Taurine drinking attenuates the burden of intestinal adult worms and muscle larvae in mice with Trichinella spiralis infection

The parasitic nematode Trichinella spiralis can cause trichinellosis, which leads to pathological processes in the intestine and muscle. The intestinal invasion determines the development, subsequent course, and consequences of the disease. Gastrointestinal nematode infection, including with T. spiralis, is accompanied by a rapid and reversible expansion of mucosal mast cell and goblet cell in the intestinal epithelium, which play important roles in the host immune response to parasite and worm expulsion from the intestine. Taurine and its derivatives have anti-infection and anti-inflammatory properties. We investigated whether taurine supplementation in mice could influence the development and pathological processes of infection with T. spiralis. Supplementing 1 % taurine in drinking water in mice infected with T. spiralis could alleviate the burden of intestinal adult worms on days 7 and 10 postinfection (all p?<?0.01) and the formation of infective muscle larvae in striated muscle during T. spiralis infection (p?<?0.01). As compared with T. spiralis infection alone, taurine treatment increased the number of goblet cells on days 7, 10, and 15 (p?<?0.01 and p?<?0.05) and alleviated intestinal mucosal mast cell hyperplasia on days 10 and 15 (all p?<?0.01). So taurine supplementation in drinking water increased infection-induced intestinal goblet cell hyperplasia and ameliorated mucosal mastocytosis. Thus, taurine can ameliorate the pathological processes of trichinellosis and may be of great value for the treatment and prevention of infection with T. spiralis and other gastrointestinal nematodes.     

Coinfection with Clonorchis sinensis modulates murine host response against Trichinella spiralis infection

Concomitant infections of different species of parasites are common in the field. Infection with one parasite species likely triggers host responses that may influence the subsequent infection of another species and alter disease outcomes. So far, the majority of studies have focused on single species parasite infection, and the mechanisms of protection induced by the first parasite infection against the secondary infection remain poorly defined. In this study, we assess the impact of trematode Clonorchis sinensis infection on the course of another tissue nematode Trichinella spiralis challenge. We observed that mice with preexisting C. sinensis infection had lower worm burden of intestinal T. spiralis than those infected with T. spiralis alone; mice with preexisting C. sinensis also had severe enteric histopathological changes and higher counts of intestinal Paneth cells in responses to T. spiralis challenge. The mRNA levels of interleukin (IL)-4, IL-10, IL-13, and tumor necrosis factor (TNF)-α from the small intestine and spleen of the different groups were analyzed using quantitative real-time polymerase chain reaction. Compared with that in mice infected with T. spiralis alone, the mRNA expression of IL-13 was significantly increased in the small intestine tissues and IL-4, IL-13, and TNF-α were significantly increased in the spleen tissues in the dually infected mice. Our findings suggest that a “preexisting” trematode infection of C. sinensis is a factor which contributes to reducing the establishment of T. spiralis adult worms in the small intestine.     

Species-specific antibody responses to the recombinant 53-kilodalton excretory and secretory proteins in mice infected with Trichinella spp

The 53-kDa proteins in larval excretory and secretory (E-S) products were expressed from five Trichinella species (T. spiralis, T. britovi, T. nativa, T. pseudospiralis, and T. papuae), using the Escherichia coli expression system, and the antibody responses to the 53-kDa recombinant proteins in mice infected with Trichinella spp. were analyzed by Western blotting. The 53-kDa protein is conserved among the five Trichinella species, with >60% similarity in amino acid sequences. The 53-kDa recombinant proteins of T. spiralis and T. pseudospiralis reacted to sera from mice infected with T. spiralis and T. pseudospiralis at 8 days postinfection (p.i.), respectively. An antibody against the 53-kDa recombinant protein of T. spiralis recognized the 53-kDa protein in the crude extracts from adult worms and 30-day p.i. muscle larvae and E-S products from muscle larvae of T. spiralis but did not recognize any proteins from T. pseudospiralis. The sera from the mice infected with T. spiralis strongly reacted with the 53-kDa recombinant protein of T. spiralis but did not react with the 53-kDa recombinant proteins of T. britovi, T. nativa, T. pseudospiralis, and T. papuae. Similarly, the sera from mice infected with T. britovi, T. nativa, T. pseudospiralis, or T. papuae strongly reacted with the 53-kDa recombinant proteins of T. britovi, T. nativa, T. pseudospiralis, or T. papuae, respectively. These results showed that the 53-kDa recombinant proteins provide early and species-specific antibody responses in mice infected with Trichinella spp.     

