The Australian dingo is an early offshoot of modern breed dogs

Dogs are uniquely associated with human dispersal and bring transformational insight into the domestication process. Dingoes represent an intriguing case within canine evolution being geographically isolated for thousands of years. Here, we present a high-quality de novo assembly of a pure dingo (CanFam_DDS). We identified large chromosomal differences relative to the current dog reference (CanFam3.1) and confirmed no expanded pancreatic amylase gene as found in breed dogs. Phylogenetic analyses using variant pairwise matrices show that the dingo is distinct from five breed dogs with 100% bootstrap support when using Greenland wolf as the outgroup. Functionally, we observe differences in methylation patterns between the dingo and German shepherd dog genomes and differences in serum biochemistry and microbiome makeup. Our results suggest that distinct demographic and environmental conditions have shaped the dingo genome. In contrast, artificial human selection has likely shaped the genomes of domestic breed dogs after divergence from the dingo

Lactobacilli can attenuate inflammation in mouse macrophages exposed to polyethylene particles in vitro

Abstract Objective It is well established that polyethylene (PE) wear particles induce macrophage production of cytokines and mediators associated with the pathogenesis of inflammatory osteolysis. The objective of this study was to examine the potential of three Lactobacillus strains to attenuate the TNF-α cytokine response of macrophages exposed to Ceridust 3615 PE particles. An in vitro experimental model using the RAW 246.7 macrophage cell line and PE particles was utilized. Results Lactobacillus strains were found to modulate the cytokines in a strain and dose specific manner. Only the Lactobacillus acidophilus strain that was tested was able to attenuate PE particle-induced TNF-α production by RAW 246.7 macrophages. This effect was independent of IL-10 cytokine levels since all three strains of lactobacilli yielded comparable levels of IL-10. It was concluded that some, but not all, Lactobacillus strains may be useful in reducing the risk of inflammatory osteolysis and that further studies in appropriate in vivo models are warranted. Furthermore, this in vitro model can be used to evaluate the inflammatory potential of new materials being tested for use as joint implants

<i>Lactobacillus fermentum</i> PC1 has the Capacity to Attenuate Joint Inflammation in Collagen-Induced Arthritis in DBA/1 Mice

Lactobacillus strains have shown efficacy in attenuating inflammation. This study evaluated the potential of Lactobacillus fermentum PC1 for the treatment of rheumatoid arthritis (RA) using a murine model of collagen-induced arthritis. On Day 1, healthy DBA/1 mice (six to eight weeks of age) were immunized, with 100 &#956;g of Chicken Type 11 collagen emulsified in complete Freund&#8217;s adjuvant (CFA) by intradermal injection, at the base of the tail. On Day 21, the mice were immunized intraperitoneally with 100 &#956;g of Bovine Type11 collagen in phosphate buffered saline (PBS). On Day 28, the mice were immunized intraperitoneally with 50 &#956;g of lipopolysaccharide (LPS). Viable L. fermentum PC1 (1 &#215; 109 colony forming units) was given daily from Day two until the end of the experiment. From Day 21 onwards, the mice were monitored daily for clinical signs of arthritis. On Day 44, the experiment was terminated. Paws were obtained for histology and serum for cytokine assays. L. fermentum PC1-fed mice had significantly reduced paw inflammation as well as decreased synovial infiltration and less cartilage damage. Circulating serum cytokine profiles revealed decreased IL-12 and increased anti-inflammatory cytokines, namely IL-4 and IL-10. Thus, early administration of L. fermentum PC1 could prove to be a valuable therapeutic agent in the management of RA

