Composition and Stability of the Microbial Community inside the Digestive Tract of the Aquatic Crustacean <Emphasis Type="Italic">Daphnia magna</Emphasis>

Small filter-feeding zooplankton organisms like the cladoceran Daphnia spp. are key members of freshwater food webs. Although several interactions between Daphnia and bacteria have been investigated, the importance of the microbial communities inside Daphnia guts has been studied only poorly so far. In the present study, we characterised the bacterial community composition inside the digestive tract of a laboratory-reared clonal culture of Daphnia magna using 16S rRNA gene libraries and terminal-restriction length polymorphism fingerprint analyses. In addition, the diversity and stability of the intestinal microbial community were investigated over time, with different food sources as well as under starvation stress and death, and were compared to the community in the cultivation water. The diversity of the Daphnia gut microbiota was low. The bacterial community consisted mainly of Betaproteobacteria (e.g. Limnohabitans sp.), few Gammaproteobacteria (e.g. Pseudomonas sp.) and Bacteroidetes that were related to facultatively anaerobic bacteria, but did not contain typical fermentative or obligately anaerobic gut bacteria. Rather, the microbiota was constantly dominated by Limnohabitans sp. which belongs to the Lhab-A1 tribe (previously called R-BT065 cluster) that is abundant in various freshwaters. Other bacterial groups varied distinctly even under constant cultivation conditions. Overall, the intestinal microbial community did not reflect the community in the surrounding cultivation water and clustered separately when analysed via the Additive Main Effects and Multiplicative Interaction model. In addition, the microbiota proved to be stable also when Daphnia were exposed to bacteria associated with a different food alga. After starvation, the community in the digestive tract was reduced to stable members. After death of the host animals, the community composition in the gut changed distinctly, and formerly undetected bacteria were activated. Our results suggest that the Daphnia microbiota consists mainly of an aerobic resident bacterial community which is indigenous to this habitat.     

The Development and Stability of the Genus <Emphasis Type="Italic">Bacteriodes</Emphasis> from Human Gut Microbiota in HFA Mice Model

Human flora-associated (HFA) mice are frequently applied in studying the ecology and metabolism of human gut microbiota. However, the development and stability of the genus Bacteriodes, a prominent bacteria group of human gut microbiota, in HFA mice have not yet fully been examined. In this study, PCR-denaturing gradient gel electrophoresis (DGGE) analysis was employed to monitor the Bacteriodes community in the fecal microbiota of six HFA Kunming mice during a period of 3 weeks. Based on the DGGE banding patterns, the majority of prominent bands in the HFA mice DGGE profile were also typical bands in the human DGGE profile, despite the absence of three bands (corresponding to two different B. thetaiotaomicron strains and one B. intestinalis strain) from the human DGGE profile. The Dice coefficient of similarity for the fecal microbiota of HFA mice in comparison to the human donor sample ranged between 74 ± 6% and 81 ± 7%. The phylogeny of bands in the DGGE profile showed that the dominant Bacteriodes species in the fecal microbiota of HFA mice were B. thetaiotaomicron, representing 66.7% of all bands. Our results indicate that the genus Bacteriodes in the fecal microbiota of HFA mice was selected from the human donor and could remain relatively stable over time.     

Engineering bacterial thiosulfate and tetrathionate sensors for detecting gut inflammation

There is a groundswell of interest in using genetically engineered sensor bacteria to study gut microbiota pathways, and diagnose or treat associated diseases. Here, we computationally identify the first biological thiosulfate sensor and an improved tetrathionate sensor, both two‐component systems from marine Shewanella species, and validate them in laboratory Escherichia coli. Then, we port these sensors into a gut‐adapted probiotic E. coli strain, and develop a method based upon oral gavage and flow cytometry of colon and fecal samples to demonstrate that colon inflammation (colitis) activates the thiosulfate sensor in mice harboring native gut microbiota. Our thiosulfate sensor may have applications in bacterial diagnostics or therapeutics. Finally, our approach can be replicated for a wide range of bacterial sensors and should thus enable a new class of minimally invasive studies of gut microbiota pathways.     

