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1.
New Phytol ; 2024 Jun 30.
Article in English | MEDLINE | ID: mdl-38946157

ABSTRACT

Fusarium diseases pose a severe global threat to major cereal crops, particularly wheat. Existing biocontrol strains against Fusarium diseases are believed to primarily rely on antagonistic mechanisms, but not widely used under field conditions. Here, we report an endophytic fungus, Purpureocillium lilacinum YZ1, that shows promise in combating wheat Fusarium diseases. Under glasshouse conditions, YZ1 inoculation increased the survival rate of Fusarium graminearum (Fg)-infected wheat seedlings from 0% to > 60% at the seedling stage, and reduced spikelet infections by 70.8% during anthesis. In field trials, the application of YZ1 resulted in an impressive 89.0% reduction in Fg-susceptible spikelets. While a slight antagonistic effect of YZ1 against Fg was observed on plates, the induction of wheat systemic resistance by YZ1, which is distantly effective, non-specific, and long-lasting, appeared to be a key contributor to YZ1's biocontrol capabilities. Utilizing three imaging methods, we confirmed YZ1 as a potent endophyte capable of rapid colonization of wheat roots, and systematically spreading to the stem and leaves. Integrating dual RNA-Seq, photosynthesis measurements and cell wall visualization supported the link between YZ1's growth-promoting abilities and the activation of wheat systemic resistance. In conclusion, endophytes such as YZ1, which exhibits non-antagonistic mechanisms, hold significant potential for industrial-scale biocontrol applications.

2.
Chemosphere ; 362: 142696, 2024 Jun 24.
Article in English | MEDLINE | ID: mdl-38925517

ABSTRACT

Polyhydroxyalkanoates (PHAs) are important candidates for replacing petroleum-based plastics. This transition is urgent for the development of a biobased economy and to protect human health and natural ecosystems. PHAs are biobased and biodegradable polyesters that when blended with other polymers, such as poly(butylene adipate-co-terephthalate) (PBAT), acquire remarkable improvements in their properties, which allow them to comply with the requirements of packaging applications. However, the biodegradation of such blends should be tested to evaluate the impact of those polymers in the environment. For instance, PBAT is a compostable aliphatic-aromatic copolyester, and its biodegradation in natural environments, such as soil, is poorly studied. In this work, we evaluated the biodegradation of a bilayer film composed of PHB and PBAT, by a soil microbiome. The bilayer film reached 47 ± 1 % mineralization in 180 days and PHB was no longer detected after this period. The increased crystallinity of the PBAT residue was a clear sign of biodegradation, indicating that the amorphous regions were preferentially biodegraded. Seven microorganisms were isolated, from which 4 were closely related to microorganisms already known as PHB degraders, but the other 3 species, closely related to Streptomyces coelicoflavus, Clonostachys rosea and Aspergillus insuetus, were found for the first time as PHB degraders. Most remarkably, two fungi closely related to Purpureocillium lilacinum and Aspergillus pseudodeflectus (99.83 % and 100 % identity by ITS sequencing) were isolated and identified as PBAT degraders. This is very interesting due to the rarity of isolating PBAT-degrading microorganisms. These results show that the bilayer film can be biodegraded in soil, at mesophilic temperatures, showing its potential to replace synthetic plastics in food packaging.