Interleukin-25 (IL-25) Promotes Efficient Protective Immunity against Trichinella spiralis Infection by Enhancing the Antigen-Specific IL-9 Response

Mammalian hosts often develop distinct immune response against the diverse parasitic helminths that have evolved for immune evasion. Interleukin-25 (IL-25), an IL-17 cytokine family member, plays a key role in initiating the protective immunity against several parasitic helminths; however, the involvement and underlying mechanisms by which IL-25 mediates immune response against Trichinella spiralis infection have not been investigated. Here we showed that IL-25 functions in promoting protective immunity against T. spiralis infection. Mice treated with IL-25 exhibited a lower worm burden and fewer muscle larvae in the later stage of T. spiralis infection. In contrast, mice treated with neutralizing antibody against IL-25 failed to expel T. spiralis effectively. During T. spiralis infection, intestinal IL-25 expression was rapidly elevated before the onset of IL-4 and IL-9 induction. While antigen-specific Th2 and Th9 immune responses were both developed during T. spiralis infection, an antigen-specific Th9 response appeared to be transiently induced in the early stage of infection. Mice into which antigen-specific T cells deficient in IL-9 were transferred were less effective in worm clearance than those given wild-type T cells. The strength of the antigen-specific Th9 immune response against T. spiralis could be enhanced or attenuated after treatment with IL-25 or neutralizing antibody against IL-25, respectively, correlating positively with the levels of intestinal mastocytosis and the expression of IL-9-regulated genes, including mast cell- and Paneth cell-specific genes. Thus, our study demonstrates that intestinal IL-25 promotes protective immunity against T. spiralis infection by inducing antigen-specific Th9 immune response.     

Effect of Trichinella spiralis intervention on TNBS-induced experimental colitis in mice

Inflammatory bowel disease (IBD), including ulcerative colitis and Crohn’s disease (CD), are chronic autoimmune diseases with a high recurrence rate. Epidemiological data have shown that the incidence of IBD increases annually because of improved sanitary conditions and reduced parasitic infection rates. In this experiment, experimental colitis was induced in mice by administering 2,4,6-trinitrobenzene sulfonic acid (TNBS) 28?days after they were infected with Trichinella spiralis to confirm that T. spiralis infection could alleviate the severity of TNBS-induced colitis.Thirty-six male BALB/c mice aged 6–8 weeks were randomly divided into four groups: control group (with 50% ethanol, Control), T. spiralis-infected group (TS-Control), TNBS-induced colitis model group (Colitis), and T. spiralis-pre-infected and TNBS-induced colitis group (TS-Colitis). The mice were sacrificed 3, 7, and 14?days after the model was established. Changes in various colitis indicators to investigate the effect of T. spiralis infection on TNBS-induced murine CD model.Results showed that the weight, DAI score, and macroscopic and microscopic colon damage in the TS-Colitis significantly decreased compared with those in the Colitis. ELISA revealed that the IFN-γ expression decreased and the IL-4 expression increased in the TS-Colitis compared with those in the Colitis. Western Blotting results revealed that the NF-κB expression increased in the Colitis and higher than those in the TS-Colitis. And Flow cytometry results revealed that the percentage of CD4+CD25+Foxp3+ Treg cells significantly increased in the TS-Colitis.T. spiralis-infected mice induced Th2 immune responses and balanced Th1 immune responses stimulated by TNBS to ameliorate intestinal inflammation.     