Membrane filter method to study the effects of Lactobacillus acidophilus and Bifidobacterium longum on fecal microbiota

A large number of commensal bacteria inhabit the intestinal tract, and interbacterial communication among gut microbiota is thought to occur. In order to analyze symbiotic relationships between probiotic strains and the gut microbiota, a ring with a membrane filter fitted to the bottom was used for in vitro investigations. Test strains comprising probiotic nitto strains (Lactobacillus acidophilus NT and Bifidobacterium longum NT) and type strains (L. acidophilus JCM1132^T and B. longum JCM1217^T) were obtained from diluted fecal samples using the membrane filter to simulate interbacterial communication. Bifidobacterium spp., Streptococcus pasteurianus, Collinsella aerofaciens, and Clostridium spp. were the most abundant gut bacteria detected before coculture with the test strains. Results of the coculture experiments indicated that the test strains significantly promote the growth of Ruminococcus gnavus, Ruminococcus torques, and Veillonella spp. and inhibit the growth of Sutterella wadsworthensis. Differences in the relative abundances of gut bacterial strains were furthermore observed after coculture of the fecal samples with each test strain. Bifidobacterium spp., which was detected as the dominant strain in the fecal samples, was found to be unaffected by coculture with the test strains. In the present study, interbacterial communication using bacterial metabolites between the test strains and the gut microbiota was demonstrated by the coculture technique. The detailed mechanisms and effects of the complex interbacterial communications that occur among the gut microbiota are, however, still unclear. Further investigation of these relationships by coculture of several fecal samples with probiotic strains is urgently required.     

Probiotic roles of Lactobacillus sp. in swine: insights from gut microbiota

The use of lactobacilli as probiotics in swine has been gaining attention due to their ability to improve growth performance and carcass quality, prevent gastrointestinal infection and most importantly, their ‘generally recognized as safe’ status. Previous studies support the potential of lactobacilli to regulate host immune systems, enhance gut metabolic capacities and maintain balance in the gut microbiota. Research on swine gut microbiota has revealed complex gut microbial community structure and showed the importance of Lactobacillus to the host's health. However, the species‐ and strain‐specific characteristics of lactobacilli that confer probiotic benefits are still not well understood. The diversity of probiotic traits in a complex gut ecosystem makes it challenging to infer the relationships between specific functions of Lactobacillus sp. and host health. In this review, we provide an overview of how lactobacilli play a pivotal role in the swine gut ecosystem and identify key characteristics that influence gut microbial community structure and the health of pigs. In addition, based on recent and ongoing meta‐omics and omics research on the gut microbiota of pigs, we suggest a workflow combining culture‐dependent and culture‐independent approaches for more effective selection of probiotic lactobacilli.     

Stereology and computer assisted three-dimensional reconstruction as tools to study probiotic effects of Aeromonas hydrophila on the digestive tract of germ-free Artemia franciscana nauplii

Aims: Validation of stereology and three‐dimensional reconstruction for monitoring the probiotic effect of Aeromonas hydrophila on the gut development of germ‐free Artemia franciscana nauplii. Methods and Results: Germ‐free Artemia nauplii were cultured using Baker’s yeast and dead Aer. hydrophila. Live Aer. hydrophila were added on the first day to the treatment group. The gut length and volume were monitored on days two and four using stereology and three‐dimensional reconstruction. Both methods showed comparable results. Stereology was least labour intensive to estimate volumes, while three‐dimensional reconstructions rendered architectural and topographical data of the gut. Moreover, a positive effect of probiotic bacterium, Aer. hydrophila is likely. Conclusion: Slight increment in the growth of the digestive tract of A. franciscana nauplii exerted by probiotic bacteria could be detected using stereology and three‐dimensional reconstruction. Significance and Impact of the Study: The gnotobiotic Artemia rearing system is unique to investigate the effects of micro‐organisms on the development of nauplii. However, in the base of this model system, only survival counts and length measurements exist as monitoring tools. Therefore, additional tools such as stereology and three‐dimensional reconstruction are prerequisite to obtain more powerful analysis.     

Investigation of the impact of feeding Lactobacillus plantarum CRL 1815 encapsulated in microbially derived polymers on the rat faecal microbiota

Aims

The aim of this study was to evaluate the impact of the administration of microencapsulated Lactobacillus plantarum CRL 1815 with two combinations of microbially derived polysaccharides, xanthan : gellan gum (1%:0·75%) and jamilan : gellan gum (1%:1%), on the rat faecal microbiota.