3.
J Ophthalmic Inflamm Infect ; 14(1): 27, 2024 Jun 12.
Article in English | MEDLINE | ID: mdl-38867117

ABSTRACT

PURPOSE: To report a case of Purpureocillium lilacinum endophthalmitis. METHODS: The case of a fungal endophthalmitis caused by Purpureocillium lilacinum documented in an immunocompetent patient with no apparent trigger. RESULTS: A 64-year-old male with a two-month history of panuveitis in his left eye was referred to our hospital. Initially misdiagnosed as sympathetic ophthalmia due to a previous surgery on his right eye 4 months before the onset of the left ocular picture, the patient received corticosteroid treatment, leading to a rapid deterioration of the left eye condition. An urgent exploratory vitrectomy was performed to identify the underlying cause, revealing endophthalmitis. Microbiological investigation yielded Purpureocillium lilacinum as the causative agent. Despite intensive treatment, including intravitreal antibiotics and antifungals, along with another surgical intervention, clinical evolution remained unfavourable, ultimately leading to the evisceration of the affected eye. CONCLUSIONS: Purpureocillium lilacinum poses a rare yet sever threat as a causative agent of fungal endophthalmitis. Managing such cases is challenging due to the delayed identification, fungus's resistance to common antifungals, and its association with prior corticosteroid misuse in most patients. This case underscores the crucial importance of heightened clinical suspicion, early diagnosis, and the exploration of alternative treatment strategies in addressing Purpureocillium lilacinum endophthalmitis. The challenges posed by this rare fungal pathogen emphasize the need for a multidisciplinary approach and continued research to improve outcomes in these complex cases.

4.
World J Clin Cases ; 12(10): 1772-1777, 2024 Apr 06.
Article in English | MEDLINE | ID: mdl-38660073

ABSTRACT

BACKGROUND: Purpureocillium lilacinum (P. lilacinum) is a saprophytic fungus widespread in soil and vegetation. As a causative agent, it is very rarely detected in humans, most commonly in the skin. CASE SUMMARY: In this article, we reported the case of a 72-year-old patient with chronic lymphocytic leukemia who was admitted with cough and fever. Computed tomography revealed an infection in the right lower lobe. Bronchoalveolar lavage fluid culture and metagenomic next-generation sequencing were ultimately confirmed to have a pulmonary infection with P. lilacinum. She was eventually discharged with good outcomes after treatment with isavuconazole. CONCLUSION: Pulmonary infection with P. lilacinum was exceedingly rare. While currently there are no definitive therapeutic agents, there are reports of high resistance to amphotericin B and fluconazole and good sensitivity to second-generation triazoles. The present report is the first known use of isavuconazole for pulmonary P. lilacinum infection. It provides new evidence for the characterization and treatment of clinical P. lilacinum lung infections.

5.
Life (Basel) ; 14(1)2024 Jan 21.
Article in English | MEDLINE | ID: mdl-38276283

ABSTRACT

There has been a substantial increase in the number of cases of invasive fungal infections worldwide, which is associated with a growing number of immunosuppressed patients and a rise in antifungal resistance. Some fungi that were previously considered harmless to humans have become emerging pathogens. One of them is Purpureocillium lilacinum, a ubiquitous filamentous fungus commonly found in the environment, especially in the air and soil. P. lilacinum belongs to a bigger group of hyaline fungi that cause hyalohyphomycosis, a fungal infection caused by fungi with colorless hyphae. Although this is a heterogeneous group of fungi, there are similarities regarding their ubiquity, ways of transmission, affected patients, and difficulties in diagnostics and treatment. In hyalohyphomycosis, the skin is one of the most affected organs, which is why the involvement of dermatologists is crucial for the initial assessment, since the timely recognition and early diagnosis of this condition can prevent life-threatening infections and death. In this review, we covered cutaneous hyalohyphomycosis caused by P. lilacinum and other fungi in the same group, including Fusarium, Penicilium, Scedosporium, Scopulariopsis, Acremonium, and Trichoderma genera.

