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The article presents data on the detection of mould fungi in the system of Aesculus hippocastanum – Cameraria ohridella which is an invasive pest of chestnuts. Entomological and mycological studies were conducted using standard methods . The study determined the species composition, occurrence index and quantitative indicators of the micromycetes present in the leaf tissues of Aesculus hippocastanum and Cameraria ohridella caterpillars. 135 samples of the caterpillars of three generations of the insects, the mines and the healthy plant leaves collected at the permanent observation points in Saratov city (Astrakhanskaya St., Bolshaya Sadovaya St., Naberezhnaya Kosmonavtov St.) were analysed. 18 species of the fungi of the genera Alternaria, Aspergillus, Cladosporium, Fusarium, Penicillium and Rhizopus were identifi ed. Numerical indicators of fungi ranged from 102 to 105 CFU/g. There was an increase in the quantitative indicators and species diversity of micromycetes during the vegetation period. Aspergillus was the dominant genus of fungi in the fi rst generation of insects, Alternaria dominated in the second generation, and Cladosporium dominated in the third generation.Thus, Ochrid myner can be a natural reservoir of various fungi, including phytopathogenic ones.

Lactulose is a prebiotic that has found a wide application in medicine and food industry. Commercial lactulose is usually synthesized by isomerization in alkaline media at high temperatures. Enzymatic methods offer a more sustainable alternative and require more moderate processing conditions. This review covers 44 years of scientific publications (1978–2022) on the enzymatic synthesis and purification of lactulose. The materials were retrieved from Scopus, Web of Science, PubMed, and Elibrary databases. The enzymatic approach to lactose-to-lactulose conversion has two methods: isomerization (direct) and transgalactosylation (via hydrolysis). Isomerization exploits cellulose-2-epimerases, but their safety status is still rather vague. As a result, cellulose-2-epimerases are not commercial. Epilactose is a by-product of isomerization. Transgalactosylation involves β-galactosidases with an official international safety status (GRAS). It is available on the market, and its action mechanism is well understood. This article systematizes various data on the conditions for obtaining the maximal yields of lactulose by different enzymes. The Kluyveromyces lactis yeast and the Aspergillus oryzae mold are the main sources of β-galactosidases in lactulose production. The yield can reach 30% if the processing conditions are optimal. Fructose remains the main problem in the production process. No scientific publications revealed a direct relationship between the maximal yields of lactulose and the molar fructose-tolactose ratios. Cellobiose epimerases make it possible to achieve high yields of lactulose (70–80%). However, these enzymes are associated with genetic engineering and mutagenesis, which challenges their safety status. The most promising trends in lactulose biotechnology include secondary dairy raw materials, immobilized enzymes, membrane reactors, complex production processes, lactose-to-lactulose conversion, and purification of final product.

Aspergiloza pulmonară asociată COVID-19 (APAC) a fost raportată recent ca și o potențială complicație infecțioasă,care afectează pacienții critici cu infecție SARS-CoV-2. Având în vedere numărul mare de cazuri și rata ridicată a letalitățiiraportate în literatura de specialitate, este argumentată necesitatea mai multor date și studii referitoare la interacțiunea dintreSARS-CoV-2 și Aspergillus spp., precum și a celor de evaluare a schemelor terapeutice administrate și a eficacității acestora.Prezentăm cazul unui pacient diagnosticat cu o formă severă de APAC, fiind printre puținele cazuri publicate ale pacienților care au supraviețuit, cu o monitorizare a evoluției leziunilor pulmonare pentru o perioadă de 6 luni.

Infecţia cu noul coronavirus SARS-CoV-2 rămâne una dintre cele mai importante pandemii până în prezent, devenind o ecuaţie cu mai multe necunoscute, una dintre necunoscute fiind şi infecţiile fungice asociate. Micozele pulmonare au o incidenţă şi mortalitate sporită, cel mai frecvent germene identificat printre cazurile de COVID-19 fiind Aspergillus spp. Datorită discordanţelor în definiţie şi în criteriile diagnostice, dar şi datorită caracterului nespecific al manifestărilor clinico-imagistice, aspergiloza pulmonară asociată COVID-19 rămâne o provocare în plan diagnostic şi de conduită

