Occurrence of Bifidobacteriaceae in human hypochlorhydria stomach.

Microbial Ecology in Health and Disease Pub Date : 2014-01-09 eCollection Date: 2014-01-01 DOI:10.3402/mehd.v25.21379
Paola Mattarelli, Giovanni Brandi, Carlo Calabrese, Fabio Fornari, Gian Maria Prati, Bruno Biavati, Barbara Sgorbati
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引用次数: 25

Abstract

Background: The human stomach, when healthy, is not a suitable host for microorganisms, but in pathological conditions such as gastritis, when gastric acid secretion is impaired, microbial overgrowth can be observed. Apart from Helicobacter pylori, the composition of microbiota, resident or exogenously introduced during neutral/high pH conditions, has not been investigated thoroughly. Thus, it is possible that Bifidobacteriaceae, important autochthonous and beneficial bacteria of human gastrointestinal microbiota, could over-colonize the stomach of hypochlorhydria patients suffering from autoimmune atrophic gastritis (AAG) or omeprazole-treated (OME) gastritis. This prompted us to characterize the Bifidobacteriaceae in such patients' gastric microbiota and to study its abnormal colonization.

Methods: Samples of gastric juices, and antrum and corpus mucosa from 23 hypochlorhydria patients (13 AAG and 10 OME) and from 10 control volunteers with base-line normochlorhydria, were cultivated in Brain Heart Infusion (BHI) and selective Bifidobacterium-Tryptone-Phytone-Yeast extract (Bif-TPY) media. The isolates were characterized by the fructose-6-phosphate phosphoketolase (F6PPK) test, electrophoresis of cellular proteins, the fermentation test, guanine-cytosine% DNA content, and DNA-DNA hybridization. Negative F6PPK isolates were characterized by order-specific polymerase chain reaction (PCR).

Results: A total of 125 isolates, assigned to the Bifidobacteriaceae family on the basis of their morphology, were obtained from AAG and OME patients, but not from normal subjects. Of these isolates, 55 were assigned to the Bifidobacteriaceae family on the basis of their fructose-6-phosphoketolase (PPK) activity, PPK being the key taxonomic enzyme of this family. The remaining 70 isolates, which were PPK-negative, were attributed to the Actinomycetales order following specific primer PCR analysis. We observed a significantly higher abundance of Bifidobacteriaceae (Bifidobacterium dentium, Scardovia inopinata, and Parascardovia denticolens) in OME group than the AAG group. Furthermore, the Actinomycetales distribution was homogeneous for both hypochlorhydria patient groups.

Conclusions: This study suggests that the Bifidobacteriaceae species, typically found in the oral cavity, readily colonizes the hypochlorhydria stomach of OME patients. The clinical relevance and the mechanism underlying this Bifidobacteriaceae presence in OME gastritis requires further functional studies.

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双歧杆菌科在人低氯酸胃中的发生。
背景:人的胃在健康时并不是微生物的适宜宿主,但在胃炎等病理状态下,当胃酸分泌受损时,可以观察到微生物过度生长。除了幽门螺杆菌外,在中性/高pH条件下,微生物群的组成,无论是常驻的还是外源的,尚未得到彻底的调查。因此,双歧杆菌科是人类胃肠道微生物群中重要的原生有益菌,可能在患有自身免疫性萎缩性胃炎(AAG)或奥美拉唑治疗(OME)胃炎的低氯血症患者的胃中过度定植。这促使我们对这类患者胃微生物群中的双歧杆菌科进行表征,并研究其异常定植。方法:将23例低氯血症患者(AAG 13例,OME 10例)和10例基线正常氯血症对照者的胃液、胃窦和胃体粘膜样本在脑心脏输注(BHI)和选择性双歧杆菌-色氨酸-植氨酸-酵母提取物(Bif-TPY)培养基中培养。采用果糖-6-磷酸磷酸酮醇酶(F6PPK)试验、细胞蛋白电泳、发酵试验、鸟嘌呤-胞嘧啶% DNA含量、DNA-DNA杂交等方法对分离菌株进行鉴定。F6PPK阴性分离株经序列特异性聚合酶链反应(PCR)鉴定。结果:从AAG和OME患者中分离到125株双歧杆菌,根据其形态划分为双歧杆菌科,但未从正常受试者中分离到。其中55株菌株根据其果糖-6-磷酸酮醇酶(PPK)活性被归为双歧杆菌科,PPK是双歧杆菌科的关键分类酶。其余70株均为ppk阴性,经特异性引物PCR分析属于放线菌目。我们观察到OME组的双歧杆菌科(牙双歧杆菌,inopinata Scardovia和牙副cardovia denticolens)的丰度显著高于AAG组。此外,两组低氯血症患者的放线菌分布均匀。结论:本研究表明,双歧杆菌科细菌通常存在于口腔中,很容易在OME患者的次氯酸胃中定植。临床相关性和这种双歧杆菌科存在于OME胃炎的机制需要进一步的功能研究。
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