6.
Braz. j. biol ; 84: e253451, 2024. tab, graf
Article in English | LILACS, VETINDEX | ID: biblio-1345553

ABSTRACT

Abstract Root-knot nematode Meloidogyne incognita is among the biotic factors which has greatly affected both the yield and the quality of the tomato crop. The egg parasitic nematode, Purpureocillium lilacinum (Pl) is considered as one of the most promising agents in controlling and overcoming this plant pathogen. The nematicidal effect of the native isolate Pl AUMC 10149 on second stage juvenile's survival and egg hatching of M. incognita at different times of exposure was tested in vitro. The obtained data showed that Pl gave a maximum percentage of J2 mortality (97.6%) and egg hatching inhibition (79.8%) after 72 hours of exposure. The potentiality of Pl as well as Bio-Nematon to control M. incognita infecting tomato was conducted using different times of application in vivo. Nine treatments with five replicates were used for such bioagents compared with the nematicide Oxamyl. Each seedling was inoculated with 1000 J2s of nematode/pot and 10 mL of Pl (1x1010 CFU/mL) or Bio-Nematon spore suspension (1x108 CFU/mL) 10mL/pot. The results indicated that the most effective treatments in reducing nematode population, number of galls and egg masses of M. incognita in plant roots was performed with treatment by Pl pre-planting and post-infection with Pl (Rf 1.9) giving a significant enhancement in plant length (64.9%), fresh weight (72.52%) and shoot dry weight (163.41%) without negatively impacting environment. Therefore, the present study confirmed that using P. lilacinum AUMC 10149 can be used as a practical supplement to environmentally friendly disease management of root-knot nematodes in Egypt.


Resumo O nematoide-das-galhas Meloidogyne incognita está entre os fatores bióticos que afetaram enormemente a produção e a qualidade da cultura do tomate. O nematoide parasita de ovos, Purpureocillium lilacinum (Pl), é considerado um dos mais promissores agentes no controle e superação desse fitopatógeno. O efeito nematicida do isolado nativo Pl AUMC 10149 na sobrevivência de juvenis de segundo estágio e na eclosão dos ovos de M. incognita em diferentes momentos de exposição foi testado in vitro. Os dados obtidos mostraram que o Pl deu um percentual máximo de mortalidade de J2 (97.6%) e inibição da eclosão dos ovos (79.8%) após 72 horas de exposição. A potencialidade de Pl e de Bio-Nematon para controlar M. incognita infectando tomate foi conduzida em diferentes tempos de aplicação in vivo. Nove tratamentos com cinco repetições foram usados ​​para tais bioagentes em comparação com o nematicida Oxamyl. Cada muda foi inoculada com 1.000 J2s de nematoide / vaso e 10 mL de Pl (1×1010 CFU/mL). Ou suspensão de esporos Bio-Nematon (1×108 CFU/mL) 10mL/pot. Os resultados indicaram que os tratamentos mais eficazes na redução da população de nematoides, número de galhas e desovas de M. incognita nas raízes das plantas foram realizados com Pl pré-plantio e pós-infecção com Pl (Rf 1.9), dando um aumento significativo no comprimento da planta (64.9%), massa fresca (72.52%) e massa seca da parte aérea (163.41%) sem impactar negativamente o meio ambiente. Portanto, o presente estudo confirmou que o uso de P. lilacinum AUMC 10149 pode ser usado como um suplemento prático para o manejo ecologicamente correto de nematoides-das-galhas no Egito.


Subject(s)
Animals , Tylenchoidea/pathogenicity , Solanum lycopersicum/parasitology , Biological Control Agents , Hypocreales
7.
Braz. j. biol ; 842024.
Article in English | LILACS-Express | LILACS, VETINDEX | ID: biblio-1469255

ABSTRACT

Abstract Root-knot nematode Meloidogyne incognita is among the biotic factors which has greatly affected both the yield and the quality of the tomato crop. The egg parasitic nematode, Purpureocillium lilacinum (Pl) is considered as one of the most promising agents in controlling and overcoming this plant pathogen. The nematicidal effect of the native isolate Pl AUMC 10149 on second stage juveniles survival and egg hatching of M. incognita at different times of exposure was tested in vitro. The obtained data showed that Pl gave a maximum percentage of J2 mortality (97.6%) and egg hatching inhibition (79.8%) after 72 hours of exposure. The potentiality of Pl as well as Bio-Nematon to control M. incognita infecting tomato was conducted using different times of application in vivo. Nine treatments with five replicates were used for such bioagents compared with the nematicide Oxamyl. Each seedling was inoculated with 1000 J2s of nematode/pot and 10 mL of Pl (1x1010 CFU/mL) or Bio-Nematon spore suspension (1x108 CFU/mL) 10mL/pot. The results indicated that the most effective treatments in reducing nematode population, number of galls and egg masses of M. incognita in plant roots was performed with treatment by Pl pre-planting and post-infection with Pl (Rf 1.9) giving a significant enhancement in plant length (64.9%), fresh weight (72.52%) and shoot dry weight (163.41%) without negatively impacting environment. Therefore, the present study confirmed that using P. lilacinum AUMC 10149 can be used as a practical supplement to environmentally friendly disease management of root-knot nematodes in Egypt.


