Molecular Epidemiology of Underreported Emerging Zoonotic Pathogen Streptococcus suis in Europe.

Streptococcus suis, a zoonotic bacterial pathogen circulated through swine, can cause severe infections in humans. Because human S. suis infections are not notifiable in most countries, incidence is underestimated. We aimed to increase insight into the molecular epidemiology of human S. suis infections in Europe. To procure data, we surveyed 7 reference laboratories and performed a systematic review of the scientific literature. We identified 236 cases of human S. suis infection from those sources and an additional 87 by scanning gray literature. We performed whole-genome sequencing to type 46 zoonotic S. suis isolates and combined them with 28 publicly available genomes in a core-genome phylogeny. Clonal complex (CC) 1 isolates accounted for 87% of typed human infections; CC20, CC25, CC87, and CC94 also caused infections. Emergence of diverse zoonotic clades and notable severity of illness in humans support classifying S. suis infection as a notifiable condition.

Starkeya nomas sp. nov., a prosthecate and budding bacterium isolated from an immunocompromized patient.

Strain HF14-78462T is an environmental bacterium found in clinical samples from an immunocompromized patient in 2014 at Hospital Universitari i Politècnic La Fe (Valencia, Spain). Phenotypically, strain HF14-78462T cells were Gram-stain-negative, aerobic, non-spore forming and non-motile small rods which formed mucous and whitish-translucent colonies when incubated at 20-36 °C. Phylogenetic analyses based on the 16S rRNA genes and the whole genomes of closest sequenced relatives confirmed that strain HF14-78462T is affiliated with the genus Starkeya. The strain was oxidase, catalase and urease positive; but indole, lysine decarboxylase, ornithine decarboxylase and DNase negative, did not produce H2S and was able to utilize a wide variety of carbon sources including acetamide, adonitol, amygdalin, l-arabinose, citric acid, glucose, mannitol and melibiose. Unlike Starkeya novella and Starkeya koreensis, strain HF14-78462T failed to grow in thiosulphate-oxidizing media and had a narrower temperature growth range. Its genome was characterized by a size of 4.83 Mbp and a C+G content of 67.75 mol%. Major fatty acids were C18:1 ω7c, cyclo C19 : 0 and C16 : 0, its polar acids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol and an aminophospholipid; while the ubiquinones were Q9 (1.8 %) and Q10 (98.2 %). Digital DNA-DNA hybridization values were 41 and 41.4 against S. novella and S. koreensis, respectively, while average nucleotide identity values were around 84 %. Phenotypic, average nucleotide identity and phylogenomic comparative studies suggest that strain HF14-78462T is a new representative of the genus Starkeya and the name Starkeya nomas sp. nov. is proposed. The type strain is HF14-78462T (=CECT 30124T=LMG 31874T).

Implication of the Polymeric Phenolic Fraction and Matrix Effect on the Antioxidant Activity, Bioaccessibility, and Bioavailability of Grape Stem Extracts.

The bioaccessibility and bioavailability of phenolics compounds of two grape stem extracts with different composition were studied. High polymeric extract (HPE) presented a higher content of total phenolics (TPC), procyanidins, hemicelluloses, proteins, and ashes, whereas low procyanidin extract (LPE) showed a higher fat, soluble sugars, and individual phenolic compounds content. Corresponding to its higher total phenolics content, HPE possesses a higher antioxidant activity (TEAC value). The digestion process reduced the antioxidant activity of the HPE up to 69%, due to the decrease of TPC (75%) with a significant loss of polymeric compounds. LPE antioxidant activity was stable, and TPC decreased by only 13% during the digestion process. Moreover, a higher antioxidant phenolic compounds bioavailability was shown in LPE in contrast to HPE. This behaviour was ascribed mainly to the negative interaction of polymeric fractions and the positive interaction of lipids with phenolic compounds. Therefore, this study highlights the convenience of carrying out previous studies to identify the better extraction conditions of individual bioavailable phenolic compounds with antioxidant activity, along with those constituents that could increase their bioaccessibility and bioavailability, such as lipids, although the role played by other components, such as hemicelluloses, cannot be ruled out.

