Buommino Elisabetta, Nocera Francesca Paola, Parisi Annamaria, Rizzo Antonietta, Donnarumma Giovanna, Mallardo Karina, Fiorito Filomena, Baroni Adone, De Martino Luisa
Department of Experimental Medicine, Second University of Naples, Italy.
Department of Veterinary Medicine and Animal Production, University of Naples "Federico II", Naples, Italy.
New Microbiol. 2016 Jul;39(3):216-223. Epub 2016 Jun 10.
Malassezia pachydermatis is a yeast belonging to the microbiota of the skin and mucous membranes of dog and cat, but it can also act as pathogen, causing dermatitis. The aim of this work was to evaluate the genetic variability of M. pachydermatis strains isolated from symptomatic dogs and cats and determine a correlation between genotype and phenotype. For this purpose eleven strains of M. pachydermatis were molecularly classified by nested-polymerase chain reaction (nested-PCR) based on ITS-1 and ITS-2 regions, specific for fungal rRNA genes. Furthermore, random amplification of polymorphic DNA (RAPD) was applied for genetic typing of M. pachydermatis isolates identifying four different genotypes. Strains belonging to genotype 1 produced the highest amount of biofilm and phospholipase activity. The inflammatory response induced by M. pachydermatis strains in immortalized human keratinocytes (HaCat cells) was significantly different when we compared the results obtained from each strain. In particular, HaCat cells infected with the strains belonging to genotypes 1 and 2 triggered the highest levels of increase in TLR-2, IL-1β, IL-6, IL-8, COX-2 and MMP-9 expression. By contrast, cells infected with the strains of genotype 3 and those of genotype 4 did not significantly induce TLR-2 and cytokines. The results obtained might suggest a possible association between genotype and virulence factors expressed by M. pachydermatis strains. This highlights the need for a more accurate identification of the yeast to improve the therapeutic approach and to monitor the onset of human infections caused by this emergent zoonotic pathogen.
厚皮马拉色菌是一种酵母菌,属于犬猫皮肤和黏膜微生物群,但它也可作为病原体,引起皮炎。本研究的目的是评估从有症状的犬猫中分离出的厚皮马拉色菌菌株的遗传变异性,并确定基因型与表型之间的相关性。为此,基于真菌rRNA基因特异的ITS-1和ITS-2区域,通过巢式聚合酶链反应(巢式PCR)对11株厚皮马拉色菌进行了分子分类。此外,应用随机扩增多态性DNA(RAPD)对厚皮马拉色菌分离株进行基因分型,鉴定出四种不同的基因型。属于基因型1的菌株产生的生物膜量和磷脂酶活性最高。当我们比较从各菌株获得的结果时,厚皮马拉色菌菌株在永生化人角质形成细胞(HaCat细胞)中诱导的炎症反应存在显著差异。特别是,感染基因型1和2菌株的HaCat细胞引发了TLR-2、IL-1β、IL-6、IL-8、COX-2和MMP-9表达的最高水平增加。相比之下,感染基因型3和4菌株的细胞未显著诱导TLR-2和细胞因子。所获得的结果可能表明厚皮马拉色菌菌株的基因型与表达的毒力因子之间可能存在关联。这凸显了更准确鉴定该酵母菌的必要性,以改进治疗方法并监测由这种新出现的人畜共患病原体引起的人类感染的发生。