Resumo O nematoide-das-galhas Meloidogyne incognita está entre os fatores bióticos que afetaram enormemente a produção e a qualidade da cultura do tomate. O nematoide parasita de ovos, Purpureocillium lilacinum (Pl), é considerado um dos mais promissores agentes no controle e superação desse fitopatógeno. O efeito nematicida do isolado nativo Pl AUMC 10149 na sobrevivência de juvenis de segundo estágio e na eclosão dos ovos de M. incognita em diferentes momentos de exposição foi testado in vitro. Os dados obtidos mostraram que o Pl deu um percentual máximo de mortalidade de J2 (97.6%) e inibição da eclosão dos ovos (79.8%) após 72 horas de exposição. A potencialidade de Pl e de Bio-Nematon para controlar M. incognita infectando tomate foi conduzida em diferentes tempos de aplicação in vivo. Nove tratamentos com cinco repetições foram usados para tais bioagentes em comparação com o nematicida Oxamyl. Cada muda foi inoculada com 1.000 J2s de nematoide / vaso e 10 mL de Pl (1×1010 CFU/mL). Ou suspensão de esporos Bio-Nematon (1×108 CFU/mL) 10mL/pot. Os resultados indicaram que os tratamentos mais eficazes na redução da população de nematoides, número de galhas e desovas de M. incognita nas raízes das plantas foram realizados com Pl pré-plantio e pós-infecção com Pl (Rf 1.9), dando um aumento significativo no comprimento da planta (64.9%), massa fresca (72.52%) e massa seca da parte aérea (163.41%) sem impactar negativamente o meio ambiente. Portanto, o presente estudo confirmou que o uso de P. lilacinum AUMC 10149 pode ser usado como um suplemento prático para o manejo ecologicamente correto de nematoides-das-galhas no Egito.

8.
Med Mycol Case Rep ; 42: 100609, 2023 Dec.
Article in English | MEDLINE | ID: mdl-37767185

ABSTRACT

We hereby make the first report of a case of mycosis caused by Purpureocillium lilacinum in CARD9 deficiency. A 40-year-old woman complained of lymph node swellings in the left cervical area. She also had chronic mucocutaneous candidiasis (CMC), and was found to have CARD9 deficiency. Lymphadenitis by P. lilacinum was confirmed. The diagnosis was difficult, as culturing the biopsy specimen at a cautiously selected temperature (25 °C) and genetic analysis were both required. Oral administration of voriconazole improved her lymphadenopathy.

