Probiotics such as Lactobacillus and Bifidobacterium have been successfully used to promote growth and prevent diseases. Previous reports have demonstrated that Bacillus subtilis (B. subtilis) was a potential probiotic for animals. In this research, 180 B. subtilis were isolated from the soil, identified and investigated in vitro. Furthermore, five B. subtilis were selected and mixed to investigate their effect on growth performance, immune response, intestine microbiota and disease resistance in rabbits. Rabbits with a diet of 106 CFU g-1 mixed B. subtilis exhibited the best growth performance, and higher serum IgG and IgA than controls (P < 0.05). Moreover, dairy with B. subtilis can promote the balance of intestinal flora. The major pro-inflammatory factor and β-defensin were up-regulated compared with the controls. After 7 weeks of feeding, the survival rate of the rabbits fed with B. subtilis was significantly higher than those in the control groups post-infected with Escherichia coli. At the same time, this study detected higher expression of β-defensin and reduced bacteria contents of the heart and cecal contents from the diet mixed with B. subtilis compared with the control groups. In conclusion, dietary supplementation with B. subtilis for rabbits could improve growth performance, serum immunoglobulin, intestinal homeostasis and immune organ index, and enhance innate immune response as well as disease resistance. These findings showed that the induction of β-defensin by B. subtilis might be an interesting new therapeutic strategy to strengthen innate defence mechanisms.

/pdf/bacillus-subtilis-improves-immunity-and-disease-resistance-khkw29vk3z.pdf

Bacillus subtilis improves immunity and disease resistance in rabbits

Assessment of probiotic properties in lactic acid bacteria isolated from wine.

Rapid and accurate molecular diagnosis is a prerequisite for precision medicine, food safety, and environmental monitoring. The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas)-based detection, as a cutting-edged technique, has become an immensely effective tool for molecular diagnosis because of its outstanding advantages including attomolar level sensitivity, sequence-targeted single-base specificity, and rapid turnover time. However, the CRISPR/Cas-based detection methods typically require a pre-amplification step to elevate the concentration of the analyte, which may produce non-specific amplicons, prolong the detection time, and raise the risk of carryover contamination. Hence, various strategies for target amplification-free CRISPR/Cas-based detection have been developed, aiming to minimize the sensitivity loss due to lack of pre-amplification, enable detection for non-nucleic acid targets, and facilitate integration in portable devices. In this review, the current status and challenges of target amplification-free CRISPR/Cas-based detection are first summarized, followed by highlighting the four main strategies to promote the performance of target amplification-free CRISPR/Cas-based technology. Furthermore, we discuss future perspectives that will contribute to developing more efficient amplification-free CRISPR/Cas detection systems.

Amplification-free CRISPR/Cas detection technology: challenges, strategies, and perspectives.

Over the past decades the use of probiotics in food has increased largely due to the manufacturer’s interest in placing “healthy” food on the market based on the consumer’s ambitions to live healthy. Due to this trend, health benefits of products containing probiotic strains such as lactobacilli are promoted and probiotic strains have been established in many different products with their numbers increasing steadily. Probiotics are used as starter cultures in dairy products such as cheese or yoghurts and in addition they are also utilized in non-dairy products such as fermented vegetables, fermented meat and pharmaceuticals, thereby, covering a large variety of products. To assure quality management, several pheno-, physico- and genotyping methods have been established to unambiguously identify probiotic lactobacilli. These methods are often specific enough to identify the probiotic strains at genus and species levels. However, the probiotic ability is often strain dependent and it is impossible to distinguish strains by basic microbiological methods. Therefore, this review aims to critically summarize and evaluate conventional identification methods for the genus Lactobacillus, complemented by techniques that are currently being developed.

