Genetic diversity and population structure of Penicillium roqueforti isolates from Turkish blue cheeses.

Blue cheeses, including renowned mold-ripened varieties such as Roquefort (France), Gorgonzola (Italy), Stilton (UK), Danablue (Denmark), and Cabrales (Spain), owe their distinct blue-green color and unique flavor to the fungal species Penicillium roqueforti. In Turkey, traditional cheeses similar to blue cheeses, namely mold-ripened Tulum and Civil, employ production techniques distinct from their European counterparts. Notably, mold-ripening in Turkish cheeses is spontaneous and does not involve starter cultures. Despite P. roqueforti being recognized for its distinct genetic populations sourced from various blue cheeses and non-cheese origins globally, the characteristics of the P. roqueforti population within Turkish cheeses remain unexplored. This study aimed to unravel the genetic characteristics and population structure of P. roqueforti from Turkish mold-ripened cheeses. Analysis of mold-ripened Civil (n = 22) and Tulum (n = 8) samples revealed 66 P. roqueforti isolates (76.6 % of total fungal isolates). Subsequently, these isolates (n = 66) and those from previous studies (Tulum n = 53, Golot n = 1) were used to assess genetic characteristics and mating genotypes. All 120 isolates harbored horizontal transfer regions (Wallaby and CheesyTer) and predominantly possessed the MAT1-2 mating genotype, similar to global blue cheese populations. However, most lacked the mpaC deletion associated with such populations. Analysis of the population with three polymorphic microsatellite markers revealed 36 haplotypes (HTs). Some cheeses contained isolates with different HTs or opposite mating genotypes, aligning with spontaneous fungal growth. Tulum and Civil isolates exhibited similar population diversity without forming distinct subgroups. Phylogenetic analysis of 20 selected isolates showed 75 % aligning with global blue cheese isolates, while 25 % formed unique clades. Overall, Turkish P. roqueforti isolates share genetic similarities with global populations but exhibit unique characteristics, suggesting potential new clades deserving further investigation. This research illuminates the characteristics of P. roqueforti isolates from Turkish cheeses, contributing to the knowledge of the global intraspecific diversity of the P. roqueforti species.

Effects of temperature, pH, and relative humidity on the growth of Penicillium paneum OM1 isolated from pears and its patulin production.

Patulin is a mycotoxin produced by several species of Penicillium sp., Aspergillus sp., and Byssochlamys sp. on apples and pears. Most studies have been focused on Penicillium expansum, a common postharvest pathogen, but little is known about the characteristics of Penicillium paneum. In the present study, we evaluated the effects of temperature, pH, and relative humidity (RH) on the growth of P. paneum OM1, which was isolated from pears, and its patulin production. The fungal strain showed the highest growth rate at 25 °C and pH 4.5 on pear puree agar medium (PPAM) under 97 % RH, while it produced the highest amount of patulin at 20 °C and pH 4.5 on PPAM under 97 % RH. Moreover, RT-qPCR analysis of relative expression levels of 5 patulin biosynthetic genes (patA, patE, patK, patL, and patN) in P. paneum OM1 exhibited that the expression of the 4 patulin biosynthetic genes except patL was up-regulated in YES medium (patulin conducive), while it was not in PDB medium (patulin non-conducive). Our data demonstrated that the 3 major environmental parameters had significant impact on the growth of P. paneum OM1 and its patulin production. These results could be exploited to prevent patulin contamination by P. paneum OM1 during pear storage.

Mycobiota contaminating some market cake samples with reference to their toxin and enzyme.

Fungi can spoil the majority of baked products. Spoilage of cake during storage is commonly associated with fungi. Therefore, this study aimed to assess the quality of different types of cakes sold in the market. The most predominant fungal genera in the tested cake samples (14 samples) were Aspergillus spp., and Penicillium spp. On Potato Dextrose Agar (PDA), the medium fungal total count was 43.3 colonies /g. Aspergillus was the most dominant genus and was isolated from six samples of cake. Aspergillus was represented by 3 species namely, A. flavus, A. niger, and A. nidulans, represented by 13.32, 19.99, and 3.33 colonies /g respectively. On Malt Extract Agar (MEA) Medium, the fungal total count was 123.24 colonies / g. Aspergillus was the most dominant isolated genus from 11 samples of cake and was represented by 5 species, namely, A. flavus, A. niger, A. ochraceous, A. terreus, and A. versicolor (26. 65, 63.29, 3.33, 6.66, and 3.33 colonies / g , respectively). Twenty-four isolates (88.88 %) of the total tested twenty-seven filamentous fungi showed positive results for amylase production. Ten isolates (37.03%) of the total tested filamentous fungi showed positive results for lipase production, and finally eleven isolates (40.74 %) of the total fungal isolates showed positive results for protease production. Aflatoxins B1, B2, G1, G2, and ochratoxin A were not detected in fourteen collected samples of cake. In this study, clove oil was the best choice overpeppermint oil and olive oil for preventing mold development when natural agents were compared. It might be due to the presence of a varietyof bioactive chemical compounds in clove oil, whose major bioactive component is eugenol, which acts as an antifungal reagent. Therefore, freshly baked cake should be consumed within afew days to avoid individuals experiencing foodborne illnesses.