9.
BMC Microbiol ; 23(1): 244, 2023 08 31.
Article in English | MEDLINE | ID: mdl-37653404

ABSTRACT

BACKGROUND: Meloidogyne incognita greatly restricts the production of protected vegetables in China. Application of biocontrol agent Purpureocillium lilacinum is an important practice to control the nematode; however, instability usually occurs especially in heavily infested field. This study aimed to illustrate the high efficiency of P. lilacinum agent with fumigant Dazomet in vitro. RESULTS: P. lilacinum YES-2-14 showed strong parasitic and nematicidal activities to M. incognita. Pre-treatment with Dazomet significantly enhanced the biocontrol effects of the fungus. After fumigation with Dazomet at a dosage of 7.5 mg kg- 1 soil, parasitism of YES-2-14 on M. incognita eggs increased by more than 50%. Meanwhile, when P. lilacinum fermentation filtrate treated following Dazomet fumigation at 10 and 20 mg kg- 1 soil, the mortalities of second-stage juveniles (J2s) increased by 110.2% and 72.7%, respectively. Both Dazomet and P. lilacinum significantly reduced the penetration ability of J2s to tomato roots. When P. lilacinum filtrate used alone, the J2s penetrating into the young roots decreased by 48.8% at 4 dpi; while in the combined treatment, almost no J2 was detected within the roots at 4 dpi and the number of knots reduced by more than 99% at 45 dpi, indicating a synergistic effect of the biocontrol fungus and fumigant. CONCLUSIONS: Pre-treatment with Dazomet greatly increased the biocontrol efficacy of P. lilacinum to M. incognita. This research provides insight into the efficient management of plant parasitic nematodes and effective use of biocontrol agents.


Subject(s)
Tylenchoidea , Animals , China , Soil
10.
Appl Microbiol Biotechnol ; 107(19): 6057-6070, 2023 Oct.
Article in English | MEDLINE | ID: mdl-37526695

ABSTRACT

Poly(butylene adipate-co-terephthalate) (PBAT), a promising biodegradable aliphatic-aromatic copolyester material, can be applied as an alternative material to reduce the adverse effects of conventional plastics. However, the degradation of PBAT plastics in soil is time-consuming, and effective PBAT-degrading microorganisms have rarely been reported. In this study, the biodegradation properties of PBAT by an elite fungal strain and related mechanisms were elucidated. Four PBAT-degrading fungal strains were isolated from farmland soils, and Purpureocillium lilacinum strain BA1S showed a prominent degradation rate. It decomposed approximately 15 wt.% of the PBAT films 30 days after inoculation. Scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and Liquid chromatography mass spectrometry (LC‒MS) were conducted to analyze the physicochemical properties and composition of the byproducts after biodegradation. In the presence of PBAT, the lipolytic enzyme activities of BA1S were remarkably induced, and its cutinase gene was also significantly upregulated. Of note, the utilization of PBAT in BA1S cells was closely correlated with intracellular cytochrome P450 (CYP) monooxygenase. Furthermore, CreA-mediated carbon catabolite repression was confirmed to be involved in regulating PBAT-degrading hydrolases and affected the degradation efficiency. This study provides new insight into the degradation of PBAT by elite fungal strains and increases knowledge on the mechanism, which can be applied to control the biodegradability of PBAT films in the future. KEY POINTS: • Purpureocillium lilacinum strain BA1S was isolated from farmland soils and degraded PBAT plastic films at a prominent rate. • The lipolytic enzyme activities of strain BA1S were induced during coculture with PBAT, and the cutinase gene was significantly upregulated during PBAT degradation. • CreA-mediated carbon catabolite repression of BA1S plays an essential role in regulating the expression of PBAT-degrading hydrolases.


Subject(s)
Plastics , Polyesters , Polyesters/metabolism , Adipates , Soil , Hydrolases
11.
J Dermatol ; 50(8): 1088-1092, 2023 Aug.
Article in English | MEDLINE | ID: mdl-37082787

ABSTRACT

Hyalohyphomycosis is a rare infection caused by a group of fungi that are devoid of pigments in their cell walls. As one of the main pathogens of hyalohyphomycosis, Purpureocillium lilacinum (former Paecilomyces lilacinus) is known for its intrinsic resistance to various antifungal agents. Here, we report three cases that coincide with a history of farming and all of them suffered from cutaneous hyalohyphomycosis caused by P. lilacinum. They shared a clinical presentation consisting of erythematous-to-violaceous painful plaques with overlying pustules on one of their forearms. Hyphae and fungal elements were highlighted by periodic acid Schiff or Gomori methenamine silver staining in their skin biopsies. Fungal cultures of their skin tissues yielded P. lilacinum, which was confirmed by both morphological and molecular characteristics. All patients responded well to oral terbinafine or itraconazole treatment. In this report, we also reviewed previously reported cases associated with either P. lilacinum or other Paecilomyces spp. infections in Taiwan.