Methods and Results

A 10‐day feeding study was performed for each polymer combination in groups of 16 rats fed either with placebo capsules, free or encapsulated Lact. plantarum or water. The composition of the faecal microbiota was analysed by fluorescence in situ hybridization and temporal temperature gradient gel electrophoresis. Degradation of placebo capsules was detected, with increased levels of polysaccharide‐degrading bacteria. Xanthan : gellan gum capsules were shown to reduce the Bifidobacterium population and increase the Clostridium histolyticum group levels, but not jamilan : gellan gum capsules. Only after administration of jamilan : gellan gum‐probiotic capsules was detected a significant increase in Lactobacillus‐Enterococcus group levels compared to controls (capsules and probiotic) as well as two bands were identified as Lact. plantarum in two profiles of ileum samples.

Conclusions

Exopolysaccharides constitute an interesting approach for colon‐targeted delivery of probiotics, where jamilan : gellan gum capsules present better biocompatibility and promising results as a probiotic carrier.

Significance and Impact of Study

This study introduces and highlights the importance of biological compatibility in the encapsulating material election, as they can modulate the gut microbiota by themselves, and the use of bacterial exopolysaccharides as a powerful source of new targeted‐delivery coating material.     

Roles of Probiotic Lactobacilli Inclusion in Helping Piglets Establish Healthy Intestinal Inter-environment for Pathogen Defense

The gastrointestinal tract of pigs is densely populated with microorganisms that closely interact with the host and with ingested feed. Gut microbiota benefits the host by providing nutrients from dietary substrates and modulating the development and function of the digestive and immune systems. An optimized gastrointestinal microbiome is crucial for pigs’ health, and establishment of the microbiome in piglets is especially important for growth and disease resistance. However, the microbiome in the gastrointestinal tract of piglets is immature and easily influenced by the environment. Supplementing the microbiome of piglets with probiotic bacteria such as Lactobacillus could help create an optimized microbiome by improving the abundance and number of lactobacilli and other indigenous probiotic bacteria. Dominant indigenous probiotic bacteria could improve piglets’ growth and immunity through certain cascade signal transduction pathways. The piglet body provides a permissive habitat and nutrients for bacterial colonization and growth. In return, probiotic bacteria produce prebiotics such as short-chain fatty acids and bacteriocins that benefit piglets by enhancing their growth and reducing their risk of enteric infection by pathogens. A comprehensive understanding of the interactions between piglets and members of their gut microbiota will help develop new dietary interventions that can enhance piglets’ growth, protect piglets from enteric diseases caused by pathogenic bacteria, and maximize host feed utilization.     

Bacteria abundance and diversity of different life stages of Plutella xylostella (Lepidoptera: Plutellidae), revealed by bacteria culture‐dependent and PCR‐DGGE methods

Microbial abundance and diversity of different life stages (fourth instar larvae, pupae and adults) of the diamondback moth, Plutella xylostella L., collected from field and reared in laboratory, were investigated using bacteria culture‐dependent method and PCR‐DGGE analysis based on the sequence of bacteria 16S rRNA V3 region gene. A large quantity of bacteria was found in all life stages of P. xylostella. Field population had higher quantity of bacteria than laboratory population, and larval gut had higher quantity than pupae and adults. Culturable bacteria differed in different life stages of P. xylostella. Twenty‐five different bacterial strains were identified in total, among them 20 strains were presented in larval gut, only 8 strains in pupae and 14 strains in adults were detected. Firmicutes bacteria, Bacillus sp., were the most dominant species in every life stage. 15 distinct bands were obtained from DGGE electrophoresis gel. The sequences blasted in GenBank database showed these bacteria belonged to six different genera. Phylogenetic analysis showed the sequences of the bacteria belonged to the Actinobacteri, Proteobacteria and Firmicutes. Serratia sp. in Proteobacteria was the most abundant species in larval gut. In pupae, unculturable bacteria were the most dominant species, and unculturable bacteria and Serratia sp. were the most dominant species in adults. Our study suggested that a combination of molecular and traditional culturing methods can be effectively used to analyze and to determine the diversity of gut microflora. These known bacteria may play important roles in development of P. xylostella.     