Characterization of Streptococcus pyogenes from Animal Clinical Specimens, Spain.

Streptococcus pyogenes appears to be almost exclusively restricted to humans, with few reports on isolation from animals. We provide a detailed characterization (emm typing, pulsed-field gel electrophoresis [PFGE], and multilocus sequence typing [MLST]) of 15 S. pyogenes isolates from animals associated with different clinical backgrounds. We also investigated erythromycin resistance mechanisms and phenotypes and virulence genes. We observed 2 emm types: emm12 (11 isolates) and emm77 (4 isolates). Similarly, we observed 2 genetic linages, sequence type (ST) 26 and ST63. Most isolates exhibited the M macrolide resistance phenotype and the mefA/ermB genotype. Isolates were grouped into 2 clones on the basis of emm-MLST-PFGE-virulence gene profile combinations: clone 1, characterized by the combined genotype emm12-ST36-pulsotype A-speG; and clone 2, characterized by the genotype emm77-ST63-pulsotype B-speC. Our results do not show conclusively that animals may represent a new reservoir of S. pyogenes but indicate the ability of human-derived S. pyogenes isolates to colonize and infect animals.

Pseudonocardia carboxydivorans in human cerebrospinal fluid: a case report in a patient with traumatic brain injury.

BACKGROUND: Members of the genus Pseudonocardia have been widely reported and recovered from several ecosystems, such as soil samples and plant samples. Pseudonocardia bacteria colonize the microbial communities on the integument of fungus gardening ant species. We present the first documented case of Pseudonocardia carboxydivorans isolated in human cerebrospinal fluid (CSF). To the best of our knowledge, this is the first report of an human infection by P. carboxydivorans. CASE PRESENTATION: A patient, who suffered a traumatic brain injury a month before, was admitted to this hospital due to gait alteration and cognitive disturbances. Culture of cerebrospinal fluid showed ramified, not acid-fast, Gram positive bacilli. The bacterium was identified by molecular methods as P. carboxydivorans. CONCLUSION: This is the first documented case of isolating P. carboxydivorans in human CSF in a case of probable meningitis. Further research is needed in order to determine its pathogenic role in human infections.

First report of human infection by Christensenella minuta, a gram-negative, strickly anaerobic rod that inhabits the human intestine.

Christensenella minuta is a Gram-negative strictly anaerobic short rod that inhabits the human gut. This bacterium was isolated in a mixed infection with Desulfovibrio desulfuricansfrom the blood of a patient with a diagnosis of acute appendicitis. The strain was identified by 16S rRNA sequence analysis. As far as we know, this is the first time C.minuta has been isolated from a human clinical specimen.

Molecular characterization of invasive Streptococcus dysgalactiae subsp. equisimilis. Multicenter study: Argentina 2011-2012.

Streptococcus dysgalactiae subsp. equisimilis (SDSE) has virulence factors similar to those of Streptococcus pyogenes. Therefore, it causes pharyngitis and severe infections indistinguishable from those caused by the classic pathogen. The objectives of this study were: to know the prevalence of SDSE invasive infections in Argentina, to study the genetic diversity, to determine the presence of virulence genes, to study antibiotic susceptibility and to detect antibiotic resistance genes. Conventional methods of identification were used. Antibiotic susceptibility was determined by the disk diffusion and the agar dilution methods and the E-test. Twenty eight centers from 16 Argentinean cities participated in the study. Twenty three isolates (16 group G and 7 group C) were obtained between July 1 2011 and June 30 2012. Two adult patients died (8.7%). Most of the isolates were recovered from blood (60.9%). All isolates carried speJ and ssa genes. stG62647, stG653 and stG840 were the most frequent emm types. Nineteen different PFGE patterns were detected. All isolates were susceptible to penicillin and levofloxacin, 6 (26.1%) showed resistance or reduced susceptibility to erythromycin [1 mef(A), 3 erm(TR), 1 mef(A)+erm(TR) and 1 erm(TR)+erm(B)] and 7 (30.4%) were resistant or exhibited reduced susceptibility to tetracycline [2 tet(M), 5 tet(M)+tet(O)]. The prevalence in Argentina was of at least 23 invasive infections by SDSE. A wide genetic diversity was observed. All isolates carried speJ and ssa genes. Similarly to other studies, macrolide resistance (26.1%) was mainly associated to the MLSB phenotype.