Subject(s)
Hyalohyphomycosis , Paecilomyces , Humans , Taiwan , Antifungal Agents/therapeutic use , Hyalohyphomycosis/microbiology , Cellulitis/drug therapy
13.
J Comp Pathol ; 201: 16-22, 2023 Feb.
Article in English | MEDLINE | ID: mdl-36646035

ABSTRACT

Entomopathogenic fungi, widely available biological agents used to control agricultural pests, are sporadically reported to cause focal or disseminated infection in reptiles and mammals, including humans. This study summarizes the clinical presentation, histopathological and molecular findings by panfungal polymerase chain reaction and sequencing of four cases of hypocrealean fungal infections in captive common green iguanas (Iguana, iguana). One case of granulomatous pneumonia, hepatitis and serositis was related to Metarhizium flavoviride complex infection. Two disseminated fungal infection cases, with scarce inflammatory cell infiltration, were caused by Beauveria bassiana while there was one case of multifocal granulomatous and necrotizing pneumonia by Purpureocillium spp. To the best of our knowledge, this is the first report of fatal mycosis infection due to entomopathogenic fungi in captive common green iguanas.


Subject(s)
Beauveria , Hypocreales , Iguanas , Mycoses , Animals , Humans , Brazil , Mycoses/microbiology , Mycoses/veterinary , Mammals
14.
Front Cell Infect Microbiol ; 12: 1033969, 2022.
Article in English | MEDLINE | ID: mdl-36530440

ABSTRACT

Since 1998, the state of Rio de Janeiro, Brazil, has become a public health problem regarding sporotrichosis, a disease caused by Sporothrix spp. involving contact with infected cats. Efforts to isolate these species from environmental sources are not always successful. In our study, soil from residences situated in cities of Rio de Janeiro where cats with sporotrichosis live was collected and cultured an attempt to isolate Sporothrix spp. but it was not successful. However, other saprophytic fungal species were isolated from soil and identified and among them Purpureocillium lilacinum was the most frequent. From there, we decided to study the in vitro interaction of this species with S. brasiliensis, the principal agent that causes sporotrichosis in this state. The results showed that ten isolates of P. lilacinum inhibited the radial mycelial growth of S. brasiliensis with different percentage of inhibition. The interaction between them revealed the pattern described as overgrowth by antagonist. In conclusion, our data suggest that fungal species with very fast growth and capable of producing metabolites could hinder the growth of Sporothrix spp., it also opens the way for the identification of secondary metabolites with biological activity that could be tested against pathogenic fungi.


Subject(s)
Sporothrix , Sporotrichosis , Sporotrichosis/veterinary , Sporotrichosis/microbiology , Brazil , Soil
15.
J Nematol ; 54(1): 20220042, 2022 Feb.
Article in English | MEDLINE | ID: mdl-36457368

ABSTRACT

Relative efficacy of various approaches for management of Meloidogyne incognita and the soilborne fungus Fusarium oxysporum f. sp. cucumerinum has been tested in cucumber under protected cultivation conditions for two seasons. Management practices, namely, chemicals (fumigant, nonfumigant, and fungicide), organic amendments (neem cake, leaves, and oil opted as soil and seed treatment), and biocontrol agents (egg-parasitic fungus and Purpureocillium lilacinum), were combined for the management of the disease complex in a randomized block design. Two significant parameters were measured: plant growth parameters (shoot length, dry shoot weight, dry root weight, and yield) and disease parameters (galls per plant, final nematode population, egg masses per plant, and fungal incidence). All treatments significantly improved plant growth parameters and reduced nematode reproduction as compared to untreated check. The integration of formalin and neem oil seed treatment favors the low root galling index compared to all other treatments in both the seasons. Formalin and neem oil seed treatment reduced the nematode population and fungal incidence, and increased the yield of cucumber during both the seasons.