The effect of intestinal trichinellosis on oral bioavailability of albendazole in mice

The effect of Trichinella spiralis infection on the pharmacokinetic profile of orally administered albendazol has been investigated in mice during the intestinal phase of the disease. Swiss CD-1 mice were orally infected with 300 ± 50 muscle larvae of T. spiralis and then treated with albendazole (ABZ) formulated in hydroxy-propyl-beta-cyclodextrins at the dose of 10 mg/kg given orally on days 0, 5, 10 and 22 post-infection (p.i.). Blood samples were taken at 0.25, 0.5, 0.75, 1, 5, 6 and 24 h post-treatment (p.t.). Adult worm establishment as well as the histopathological alterations induced in the small intestine was assessed on days 0, 5, 10 and 22 p.i. The area under the blood concentrations to time curve (AUC) for ABZ sulphoxide was not significantly higher in infected mice than in control during the first step of intestinal infection (day 5 p.i.), whereas in the late step (day 10 p.i.), it was significantly lower. On day 22 p.i., the AUC showed similar values in both groups. The histopathological analysis showed a transient acute inflammatory reaction that varied from moderate to severe as the infection progressed from the early to the late intestinal stage. After intestinal infection, the inflammation was mild or absent with no signs of chronic effects. The histopathological studies correlated with the pharmacokinetics of ABZ and show that after transient inflammation induced by intestinal T. spiralis infection, the mucosa is restored to allow absorption of ABZ up to levels comparable to those observed in non-infected controls.     

Construction and use of a Trichinella spiralis phage display library to identify the interactions between parasite and host enterocytes

Although it has been known for many years that Trichinella spiralis initiates infection by penetrating the columnar epithelium of the small intestine, the mechanisms by which T. spiralis infective larvae recognize and invade the intestinal epithelial cells (IECs) are unknown. It is speculated that the molecular interactions between the parasite and host enterocytes may mediate the recognition and invasion of IECs by T. spiralis. However, no Trichinella proteins that interact with the enterocytes have been identified previously. The aim of this study was to identify Trichinella proteins that bind to IECs by screening a T7 phage display cDNA library constructed using messenger RNA from T. spiralis intestinal infective larvae. Following five rounds of biopanning, sequencing, and bioinformatics analysis, ten T. spiralis proteins (Tsp1–Tsp10) with significant binding to normal mouse IECs were identified. The results of the protein classification showed that six proteins (Tsp1, calcium-transporting ATPase 2 protein; Tsp4, ovochymase-1; Tsp6, T-complex protein 1 subunit eta; Tsp7, glycosyl hydrolase family 47; Tsp8, DNA replication licensing factor MCM3; and Tsp10, nudix hydrolase) of these T. spiralis proteins were annotated with putative molecular functions. Out of the six proteins, five have catalytic activity, four have binding activity, and one has transporter activity. Anti-Tsp10 antibodies prevented the in vitro partial invasion of IECs by infective larvae and the mice immunized with the recombinant phage T7-Tsp10 showed a 62.8 % reduction in adult worms following challenge with T. spiralis muscle larvae. Although their biological functions are not yet fully known, these proteins might be related to the larval invasion of host enterocytes. Future experiments will be necessary to ascertain whether these proteins play important roles in the recognition and invasion of host enterocytes. The construction and biopanning of Trichinella phage display libraries provide a novel approach for searching for candidate genes that are related to invasion and for identifying protein interactions between parasite and host.     

Changes in the activity and kinetics of mouse intestinal glutathione transferase during experimental trichinellosis

The glutathione S-transferases (GSTs) are a group of multifunctional enzymes, which play a critical role in cellular detoxification. Our investigations deal with the contribution of GST in the biochemical defense against Trichinella spiralis infection. The aim of this study was to examine the effect of T. spiralis infection on the total activity and kinetic properties of cytosolic GST in the intestine during the intestinal phase of experimental trichinellosis in mice. Our results showed a statistically significant increase (relative to the uninfected control) in the total GST activity both in the small and large intestines of the infected mice. Moreover, we observed changes in the kinetics of substrate saturation of GST. Trichinellosis in the small intestine caused a 12-fold decrease in the low K _m value and a sixfold increase in the high K _m value. In the large intestine, infection with T. spiralis caused only a fivefold increase in the low K _m value, whereas the high K _m value remained unchanged. We suggest that GST from the mouse small intestine could be involved in the detoxification of parasite excretory–secretory products released to the host intestine during trichinellosis and that these products influence the quaternary structure of this enzyme.     

