Paecilomyces/Purpureocillium keratitis: A consecutive study with a case series and literature review.

Paecilomyces/Purpureocillium species is an emerging pathogen of fungal keratitis; the risk factor, clinical course, and outcome of Paecilomyces/Purpureocillium keratitis need more exploration. We retrospectively reviewed 12 patients with culture-proven Paecilomyces/Purpureocillium keratitis in our hospital from 2003 to 2017 and combined them with 50 previous cases reported after the review conducted by Yuan et al. in 2009. Clinical features between the previously and newly reported cases were compared using the publication by Yuan et al. as a cutoff point. The mean age of the 62 newly reported patients with Paecilomyces/Purpureocillium keratitis was 52.7 years. Of these, contact lens wear was the most common predisposing factor (n = 32, 52%), followed by a preexisting corneal disease or previous ocular surgery (n = 12, 19%), and trauma (n = 8, 13%). Fifty eyes (81%) were treated with voriconazole, of which 31 (63%) were medically cured. Twenty-one of 62 eyes (34%) required therapeutic surgery. Compared with the 42 patients reported by Yuan et al., the patients were younger (P = .025); a higher proportion of the patients were contact lens wearers (P = .005); more patients were treated with voriconazole (P = .000); fewer patients required therapeutic surgery (P = .000) in recent reports. Contact lens use has become the major risk factor for Paecilomyces/Purpureocillium keratitis. The surgical rate has been significantly lower in recent publications, probably because of the prevalent use of voriconazole.

[Role of erythrocytes in mechanisms of nonspecific protection of blood in infection caused by the fungus of genus Paecilomyces.]

Paecilomyces variotii is a commonly occurring species in air and food, and it is also associated with many types of human infections. Tissue forms of the fungus Paecilomyces variotii or their cytoskeletons were revealed in the cytoplasm of erythrocytes in patients with allergy and bronchial asthma in paecilomycosis. Our study was aimed at investigating the role of red blood cells in the mechanisms of the nonspecific protection of the host in conditions of chronic persistent infection of the blood with the fungus of the genus Paecilomyces. We examined a total of eighty-four 16-to-72-year-old patients (39 men and 45 women) presenting with activation of paecilomyces infection in blood. We used laboratory, biochemical, allergic-and-immunological and microbiological methods of study. Fungal cultures were identified phenotypically and by means of phylogenetic analysis.Our findings are suggestive of a new type of the oxygen-dependent mechanism of cytotoxicity of erythrocytes, which is caused by permanent formation of reactive oxygen species as a result of non-enzymatic oxidation of haemoglobin to methaemoglobin. The resulting superoxide anion radical (O2-), hydrogen peroxide (H2O2), and hydroxyl radical (OH-) exhibit a powerful bactericidal action which is, probably, activated when the fungal cells are captured and immersed in the erythrocyte cytoplasm or in a closed cavity formed by RBCs around large fungal cells. In conditions of chronic blood infection with tissue forms of fungi of the genus Paecilomyces oxygen-dependent cytotoxicity of erythrocytes is the main mechanism of readjustment of blood from the infectious agent of Paecilomycosis.

Potential of endophytic fungi as biocontrol agents of Duponchelia fovealis (Zeller) (Lepidoptera: Crambidae) / Fungos endofíticos para o biocontrole de Duponchelia fovealis (Zeller) (Lepidoptera: Crambidae)

Abstract This study reports the first assessment of endophytic fungi isolated from strawberry leaves and selection of isolates for the control of Duponchelia fovealis, a new pest of strawberries. A total of 400 strawberry leaves of the cultivar 'Albion' were collected in four commercial farms. Leaves were disinfected, cut in fragments, and placed on Petri dishes containing potato dextrose agar media with tetracycline and incubated for 30 days. Following this time, 517 fungal colonies were isolated, and thirteen genera were identified: Cladosporium, Aspergillus, Nigrospora, Fusarium, Trichoderma, Chaetomium, Alternaria, Paecilomyces, Penicillium, Ulocladium, Bipolaris, Diaporthe, and Phoma. Eight isolates belonging to the genera Aspergillus, Diaporthe, Paecilomyces, and Cladosporium were selected for pathogenicity bioassays against third instar larvae of D. fovealis. Isolates of Paecilomyces induced the highest mortality rates.