16.
Arch Microbiol ; 204(11): 673, 2022 Oct 18.
Article in English | MEDLINE | ID: mdl-36255493

ABSTRACT

A lectin PCL, from Purpureocillium lilacinum a saprophytic, filamentous fungus was purified from the crude extract of the mycelia using 70% ammonium sulphate precipitation followed by affinity chromatography on mucin-Sepharose 4 B column. PCL is a monomer with an apparent molecular mass of 18.5 kDa as revealed by SDS-PAGE under both reducing and non-reducing conditions. PCL is a blood group non-specific lectin and has highest affinity towards chitin, mucin, asialomucin, fetuin with a MIC of 0.15 µg/mL and also recognizes L-fucose, galactose, lactose, N-acetyl galactosamine, hyaluronic acid. PCL is stable up to 60 °C and within the pH range 4-8. To understand its role in pathogenesis, effect of PCL was evaluated on human corneal epithelial cells (HCECs). PCL showed strong glycan mediated binding to HCECs and PCL showed proinflammatory response at lower concentrations by stimulating secretion of IL-6, 8. In contrast PCL at higher concentrations revealed opposite effect of HCECs growth inhibition. All these results collectively support the involvement of PCL in mediating host pathogen interactions possibly leading to pathogenesis. In addition, considering the entomopathogenic effect of Purpureocillium lilacinum, PCL may be attributed for this beneficiary effect, which needs to be explored.


Subject(s)
Blood Group Antigens , Keratitis , Humans , Lectins , Fucose , Galactose , Lactose , Ammonium Sulfate/metabolism , Sepharose , Hyaluronic Acid , Interleukin-6 , Keratitis/microbiology , Chitin/metabolism , Fetuins , Mucins/metabolism , Complex Mixtures , Galactosamine
17.
Biocontrol Sci ; 27(3): 117-130, 2022.
Article in English | MEDLINE | ID: mdl-36216563

ABSTRACT

Purpureocillium lilacinum has been recently found to contaminate a 20% (200,000 µg/mL) aqueous solution of polyhexamethylene biguanide hydrochloride (PHMB) . We aimed to elucidate the mechanism underlying the resistance of P. lilacinum to PHMB. First, we induced the PHMB-resistant (IR) strains IFM 67050 (IR) and IFM 65838 (IR) from the type strain P. lilacinum CBS 284.36T via cultivation in a medium containing high concentrations of PHMB. We then analyzed the DNA sequences via Illumina sequencing to evaluate the presence of genetic mutations in IFM 65838 (IR) . Further, we established an IFM 65838 (IR) uridine/uracil auxotrophic strain, and using the orotidine-5'-decarboxylase gene, pyrG as a selection marker, we tried to knockout a mutant gene in IFM 65838 (IR) using the CRISPR-Cas9 genome-editing technique. The growth rates of IFM 67050 (IR) and IFM 65838 (IR) in medium containing PHMB increased, and the minimum inhibitory concentrations (MICs) against PHMB also increased. Based on the DNA sequence analysis, we found a nonsynonymous point mutation in the gene PLI-008146 (G779A) in IFM 67050 (IR) and IFM 65838 (IR) . This point mutation leads to site combinations of splicing changes that cause partial sequences deletion (p.Y251_G281del) in the ΔPLI-008146 locus of IFM 65838 (IR) , and deletion sequences include partial adenosine/AMP deaminase motif (PF00962) orthologous to adenosine deaminase (ADA) (GeneBank: OAQ82383.1) . Furthermore, the mutant gene ΔPLI-008146 was successfully knocked out from the resistanceinduced strain using a novel CRISPR-Cas9 gene transformation method. A considerable reduction in growth rate and MIC against PHMB was observed in the absence of the mutant gene. Therefore, ADA may represent an important resistance factor in PHMB-resistant P. lilacinum.