Introduction. In children with rheumatic diseases, severe fungal infections (invasive mycoses – IM) are not well understood.Objectives. To analyze risk factors, disease course of IM in children with systemic rheumatic diseases.Materials and methods. For diagnosis of IM were used criteria EORTC/MSGERC, 2019. We reviewed the literature over the past 15 years on IM in children with rheumatic diseases from the international databases Pubmed and Web of Science.Results. In retrospective multicenter study were included 8 children with IM and systemic rheumatic diseases: ANCA-associated vasculitis (n=4), systemic lupus erythematosus (n=3), juvenile rheumatoid arthritis (n=1). Median age was 13,5 (8-17) y., boys – 67%. Invasive aspergillosis was diagnosed in 5 patients and invasive candidiasis – 3. The risk factors of invasive mycoses were high rheumatic disease activity (100%), corticosteroids (prednisolone ≥ 0,3 mg/kg/d) use for ≥21 d (87,5%), immunosuppressive therapy (87,5%), recent (≤ 2 weeks) pulse steroid therapy (75%), hemophagocytic lymphohistiocytosis (62,5%), prolonged (≥ 10 days) severe neutropenia (≤ 0,5х109/l) (62,5%), and prolonged (≥10 days lymphopenia (≤ 1,0х109/l) (37,5%). In patients with invasive aspergillosis the involved organ was the lung, in patients with invasive candidiasis a candidemia was diagnoses. All patients received antifungal therapy. The overall 30 days survival rate was 37,5%.Сonclusions. Children with high rheumatic diseases activity and intensive treatment with immunosuppressive agents should be considered as patients with a high risk of invasive mycoses with a high mortality.

Objective: to study risk factors for invasive aspergillosis (IA), its etiology, clinical manifestations, and treatment efficiency in patients with rheumatic diseases (RD).Patients and methods. The first study of proven and probable IA (EORT/MSGERC, 2019) was conducted in 18 patients with RD, who accounted for 3% of all adult IA patients (n=699) included in the 1998–2020 registry of the Department of Clinical Mycology, Allergology, and Immunology, I.I. Mechnikov North-Western State Medical University (Group 1). This group comprised 56% women; the median age was 59 [21; 75] years. Group 2 (a comparison group) included 610 adult hematology patients with IA (median age, 45 [18; 79] years; 42% women). A prospective case-control study was conducted to identify risk factors for IA in patients with RD: 36 rheumatic patients without IA (median age, 58 (18–79) years; 61% women) (a control group).Results and discussion. Patients with RD were found to often develop IA in the presence of anti-neutrophilic cytoplasmic antibody-associated vasculitis (granulomatosis with polyangiitis and microscopic polyangiitis) and systemic lupus erythematosus (50 and 16%, respectively). It was shown for the first time that the likelihood of IA in patients with RD increases with prolonged (median 14 days) lymphocytopenia during RD treatment (odds ratio 13.0; 95% confidence interval, 3.3–50.3). The main causative agents of IA were A. fumigatus (50%) and A. niger (29%). IA was more severe in Group 1 than in Group 2: in the resuscitation and intensive care units, there were 44 and 18%, respectively (p=0.01). Group 1 versus Group 2 more frequently had respiratory failure (61 and 37%, respectively; p=0.03), hemoptysis (28 and 7%; p=0.0001), chest pain (17 and 7%; p=0.04), and cardiac involvement (11 and 1%; p=0.0001), and less frequently had fever (67 and 85%; p=0.01). The common site of IA was the lung (83%); the characteristic feature detected by computed tomography (CT) is pulmonary cavitation (44%). Antifungal therapy was used in 89% of Group 1 patients; the overall 12-week survival was 69%.Conclusion. In patients with RD, it is difficult to differentiate between the progression of the underlying disease, adverse drug reactions, infectious complications, or a combination of these disorders due to the similarity of their clinical manifestations. When RD patients with infectious syndrome and respiratory failure develop prolonged lymphocytopenia during combination therapy, AI should be suspected and lung CT, bronchoscopy, and mycological examination of the material obtained by bronchoalveolar lavage be done.

Aspergillus is a saprophytic mold and its natural habitat is the soil. It is found worldwide indoors and outdoors in potted soil, compost, freshly cut grasses, decaying vegetation and in sewers. Aspergillus produces a bountiful number of spores and releases 2-3 micron sized spores into the air daily. It grows best at 37-40 °C, which is similar to the temperature in the lungs. These spores will remain airborne for a long period of time. It is estimated that humans inhale hundreds of spores daily. Several fungi other than aspergillus have been known to be implicated. Hence, the term allergic bronchopulmonary mycoses would be more appropriate unless the specific fungus is identified - which could be candida, helminthosporium, curvularia, bipolaris, cladosporium, or others. The review article is focused on the prototype allergic bronchopulmonary aspergillosis, its epidemiology, pathogenesis, diagnosis and treatment. Bronchopulmonary aspergillosis should be considered in patients with poorly controlled asthma despite appropriate routine therapy and environmental control. The need for frequent courses of corticosteroids with temporary improvement should raise the index of suspicion and appropriate evaluation be done. Early recognition and prompt initiation of appropriate corticosteroid treatment regimen would reduce the risk of development or progression of bronchiectasis and lung tissue damage. Regular follow up and monitoring serum total IgE level can predict exacerbations and should prompt corticosteroid treatment. Long term follow-up is important as relapses can occur years of remission.