Potential probiotics of the Far Eastern trepang <Emphasis Type="Italic">Apostychopus japonicus</Emphasis> producing digestive enzymes

Data were obtained on species diversity of the digestive tract microflora of the Far Eastern trepang Apostychopus japonicus. The probiotic characteristics of microbial isolates were investigated related to their capacity for synthesis of digestive enzymes (amylase, chondroitin sulfatase, chitinase, and alginate lyase). The Pseudomonas stutzeri strain exhibiting high activity of all the investigated enzymes was found to be preferable among the potential probiotics used for cultivation of the Far Eastern trepang. Preparations based on mixed cultures of a Bacillus pumilus strain with high chondroitin sulfatase and chitinase activities with strains of B. coagulans and B. megaterium K13 with high activity of amylases and alginate lyases are also promising.     

Experimental addition of <Emphasis Type="Italic">Eleutherococcus senticosus</Emphasis> and probiotic to the canine diet

There is a current trend to support pet health through the addition of natural supplements to their diet, taking into account the high incidence of medical conditions related to their immune system and gastrointestinal tract. This study investigates effects of the plant Eleutherococcus senticosus as a dietary additive on faecal microbiota, faecal characteristics, blood serum biochemistry and selected parameters of cellular immunity in healthy dogs. A combination of the plant with the canine-derived probiotic strain Lactobacillus fermentum CCM 7421 was also evaluated. Thirty-two dogs were devided into 4 treatment groups; receiving no additive (control), dry root extract of E. senticosus (8 mg/kg of body weight), probiotic strain (10⁸ CFU/mL, 0.1 mL/kg bw) and the combination of both additives. The trial lasted 49 days with 14 days supplementation period. Results confirm no antimicrobial effect of the plant on the probiotic abundance either in vitro (cultivation test) or in vivo. The numbers of clostridia, lactic acid bacteria and Gram-negative bacteria as well as the concentration of serum total protein, triglyceride, glucose and aspartate aminotransferase were significantly altered according to the treatment group. Leukocyte phagocytosis was significantly stimulated by the addition of probiotic while application of plant alone led to a significant decrease.     

Molecular Monitoring of the Fecal Microbiota of Healthy Human Subjects during Administration of Lactulose and Saccharomyces boulardii

Diet is a major factor in maintaining a healthy human gastrointestinal tract, and this has triggered the development of functional foods containing a probiotic and/or prebiotic component intended to improve the host's health via modulation of the intestinal microbiota. In this study, a long-term placebo-controlled crossover feeding study in which each subject received several treatments was performed to monitor the effect of a prebiotic substrate (i.e., lactulose), a probiotic organism (i.e., Saccharomyces boulardii), and their synbiotic combination on the fecal microbiota of three groups of 10 healthy human subjects differing in prebiotic dose and/or intake of placebo versus synbiotic. For this purpose, denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA gene amplicons was used to detect possible changes in the overall bacterial composition using the universal V₃ primer and to detect possible changes at the subpopulation level using group-specific primers targeting the Bacteroides fragilis subgroup, the genus Bifidobacterium, the Clostridium lituseburense group (cluster XI), and the Clostridium coccoides-Eubacterium rectale group (cluster XIVa). Although these populations remained fairly stable based on DGGE profiling, one pronounced change was observed in the universal fingerprint profiles after lactulose ingestion. Band position analysis and band sequencing revealed that a band appearing or intensifying following lactulose administration could be assigned to the species Bifidobacterium adolescentis. Subsequent analysis with real-time PCR (RT-PCR) indicated a statistically significant increase (P < 0.05) in total bifidobacteria in one of the three subject groups after lactulose administration, whereas a similar but nonsignificant trend was observed in the other two groups. Combined RT-PCR results from two subject groups indicated a borderline significant increase (P = 0.074) of B. adolescentis following lactulose intake. The probiotic yeast S. boulardii did not display any detectable universal changes in the DGGE profiles, nor did it influence the bifidobacterial levels. This study highlighted the capacity of an integrated approach consisting of DGGE analysis and RT-PCR to monitor and quantify pronounced changes in the fecal microbiota of healthy subjects upon functional food administration.     