Resumo Este trabalho apresenta a primeira contribuição no isolamento de fungos endofíticos de folhas de morangueiro e na seleção de isolados para controle de Duponchelia fovealis, uma nova praga do morangueiro. Foram coletadas 400 folhas da cultivar 'Albion' em quatro lavouras comerciais de morangueiro. As folhas foram desinfetadas, cortadas em fragmentos e depositadas em placas de Petri contendo Ágar Batata Dextrose como meio, em conjunto com tetraciclina e incubados durante 30 dias. Um total de 517 colônias fúngicas e treze gêneros foram isolados: Cladosporium, Aspergillus, Nigrospora, Fusarium, Trichoderma, Chaetomium, Alternaria, Paecilomyces, Penicillium, Ulocladium, Bipolaris, Diaporthe e Phoma. Oito isolados pertencentes aos gêneros Aspergillus, Diaporthe, Paecilomyces e Cladosporium foram selecionados para os bioensaios de virulência contra larvas de 3º instar de D. fovealis. Isolados de Paecilomyces causaram as maiores taxas de mortalidade.

Phylogeny of Paecilomyces, the causal agent of pistachio and some other trees dieback disease in Iran.

One of the most important fungal agents of pistachio dieback disease belongs to the ascomycete genus Paecilomyces that has been identified as P. variotii. In 2012-2014, 700 plant samples from pistachio trees and 27 other plant species with dieback symptoms were collected from 10 provinces of Iran. Of the 567 pistachio samples, 277 Paecilomyces strains were obtained and from the 133 samples of other plants (except pistachio and including Pistacia mutica, Punica granatum, Prunus amygdalus, Caesalpinia gilliesii, Nerium oleander, Tamarix aphylla, Tamarix hispida and Haloxylon sp.), 23 fungal isolates were recovered and five isolates were obtained from the air of infected pistachio orchards. Based on morphology, all 305 isolates were identified as P. variotii. Physiological studies revealed that 299 isolates belong to P. formosus. Three isolates were assigned to P. variotii, while three isolates could not be assigned to any of the known species. Of the 305 isolates, 62 were selected for phylogenetic analysis based on DNA variation (ITS, ß-tubulin and calmodulin). This analysis showed that all of our isolates form a clade with P. formosus. P. formosus consists of the three former species P. formosa, P. lecythidis and P. maximus. This study shows that our isolates form a strongly supported clade with strains of P. lecythidis. So, the causal agent of dieback disease of pistachio and other examined trees is P. formosus which is closely related to the former species P. lecythidis and has some differences with the former species P. formosa and P. maximus. Based on phylogenetic studies P. formosus thus seems to be a species complex that could be divided into three separate species.

Potential of endophytic fungi as biocontrol agents of Duponchelia fovealis (Zeller) (Lepidoptera:Crambidae).

This study reports the first assessment of endophytic fungi isolated from strawberry leaves and selection of isolates for the control of Duponchelia fovealis, a new pest of strawberries. A total of 400 strawberry leaves of the cultivar 'Albion' were collected in four commercial farms. Leaves were disinfected, cut in fragments, and placed on Petri dishes containing potato dextrose agar media with tetracycline and incubated for 30 days. Following this time, 517 fungal colonies were isolated, and thirteen genera were identified: Cladosporium, Aspergillus, Nigrospora, Fusarium, Trichoderma, Chaetomium, Alternaria, Paecilomyces, Penicillium, Ulocladium, Bipolaris, Diaporthe, and Phoma. Eight isolates belonging to the genera Aspergillus, Diaporthe, Paecilomyces, and Cladosporium were selected for pathogenicity bioassays against third instar larvae of D. fovealis. Isolates of Paecilomyces induced the highest mortality rates.