Severe pneumonia with pleural effusion caused by co-infection of Paecilomyces variotii and Penicillium oxalicum in a diabetic patient.

BACKGROUND PAECILOMYCES: and Penicillium are considered as rare opportunistic pathogens in immunocompromised hosts, and pneumonia caused by Paecilomyces and Penicillium is rare. In this study, we present first case of severe pneumonia with pleural effusion caused by co-infection of Paecilomyces variotii (P. variotii) and Penicillium oxalicum (P. oxalicum) in a 66-year-old female with poorly controlled type 2 diabetes. CASE PRESENTATION: A 56-year-old woman patient presented to hospital for nausea, poor appetite, and vomiting for one day. On the second day of admission, blood culture and renal puncture fluid culture grew multidrug-resistant Escherichia coli (imipenem/cilastatin sensitive), and she received combination therapy with imipenem/cilastatin (1 g, every 8 h) and vancomycin (0.5 g, every 12 h). On the fourth day, she developed symptoms of respiratory failure. Pulmonary computed tomography (CT) showed an increase in pneumonia compared to before, with minor pleural effusion on both sides. Two fungi were isolated repeatedly from BALF culture, which were confirmed as P. variotii and P. oxalicum by Internal transcribed spacer (ITS) sequencing. Her pleural effusion was completely absorbed, pneumonia symptoms have significantly improved and discharged with receiving liposomal amphotericin B treatment for four weeks. CONCLUSIONS: It is worth noting that clinicians and laboratory personnel should not simply consider Paecilomyces and Penicillium species as contaminants, especially in immunocompromised patients. Early fungal identification and antifungal drug sensitivity are crucial for clinical drug selection and patient prognosis.

Portuguese cork industry: filling the knowledge gap regarding occupational exposure to fungi and related health effects.

Introduction: The presence of the Penicillium section Aspergilloides (formerly known as Penicillium glabrum) in the cork industry involves the risk of respiratory diseases such as suberosis. Methods: The aim of this study was to corroborate the predominant fungi present in this occupational environment by performing a mycological analysis of 360 workers' nasal exudates collected by nasal swabs. Additionally, evaluation of respiratory disorders among the cork workers was also performed by spirometry. Results: Penicillium section Aspergilloides was detected by qPCR in 37 out of the 360 nasal swabs collected from workers' samples. From those, 25 remained negative for Penicillium sp. when using culture-based methods. A significant association was found between ventilatory defects and years of work in the cork industry, with those people working for 10 or more years in this industry having an approximately two-fold increased risk of having ventilatory defects compared to those working less time in this setting. Among the workers who detected the presence of Penicillium section Aspergilloides, those with symptoms presented slightly higher average values of CFU. Discussion: Overall, the results obtained in this study show that working in the cork industry may have adverse effects on worker's respiratory health. Nevertheless, more studies are needed (e.g., using serological assays) to clarify the impact of each risk factor (fungi and dust) on disease etiology.

Antagonistic Activities of Metschnikowia pulcherrima Isolates Against Penicillium expansum on Amasya Apples.