Changes in Intestinal Fluid Transport and Immune Responses to Enterotoxins Due to Concomitant Parasitic Infection

The effect of a parasitic infection on enterotoxic diarrhea and on local and systemic formation of antibody to the toxin after immunization was studied in mice. Trichinella spiralis infection was chosen as the model, since the effects of the parasite when residing in both intestinal and extraintestinal sites can be studied. It was found that during the intestinal stage of the infection, the fluid response to cholera toxin as well as dibutyryl-cyclic adenosine 3′,5′ -monophosphate was greatly enhanced and that this was associated with a marked reduction in the absorption of fluid from the intestine. Later in the infection (migration stage), fluid accumulation in response to cholera toxin was significantly reduced, whereas absorption was normal and secretion in response to dibutyryl-cyclic adenosine 3′,5′-monophosphate was somewhat increased. Still later in the infection (muscular stage), the fluid-secretory response to cholera toxin was normal. There was a drastic depression of local formation of antitoxin of both immunoglobulin and immunoglobulin classes in mice given the first two of four oral immunizations with cholera toxin during the intestinal stage of T. spiralis infection. When the priming was given before or after the intestinal stage, the local antitoxin response was not affected. The titers of circulating antibodies were also depressed in mice given the first immunizations during the intestinal stage. In addition, significant though less pronounced depression of the serum antibody response was observed in mice primed during the extraintestinal stage.     

Development of cellular immune response of mice to infection with low doses of Trichinella spiralis, Trichinella britovi and Trichinella pseudospiralis larvae

The murine cellular immune response to the infection with ten larvae of encapsulating (Trichinella spiralis, Trichinella britovi) and non-encapsulating species (Trichinella pseudospiralis) was studied. Both T. spiralis and T. britovi stimulated the proliferation of splenic T and B lymphocytes during the intestinal phase of infection, but T. spiralis activated the proliferative response also at the muscle phase, particularly in B cells. Non-encapsulating T. pseudospiralis stimulated the proliferation of T and B cells only on day 10 post-infection (p.i.) and later at the muscle phase. The numbers of splenic CD4 and CD8 T cells of T. spiralis infected mice were significantly increased till day 10 p.i., i.e., at the intestinal phase, and then at the late muscle phase, on day 60 p.i. T. britovi infection increased the CD4 and CD8 T cell numbers only on day 30 p.i. Decreased numbers of CD4 and CD8 T cells after T. pseudospiralis infection suggest a suppression of cellular immunity. Both encapsulating Trichinella species induced the Th2 response (cytokines interleukin-5 (IL-5) and interleukin-10) at the intestinal phase and the Th2 dominant response at the advanced muscle phase. Interferon-γ (IFN-γ) production (Th1 type) started to increase with migrating newborn larvae from day 15 p.i. till the end of the experiment. IL-5 production was suppressed during the intestinal phase of T. pseudospiralis infection. The immune response to T. pseudospiralis was directed more to the Th1 response at the muscle phase, the high IFN-γ production was found on day 10 p.i. and it peaked on days 45 and 60 p.i.     

Mast cell production of IL-4 and TNF may be required for protective and pathological responses in gastrointestinal helminth infection

Expulsion of the gastrointestinal nematode Trichinella spiralis is associated with Th2 responses and intestinal inflammation, which correlate with a marked mast cell (MC) response. To address the role of MC-derived cytokines in the induction of protective responses, WBB6F1-KitW/KitW-v (W/W^v) mice were reconstituted with wild-type, tumor necrosis factor (TNF)-α^−/−, or interleukin (IL)-4^−/− bone marrow (BM) prior to infection with T. spiralis. W/W^v mice reconstituted with TNF-α^−/− or IL-4^−/− BM expelled the parasite less efficiently and showed diminished enteropathy, whereas protective responses were normal in W/W^v mice reconstituted with wild-type BM and were accompanied by intestinal pathology. MC responses were reduced in W/W^v mice reconstituted with IL-4^−/− BM and to a lesser extent when reconstituted with TNF-α^−/−. These results suggest that MC-derived IL-4 and TNF may regulate the induction of protective Th2 responses and intestinal inflammation associated with the expulsion of T. spiralis. Significantly, these studies suggest a role for MC-derived cytokines as autocrine growth factors.     