Purpureocillium lilacinum tattoo-related skin infection in a kidney transplant recipient.

Purpureocillium lilacinum is an emerging pathogenic mold among immunocompromised hosts that causes cutaneous infections related to skin breakdown. We present the first reported case of P. lilacinum tattoo-related skin infection, to our knowledge. A kidney transplant recipient recently treated for acute cellular rejection presented with skin papules overlying a tattoo. Diagnosis was confirmed on culture, histology, and 18S ribosomal RNA polymerase chain reaction. The morphological features on culture characteristic of P. lilacinum included violet colonies on malt extract agar, long tapering brush-like phialides, and elliptical conidia attached in chains. P. lilacinum has intrinsic resistance to many antifungal agents including amphotericin B, but voriconazole and posaconazole have good in vitro activity. The patient was treated with voriconazole with subsequent resolution of the papules after 3 months of therapy.

White mold on cultivated morels caused by Paecilomyces penicillatus.

Morchella (morel) includes prized edible and medical mushrooms in the world. Since 2012, commercial cultivation of morels in the field has developed rapidly in China. However, coupled with the rapid expansion of morel cultivation, diseases have been become serious threats to morel production. White mold is one of the most serious diseases on cultivated morels. This study aimed to confirm this pathogen by following Koch's postulates, and to identify it using molecular evidence. Our results indicated that healthy Morchella fruiting bodies inoculated with Paecilomyces sp. isolates produced typical white mold symptoms, and the internal transcribed spacer sequences of the Paecilomyces sp. were 99% similar to that recovered from an epitype of Paecilomyces penicillatus. Therefore, P. penicillatus was considered to be the causative agent of white mold. White mold occurred from the initial harvest to the storage and preservation process, and it produced white mold-like symptoms on the caps and stripes of Morchella. This is the first time that white mold has been reported on cultivated Morchella.

[HYPEREOSINOPHILIC SYNDROME ASSOCIATED WITH SEPSIS DUE TO PAECILOMYCES FUNGI DISSEMINATED INTO THE LIVER].

A 49 year old woman with signs of chronic sepsis, hepatomegaly, and high eosinophil count was under long-term examination including consultations with an oncologist, parasitologist, and hematologist, diagnostic laparotomy, and studies of liver biopsies. Seeding blood samples onto Saburo's medium resulted in the growth of Paecilomyces variotii Bainier colonies. Counting mature spherules of the fungus revealed 59000 spherules per 1 mcl compared with the normal value of 1000-6000 which suggested paecilomycotic etiology of sepsis. The histological study of liver biopsies demonstrated hemorrhagic foci and eosinophilic infiltrates around fungal spherules. The clinical recovery of the patient was achieved after 3 courses of pulsed terbinafine therapy (500 mg/d every other day for 14 days during a month) in combination with vitamins and i/v infusion of 100 ml of a fluconazole solution (2 mg/ml) every third day (10 procedures during a course of therapy).

Isolation and Classification of Fungal Whitefly Entomopathogens from Soils of Qinghai-Tibet Plateau and Gansu Corridor in China.

Qinghai-Tibet Plateau and Gansu Corridor of China with distinct geographic and climatic conditions are remote and less disturbed by humans, in which are likely to find some new strains of fungal entomopathogens against B-biotype whiteflies that is a very important invading pest worldwide. In this research, nineteen strains among six species of entomogenous fungi were isolated from the soil samples collected from 32 locations in Qinghai-Tibet Plateau and Gansu Corridor. From the data of isolation rates, it was indicated that the good biodiversity of entomogenous fungi was found in the soil covered good vegetations. On the contrary, no strains were isolated from the desert areas. In addition, the dominant species, Isaria fumosorosea and Metarhizium anisopliae var. anisopliae in the Qinghai-Tibet Plateau are different from the strains of other places based on ITS genetic homology analysis. It was verified that the Qinghai-Tibet Plateau area was less disturbed by human, and the fungi in this place exchanged less compared with other regional species. All of these strains showed the pathogenicity against the B-biotype whitefly with the mortality of more than 30%. However, a few strains of Paecilomyces lilacinus, Lecanicillium psalliotae, Aspergillus ustus, I. fumosorosea and M. anisopliae var. anisopliae had better virulence with LC50s of 0.36-26.44×10(6) spores/mL on post-treatment day 6-7. Especially, the L. psalliotae strain LpTS01 was the greatest virulence with LC50 of 0.36×10(6)spores/mL and LT50 of 4.23d. Our research thus presents some new insights to discover new entomopathogenic fungal strains used for B-biotype whitefly biocontrol.