Subject(s)
AMP Deaminase , Carboxy-Lyases , Adenosine , Adenosine Deaminase , Biguanides/pharmacology , Hypocreales , Uracil , Uridine
18.
Mycopathologia ; 187(5-6): 439-453, 2022 Dec.
Article in English | MEDLINE | ID: mdl-36178544

ABSTRACT

In a multicenter, prospective study of filamentous fungal keratitis in Greece, predisposing factors, etiology, treatment practices, and outcome, were determined. Corneal scrapings were collected from patients with clinical suspicion of fungal keratitis, and demographic and clinical data were recorded. Fungal identification was based on morphology, molecular methods, and matrix assisted laser desorption ionization time-of-flight mass-spectrometry. A total of 35 cases were identified in a 16-year study period. Female to male ratio was 1:1.7 and median age 48 years. Corneal injury by plant material, and soft contact lens use were the main risk factors (42.8% and 31.4%, respectively). Trauma was the leading risk factor for men (68.1%), contact lens use (61.5%) for women. Fusarium species were isolated more frequently (n = 21, 61.8%). F. solani was mostly associated with trauma, F. verticillioides and F. proliferatum with soft contact lens use. Other fungi were: Purpureocillium lilacinum (14.7%), Alternaria (11.8%), Aspergillus (8.8%), and Phoma foliaceiphila, Beauveria bassiana and Curvularia spicifera, one case each. Amphotericin B and voriconazole MIC50s against Fusarium were 2 mg/L and 4 mg/L respectively. Antifungal therapy consisted mainly of voriconazole locally or both locally and systemically, alone or in combination with liposomal AmB. Cure/improvement rate with antifungal therapy alone was 52%, keratoplasty was required in 40% of cases, and enucleation in 8%. In conclusion, filamentous fungal keratitis in Greece is rare, but with considerable morbidity. A large proportion of cases resulted in keratoplasty despite appropriate antifungal treatment.


Subject(s)
Corneal Ulcer , Eye Infections, Fungal , Fusarium , Keratitis , Humans , Male , Female , Middle Aged , Voriconazole/therapeutic use , Antifungal Agents/therapeutic use , Prospective Studies , Greece/epidemiology , Eye Infections, Fungal/drug therapy , Eye Infections, Fungal/epidemiology , Eye Infections, Fungal/microbiology , Keratitis/drug therapy , Keratitis/epidemiology , Keratitis/microbiology , Corneal Ulcer/drug therapy , Alternaria
19.
Med Mycol Case Rep ; 38: 5-8, 2022 Dec.
Article in English | MEDLINE | ID: mdl-36147049

ABSTRACT

Our case reports a 52-year-old woman who presented with Purpureocillium lilacinum skin infection after a renal transplantation. The diagnosis was difficult and this species exhibits many resistances to antifungal agents. The clinical history was marked by a relapse causes by a foreign body. Our case suggests that posaconazole may be an alternative to cure P. lilacinum infection, and that the surgical debridement, the identification and removal of a foreign body may improve the prognosis.

20.
Blood Sci ; 4(2): 89-92, 2022 Apr.
Article in English | MEDLINE | ID: mdl-35957666

ABSTRACT

Purpureocillium lilacinum (P lilacinum) is a rare pathogenic fungus, which mainly involves immunocompromised individuals. Here, we report a case of complicated multiple-organ infections involving skin, lungs, and spleen in a 63-year-old female with Evans' syndrome after 9 months of glucocorticoid treatment. Microbial examinations of skin biopsy and blood samples revealed P lilacinum infections. Posaconazole was effective in this patient. During anti-fungi treatment, she developed varicella-zoster virus infection and was diagnosed through next-generation sequencing examination. In conclusion, P lilacinum may affect different organ systems and is susceptible to posaconazole treatment. The molecular-based methods like microbial cell-free DNA sequencing could provide accurate and timely identification of a wide range of infections.

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