Modulation of gut mucosal microbiota as a mechanism of probiotics-based adjunctive therapy for ulcerative colitis

This was a pilot study aiming to evaluate the effects of probiotics as adjunctive treatment for ulcerative colitis (UC). Twenty-five active patients with UC were assigned to the probiotic (n = 12) and placebo (n = 13) groups. The probiotic group received mesalazine (60 mg kg⁻¹ day⁻¹) and oral probiotics (containing Lactobacillus casei Zhang, Lactobacillus plantarum P-8 and Bifidobacterium animalis subsp. lactis V9) twice daily for 12 weeks, while the placebo group received the same amounts of mesalazine and placebo. The clinical outcomes were assessed. The gut mucosal microbiota was profiled by PacBio single-molecule, real-time (SMRT) sequencing of the full-length 16S rRNA of biopsy samples obtained by colonoscopy. A significantly greater magnitude of reduction was observed in the UC disease activity index (UCDAI) in the probiotic group compared with the placebo group (P = 0.043), accompanying by a higher remission rate (91.67% for probiotic-receivers versus 69.23% for placebo-receivers, P = 0.034). The probiotics could protect from diminishing of the microbiota diversity and richness. Moreover, the gut mucosal microbiota of the probiotic-receivers had significantly more beneficial bacteria like Eubacterium ramulus (P < 0.05), Pediococcus pentosaceus (P < 0.05), Bacteroides fragilis (P = 0.02) and Weissella cibaria (P = 0.04). Additionally, the relative abundances of the beneficial bacteria correlated significantly but negatively with the UCDAI score, suggesting that the probiotics might alleviate UC symptoms by modulating the gut mucosal microbiota. Our research has provided new insights into the mechanism of symptom alleviation in UC by applying probiotic-based adjunctive treatment.     

Immunology and probiotic impact of the newborn and young children intestinal microflora

Human body has developed a holistic defence system, which mission is either to recognize and destroy the aggressive invaders or to evolve mechanisms permitting to minimize or restore the consequences of harmful actions. The host immune system keeps the capital role to preserve the microbial intestinal balance via the barrier effect. Specifically, pathogenic invaders such as, bacteria, parasites, viruses and other xenobiotic invaders are rejected out of the body via barriers formed by the skin, mucosa and intestinal flora. In case physical barriers are breached, the immune system with its many components comes into action in order to fence infection. The intestine itself is considered as an “active organ” due to its abundant bacterial flora and to its large metabolic activity. The variation among different species or even among different strains within a species reflects the complexity of the genetic polymorphism which regulates the immune system functions. Additionally factors such as, gender, particular habits, smoking, alcohol consumption, diet, religion, age, gender, precedent infections and vaccinations must be involved. Hormonal profile and stress seems to be associated to the integrity microbiota and inducing immune system alterations. Which bacterial species are needed for inducing a proper barrier effect is not known, but it is generally accepted that this barrier function can be strongly supported by providing benefic alimentary supplements called functional foods. In this vein it is stressed the fact that early intestinal colonization with organisms such as Lactobacilli and Bifidobacteria and possibly subsequent protection from many different types of diseases. Moreover, this benefic microflora dominated but Bifidobacteria and Lactobacilli support the concept of their ability to modify the gut microbiota by reducing the risk of cancer following their capacity to decrease β-glucoronidase and carcinogen levels. Because of their beneficial roles in the human gastrointestinal tract, LAB are referred to as “probiotics”, and efforts are underway to employ them in modern nutrition habits with so-called functional foods. Members of Lactobacillus and Bifidobacterium genera are normal residents of the microbiota in the human gastrointestinal tract, in which they developed soon after birth. But, whether such probiotic strains derived from the human gut should be commercially employed in the so-called functional foods is a matter of debate between scientists and the industrial world. Within a few hours from birth the newborn develops its normal bacterial flora. Indeed human milk frequently contains low amounts of non-pathogenic bacteria like Streptococcus, Micrococcus, Lactobacillus, Staphylococcus, Corynebacterium and Bifidobacterium. In general, bacteria start to appear in feces within a few hours after birth. Colonization by Bifidobacterium occurs generally within 4 days of life. Claims have been made for positive effects of Bifidobacterium on infant growth and health. The effect of certain bacteria having a benefic action on the intestinal ecosystem is largely discussed during the last years by many authors. Bifidobacterium is reported to be a probiotic bacterium, exercising a beneficial effect on the intestinal flora. An antagonism has been reported between B. bifidum and C. perfringens in the intestine of newborns delivered by cesarian section. The aim of the probiotic approach is to repair the deficiencies in the gut flora and restore the protective effect. However, the possible ways in which the gut microbiota is being influenced by probiotics is yet unknown.     