Purpureocillium lilacinum, potential agent for biological control of the leaf-cutting ant Acromyrmex lundii.

Many leaf-cutter ant species are well known pests in Latin America, including species of the genera Acromyrmex and Atta. An environmentally friendly strategy to reduce the number of leafcutter ants and avoid indiscriminate use of chemical pesticides is biological control. In this work we evaluated the effectiveness of a strain of the entomopathogen Purpureocillium lilacinum, against worker ants from six Acromyrmex lundii field colonies, after immersions in pure suspensions at a concentration of 1×10(6)conidiaml(-1). Survival of ants treated with P. lilacinum was significantly lower than that recorded in controls, and median lethal time (LT50) was 6-7days. P. lilacinum was responsible for 85.6% (80.6-89.7) of the mortality in inoculated ants, in which we found that the percentage of other entomopathogens that naturally infected ants decreased also, suggesting a good competitive capability of the fungus. Horizontal transmission to non-inoculated ants was also evidenced, given that 58.5% (41.9-64.2) of them died because of P. lilacinum. Moreover, we tested pathogenicity for three concentrations of this strain (1.0×10(4), 10(6) and 10(8)conidiaml(-1)) and found a significantly faster mortality of ants and greater median percentage of infection at 10(8)conidiaml(-1) of P. lilacinum. CL50 value was 2.8×10(5)conidiaml(-1). We thus propose the use of P. lilacinum as a biological control agent of leafcutter ants in crops and plantations.

The role of a phospholipase (PLD) in virulence of Purpureocillium lilacinum (Paecilomyces lilacinum).

Phospholipases are key enzymes in pathogenic fungi that cleave host phospholipids, resulting in membrane destabilization and host cell penetration. However, understanding the role of phospholipases on the virulence of the filamentous fungus Purpureocillium lilacinum has been still rather limited. In this study, pld gene was characterized. It encodes the protein phospholipase D (PLD) in P. lilacinum. This gene, 3303 bp open reading frame fragment (ORF), encodes a protein of 1100 amino acids with high similarity to the same gene from Penicillium oxalicum and Aspergillus fumigatus. Secondary structure prediction showed two PLD phosphodiesterase domains (437-464 bp and 885-912 bp). The pld gene was significantly regulated during infection of Meloidogyne incognita eggs by P. lilacinum. The expression of pld gene using RT-PCR was the highest at 36 and 48 h, which introduce evidence that the presence of M. incognita may induce the expression of the pld gene in P. lilacinum. In addition, maltose and l-alanine were found to increase the expression of pld gene. An acidic environment (pH 3.0-4.0) and moderate temperatures (27-29 °C) are favorable for pld expression in P. lilacinum.

[Eye infections caused by Purpureocillium lilacinum: A case report and literature review]. / Infecciones oculares por Purpureocillium lilacinum: presentación de un caso y revisión de la literatura.