Fournier's gangrene caused by Actinomyces funkei, Fusobacterium gonidiaformans and Clostridium hathewayi.

We report the first case of Fournier's gangrene caused by three unusual anaerobic organisms: Actinomyces funkei, Fusobacterium gonidiaformans and Clostridium hathewayi. The infection occurred in a 73-year-old man without typical risk factors for the development of Fournier's gangrene. Clinical outcome was good after prolonged antibiotic treatment and extensive debridement of the perineum. The case suggests that A. funkei, F. gonidiaformans and C. hathewayi should be considered as potential pathogens of Fournier's gangrene. Human infections caused by these organisms are very rare but can be underestimated because correct identification is very difficult, especially in polymicrobial infections such as Fournier's gangrene.

Peritonitis caused by Bifidobacterium longum: case report and literature review.

Bifidobacterium spp. rarely causes human infections. We report a case of a 42-year-old man with a history of pancolonic diverticulosis, who suffered a purulent peritonitis caused by Bifidobacterium longum secondary to intestinal perforation. Clinical outcome was good after urgent surgery and antibiotic treatment with imipenem and amoxicillin/clavulanic acid. Our case shows that Bifidobacterium spp. should be considered as a cause of peritonitis, especially in patients with risk of intestinal perforation. The review of the literature shows that these organisms can cause a wide spectrum of severe infections, especially in patients with underlying diseases. Infections caused by Bifidobacterium spp. may be overlooked or underreported since it may be considered normal microbiota.

Skin and soft-tissue infections caused by Actinobaculum schaalii: report of two cases and literature review.

Skin and soft-tissue infections (SSTIs) caused by Actinobaculum spp. are very rare. In the present study, we report two cases and review the literature. The first case was an immunocompromised patient with an extensive cellulitis secondary to an inguinal abscess, and the second case was a patient with a pilonidal abscess. Clinical outcomes of both patients were good after surgical drainage and treatment with cloxacillin. The review of the literature showed that SSTIs caused by Actinobaculum spp. are usually located on the perineal and inguinal regions and can be severe, particularly in immunocompromised patients. SSTIs caused by Actinobaculum spp. can be overlooked because identification is often difficult and they can be considered as contaminants.

Influence of culinary skills on ultraprocessed food consumption and Mediterranean diet adherence: An integrative review.

The Mediterranean diet is a diet pattern with several healthy characteristics, including a high consumption of fresh fruits, vegetables, whole-grain products, and extra virgin olive oil as the main fat source. It is also characterized by a low consumption of processed foods and high-fat meats. However, there has been a nutritional transition such that, because of globalization and economic growth, the consumption of ultraprocessed foods is increasing, comprising 14% to 57% of total energy intake. North America and Australia have the worst prognosis. Further, the younger population consumes greater amounts of ultraprocessed foods compared with adults, whereas the older population has the lowest rates. Among the various factors responsible for the nutritional transition, culinary skills seem an important function of diet quality, but, unfortunately, culinary skills have been little investigated by the scientific community. Higher culinary skills may be related to increased consumption of homemade meals, higher diet quality, and greater consumption of vegetables and fruits and may be associated with higher Mediterranean diet adherence. Consequently, ultraprocessed foods, ready-to-eat food, and delivered meals, as well as meals outside the home, are reduced. Unfortunately, the process of generational transmission of cooking skills is progressively weakening, aggravated by the reduction of free time for home cooking. Still, even though there are few interventions conducted to improve culinary skills, especially those focused on children, the available results suggest that there have been positive changes in the direction of a healthier dietary pattern. This integrative review analyzes consumption levels of ultraprocessed foods. We then consider the extent to which culinary skills affect ultraprocessed food consumption levels and the relative impact on obesity.