Postharvest fungal diseases cause serious fruit losses and food safety issues worldwide. The trend in preventing food loss and waste has shifted to environmentally friendly and sustainable methods, such as biological control. Penicillium expansum is a common postharvest contaminant fungus that causes blue mould disease and patulin formation on apples. This study aimed to provide biocontrol using Metschnikowia pulcherrima isolates against P. expansum, and to understand their antagonistic action mechanisms. In vitro, 38.77-51.69% of mycelial growth inhibition of P. expansum was achieved by M. pulcherrima isolates with the dual culture assay, while this rate was 69.45-84.89% in the disc diffusion assay. The disease symptoms of P. expansum on wounds were reduced by M. pulcherrima, on Amasya apples. The lesion diameter, 41.84 mm after 12 d of incubation in control, was measured as 24.14 mm when treated with the most effective M. pulcherrima DN-MP in vivo. Although the antagonistic mechanisms of M. pulcherrima isolates were similar, there was a difference between their activities. In general, DN-HS and DN-MP isolates were found to be more effective. In light of all these results, it can be said that M. pulcherrima isolates used in the study have an antagonistic effect against the growth of P. expansum both in vitro and in vivo in Amasya apples, therefore, when the appropriate formulation is provided, they can be used as an alternative biocontrol agent to chemical fungicides in the prevention of postharvest diseases.

The Fungal Diversity and Potential Pathogens Associated with Postharvest Fruit Rot of 'Huangguan' Pear (Pyrus bretschneideri) in Hebei Province, China.

Postharvest fruit rot caused by pathogens is a serious problem in the pear industry. This study investigated the fungal diversity and main pathogens and identified a new pathogen in the stored 'Huangguan' pear (Pyrus bretschneideri Rehd.), the dominant pear variety in northern China. We sampled 20 refrigeration houses from five main producing regions in Hebei Province and used Illumina sequencing technology to detect the fungal composition. Alternaria (56.3%) was the most abundant fungus, followed by Penicillium (9.2%) and Monilinia (6.2%). We also isolated and identified nine strains of Alternaria and four strains of Penicillium. Moreover, we observed a new postharvest fruit disease in 'Huangguan' pear caused by Stemphylium eturmiunum, which was confirmed by phylogenetic analysis by combining the sequences of three conserved genes, including internal transcribed spacer, gapdh, and calmodulin. This study marks the first documentation of S. eturmiunum causing fruit rot in 'Huangguan' pears, offering valuable insights for identifying and controlling this newly identified postharvest disease.

Ochratoxin A Contamination of California Pistachios and Identification of Causal Agents.

Ochratoxin A (OTA) is a potent mycotoxin produced by Aspergillus and Penicillium spp., which contaminates many crops, including pistachios. Pistachios contaminated with OTA may be subjected to border rejections resulting in significant economic losses to the United States agricultural revenues. The current study examined prevalence of OTA in California-grown pistachios and identified its causal agents. OTA was detected in 20% of samples from 2018 to 2021 (n = 809), with 18% of samples exceeding the European Union regulatory limit of 5 µg/kg. Fungi potentially responsible for OTA contamination were isolated from leaves, nuts, and soil collected from 14 pistachio orchards across California. A total of 1,882 isolates of Aspergillus section Nigri and 85 isolates of section Circumdati were recovered. Within section Nigri, 216 (11.5%) isolates were identified as potential OTA producers using a boscalid-resistance assay. Phylogenetic analyses of partial gene sequences for ß-tubulin and calmodulin genes resolved section Circumdati into four species: A. ochraceus (33%), A. melleus (28%), A. bridgeri (21%), and A. westerdijkiae (19%). A. westerdijkiae produced the highest levels of OTA in inoculated pistachios (47 µg/g), followed by A. ochraceus (9.6 µg/g) and A. melleus (3.3 µg/g). A. bridgeri did not produce OTA. OTA production by section Circumdati was optimal from 20 to 30°C. All 216 boscalid-resistant isolates from section Nigri were identified as A. tubingensis, and representative isolates (n = 130) produced 3.8 µg/kg OTA in inoculated pistachios. This is the first detailed report on OTA contamination and causal fungi in California pistachios and will be helpful in devising effective management strategies.

Wide distribution of resistance to the fungicides fludioxonil and iprodione in Penicillium species.

Fludioxonil and iprodione are effective fungicides widely used for crop protection and are essential for controlling plant pathogenic fungi. The emergence of fungicide-resistant strains of targeted pathogens is regularly monitored, and several cases have been reported. Non-targeted fungi may also be exposed to the fungicide residues in agricultural fields. However, there are no comprehensive reports on fungicide-resistant strains of non-targeted fungi. Here, we surveyed 99 strains, representing 12 Penicillium species, that were isolated from a variety of environments, including foods, dead bodies, and clinical samples. Among the Penicillium strains, including non-pathogenic P. chrysogenum and P. camembertii, as well as postharvest pathogens P. expansum and P. digitatum, 14 and 20 showed resistance to fludioxonil and iprodione, respectively, and 6 showed multi-drug resistance to the fungicides. Sequence analyses revealed that some strains of P. chrysogenum and Penicillium oxalicum had mutations in NikA, a group III histidine kinase of the high-osmolarity glycerol pathway, which is the mode of action for fludioxonil and iprodione. The single nucleotide polymorphisms of G693D and T1318P in P. chrysogenum and T960S in P. oxalicum were only present in the fludioxonil- or iprodione-resistant strains. These strains also exhibited resistance to pyrrolnitrin, which is the lead compound in fludioxonil and is naturally produced by some Pseudomonas species. This study demonstrated that non-targeted Penicillium strains distributed throughout the environment possess fungicide resistance.