BACKGROUND: Purpureocillium lilacinum eye infections (previously called Paecilomyces lilacinus) make up a significant percentage of the recorded cases of infection by this fungus, and is considered as an emerging pathogen. AIMS: To report a case of ocular mycosis in a patient aged 70, with a double corneal transplantation in the right eye, and exhibiting a poor response to antifungal and surgical treatment. METHODS: Corneal ring and ocular tissues obtained by surgical procedures were cultured in common mycological media. Molecular identification of the isolated fungus was obtained. RESULTS: Colonies of a filamentous fungus were obtained, and according to the macroscopic and microscopic morphology it was identified as P. lilacinum. The identification was confirmed by molecular methods in a reference laboratory. CONCLUSIONS: Eye infections due to P. lilacinum are rare but serious diseases that requires rapid diagnostic and therapeutic measures to enable visual function to recover.

The entomopathogenic fungal endophytes Purpureocillium lilacinum (formerly Paecilomyces lilacinus) and Beauveria bassiana negatively affect cotton aphid reproduction under both greenhouse and field conditions.

The effects of two entomopathogenic fungal endophytes, Beauveria bassiana and Purpureocillium lilacinum (formerly Paecilomyces lilacinus), were assessed on the reproduction of cotton aphid, Aphis gossypii Glover (Homoptera:Aphididae), through in planta feeding trials. In replicate greenhouse and field trials, cotton plants (Gossypium hirsutum) were inoculated as seed treatments with two concentrations of B. bassiana or P. lilacinum conidia. Positive colonization of cotton by the endophytes was confirmed through potato dextrose agar (PDA) media plating and PCR analysis. Inoculation and colonization of cotton by either B. bassiana or P. lilacinum negatively affected aphid reproduction over periods of seven and 14 days in a series of greenhouse trials. Field trials were conducted in the summers of 2012 and 2013 in which cotton plants inoculated as seed treatments with B. bassiana and P. lilacinum were exposed to cotton aphids for 14 days. There was a significant overall effect of endophyte treatment on the number of cotton aphids per plant. Plants inoculated with B. bassiana had significantly lower numbers of aphids across both years. The number of aphids on plants inoculated with P. lilacinum exhibited a similar, but non-significant, reduction in numbers relative to control plants. We also tested the pathogenicity of both P. lilacinum and B. bassiana strains used in the experiments against cotton aphids in a survival experiment where 60% and 57% of treated aphids, respectively, died from infection over seven days versus 10% mortality among control insects. Our results demonstrate (i) the successful establishment of P. lilacinum and B. bassiana as endophytes in cotton via seed inoculation, (ii) subsequent negative effects of the presence of both target endophytes on cotton aphid reproduction using whole plant assays, and (iii) that the P. lilacinum strain used is both endophytic and pathogenic to cotton aphids. Our results illustrate the potential of using these endophytes for the biological control of aphids and other herbivores under greenhouse and field conditions.

Role of Karanja deoiled cake based medium in production of protease and fatty acids by Paecilomyces lilacinus 6029.

Protease and fatty acids are known to serve as pathogenic factors against root-knot nematodes. Here, we utilized Karanja deoiled cake as a nitrogen source for the first time in induction of protease by Paecilomyces lilacinus. Fatty acids, especially butyric acid, have also been detected in the same fungal culture filtrate.

[The epizootology and epidemiology of echinococcosis and paecilomycosis].

Fungi of the genus Paecilomyces, a causative agent of the fungal disease paecilomycosis, have been found in the blood of productive animals (sheep, cattle, pigs, goats) that are carriers of Echinococcus larvocysts. In the Republic of Uzbekistan, the echinococcosis affection of sheep, cattle, pigs, and goats continues to remain high. Domestic fowls (chickens, turkeys, and gooses) have been established to be blood carriers of Paecilomyces fungi, which is also true for wild birds, such as my-lady's-belts, quails, chukars, and crows. The degree of fungal affection has been also established for birds.

[Study of the pathogenesis of paecilomycosis-complicated echinococcosis].

Experiments were carried out on 35 lambs infected with echinococcus eggs taken from canine donors, 5 lambs with echinococcosis, 5 lambs with echinococcosis + paecilomycosis, 5 lambs with coenurosis, 5 with cysticercosis ovis, 5 with alveococcosis, 5 with paecilomycosis, inoculated by the material taken from patients; and 5 healthy lambs formed a control group. Cystic hydatidosis was found to induce substantial cardiovascular changes.