Sourdough bread enriched with exopolysaccharides and gazpacho by-products modulates in vitro the microbiota dysbiosis.

Sourdough bread enriched with soluble fiber (by in-situ exopolysaccharides production) and insoluble fiber (by gazpacho by-products addition) showed prebiotic effects an in vitro dynamic colonic fermentation performance with obese volunteer's microbiota. Bifidobacterium population was maintained whereas Lactobacillus increased throughout the colonic sections. Conversely, Enterobacteriaceae and Clostridium groups clearly decreased. Specific bacteria associated with beneficial effects increased in the ascending colon (Lactobacillus fermentum, Lactobacillus paracasei, Bifidobacterium longum and Bifidobacterium adolescentis) whereas Eubacterium eligens, Alistipes senegalensis, Prevotella copri and Eubacterium desmolans increased in the transversal and descending colon. Additionally, Blautia faecis and Ruminococcus albus increased in the transversal colon, and Bifidobacterium longum, Roseburia faecis and Victivallis vadensis in the descending colon. Bifidobacterium and Lactobacillus fermented the in-situ exopolysaccharides and released pectins from gazpacho by-products, as well as cellulosic degraded bacteria. This increased the short and medium chain fatty acids. Acetic acid, as well as butyric acid, increased throughout the colonic tract, which showed greater increases only in the transversal and descending colonic segments. Conversely, propionic acid was slightly affected by the colonic fermentation. These results show that sourdough bread is a useful food matrix for the enrichment of vegetable by-products (or other fibers) in order to formulate products with microbiota modulatory capacities.

Fiber from elicited butternut pumpkin (Cucurbita moschata D. cv. Ariel) modulates the human intestinal microbiota dysbiosis.

Elicited pumpkin was evaluated as a potential daily consumption product able to modulate the gut microbiota. An in vitro dynamic colonic fermentation performance with microbiota from obese volunteers was used. Prebiotic effects were observed after the pumpkin treatment. Bifidobacterium abundance was maintained during the treatment period whereas Lactobacillus increased in the transversal and descending colon. Conversely, Enterobacteriaceae and Clostridium groups were more stable, although scarce decreasing trends were observed for same species. Increments of Lactobacillus acidophilus and Limosilactobacillus fermentum (old Lactobacillus fermentum) were observed in the whole colonic tract after the treatment period. However, modulatory effects were mainly observed in the transversal and descending colon. Diverse bacteria species were increased, such as Akkermansia muciniphila, Bacteroides dorei, Cloacibacillus porcorum, Clostridium lactatifermentans, Ruminococcus albus, Ruminococcus lactaris, Coprococcus catus, Alistipes shahii or Bacteroides vulgatus. The prebiotic effect of the elicited pumpkin was provided by the fiber of the pumpkin, suggesting a release of pectin molecules in the transversal and distal colonic tract through low cellulosic fiber degradation, explaining the increases in the total propionic and butyric acid in these colonic sections. Also, a possible modulatory role of carotenoids from the sample was suggested since carotenes were found in the descending colon. Hence, the results of this research highlighted pumpkin as a natural product able to modulate the microbiota towards a healthier profile.

Burkholderia cepacia complex infection in an Adult Cystic Fibrosis unit in Madrid.