In vitro and in vivo evaluation of the probiotic attributes of Lactobacillus kefiranofaciens XL10 isolated from Tibetan kefir grain

Lactobacillus kefiranofaciens XL10, with a high yield of extracellular polysaccharide (EPS), is isolated from Tibetan kefir grain and benefits the health of human beings and has been considered to exhibit probiotic potential in vitro and in vivo. The probiotic function of the strain was studied extensively, viz., acid and bile salt tolerances, cell surface hydrophobicity and autoaggregation, the modulation of gut microbiota, and the distribution and colonization of XL10 in the mouse intestinal tract after oral administration. XL10 could survive 3-h incubation at pH 3.5 and exhibited cell surface hydrophobicity of ∼79.9% and autoaggregation of ∼27.8%. After continuous oral administration of XL10 for 2 weeks, the Bifidobacteriaceae family increased, accompanied by an observable decline in Proteobacteria phyla in the tested mice. Butyrivibrio and Pseudobutyrivibrio, recognized as butyric acid-producing bacteria, could also be detected at day 7 and day 14, respectively. The most abundant community in the mouse gut had formed by day 14. Additionally, we found that XL10 successfully adhered to the mucous tissue and colonized the ileum of the mice based on fluorescence imaging, flow cytometry, and qPCR. Our results suggested that XL10 has excellent probiotic properties and represents an alternative for exploitation in the development of novel functional foods.

     

Bifidobacterium longum supplementation improves age-related delays in fracture repair

Age-related delays in bone repair remains an important clinical issue that can prolong pain and suffering. It is now well established that inflammation increases with aging and that this exacerbated inflammatory response can influence skeletal regeneration. Recently, simple dietary supplementation with beneficial probiotic bacteria has been shown to influence fracture repair in young mice. However, the contribution of the gut microbiota to age-related impairments in fracture healing remains unknown. Here, we sought to determine whether supplementation with a single beneficial probiotic species, Bifidobacterium longum (B. longum), would promote fracture repair in aged (18-month-old) female mice. We found that B. longum supplementation accelerated bony callus formation which improved mechanical properties of the fractured limb. We attribute these pro-regenerative effects of B. longum to preservation of intestinal barrier, dampened systemic inflammation, and maintenance of the microbiota community structure. Moreover, B. longum attenuated many of the fracture-induced systemic pathologies. Our study provides evidence that targeting the gut microbiota using simple dietary approaches can improve fracture healing outcomes and minimize systemic pathologies in the context of aging.     

Composition of Acridid gut bacterial communities as revealed by 16S rRNA gene analysis

The gut bacterial community from four species of feral locusts and grasshoppers was determined by denaturing gradient gel electrophoresis (DGGE) analysis of bacterial 16S rRNA gene fragments. The study revealed an effect of phase polymorphism on gut bacterial diversity in brown locusts from South Africa. A single bacterial phylotype, consistent with Citrobacter sp. dominated the gut microbiota of two sympatric populations of Moroccan and Italian locusts in Spain. There was evidence for Wollbachia sp. in the meadow grasshopper caught locally in the UK. Sequence analysis of DGGE products did not reveal evidence for unculturable bacteria and homologies suggested that bacterial species were principally Gammaproteobacteria from the family Enterobacteriaceae similar to those recorded previously in laboratory reared locusts.     