Rare isotachin-derived from the Dasymaschalon rostratum fungus Penicillium tanzanicum ZY-5.

Four rare isotachin-derived, isotachins E-H (1-4), together with two known biogenetically related isotachin derivatives (5 and 6) were isolated from the solid rice fermentation of a fungus Penicillium tanzanicum ZY-5 obtained from a medicinal plant Dasymaschalon rostratum collected from the Changjiang County, Hainan Province, China. Their structures were elucidated using comprehensive spectroscopic methods. The single-crystal X-ray diffraction of compound 5 was determined. Compounds 1-4 have a trans-3-(methylthio)-acrylic acid fragment, which are rare in nature. The inhibitory activities of all compounds against the nitric oxide (NO) production induced by lipopolysaccharide in mouse macrophage RAW 264.7 cells in vitro were evaluated.

Occurrence and Characterization of Penicillium Species Isolated from Post-Harvest Apples in Lebanon.

The apple is one of the most important fruit tree crops in the Mediterranean region. Lebanon, in particular, is among the top apple producer countries in the Middle East; however, recently, several types of damage, particularly rot symptoms, have been detected on fruits in cold storage. This study aims to identify the causal agents of apple decay in Lebanese post-harvest facilities and characterize a set of 39 representative strains of the toxigenic fungus Penicillium. The results demonstrated that blue mould was the most frequent fungal disease associated with apples showing symptoms of decay after 3-4 months of storage at 0 °C, with an average frequency of 76.5% and 80.6% on cv. Red and cv. Golden Delicious apples, respectively. The morphological identification and phylogenetic analysis of benA gene showed that most Penicillium strains (87.2%) belong to P. expansum species whereas the remaining strains (12.8%) belong to P. solitum. Furthermore, 67.7% of P. expansum strains produced patulin when grown on apple puree for 14 days at 25 °C with values ranging from 10.7 mg kg-1 to 125.9 mg kg-1, whereas all P. solitum did not produce the mycotoxin. This study highlights the presence of Penicillium spp. and their related mycotoxin risk during apple storage and calls for the implementation of proper measures to decrease the risk of mycotoxin contamination of apple fruit products.

Fungal and Bacterial Biodeterioration of Outdoor Canvas Paintings: The Case of the Cloisters of Quito, Ecuador.

The historic center of Quito, Ecuador, was one of the first World Cultural Heritage Sites declared by UNESCO in 1978. There are numerous religious buildings built during the Spanish colonial period reflecting the cultural heritage in this area. Between them, the cloisters of San Francisco, Santo Domingo, and Santa Clara should be highlighted. The specific problems of conservation of the outdoor canvas paintings are not well known at the moment. The objective of this paper is to achieve a conservation study of the canvas paintings exhibited in these three cloisters of the historic center of Quito in order to identify the microbial agents and the main bioclimatic parameters of deterioration. For this, a study of the state of conservation of five canvas paintings has been carried out, as well as a sampling and identification of the main microorganisms present on the obverse and reverse of the works, employing diverse techniques, traditional and biomolecular ones. An analysis of climatic conditions has also been achieved in the cloister of San Francisco. The results of the study indicate that the exhibition conditions in the cloisters are really problematic for the conservation of paintings. Important biodeteriorating agents have been isolated, including fungi and bacteria species belonging, among others, to the genera Bacillus, Penicillium, Alternaria, Mucor, and Aspergillus. We have also researched its relationship with the deterioration state of the artworks and the exhibition conditions in each case, proposing guidelines for the proper conservation of this important World Cultural Heritage.

Cold-active catalase from the psychrotolerant fungus Penicillium griseofulvum.