<Emphasis Type="Italic">Trichinella spiralis</Emphasis> reinfection: macrophage activity in BALB/c mice

The role of macrophages and their products—nitric oxide (NO) and superoxide anion (O₂ ⁻)—were examined in BALB/c mice reinfected with Trichinella spiralis. Mice were infected twice with 400 T. spiralis larvae; the secondary infection was performed 60 days after the primary one. Adult T. spiralis in intestines were detected from 5 to 20 days postprimary infection (dpi), and postreinfection (dpri), but the intensity of worm expulsion was greater and quicker after the reinfection. The highest muscle larvae numbers were detected from 60 dpi till 90 dpi (30 dpri), and thereafter, a great reduction was noted. The high numbers of macrophages in the peritoneal cavity of infected mice were obtained at 20 dpi, but the highest numbers of these cells from 10 to 30 dpri were observed. The production of NO by macrophages in infected mice was suppressed at 5 dpi, and then, NO release increased until 60 dpi. On the contrary, the long-lasting (5–30 dpri) suppression of NO production after T. spiralis reinfection was observed. The levels of NO in plasma and urine were lower in infected mice till 20 dpi; there were no differences in plasma and urine NO level after the reinfection in comparison to control uninfected animals. The production of O₂ ⁻ in peritoneal macrophages was inhibited during the first 2 weeks after infection, but the reinfection caused great increase in O₂ ⁻ production lasting 30 days.     

A murine model of tuberculosis/type 2 diabetes comorbidity for investigating the microbiome,metabolome and associated immune parameters

Tuberculosis (TB) is one of the deadliest infectious diseases in the world. The metabolic disease type 2 diabetes (T2D) significantly increases the risk of developing active TB. Effective new TB vaccine candidates and novel therapeutic interventions are required to meet the challenges of global TB eradication. Recent evidence suggests that the microbiota plays a significant role in how the host responds to infection, injury and neoplastic changes. Animal models that closely reflect human physiology are crucial in assessing new treatments and to decipher the underlying immunological defects responsible for increased TB susceptibility in comorbid patients. In this study, using a diet‐induced murine T2D model that reflects the etiopathogenesis of clinical T2D and increased TB susceptibility, we investigated how the intestinal microbiota may impact the development of T2D, and how the gut microbial composition changes following a very low‐dose aerosol infection with Mycobacterium tuberculosis (Mtb). Our data revealed a substantial intestinal microbiota dysbiosis in T2D mice compared to non‐diabetic animals. The observed differences were comparable to previous clinical reports in TB patients, in which it was shown that Mtb infection causes rapid loss of microbial diversity. Furthermore, diversity index and principle component analyses demonstrated distinct clustering of Mtb‐infected non‐diabetic mice vs. Mtb‐infected T2D mice. Our findings support a broad applicability of T2D mice as a tractable small animal model for studying distinct immune parameters, microbiota and the immune‐metabolome of TB/T2D comorbidity. This model may also enable answers to be found to critical outstanding questions about targeted interventions of the gut microbiota and the gut‐lung axis.     

Synergy between intraepithelial lymphocytes and lamina propria T cells drives intestinal inflammation during infection

Oral infection of C57BL/6 mice with Toxoplasma gondii triggers severe necrosis in the ileum within 7–10 days of infection. Lesion development is mediated by Th-1 cytokines, CD4⁺ T cells, and subepithelial bacterial translocation. As such, these features share similarity to Crohn's disease. Recently, we uncovered a role for intraepithelial lymphocytes (IELs) in mediating pathology after Toxoplasma infection. We show here that αβ and not γδ T-cell IELs mediate intestinal damage. By adoptive transfer of mucosal T cells into naive Rag1^−/− mice, we demonstrate that IELs do not function alone to cause inflammatory lesions, but act with CD4⁺ T lymphocytes from the lamina propria (LP). Furthermore, recipient mice pretreated with broad-spectrum antibiotics to eliminate intestinal flora resisted intestinal disease after transfer of IELs and LP lymphocytes. Our data provide valuable new insights into the mechanisms of intestinal inflammation, findings that have important implications for understanding human inflammatory bowel disease.     