The aim: to assess the incidence of various forms of pulmonary aspergillosis in patients with cystic fibrosis.Materials and methods. In 2014-2017 yy. in prospective study in different regions of Russia were included 190 patients with cystic fibrosis aged 1 to 37 years. Children – 130, adults – 60. All patients underwent allergy (skin tests with fungal allergens, total IgE level, specific IgE to fungal allergens) and mycology (microscopy and cultural investigations of respiratory biomaterials) testing. Chest computed tomography was performed according to the indications.Results. The incidence of fungal sensitization in patients with cystic fibrosis was 57%, to Aspergillus spp. – 27%. The incidence of allergic bronchopulmonary aspergillosis was 5,7%, chronic lung aspergillosis – 4,2%, invasive aspergillosis developed in one patient (0,5%) during immunosuppressive therapy after liver transplantation.Conclusion. The incidence of pulmonary aspergillosis in patients with cystic fibrosis in the Russian Federation was first determined (10.5%). Mycology testing is indicated for patients with cystic fibrosis for early treatment of different variants of pulmonary aspergillosis.

In the retrospective multicenter study during 2007–2017 we included 59 oncohematological patients with mucormycosis and 541 patients with invasive aspergillosis. Our study showed that mucomorhycosis more often developed in children and adolescents (p = 0.001), and after «graft versus host» disease development (p = 0.0001). Patients with mucormycosis were more immunosuppressed: severe neutropenia was in 88 % vs. 82 %, median duration of neutropenia ‒ 30 days vs. 14 days, p = 0.0001, lymphocytopenia – 77 % vs. 65 %, median duration of lymphocytopenia – 25 days vs. 14 days, p = 0.001. The main sites of infection were lungs, nevertheless in patients with mucormycosis it was less frequent (73 % vs. 97 %, p = 0.02), but more frequent were ≥2 organs involvement (42 % vs. 8 %, p = 0.001) and paranasal sinuses involvement (15 % vs. 6 %, p = 0.04). Typical clinical features of mucomorhycosis were localized pain syndrome (53 % vs. 5 %, p = 0.0001), hemoptysis (32 % vs. 6 %, p = 0.001), on lung computed tomography scan – pleural effusion (53 % vs. 7 %, p = 0.003), lesions with destruction (38 % vs. 8 %, p = 0.0001) and “a reverse halo” symptom (17 % vs. 3 %). The overall 12-week survival was significantly lower in patients with mucormycosis (49 % vs. 81 %, p = 0.0001). In both groups unfavorable prognosis factors were: ≥2 organs involvement (p = 0.0009) and concomitant bacterial or viral infection (p = 0.001 and p = 0.008 respectively). In mucormycosis patients favorable prognosis factor was remission of underlying disease (p = 0.006), in invasive aspergillosis patients – early bronchoscopy (p = 0.003), voriconazole use (p = 0.0007) and secondary antifungal prophylaxis (p = 0.0001).

Objective. Data analysis of the register of patients with invasive aspergillosis (IA), which was founded in Saint Petersburg (1998–2017), and clinical case description of successful treatment of IA and mucormycosis with lungs involvement in a patient with Hodgkin’s lymphoma.Materials and methods. In the study were included 29 oncohematological patients with IA and mucormycosis. In control group were included 483 oncohematological patients with IA. We used criteria EORTS/MSG, 2008 for IA and mucormycosis diagnosis.Results. We identified that the combination of IA and mucormycosis significantly often develops in patients with acute lymphoblastic leukemia (32 %, р = 0.001), and allogeneic hematopoietic stem cells transplants (allo-HSCT) recipients (52 %, р = 0.001). In mixed-infection Aspergillus nidulans was frequent IA etiological agent (11 %, р = 0.001). The main mucormycosis etiological agents were Rhizopus spp. (45 %), Lichtheimia corуmbifera (20 %). The main sites of the localization were lungs (76 %), disseminated process and paranasal sinuses involvement were identified more frequently (45 % and 17 % (р = 0.0001; р = 0.002), respectively). Typical clinical feature of IA and mucormycosis combinations was hemoptysis (24 %, р = 0.008), radiological signs – lesions with cavities destruction (38 %), hydrothorax (29 %) and a “reverse halo” symptom (17 %). Antifungal therapy received 76 % of patients, surgery – 34 %.Conclusion. Mucormycosis was revealed in 5.7 % of patients with IA. The main risk factors for co-infection are allo-HSCT, long-term agranulocytosis, lymphocytopenia and glucocorticosteroid therapy. Overall 12 weeks survival in patients with mixed-infection was 38 %, significantly lower than in patients with IA (р = 0.005). An unfavorable prognosis factor was dissemination of mycotic infection (р = 0.009).

