Microbial interactions may impact patient's diagnosis, prognosis and treatment. Sporotrichosis is a hyperendemic neglected zoonosis in Brazil, caused by Sporothrix brasiliensis. Four pairs of clinical isolates of Sporothrix were recovered from four diseased cats (CIM01-CIM04, two isolates per animal) raising the possibility of coinfection in a sporotrichosis hyperendemic area, Brazil. Each isolate of the pair had distinct pigmentation in mycological culture, and was designated as "Light" or "Dark", for low and high pigmentation, respectively. Dark isolates reacted strongly with monoclonal antibodies to melanin (p ≤ 0.05) by both ELISA and FACS quantitation, and displayed a ring pattern with some regions exhibiting higher punctuated labeling at cell wall by immunofluorescence. In turn, Light isolates reacted less intensely, with few and discrete punctuated labeling at the cell wall. PCR identified all isolates as S. brasiliensis, MAT1-2 idiomorph. Sequencing of β-tubulin and calmodulin genes followed by phylogenetic analysis placed all eight isolates within the same cluster as others from the Brazilian hyperendemic area. The ability of these strains to stimulate cytokine production by human PBMCs (Peripheral blood mononuclear cells) was also analyzed. CIM01 and CIM03 Light and Dark isolates showed similar cytokine profiles to the control strain, while CIM02 and CIM04 behaved differently (p < 0.001), suggesting that differences in the surface of the isolates can influence host-fungus interaction. MICs for amphotericin B, terbinafine, caspofungin, micafungin, itraconazole, fluconazole, and voriconazole were obtained (CLSI M38-A2/M27-A3). Pairwise comparisons showed distinct MICs between Sporothrix Light and Dark isolates, higher than at least two-fold dilutions, to at least one of the antifungals tested. Isolates from the same pair displayed discrepancies in relation to fungistatic or fungicidal drug activity, notably after itraconazole exposure. Since S. brasiliensis Light and Dark isolates show disparate phenotypic parameters it is quite possible that coinfection represents a common occurrence in the hyperendemic area, with potential clinical implications on feline sporotrichosis dynamics. Alternatively, future studies will address if this specie may have, as reported for other fungi, broad phenotypic plasticity.

中文翻译：

---

在巴西孢子丝菌病高流行区由不同的巴西孢子丝菌共感染家猫。

微生物相互作用可能会影响患者的诊断、预后和治疗。孢子丝菌病是巴西的一种高度地方性被忽视的人畜共患病，由巴西孢子丝菌引起。从四只患病猫（CIM01-CIM04，每只动物两个分离物）中回收了四对孢子丝菌的临床分离物，这增加了在巴西孢子丝菌病高流行区合并感染的可能性。该对中的每个分离物在真菌培养中具有不同的色素沉着，并且分别为低色素沉着和高色素沉着指定为“浅”或“深”。通过 ELISA 和 FACS 定量，深色分离物与黑色素单克隆抗体 (p ≤ 0.05) 发生强烈反应，并显示出环状模式，某些区域通过免疫荧光在细胞壁上表现出更高的间断标记。反过来，光分离物反应不那么强烈，在细胞壁上有很少和离散的标点符号。PCR 将所有分离株鉴定为 S. brasiliensis，MAT1-2 独特型。对 β-微管蛋白和钙调蛋白基因进行测序，然后进行系统发育分析，将所有 8 个分离株与来自巴西高流行区的其他分离株置于同一簇内。还分析了这些菌株刺激人 PBMC（外周血单核细胞）产生细胞因子的能力。CIM01 和 CIM03 浅色和深色分离物显示出与对照菌株相似的细胞因子谱，而 CIM02 和 CIM04 表现不同（p < 0.001），表明分离物表面的差异会影响宿主-真菌的相互作用。获得了两性霉素 B、特比萘芬、卡泊芬净、米卡芬净、伊曲康唑、氟康唑和伏立康唑的 MIC (CLSI M38-A2/M27-A3)。成对比较显示，Sporothrix Light 和 Dark 分离株与至少一种所测试的抗真菌剂之间的 MIC 明显高于至少两倍的稀释度。来自同一对的分离株在抑真菌或杀真菌药物活性方面表现出差异，尤其是在接触伊曲康唑后。由于 S. brasiliensis Light 和 Dark 分离株显示出不同的表型参数，很可能在高流行区共感染很常见，对猫的孢子丝菌病动力学具有潜在的临床意义。或者，未来的研究将解决该物种是否如其他真菌报道的那样具有广泛的表型可塑性。来自同一对的分离株在抑真菌或杀真菌药物活性方面表现出差异，尤其是在接触伊曲康唑后。由于 S. brasiliensis Light 和 Dark 分离株显示出不同的表型参数，很可能在高流行区共感染很常见，对猫的孢子丝菌病动力学具有潜在的临床意义。或者，未来的研究将解决该物种是否如其他真菌报道的那样具有广泛的表型可塑性。来自同一对的分离株在抑真菌或杀真菌药物活性方面表现出差异，尤其是在接触伊曲康唑后。由于 S. brasiliensis Light 和 Dark 分离株显示出不同的表型参数，因此很可能共感染是高流行区的常见事件，对猫的孢子丝菌病动力学具有潜在的临床意义。或者，未来的研究将解决该物种是否如其他真菌报道的那样具有广泛的表型可塑性。对猫孢子丝菌病动力学具有潜在的临床意义。或者，未来的研究将解决该物种是否如其他真菌报道的那样具有广泛的表型可塑性。对猫孢子丝菌病动力学具有潜在的临床意义。或者，未来的研究将解决该物种是否如其他真菌报道的那样具有广泛的表型可塑性。