacy of topical or systemic interventions for treatment or prevention of canine AD. Studies had to report assessments of either pruritus or skin lesions, or both. Studies were selected and data extracted by two reviewers, with discrepancies resolved by a third arbitrator. Missing data were requested from study authors of recently published trials. Pooling of results and meta-analyses were performed for studies reporting similar interventions and outcome measures. A total of 49 RCTs were selected, which had enrolled 2126 dogs. This review found some evidence of efficacy of topical tacrolimus (3 RCTs), topical triamcinolone (1), oral glucocorticoids (5), oral ciclosporin (6), subcutaneous recombinant γ-interferon (1) and subcutaneous allergen-specific immunotherapy (3) to decrease pruritus and/or skin lesions of AD in dogs. One high-quality RCT showed that an oral essential fatty acid supplement could reduce prednisolone consumption by approximately half. Additional RCTs of high design quality must be performed to remedy previous flaws and to test interventions for prevention of flares of this disease.     

青贮饲料乳酸菌添加剂的应用现状

青贮饲料乳酸菌添加剂对改善青贮饲料品质具有显著作用。为了明确目前国内外对乳酸菌饲料添加剂的研究及应用情况,本文对近年来青贮饲料乳酸菌添加剂的发酵特性,筛选鉴定及应用等方面的研究做了系统的归纳和阐述。目前研究已明确了青贮饲料乳酸菌添加剂的发酵原理和青贮的过程,研究内容主要包括以下几个方面:1)乳酸菌添加剂本身、环境因素及饲料作物特性这3个方面对发酵产生的影响;2)青贮饲料乳酸菌添加剂的筛选和优良乳酸菌添加至青贮饲料的研究;3)乳酸菌菌株单独添加、与酶制剂或其他功能性微生物混合添加应用研究。综合以上研究,认为未来我国可以对不同生态环境中的乳酸菌资源进行发掘、筛选,并根据草牧业发展的实际需求以及我国现有的农艺,加强乳酸菌加工利用工艺的研发。     

Impact of lactic acid fermentation in the large intestine on acute lactic acidosis in cattle

Microbial and fermentation changes in the ingesta of the large intestine and their influence on the pathogenesis of acute lactic acidosis were studied in 4 cows fitted with permanent cannulas in the ileum and cecum. Feed mixture containing 65% of maize was infused into the cecum for several days in amounts of 2 and 4 kg per day. The daily amount was divided in 8 equal portions and given with 3 l of warm physiologic saline solution. During the period of ad libitum feeding of hay, the pH values in cecal digesta were 7.4 to 7.6 and the amount of total volatile fatty acids 40-60 mmol/kg with high molar percentage (87-90 mol%) of acetic acid. As to lactic acid only the L(+) lactic isomer was found in a concentration of about 0.4 mmol/kg. Infusion of low amounts of starch induced mild lactic acid fermentation in the cecum associated with a pronounced increase in the concentration of L(+) and D (-) lactic acid to peak levels of 80 +/- 10 mmol/kg and 7 +/- 1 mmol/kg, respectively. Lactic acid fermentation ceased within 2 to 3 days indicating that the gut microflora had adapted to the starch infusion. Slight decreases of blood pH and bicarbonates in blood as well as a moderate increase of netto acid-base excretion in urine indicated mild changes of acid-base balance, but clinically no pathological symptoms were observed. Higher amounts of infused starch caused pronounced lactic acid production in the large intestine which persisted throughout the experiment. Peak L(+) and D(-) lactic acid concentration in cecal digesta reached on the average 137 +/- 16 mmol/kg and 45 +/- 7 mmol/kg respectively. Significant decreases of blood pH values from 7.41 +/- 0.02 to 7.18 +/- 0.08 (P < 0.001), actual bicarbonate from 28.2 +/- 3.2 to 11.0 +/- 2.6 mmol/l (P < 0.001) and base excess from 3.9 +/- 3.6 to -15.2 +/- 3.8 mmol/l (P < 0.001) were observed. D (-) lactic acid concentration in blood increased to 3.2 +/- 0.4 mmol/l, but L(+) lactic acid values remained unchanged under 1 mmol/l. Clear clinical symptoms of indigestion and intoxication characterized by severe inappetence, ruminal stasis and general weakness were also observed. Typical clinical symptoms of disease as well as blood and urine changes in acid-base balance indicated that lactic acid fermentation in the large intestine contributes considerably to the pathogenesis of acute ruminant lactic acidosis.     

Significance of fecal volatile fatty acids in shedding of Escherichia coli O157 from calves: experimental infection and preliminary use of a probiotic product

Cattle have been recognized as a principal reservoir of Escherichia coli O157:H7. This organism appears to be confined to the gastrointestinal tract and is shed in feces. A probiotic product containing lactic acid-producing Streptococcus bovis LCB6 and Lactobacillus gallinarum LCB 12 isolated from adult cattle was developed, and a preliminary experiment was conducted to evaluate its effect on the elimination of E. coli O157 from experimentally infected calves. Eight 4-month-old Holstein calves were orally challenged with E. coli O157 and the probiotic product was administered against four calves continued fecal shedding of E. coli O157 by the 7th day after infection. Fecal shedding of E. coli O157 was completely inhibited and re-shedding was not detected in any of the animals. Remarkable increase of VFAs, especially that of acetic acid in feces after the administration of probiotic bacteria correlated with the diminution of E. coli O157. Four calves that had spontaneously ceased fecal shedding of E. coli O157 by the 7th day exhibited a high concentration of VFAs in feces before and after experimental infection. Although our results are preliminary and obtained from calves under limited conditions, the possible application of probiotic product to reduce fecal shedding of E. coli O157 from cattle is suggested.     

Preliminary investigation of the probiotic potential of Lactobacillus rhamnosus strain GG in horses: fecal recovery following oral administration and safety

This study was designed to evaluate whether Lactobacillus rhamnosus strain GG (LGG), an extensively studied probiotic organism in humans, can colonize the intestines of adult horses and foals. Lactobacillus rhamnosus strain GG was administered to adult horses at doses of 1 x 10(9) CFU/50kg bodyweight (BW)/day (group 1, 7 horses), 1 x 10(10) colony forming units/ 50kg BW/day (group 2, 7 horses) and 5 x 10(10) colony forming units/50kg BW/day (group 3, 7 horses) for 5 d. Foals received 2 x 10(10) colony forming units/50kg BW/day (group 1, 7 foals) or 1 x 10(11) colony forming units/50kg BW/day (group 2, 7 foals) for 5 d. Fecal levels of L. rhamnosus strain GG in adult horses were low and variable in the 2 lower dose groups. Even in the high dose group, colonization was relatively low. In contrast, more consistent intestinal colonization was present in foals, and colonization persisted for up to 9 d following cessation of administration. No adverse effects were observed in any animal. Clinical studies evaluating this probiotic are indicated in foals. The presence of this organism in the feces of adult horses may only represent passive movement through the intestinal tract, not actual colonization. Consistent intestinal colonization in adults was only achieved with a prohibitively high dose.