The effectors responsible for gastrointestinal nematode parasites, Trichinella spiralis, expulsion in rats

Helper T (Th2) cells type 2 have a central role in host protective responses to gastrointestinal nematode parasite, Trichinella spiralis infection, but the actual effector mechanisms involved in parasite expulsion are still uncertain. Recent evidences suggest that mast cell recruitment and activation may associate with parasite elimination from host intestines in mice. On the other hand, IgE production may induce defensive responses to primary infection with the helminth in rats. The differences of host effector mechanisms to the same experimental infections might disturb our understanding of the host protective mechanisms to gastrointestinal nematode parasite infection. In order to redefine these differences, we investigated in detail the relationship between intestinal immune responses and worm expulsion following T. spiralis infection among several rat strains including mutants. As a result, there were significant correlations of parasite expulsion with mast cell hyperplasia in addition to serum IgE level. Moreover, mast cell-deficient and dysfunction rats showed delayed worm elimination from their gut. Therefore, the present study suggests that mast cells should also be one of the prominent effector cells involved in T. spiralis parasite expulsion in rats as well as mice.     

Interleukin-9 is involved in host protective immunity to intestinal nematode infection

Murine studies have demonstrated that, as with other nematodes, infection with the intestinal nematode Trichinella spiralis is associated with a pronounced intestinal mastocytosis, eosinophilia and an elevation in serum levels of total IgE. Both interleukin (IL)-4 and IL-5 are clearly important in the generation of IgE responses and eosinophilia, respectively, but the control of mucosal mastocytosis in vivo is not as well defined. Mucosal mast cells appear to be particularly important with regard to T. spiralis infections as there is good evidence to suggest their involvement in expulsion of the parasite from the host. In this study we examined the effect of the overproduction of the Th2 cytokine IL-9 on infection with this nematode. We demonstrate that naive IL-9-transgenic mice have an intense intestinal mastocytosis and high serum levels of mouse mast cell protease-1. Moreover, upon infection high titers of parasite-specific IgG1 were observed with a heightened mast cell response, which was associated with the rapid expulsion of T. spiralis from the gut. Furthermore, as depression of this mast cell response, using anti-c-kit antibodies, resulted in the inability of these mice to expel the parasite, this study clearly demonstrates an activity of IL-9 on mucosal mastocytosis and the host protective immune response in vivo.     

Microbiome Diversity in the Bronchial Tracts of Patients with Chronic Obstructive Pulmonary Disease

Culture of bacteria from bronchial secretions in respiratory patients has low sensitivity and does not allow for complete assessment of microbial diversity across different bronchial compartments. In addition, a significant number of clinical studies are based on sputum samples, and it is not known to what extent they describe the real diversity of the mucosa. In order to identify previously unrecognized lower airway bacteria and to investigate the complexity and distribution of microbiota in patients with chronic obstructive pulmonary disease (COPD), we performed PCR amplification and pyrosequencing of the 16S rRNA gene in patients not showing signs or symptoms of infection. Four types of respiratory samples (sputum, bronchial aspirate, bronchoalveolar lavage, and bronchial mucosa) were taken from each individual, obtaining on average >1,000 16S rRNA sequences per sample. The total number of genera per patient was >100, showing a high diversity, with Streptococcus, Prevotella, Moraxella, Haemophilus, Acinetobacter, Fusobacterium, and Neisseria being the most commonly identified. Sputum samples showed significantly lower diversity than the other three sample types. Lower-bronchial-tree samples, i.e., bronchoalveolar lavage and bronchial mucosa, showed a very similar bacterial compositions in contrast to sputum and bronchial aspirate samples. Thus, sputum and bronchial aspirate samples are upper bronchial tree samples that are not representative of the lower bronchial mucosa flora, and bronchoalveolar lavage samples showed the results closest to those for the bronchial mucosa. Our data confirm that the bronchial tree is not sterile in COPD patients and support the existence a different microbiota in the upper and lower compartments.     