INTRODUCTION: Burkholderia cepacia complex have emerged as significant pathogens in cystic fibrosis (CF) patients due to the risk of cepacia syndrome and the innate multi-resistance of the microorganisms to antibiotics. The aim of this study was to describe the antimicrobial susceptibility profiles, the genotypes and subtypes of BCC, and the clinical evolution of CF patients with BCC. METHODS: The lung function and Brasfield and Shwachman score were assessed in 12 patients. BCC were identified and susceptibility was studied by MicroScan (Siemens). Species and genospecies of BCC were confirmed by molecular methods in a Reference Centre (Majadahonda). RESULTS: BCC were identified in 12 of 70 patients (17.1%) over a ten year period. The mean age to colonization by BCC was 24.4 years (SD: 7.71). B. cenocepacia was isolated in 4 patients (33.3%), B. contaminans was isolated in 3 patients (25%), both B. vietnamiensis and B. stabilis were isolated in 2 patients (16.7%), and B. cepacia, B. multivorans and B. late were isolated in one patient (8.3%). Among the B. cenocepacia, subtype IIIa was identified in two strains, and subtype IIIb was identified in the other two strains. There was susceptibility to meropenem in 90% of BCC, 80% to cotrimoxazole, 60% to minocycline, 50% to ceftazidime, and 40% to levofloxacin. CONCLUSIONS: B. cenocepacia was the most prevalent species among the BCC isolated in CF adult patients, and subtypes IIIa and IIIb were identified in the 50% of the strains. Meropenem and cotrimoxazole showed the best activity.

Impact of a retrograded starch ingredient obtained from Negro Jamapa beans (Phaseolus vulgaris L. Negro Jamapa) on glucose metabolism and oxidative stress in induced diabetic lab rats model.

A retrograded starch ingredient obtained from Negro Jampa beans (Phaseolus vulgaris L. Negro Jamapa), applying a debranching process (18 U/g for 12 h) and retrogradation (2 °C for 3 days), was evaluated as a potential functional ingredient to improve glucose homeostasis in a diabetic animal model. The obtained ingredient was reduced in rapid digestible starch amount compared to its related isolated native starch (34.1 % and 53.6 %, respectively) and resistant (33.4 % and 22.3 %, respectively). Therefore, a reduced total digestibility was determined for the obtained ingredient compared to native starch (51.5 % and 79 %, respectively). As consequence, diabetic animals fed with functional ingredient replacement (30 %) showed a lower and attenuated postprandial glucose levels, reducing the hyperglycaemic condition, compared with the non-treated animals (r2 = 0.9775; p ≤ 0.05), reducing the glucose serum levels 73 % compared (17.21 vs 23.6 mmol/L, respectively). Also, significant improvement on weight gain (49.75 ± 34.1 g) compared to non-treated (18.14 ± 45.52 g), as well as lower insulin resistance index and improved oxidative stress status was determined for the treated group. These results highlight the potential of retrograded starch obtained from Negro Jamapa beans as a functional ingredient focus on the improvement of the glucose homeostasis and diabetic condition.

The microbiota-metabolic syndrome axis as a promoter of metabolic osteoarthritis.

The relation between obesity and osteoarthritis (OA) development has been traditionally explained as consequence of the excessive joint effort derived of overweight. However, in the last two decades a metabolic OA has been suggested through diverse molecular mechanism implying metabolic syndrome, although more investigation must be conducted to elucidate it. Metabolic syndrome is responsible of the release of diverse inflammatory cytokines, specially the increased adipokine in obesity, causing a chronic low-grade inflammatory status that alters the joint homeostasis. In this scenario, the microbiota dysbiosis contribute by worsening the low-grade chronic inflammation or causing metabolic disorders mediated by endotoxemia generated by an increased lipopolysaccharides intake. This results in joint inflammation and cartilage degradation, which contributes to the development of OA. Also, the insulin resistance provoked by type 2 Diabetes contributes to the OA development. When intake patterns are considered, some coincidences can be pointed between the food patterns associated to the metabolic syndrome and the food patterns associated to OA development. Therefore, these coincidences support the idea of a molecular mechanism of the OA development caused by the molecular mechanism generated under the metabolic syndrome status. This review points the relation between metabolic syndrome and OA, showing the connected molecular mechanisms between both pathologies as well as the shared dietary patterns that promote or prevent both pathologies.