/pdf/timely-approaches-to-identify-probiotic-species-of-the-genus-3skc2g0dhc.pdf

Timely approaches to identify probiotic species of the genus Lactobacillus

Bile salt hydrolase (BSH) activity, hypo-cholesterolemic effect, and probiotic properties have been reported for Enterococcus strains isolated from animal and human gut and fermented foods but not for strains isolated from environmental niches, like aquatic and terrestrial plants, soil, and water. The present study is the first report on isolation of Enterococcus faecium from rhizospheric soils that harbor the bsh gene, remove cholesterol in vitro, and possess essential and desirable probiotic attributes. Fifteen samples were collected from different sites located in northern, southern, and central regions of India, of which five yielded pure colonies that were named LR2, LR3, ER5, LR13, and VB1. These were identified by 16S rRNA gene sequencing as E. faecium and evaluated for BSH activity, cholesterol-lowering potential in vitro, and probiotic properties. Our results indicated that all the strains were capable of surviving the harsh conditions of the gastrointestinal tract and did not harbor any of the virulence genes. Though all strains showed the presence of bsh and potential for cholesterol removal, E. faecium strain LR13 showed a remarkable cholesterol removal capability and vancomycin susceptibility and possessed most of the desirable and essential attributes of a probiotic. Hence, it seems to be a fairly promising probiotic candidate that needs to be further evaluated in in vivo studies, especially for its hypo-cholesterolemic potential.

/pdf/evaluation-of-bile-salt-hydrolases-cholesterol-lowering-1gp94o9g75.pdf

Evaluation of Bile Salt Hydrolases, Cholesterol-Lowering Capabilities, and Probiotic Potential of Enterococcus faecium Isolated From Rhizosphere.

Aptamer-based Cas14a1 biosensor for amplification-free live pathogenic detection.

The aims of this study were to identify and characterize lactic acid bacteria (LAB) isolated from healthy forest musk deer (FMD) feces which have good potential to develop orally delivered probiotics applied in FMD. Eight LAB isolates were obtained from feces samples and were identified as Lactobacillus acidipiscis, Lactobacillus plantarum, Enterococcus faecium, Enterococcus mundtii, Enterococcus durans and Leuconostoc fallax using morphology, physiology, biochemical and 16s rDNA PCR methods. Five strains were selected after safety and function tests which showed no hemolytic activity, no harmful indole and hydrogen sulphide produced, safety to KM mouse, growth inhibition toEscherichia coli (ATCC25922), Salmonella pullorum (CVCC527) and Staphylococcus aureus (ATCC29213), and no acquired resistance genes detected. The growth of five strains could not be inhibited by 0.9% pig bile. Two Lactobacillus strains could survive at as low as pH 2.0. One L. acidipiscis strain and one E. durans strain could survive at as high as 70°C. Five strains identified as L. acidipiscis, L. plantarum, Enterococcus faeciumand L. fallax could be well suitable for developing an orally delivered probiotic. To our knowledge, this is the first time to isolate and identify LAB from forest musk deer feces and the characterized strains have good potential for developing an orally delivered probiotic to improve health condition of forest musk deer.

	 

	Key words: Lactic acid bacteria (LAB), probiotics, Lactobacillus, Enterococcus,Leuconostoc.

Identification and characterization of lactic acid bacteria from forest musk deer feces

A self-powered biosensing system based on triboelectric nanogenerator for rapid bacterial DNA detection

Ka/V-band LEO high-throughput satellite and integrated satellite-terrestrial network experiment system: First two years flight results

Rapid, efficient, specific and sensitive diagnostic techniques are critical for selecting appropriate treatments for drug-resistant bacterial infections. To address this challenge, we have developed a novel diagnostic method, called the Dual-Cas Tandem Diagnostic Platform (DTDP), which combines the use of Cas9 nickase (Cas9n) and Cas12a. DTDP works by utilizing the Cas9n-sgRNA complex to create a nick in the target strand's double-stranded DNA (dsDNA). This prompts DNA polymerase to displace the single-stranded DNA (ssDNA) and leads to cycles of DNA replication through nicking, displacement, and extension. The ssDNA is then detected by the Cas12a-crRNA complex (which is PAM-free), activating trans-cleavage and generating a fluorescent signal from the fluorescent reporter. DTDP exhibits a high sensitivity (1 CFU/mL or 100 ag/μL), high specificity (specifically to MRSA in nine pathogenic species), and excellent accuracy (100%). The dual RNA recognition process in our method improves diagnostic specificity by decreasing the limitations of Cas12a in detecting dsDNA protospacer adjacent motifs (PAMs) and leverages multiple advantages of multi-Cas enzymes in diagnostics. This novel approach to pathogenic microorganism detection has also great potential for clinical diagnosis.

Development of an inducible Cas9 nickase and PAM-free Cas12a platform for bacterial diagnostics.

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