We present the results of a multicenter study of 445 patients with “proven” and “probable” invasive aspergillosis (EORTC/MSG, 2008). Invasive aspergillosis usually occurs in patients with hematological malignancies (88 %), main underlying diseases were acute myeloid and acute lymphoblastic leukemia. The risk factors: prolonged agranulocytosis (64 %), cytostatic chemotherapy (57 %), corticosteroid treatment (45 %), and allogeneic hematopoietic stem cells transplantation (29 %). The pathogens – A. fumigatus (42 %), A. niger (33 %), and A. flavus (21 %). The main site of infection were lungs (86 %). 12 week overall survival was 83 %. Bronchoscopy use for the early diagnosis (p = 0.01), adequatetherapy with voriconazole (p = 0.002) and secondary antifungal prophylaxis (p = 0.0003) were positive prognostic factors for survival of patients with invasive aspergillosis.

Exchanges of morphology, speed of radial growth, and accommodation of inorganic polyphosphates in cells of microscopic fungi of the genera Alternaria, Aspergillus, Cladosporium and Penicillum in condition of carbon limitation are characterized. Results obtained confirm existence of various ways of realization of adoptive responses of microscopic fungi to this stress factor. [ABSTRACT FROM AUTHOR]

The analysis of literature and water data on the presence of microbiological safety indicators of micromycetes was conducted, that are able to exist and reproduce in tap water entering the consumer. Microscopic fungi (Penicillium, Rhodotorula, Fusarium, Cladosporium, Aspergillus) in the amount of 10 to 303,3 CFU/dm3 were detected in the samples of tap drinking water studied. At the same time, tap water in 50.0% of cases is contaminated with pathogenic micromycetes, the structure of which is represented by: Penicillium spp. (40.0%), Aspergillus (28.75%), Rhodotorula spp. (17.5%), Fusarium (10.0%), Cladosporium spp. (3.75%). In terms of quantitative traits, micromycetes of the genus Rhodotorula dominated in drinking water - 150.0±23.3 CFU/dm3, Penicillium, on average, was detected in the amount of 87.0±7.7 CFU/m3, Fusarium - 60.0±6.4 CFU/dm3, Aspergillus - 22.0±4.9 CFU/dm3, and Cladosporium - 10.0±3.3 CFU/dm3. The presence of micromycetes in water was monitored depending on the season of the year. A direct strong correlation was established between the average monthly air temperature and the total frequency of detection in drinking tap water of all types of micromycetes. The results indicate that the water samples do not meet the requirements of DSTU 4808-2007 for the presence of micromycetes. At the same time, according to the requirements of DSanPiN 2.2.4-171-10 and Council Directive 98/83/EC, microscopic fungi are not regulated at all in tap water. Therefore, we propose to revise the standards, namely, to include micromycetes in the indicators of microbiological safety, which will make it possible to improve water quality control.