Microbiota dysbiosis caused by dietetic patterns as a promoter of Alzheimer's disease through metabolic syndrome mechanisms.

Microbiota dysbiosis and metabolic syndrome, consequences of a non-adequate diet, generate a feedback pathogenic state implicated in Alzheimer's disease development. The lower production of short chain fatty acids (SCFAs) under dysbiosis status leads to lipid homeostasis deregulation and decreases Angptl4 release and AMPK activation in the adipose tissue, promoting higher lipid storage (adipocyte hypertrophy) and cholesterol levels. Also, low SCFA generation reduces GPR41 and GPR43 receptor activation at the adipose tissue (increasing leptin release and leptin receptor resistance) and intestinal levels, reducing the release of GLP-1 and YPP. Therefore, lower satiety sensation and energy expenditure occur, promoting a weight gaining environment mediated by higher food intake and lipid storage, developing dyslipemia. In this context, higher glucose levels, together with higher free fatty acids in the bloodstream, promote glycolipotoxicity, provoking a reduction in insulin released, insulin receptor resistance, advanced glycation products (AGEs) and type 2 diabetes. Intestinal dysbiosis and low SCFAs reduce bacterial biodiversity, increasing lipopolysaccharide (LPS)-producing bacteria and intestinal barrier permeability. Higher amounts of LPS pass to the bloodstream (endotoxemia), causing a low-grade chronic inflammatory state characterized by higher levels of leptin, IL-1ß, IL-6 and TNF-α, together with a reduced release of adiponectin and IL-10. At the brain and neuronal levels, the generated insulin resistance, low-grade chronic inflammation, leptin resistance, AGE production and LPS increase directly impact the secretase enzymes and tau hyperphosphorylation, creating an enabling environment for ß-amyloid senile plaque and tau tangled formations and, as a consequence, Alzheimer's initiation, development and maintenance.

Molecular epidemiology, antimicrobial susceptibilities and resistance mechanisms of Streptococcus pyogenes isolates resistant to erythromycin and tetracycline in Spain (1994-2006).

BACKGROUND: Group A Streptococcus (GAS) causes human diseases ranging in severity from uncomplicated pharyngitis to life-threatening necrotizing fasciitis and shows high rates of macrolide resistance in several countries. Our goal is to identify antimicrobial resistance in Spanish GAS isolates collected between 1994 and 2006 and to determine the molecular epidemiology (emm/T typing and PFGE) and resistance mechanisms of those resistant to erythromycin and tetracycline. RESULTS: Two hundred ninety-five out of 898 isolates (32.8%) were erythromycin resistant, with the predominance of emm4T4, emm75T25, and emm28T28, accounting the 67.1% of the 21 emm/T types. Spread of emm4T4, emm75T25 and emm28T28 resistant clones caused high rates of macrolide resistance. The distribution of the phenotypes was M (76.9%), cMLSB (20.3%), iMLSB (2.7%) with the involvement of the erythromycin resistance genes mef(A) (89.5%), msr(D) (81.7%), erm(B) (37.3%) and erm(A) (35.9%).Sixty-one isolates were tetracycline resistant, with the main representation of the emm77T28 among 20 emm/T types. To note, the combination of tet(M) and tet(O) tetracycline resistance genes were similar to tet(M) alone reaching values close to 40%. Resistance to both antibiotics was detected in 19 isolates of 7 emm/T types, being emm11T11 and the cMLSB phenotype the most frequent ones. erm(B) and tet(M) were present in almost all the strains, while erm(A), mef(A), msr(D) and tet(O) appeared in less than half of them. CONCLUSIONS: Spanish GAS were highly resistant to macrolides meanwhile showed minor resistance rate to tetracycline. A remarkable correlation between antimicrobial resistance and emm/T type was noticed. Clonal spread of emm4T4, emm75T25 and emm28T28 was the main responsable for macrolide resistance where as that emm77T28 clones were it to tetraclycline resistance. A wide variety of macrolide resistance genes were responsible for three macrolide resistance phenotypes.