Evaluation of the adjuvant and anti-inflammatory effects of L. fermentum PC1

The aim of this thesis was to examine the adjuvant and anti-inflammatory properties of L. fermentum PC1 using in vivo animal models and tissue culture cell lines. L. fermentum PC2 and L. acidophilus L10 were used as comparative strains for interpretation of the L fermentum PC1 properties.L. fermentum PC1 functioned efficiently as an oral adjuvant in a murine model. The magnitude and nature of the response was influenced by the dosing regime and dose of adjuvant, type of co-administered antigen and host genetics. In addition, L. fermentum PC1 was shown to redirect a Th2 response polarized by cholera toxin (CT), and to a certain degree maintained a Th1 biased response after priming with L. fermentum PC1 and subsequent boosting with CT.The possible cellular mechanism(s) whereby L. fermentum PC1 functioned as an adjuvant include: by production of cytokine from dendritic cells, macrophages and epithelial cells; upregulated expression of co-stimulatory molecules on dendritic cells; enhanced uptake of by-stander soluble antigen; and enhanced intracellular bacterial killing by macrophages; and enhanced capacity of dendritic cells to activate naïve T cells.The anti-inflammatory capacity of L. fermentum PC1 primed macrophages was examined in vitro using two different inflammatory stimulators: LPS and ultra high molecular weight polyethylene wear particles (WP). L. fermentum PC1 was able to down-regulate LPS, but not the WP induced inflammation. L. fermentum PC1 also significantly down regulated S. Typhimurium induced inflammation in epithelial cells. This effect was mediated by factors present in both the cell wall extracts and spent culture medium of L. fermentum PC1. Characterization of the anti-inflammatory molecules(s) revealed it to be of a molecular weight of Mr 1,000 &#150; 6,000, heat stable, with optimum functionality at pH 4-5, and contains carbohydrate and protein moieties.It was concluded L. fermentum PC1 functioned as an adjuvant and factors were identified that influenced an optimum response. Significant insight into the cellular mechanisms behind its adjuvanticity has been obtained. In addition, the anti-inflammatory function of L. fermentum PC1 appears to be dependent on the nature of the inflammatory stimuli and is mediated by a soluble factor

An experimental model for the spatial structuring and selection of bacterial communities

Community-level selection is an important concept in evolutionary biology and has been predicted to arise in systems that are spatially structured. Here we develop an experimental model for spatially-structured bacterial communities based on coaggregating strains and test their relative fitness under a defined selection pressure. As selection we apply protozoan grazing in a defined, continuous culturing system. We demonstrate that a slow-growing bacterial strain Blastomonas natatoria 2.1, which forms coaggregates with Micrococcus luteus, can outcompete a fast-growing, closely related strain Blastomonas natatoria 2.8 under conditions of protozoan grazing. The competitive benefit provided by spatial structuring has implications for the evolution of natural bacterial communities in the environment

Probiotic supplementation in neonates with congenital gastrointestinal surgical conditions: a pilot randomised controlled trial

OBJECTIVE: To evaluate whether probiotic supplementation attenuates gut-dysbiosis in neonates with congenital gastrointestinal surgical conditions (CGISC). METHODS: Sixty-one neonates (≥35 weeks gestation) with CGISC were randomised to receive daily supplementation with a triple-strain bifidobacterial probiotic (n = 30) or placebo (n = 31) until discharge. Stool microbiota was analysed using 16S ribosomal RNA gene sequencing on samples collected before (T1), 1 week (T2), and 2 weeks (T3) after supplementation and before discharge (T4). The primary outcome was the sum of the relative abundance of potentially pathogenic families of Clostridiaceae, Enterobacteriaceae, Enterococcaceae, Pseudomonaceae, Staphylococcaeae, Streptococcaceae, and Yersiniaceae at T3. RESULTS: The median gestational age [38 weeks (IQR: 37.1–38.9)] was similar in both groups. The probiotic group had lower rates of caesarean deliveries (40% versus 70%, p = 0.02). The relative abundance of potentially pathogenic families was lower in the probiotic group compared to placebo at T3 [(median: 50.4 (IQR: 26.6–67.6) versus 67.1 (IQR: 50.9–96.2); p = 0.04). Relative abundance of Bifidobacteriaceae was higher in the probiotic group at T3 [(median: 39.8 (IQR: 24.9–52.1) versus 0.03 (IQR 0.02–2.1); p < 0.001). Stratified analysis continued to show a higher abundance of Bifidobacteriaceae in the probiotic group, irrespective of the mode of delivery. CONCLUSIONS: Probiotic supplementation attenuated gut dysbiosis in neonates with CGISC. TRIAL REGISTRATION: http://www.anzctr.org.au (ACTRN12617001401347). IMPACT: ● Probiotic supplementation attenuates gut dysbiosis and improves stool short-chain fatty acid levels in neonates with congenital gastrointestinal surgical conditions. ● This is the second pilot RCT of probiotic supplementation in neonates with congenital gastrointestinal conditions. ● These findings will pave the way for conducting multicentre RCTs in this area

Probiotic supplementation for neonates with congenital gastrointestinal surgical conditions: guidelines for future research