Cold-active catalase (CAT) elicits great interest because of its vast prospective at the medical, commercial, and biotechnological levels. The study paper reports the production of cold-active CAT by the strain Penicillium griseofulvum P29 isolated from Antarctic soil. Improved enzyme production was achieved by optimization of medium and culture conditions. Maximum CAT was demonstrated under low glucose content (2%), 10% inoculum size, temperature 20°C, and dissolved oxygen concentration (DO) 40%. An effective laboratory technology based on changing the oxidative stress level through an increase of DO in the bioreactor was developed. The used strategy resulted in a 1.7- and 1.4-fold enhanced total enzyme activity and maximum enzyme productivity. The enzyme was purified and characterized. P. griseofulvum P29 CAT was most active at approximately 20°C and pH 6.0. Its thermostability was in the range between 5°C and 40°C.

Austalide K from the Fungus Penicillium rudallense Prevents LPS-Induced Bone Loss in Mice by Inhibiting Osteoclast Differentiation and Promoting Osteoblast Differentiation.

Osteoporosis is a chronic disease that has become a serious public health problem due to the associated reduction in quality of life and its increasing financial burden. It is known that inhibiting osteoclast differentiation and promoting osteoblast formation prevents osteoporosis. As there is no drug with this dual activity without clinical side effects, new alternatives are needed. Here, we demonstrate that austalide K, isolated from the marine fungus Penicillium rudallenes, has dual activities in bone remodeling. Austalide K inhibits the receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclast differentiation and improves bone morphogenetic protein (BMP)-2-mediated osteoblast differentiation in vitro without cytotoxicity. The nuclear factor of activated T cells c1 (NFATc1), tartrate-resistant acid phosphatase (TRAP), dendritic cell-specific transmembrane protein (DC-STAMP), and cathepsin K (CTSK) osteoclast-formation-related genes were reduced and alkaline phosphatase (ALP), runt-related transcription factor 2 (Runx2), osteocalcin (OCN), and osteopontin (OPN) (osteoblast activation-related genes) were simultaneously upregulated by treatment with austalide K. Furthermore, austalide K showed good efficacy in an LPS-induced bone loss in vivo model. Bone volume, trabecular separation, trabecular thickness, and bone mineral density were recovered by austalide K. On the basis of these results, austalide K may lead to new drug treatments for bone diseases such as osteoporosis.

Of fungi and ticks: Morphological and molecular characterization of fungal contaminants of a laboratory-reared Ixodes ricinus colony.

Establishing and maintaining tick colonies in the laboratory is essential for studying their biology and pathogen transmission, or for the development of new tick control methods. Due to their requirement for very high humidity, these laboratory-bred colonies are frequently subject to fungal contamination. In the present study, we aimed to identify the fungal species that contaminated a laboratory-reared colony of Ixodes ricinus through microscopic observation and molecular identification. We identified three different taxa isolated from the ticks: Aspergillus parasiticus, Penicillium steckii, and Scopulariopsis brevicaulis. These three species are usually regarded as environmental saprophytic molds but both direct and indirect evidence suggest that they could also be considered as entomopathogenic fungi. Although we do not have any direct evidence that the fungi isolated from I. ricinus in this study could cause lethal infections in ticks, we observed that once infected, heavy fungal growth coupled with very high mortality rates suggest that studying the entomopathogenic potential of these fungi could be relevant to biological tick control.

A novel qPCR based-method for detection and quantification of three recurrent species of Penicillium isolated from bioaerosols in mold-damaged homes.

Fungal contamination of indoor environments can cause respiratory diseases and induce damages to building materials. Among the fungal species found in mold-damaged homes, Penicillium brevicompactum, P. chrysogenum and P. crustosum can be considered as recurrent strains. In this study, we therefore propose a rapid and novel qPCR-based method in order to allow the monitoring of these three fungal species. The method developed allows the quantification of the target DNA of these three Penicillium species with a limit of quantification of 0.01 ng/µL without significant difference with spectrophotometry quantification assay for DNA concentrations between 5 and 100 ng/µL. This technique also enables the rapid detection of these three species in complex mixtures of DNA extracted from 15 bioaerosols collected in mold-damaged homes and previously cultured on agar plate. This new sensitive and specific qPCR technique can thus be easily integrated into bioaerosol studies.

Climatic conditions and farm practices affected the prevalence of Aspergillus section Flavi on different types of dairy goat's feed.