Characterization of the vaginal fungal flora in pregnant diabetic women by 18S rRNA sequencing

Pregnancy and diabetes are regarded as individual risk factors for vaginal candidiasis. The high prevalence of vaginal candidiasis in pregnant diabetic women can be explained by disruption of the balance of the vaginal normal flora. However, little is known about the overall structure and composition of the vaginal fungal flora in pregnant diabetic women. In the present study, the diversity and richness of the vaginal fungal flora in healthy non-pregnant women (group HN), healthy pregnant women (group HP), women with gestational diabetes mellitus (group GDM), and pregnant women with diabetes mellitus type I (group T1DM) were investigated using an 18S rRNA gene clone library method. Our data demonstrated that the composition of the vaginal fungal flora in the four groups could be divided into two phyla (Ascomycetes, 20/26, and Basidiomycetes, 6/26). The most predominant vaginal fungal species belonged to the Candida and Saccharomyces genera, uncultured fungi, and a large number of low-abundance taxa that were unrecorded or underrepresented in previous studies using cultivation-dependent methods. Variation in operational taxonomic units (OTUs) between the study cohorts was generally high in the clone libraries, as 9, 13, 17, and 20 phylotypes were identified in groups HN, HP, GDM, and T1DM, respectively. The Shannon indices of groups GDM and T1DM (with poorer glycemic control) were significantly higher compared to groups HN and HP (p?<?0.05). The data presented here revealed an increased diversity and varied composition of the vaginal fungal flora in pregnant diabetic women and demonstrated that poor glycemic control might be associated with disturbances in the vaginal fungal flora.     

Intestinal ameliorative effects of traditional Ogi-tutu,Vernonia amygdalina and Psidium guajava in mice infected with Vibrio cholera

BackgroundCholera, a severe acute watery diarrhea caused by Vibrio cholerae is endemic in Nigeria with most cases occurring in the rural areas. In South West Nigeria, some individuals resort to alternative treatments such as Ogi-tutu, Psidium guajava and Vernonia amygdalina during infections. The effectiveness of these alternatives in the prevention and treatment of V. cholerae infection requires experimental investigation.ObjectiveThis study was designed to investigate the ameliorative effects of Ogi-tutu, Vernonia amygdalina and Psidium guajava on intestinal histopathology of experimental mice infected with V. cholerae.MethodsPreliminary investigation of in vitro vibriocidal activities of these alternatives were carried out using agar cup diffusion assay. For ameliorative effects, adult mice were inoculated with 100 µl (106 cells) of Vibrio cholerae and dosed at 0 h (immediate prevention) and 4 h (treatment of infection) and their intestines were histopathologically evaluated.ResultsThe histopathological changes were the same irrespective of the treated groups, but the lesions varied in extent and severity. The ameliorative effects in decreasing order were V. amygdalina > P. guajava > Ogi-tutu.ConclusionV. amygdalina gave the best ameliorative effects in the prevention and treatment of V. cholerae infection.     

Metagenomic analyses reveal antibiotic-induced temporal and spatial changes in intestinal microbiota with associated alterations in immune cell homeostasis

Despite widespread use of antibiotics, few studies have measured their effects on the burden or diversity of bacteria in the mammalian intestine. We developed an oral antibiotic treatment protocol and characterized its effects on murine intestinal bacterial communities and immune cell homeostasis. Antibiotic administration resulted in a 10-fold reduction in the amount of intestinal bacteria present and sequencing of 16S rDNA segments revealed significant temporal and spatial effects on luminal and mucosal-associated communities including reductions in luminal Firmicutes and mucosal-associated Lactobacillus species, and persistence of bacteria belonging to the Bacteroidetes and Proteobacteria phyla. Concurrently, antibiotic administration resulted in reduced RELMβ production, and reduced production of interferon-γ and interleukin-17A by mucosal CD4⁺ T lymphocytes. This comprehensive temporal and spatial metagenomic analyses will provide a resource and framework to test the influence of bacterial communities in murine models of human disease.