[The agent Latrodectus and canine paecilomycotic eclampsia as a laboratory model in the survey of treatment for mycoses and parasitic diseases].

The extragent used to prepare a Latrodectus mactans hydrocarbon extract is a multicomponent system composed of alkanes, alkenes, and arenes. More than 100 compounds were identified in the hydrocarbon extract (petroleum). The petroleum matrix of Latrodectus mactans was first obtained to manufacture homeopathic remedies. The authors could prepare the first Russian homeopathic medicine from Latrodectus mactans, which proved to be effective in treating canine eclampsia. Canine experiments provide a rationale for the authors' choice as the only homeopathic remedy among thousand known drugs to treat female eclampsia. It is Latrodectus mactans that is in the list of homeopathic medicines permitted for use in accordance with Order No. 335 (Supplement 2) of the Ministry of Health and Medical Industry of Russia, issued on November 29, 1995. It is manufactured from Latrodectus mactans living in the USA.

Occurrence of pathogenic fungi to Amblyomma cajennense in a rural area of Central Brazil and their activities against vectors of Rocky Mountain spotted fever.

Two isolates of Beauveria bassiana and one of Purpureocillium lilacinum (=Paecilomyces lilacinus) were found infecting Amblyomma cajennense engorged females collected on horses (0.15% infection rate from a total of 1982 specimens) and another two isolates of P. lilacinum and one Metarhizium anisopliae detected in soils (2.1% from 144 samples) collected in typical pasture habitats of this tick in Central Brazil from October 2009 to March 2011. Fungi were isolated from soils with Rhipicephalus sanguineus as surrogate baits. No fungi were found in ticks or soils during the driest months (May to August). Testing pathogenicity of fungi all R. sanguineus females were killed regardless of the isolate and fungi sporulated abundantly on the cadavers. A. cajennense was less susceptible to infection with P. lilacinum within 20 days than R. sanguineus. All three fungal species probably act as natural antagonists of A. cajennense particularly in the rainy season and have interest for integrate control of vectors of Rocky Mountain spotted fever.

Laboratory evaluation of Isaria fumosorosea against Spodoptera littoralis.

Isaria fumosorosea (syn. Paecilomyces fumosoroseus) is potentially useful for the biological control of economically important agricultural and forest insect pests. We evaluated efficacy of two strains of this entomopathogenic fungus against last instar larvae and pupae of Egyptian cotton leafworm, Spodoptera littoralis. The first strain was Apopka 97 which is an active ingredient of commercial biopesticide PreFeRal WG (Biobest, Belgium). The second strain was isolated from Cameraria ohridella and is currently deposited under number CCM 8367 as a patent culture in the Czech Collection of Microorganisms in Brno. Blastospores of both strains were obtained after 120 hours submerged cultivation in grow media using orbital shaker. The concentration of blastospores was adjusted to 5 x 10(7) spores/ml of suspension. Soaking agent Tween 80 was added to the suspension at concentration 0.02%. Lethal effects of both fungal strains on S. littoralis were evaluated using standard dip test. Treated insects were individually placed into plastic Petri dishes (diam. 9 cm) and kept at constant laboratory conditions (temperature 23 degrees C, R.H. approx. 100%, 16L:8D photoperiod). Virulence of the strains was expressed as percentages of cumulative daily mortality corrected for mortality in the control variant. Obtained results revealed higher virulence of CCM 8367 blastospores to the last instar larvae of S. littoralis (93.1% mortality) on the 7th day after the treatment compared to Apopka 97 (65.5% mortality). Even more obvious difference was found in pupae, where corrected mortality of CCM 8367-treated pupae was 80.0% while mortality in Apopka 97-treated pupae reached only 3.3% on the 8th day after the treatment. We can conclude that the strain I. fumosorosea CCM 8367 has strong insecticidal effects on S. littoralis and has a potential to be implemented as a novel biocontrol agent.