The present work aimed to determine the prevalence of aflatoxigenic Aspergillus section Flavi on different types of dairy goat's feed samples obtained from four dairy goat's farms around the central region of Peninsular Malaysia, and to examine the effects of climatic conditions (temperature, relative humidity) of the dairy goat's farms, and their feeding and storage practices on the fungal prevalence of different types of dairy goat's feed. A total of 60 goat's feed samples were obtained, and their proximate composition and water activity were determined, following which they were cultivated on DRBC and AFPA for total fungal load and Aspergillus section Flavi load determination, respectively. Fungal isolates were identified morphologically, and toxigenicity potentials of Aspergillus section Flavi isolates were determined using CCA. The temperature and relative humidity data of all farms were obtained from the Malaysian Meteorological Department. The total fungal loads (on DRBC) of the goat's feed samples were log 0.767 to 7.071 CFU/g which included the common feed contaminants such as Aspergillus, Fusarium, and Penicillium. The Aspergillus section Flavi loads (on AFPA) were log 0.667 to 3.206 CFU/g. Farm A yielded the highest number of Aspergillus section Flavi isolates as well as the highest number of aflatoxigenic isolates. It was found that climatic conditions and different practices between farms positively influenced the fungal prevalence on goat's feed samples based on the Pearson correlation analysis. The prevalence of mycotoxigenic isolates on goat's feed warrants for urgent intervention to ensure that goats are being fed with nutritionally adequate and safe feed. The presence of aflatoxigenic Aspergillus section Flavi isolates indicates the risk of aflatoxin B1 contamination on the goat's feed, aflatoxicosis development in the goats, and aflatoxin M1 bio-transformation in the goat's milk. This is a potential threat to the flourishing goat's milk industry in Malaysia.

Characterization of Penicillium crustosum L-asparaginase and its acrylamide alleviation efficiency in roasted coffee beans at non-cytotoxic levels.

This work aims at isolating a fungal source for L-asparaginase production to be applied in reducing acrylamide levels in coffee beans at non-cytotoxic levels. An L-asparaginase-producing fungus was isolated from an agricultural soil sample and identified as Penicillium crustosum NMKA 511. A maximum L-asparaginase activity of 19.10 U/mL was obtained by the above-mentioned fungus when grown under optimum conditions (i.e. 16.96 g/L sucrose as carbon source, 1.92 g/L peptone as nitrogen source, pH 7.7 and 33.5 °C). Further, the produced L-asparaginase was purified and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that P. crustosum L-asparaginase was a heterodimer enzyme with molecular weights of approximately 41.3 and 44.4 kDa. Also, the purified P. crustosum L-asparaginase was specific towards L-asparagine and showed negligible and no effects towards L-glutamine and D-asparagine, respectively. Additionally, the purified L-asparaginase reduced the acrylamide levels by 80.7% and 75.8% in light and dark roasted coffee beans, respectively. The amount of L-asparaginase used to reduce acrylamide was considered safe when cell viability reached 94.6%.

Linking Pélardon artisanal goat cheese microbial communities to aroma compounds during cheese-making and ripening.

Pélardon is an artisanal French raw goat's milk cheese, produced using natural whey as a backslop. The aim of this study was to identify key microbial players involved in the acidification and aroma production of this Protected Designation of Origin cheese. Microbial diversity of samples, collected from the raw milk to 3-month cheese ripening, was determined by culture-dependent (MALDI-TOF analysis of 2877 isolates) and -independent (ITS2 and 16S metabarcoding) approaches and linked to changes in biochemical profiles (volatile compounds and acids). In parallel, potential dominant autochthonous microorganism reservoirs were also investigated by sampling the cheese-factory environment. Complex and increasing microbial diversity was observed by both approaches during ripening although major discrepancies were observed regarding Lactococcus lactis and Lacticaseibacillus paracasei fate. By correlating microbial shifts to biochemical changes, Lactococcus lactis was identified as the main acidifying bacterium, while L. mesenteroides and Geotrichum candidum were prevalent and associated with amino acids catabolism after the acidification step. The three species were dominant in the whey (backslop). In contrast, L. paracasei, Enterococcus faecalis, Penicillium commune and Scopulariopsis brevicaulis, which dominated during ripening, likely originated from the cheese-making environment. All these four species were positively correlated to major volatile compounds responsible for the goaty and earthy Pélardon cheese aroma. Overall, this work highlighted the power of MALDI-TOF and molecular techniques combined with volatilome analyses to dynamically follow and identify microbial communities during cheese-making and successively identify the key-players involved in aroma production and contributing to the typicity of Pélardon cheese.