Wild bacterial probiotics fed to larvae of mass‐reared Queensland fruit fly [Bactrocera tryoni (Froggatt)] do not impact long‐term survival,mate selection,or locomotor activity

Queensland fruit fly [Bactrocera tryoni (Froggatt), Diptera, Tephritidae] is the most devastating insect pest impacting Australian horticulture. The Sterile Insect Technique (SIT) is an important component of tephritid pest management programs. However, mass‐rearing and irradiation (to render insects sterile) may reduce the fitness and performance of the insect, including the ability of sterile males to successfully compete for wild females. Manipulation of the gut microbiome, including the supplementation with bacterial probiotics shows promise for enhancing the quality of mass‐reared sterile flies, however there are fewer published studies targeting the larval stage. In this study, we supplemented the larval stage of mass‐reared B. tryoni with bacterial probiotics. We tested several individual bacteria that had been previously isolated and characterized from the gut of wild B. tryoni larvae including Asaia sp., Enterobacter sp., Lactobacillus sp., Leuconostoc sp. We also tested a consortium of all four of these bacterial isolates. The fitness parameters tested included adult survival in field cages, laboratory mate selection of bacteria supplemented males by bacteria nonsupplemented females, and laboratory locomotor activity of adult flies. None of the bacterial probiotic treatments in the current study was significantly different to the control for field survival, mate selection or locomotor activity of adult B. tryoni, which agree with some of the other studies regarding bacterial probiotics fed to the larval stage of tephritids. Future work is needed to determine if feeding the same, and/or other probiotics to adults, as opposed to larvae can positively impact survival, mating performance, mating competitiveness and locomotor activity of B. tryoni. The bacterial group(s) and function of bacterial species that increase fitness and competitiveness is also of interest to tephritid mass‐rearing programs.     

Microbial shifts of the silkworm larval gut in response to lettuce leaf feeding

Silkworm (Bombyx mori L.) larvae were used as an ideal animal protein source for astronauts in the bioregenerative life support system (BLSS). Here, we compared the difference in bacterial communities of the silkworm larval gut between the BLSS rearing way (BRW) and the traditional rearing way (TRW) through culture-dependent approach, 16S rRNA gene analysis, and denaturing gradient gel electrophoresis (DGGE). The culture-dependent approach revealed that the numbers of gut bacteria of silkworm in the BRW significantly decreased compared with that of the TRW. The analysis of clone libraries showed that the gut microbiota in the BRW was significantly less diverse than that in the TRW. Acinetobacter and Bacteroides were dominant populations in the BRW, and Bacillus and Arcobacter dominated in the TRW. DGGE profiles confirmed the difference of silkworm gut bacterial community between two rearing ways. These results demonstrate that gut bacteria change from the BRW contributes to the decrease of silkworm physiological activity. This study increases our understanding of the change of silkworm gut microbiota in response to lettuce leaf feeding in the BRW. We could use the dominant populations to make probiotic products for nutrient absorption and disease prevention in the BLSS to improve gut microecology, as well as the yield and quality of animal protein.     

Phenological asynchrony between host plant and gypsy moth reduces insect gut microbiota and susceptibility to Bacillus thuringiensis

The phenological synchrony between the emergence of overwintering herbivorous insects and the budding of host plants is considered a crucial factor in the population dynamics of herbivores. However, the mechanisms driving the interactions between the host plant, herbivores, and their pathogens are often obscure. In the current study, an artificially induced phenological asynchrony was used to investigate how the asynchrony between silver birch Betula pendula and gypsy moth Lymantria dispar affects the immunity of the insect to bacteria, its susceptibility to the entomopathogenic bacteria Bacillus thuringiensis, and the diversity in its midgut microbiota. The lysozyme‐like activity in both the midgut and hemolymph plasma and the nonspecific esterase activity and antimicrobial peptide gene expression in the midgut were studied in both noninfected and B. thuringiensis‐infected larvae. Our results provide the first evidence that phenologically asynchronous larvae are less susceptible to B. thuringiensis infection than phenologically synchronous larvae, and our results show that these effects are related to the high basic levels and B. thuringiensis‐induced levels of lysozyme‐like activities. Moreover, a 16S rRNA analysis revealed that dramatic decreases in the diversity of the larval gut bacterial consortia occurred under the effect of asynchrony. Larvae infected with B. thuringiensis presented decreased microbiota diversity if the larvae were reared synchronously with the host plant but not if they were reared asynchronously. Our study demonstrates the significant effect of phenological asynchrony on innate immunity‐mediated interactions between herbivores and entomopathogenic bacteria and highlights the role of nonpathogenic gut bacteria in these interactions.