Our pilot RCT found that probiotic supplementation with the three-strain bifidobacterial product (B. breve M-16V, B. longum subsp. infantis M-63 and B. longum subsp. longum BB536) attenuates gut dysbiosis, increases stool short-chain fatty acid (SCFA) levels and improves the growth of head circumference in neonates with congenital gastrointestinal surgical conditions (CGISC). In this article, we have provided guidelines for designing future multicentre RCTs based on the experience gained from our pilot RCT. The recommendations include advice about sample size, potential confounders, outcomes of interest, probiotic strain selection, storage, dose, duration and microbial quality assurance, collection of stool samples, storage and analysis and reporting. Following these guidelines will increase the validity of future RCTs in this area and hence confidence in their results. IMPACT: Probiotic supplementation attenuates gut dysbiosis, increases stool short-chain fatty acid (SCFA) levels and improves the growth of head circumference in neonates with congenital gastrointestinal surgical conditions. The current review provides evidence-based guidelines to conduct adequately powered RCTs in this field.Published versionThis work was supported by the Centre for Neonatal Research and Education, Neonatal Directorate, Child and Adolescent Health Service, Western Australia. Open Access funding enabled and organized by CAUL and its Member Institutions

Primary and secondary outcomes stratified by gestational age at birth.

<p>Data represents median, 25^th–75^th percentile and minimum-maximum or N (%), unless otherwise stated.</p><p>BD = below detection limit.</p><p>Counts below the detection limit were assigned the value 4.4 [log₁₀(27,500)] for analysis.</p><p>p-values<0.025 considered significant after Bonferroni adjustment.</p><p>S2-S1: Difference between sample 2 and sample 1.</p><p>*Chi- square test was used to generate p values.</p

Effect of <i>Bifidobacterium breve</i> M-16V Supplementation on Fecal Bifidobacteria in Preterm Neonates - A Randomised Double Blind Placebo Controlled Trial

<div><p>Background</p><p>Probiotic supplementation significantly reduces the risk of necrotising enterocolitis (NEC) and all cause mortality in preterm neonates. Independent quality assessment is important before introducing routine probiotic supplementation in this cohort.</p><p>Aim</p><p>To assess product quality, and confirm that <i>Bifidobacterium breve</i> (<i>B. breve</i>) M-16V supplementation will increase fecal <i>B. breve</i> counts without adverse effects.</p><p>Methods and Participants</p><p>Strain identity (16S rRNA gene sequencing), viability over 2 year shelf-life were confirmed, and microbial contamination of the product was ruled out. In a controlled trial preterm neonates (Gestation <33 weeks) ready to commence or on feeds for <12 hours were randomly allocated to either <i>B. breve</i> M-16V (3×10⁹ cfu/day) or placebo (dextrin) supplementation until the corrected age 37 weeks. Stool samples were collected before (S1) and after 3 weeks of supplementation (S2) for studying fecal <i>B. breve</i> levels using quantitative PCR (Primary outcome). Secondary outcomes included total fecal bifidobacteria and NEC≥Stage II. Categorical and continuous outcomes were analysed using Chi-square and Mann-Whitney tests, and McNemar and Wilcoxon signed-rank tests for paired comparisons.</p><p>Results</p><p>A total of 159 neonates (Probiotic: 79, Placebo: 80) were enrolled. Maternal and neonatal demographic characteristics were comparable between the groups. The proportion of neonates with detectable <i>B. breve</i> increased significantly post intervention: Placebo: [S1:2/66 (3%), S2: 25/66 (38%), p<0.001] Probiotic: [S1: 29/74 (40%), S2: 67/74 (91%), p<0.001].</p><p>Median S1 <i>B. breve</i> counts in both groups were below detection (<4.7 log cells.g⁻¹), increasing significantly in S2 for the probiotic group (log 8.6) while remaining <4.7 log in the control group (p<0.001). There were no adverse effects including probiotic sepsis and no deaths. NEC≥Stage II occurred in only 1 neonate (placebo group).</p><p>Conclusion</p><p><i>B. breve</i> M-16V is a suitable probiotic strain for routine use in preterm neonates.</p><p>Trial Registration</p><p>Australia New Zealand Clinical Trial Registry <a href="http://www.anzctr.org.au/TrialSearch.aspx" target="_blank">ACTRN 12609000374268</a></p></div