The introduction of methods for food production using microbial synthesis, including those obtained with the help of genetically modified (GM) microorganisms, at the present stage, allows to increase production volumes and reduce the cost of food. At the same time, such products in accordance with TR CU 021/2011 "On food safety" are classified as a "novel food"» and can be placed on the market only after its risk estimation for health. The emergence of new data and research methods in the last years has made it necessary to improve the risk assessment system for this category of food. The aim of the research is to develope risk assessment approaches of food obtained by microbial synthesis on the example of the GM strain Aspergillus awamori Xyl T-15 and the enzyme preparation (EP) (a complex of glucoamylase and xylanase) produced by it. Material and methods . Outbred ICR mice (CD-1) and Wistar rats (males and females) were used in the experimental studies. Investigations of GM strain A. awamori Xyl T-15 virulence and its ability to disseminate internal organs have been carried out. Acute and subacute (during 80 days) toxicity of EP (a complex of glucoamylase and xylanase) have been studied. Results . The presented experimental data allow us to make a conclusion about the avirulence of the A. awamori Xyl T-15 strain, the lack of ability to disseminate internal organs (invasiveness). At the same time, the strain is characterized by the ability to produce mycotoxins (ochratoxin, fumonisin B2, T-2 and HT-2 toxins). The EP, a complex of glucoamylase and xylanase from A. awamori Xyl T-15, has a low oral acute toxicity for rats (LD 50 >5000 mg/kg). I ntragastric EP administration at doses of 10, 100 and 1000 mg/kg of body weight during 80 days had not revealed adversely affect on the rate of weight gain in animals, indicators of anxiety and cognitive function, and some studied biochemical indicators. At a dose of 100 mg/kg b.w. or more, there were changes in the relative mass of organs (lungs, kidneys, adrenal glands), small shifts in the parameters of erythropoiesis and leukocyte formula, at a dose of 1000 mg/kg b.w. - an increase in oxidative DNA destruction. T he most pronounced and dose-dependent was the effect of the EP on hepatocyte apoptosis. According to this indicator, the not observed adverse effect level (NOAEL) for EP is not more than 100 mg/kg b.w. in terms of protein. The main target organ for the toxic effect of EP is the liver. Conclusion . The data obtained demonstrate the necessity to conduct an additional analysis of the risks of possible negative effects of EP, namely, to study its impact on the gut microbiocenosis and the immune status of experimental animals, to analyze the presence of determinants of pathogenicity and antibiotic resistance, DNA of selective marker genes of A. awamori Xyl T-15 strain by PCR analysis and DNA sequencing methods., Competing Interests: The authors declare no overt and potential conflict of interest related to the publication of this article., (Copyright© GEOTAR-Media Publishing Group.)

The mycelial Fungi Penicillium funiculosum, P. citrinum, P. expansum, P. chrysogenum, Aspergillus ochraceus, A. alliaceus, A. luchaensis, A. flavus, and A. niger were isolated from enrichment cultures. These fungi actively destruct carbon deposits formed during exploitation of aircraft. A biotechnological method for removing fouling from parts of aircraft engines (PAE) was developed. This method is less laborious, more rapid and ecologically clean than contemporary chemical methods. Scanning microscopy was suggested to use for estimating the degree of decarbonization of PAE surfaces.

Results of studies of certain fungal extracellular ribonucleases mainly isolated from representatives of the genera Aspergillus and Penicillium are summarized. The isolation of these enzymes in highly purified states in the 1970-1980s strongly stimulated further studies of their structure, functions, and mechanisms of action. This also promoted the use of ribonucleases as catalysts in oligoribonucleotide syntheses and as objects of comparative and evolutionary biochemistry and other research works. Results of studies of the primary, secondary, and spatial structures of guanyl-specific fungal ribonucleases are reviewed. These studies revealed a high homology within the subfamilies of fungal, bacterial, and actinomycete RNases. Characteristics of the nonspecific Pb2 RNase are considered.

The immobilization of inulinase by ionexchange AB-26 and AB-17-2P has been made by adsorbtion and glutaraldehyde methods. The effect of UV-radiation, carbamide and gamma-rays on the stability of native and immobilized enzyme has been investigated. The stability of inulinase in relation to denaturation agents has been shown to increase with the immobilization of ionexchange. The character of binding with the matrix affects greatly the stability of immobilized enzyme to physical factors.

A method is suggested to determine toxigenicity of microscopic fungi from genera Aspergillus, Penicillium, Fusarium, etc. which contaminate fodder, food products and environment objects. The method is based on the property of micromycetes isolated from fodder, food products and environment objects to produce mycotoxins under optimal conditions of their cultivation on the nutrient media. Their further extraction by the organic solvent, evaporation and emulsification of the extract and its peroral introduction to young female rats cause a disease and death of animals. The suggested method may be used for recognizing and preventing mycotoxicoses of farm animals and poultry.

An analysis of 813 specimens of agricultural crops and 155 specimens of good quality products from retail trading establishments demonstrated that more than 50 per cent of them were contaminated with molds of the A. flavus species. From among 489 strains isolated the toxigenic ones comprised 5.1 per cent and according to their morphological characteristics they were referred to molds of the A. flavus and A. fumigatus and Penicillium species. Toxigenic strains were isolated only in specimens of agricultural crops, none of them having been found in the products coming from the retail trade establishments. In the grain itself and in the food products none of the fluorescent components characteristics of aflatoxins were isolated.