Stem end rot, caused by the fungal pathogen Lasiodiplodia theobromae, is one of the most economically significant post-harvest diseases affecting mangoes globally. Seventy isolates of L. theobromae were collected from mango samples exhibiting stem end rot symptoms to characterize the cultural and morphological diversity. Samples were sourced from various orchards, wholesale and retail markets across the Junagadh and Gir Somnath districts of Gujarat during the 2024 mango season. All the isolates showed significant variations in their characteristics. A majority of the isolates (65 out of 70) exhibited a fast growth rate, with a mean colony diameter exceeding 85 mm. Four isolates showed a medium growth rate, while only one isolate (LT 17) displayed a slow growth rate of 69.83 mm. Colonies displayed a wide range of colours (cottony white and greyish white to grey, dark grey, black) and textures (aerial-woolly, fluffy, velvety, flate) with margins observed as either regular smooth/flat or irregular rough. Pigmentation also differed, appearing as black, dark black, pink or absent. Sporulation levels varied significantly, categorized into excellent, good, moderate, poor and no sporulations. Furthermore, the conidial dimensions showed considerable diversity, with lengths ranging from 14.5 to 35 μm and widths from 7 to 17 μm. Conidial shapes were observed as oval, subovoid to ellipsoid and ellipsoid. Immature conidia were consistently hyaline and single-celled, maturing into dark brown, two-celled structures with a single septum. These findings confirm a high level of diversity within the L. theobromae population in the study area, which has implications for disease management.

BackgroundTwig blight (TB), caused by Pestalotiopsis-like species, induces twig dieback in Myrica rubra, leading to substantial economic losses in its production across China and posing a significant threat to M. rubra cultivation. To date, the immune response of M. rubra to Pestalotiopsis-like infections remains poorly understood.ResultsIn this study, full-length transcriptomics and untargeted metabolomics were employed to investigate the immune responses of resistant “Dingao” (DA) and susceptible “Dongkui” (DK) M. rubra cultivars to Pestalotiopsis pathogen infection at early (1DPI) and later (5DPI) stage. The main findings revealed that the resistant DA triggered a rapid and robust immune response at 1DPI, with 1,111 differentially expressed genes (DEGs), including 583 upregulated and 528 downregulated. As the immune response progressed to 5DPI, the number of DEGs in Dingao was reduced to 324 DEGs (114 upregulated and 210 downregulated). In contrast, the susceptible Dongkui cultivar exhibited a weak and delayed defense response. At 1DPI only 99 DEGs (26 upregulated and 73 downregulated) were identified in DK leaves; by 5DPI, a substantial shift occurred in Dongkui samples, with 3,414 DEGs (2,141 upregulated and 1,273 downregulated), showing a late but massive defense response. In Dingao samples, WRKY, AP2/ERF transcription factors (TFs) were the most upregulated TFs at the early immune response stage; by 5DPI, NAC and WRKY families became the most prominently expressed TFs. In Dongkui, notable TF upregulation (including WRKY, MYB and AP2/ERF) was only observed at 5DPI and there were not significantly expression changes at the early immune response stage. Further analysis using untargeted metabolomics revealed that Dongkui exhibited 280 upregulated and 12 downregulated differentially accumulated metabolites (DMs) at 1DPI; 503 and 166 DMs were found to be upregulated and regulated at 5DPI. In Dingao plants, 472 DMs were upregulated and 110 downregulated at 1DPI, whereas at 5DPI, 387 DMs were upregulated and 219 downregulated.ConclusionsThrough a combined analysis of full-length transcriptomics and untargeted metabolomics, we revealed distinct molecular and metabolic responses between resistant “Dingao” (DA) and susceptible “Dongkui” (DK) M. rubra cultivars under twig blight stress.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12870-025-07543-1.

The present study investigates the incidence, abundance, and diversity of fungi colonizing Chrysanthemum spp. flowers were collected from varied geographical locations and developmental stages. During the present investigation, a total of 41 fungal species were recorded at 10 locations. The dominant genera were Alternaria, Aspergillus, Cladosporium, Penicillium, Curvularia, Fusarium, Nigrospora, and Rhizopus, Chrysanthemum spp. The stages of colonisation of fungi were diverse at the stage of the flower (semi-open bud, open bud, bloom flower, and senescent flower). At the Bloom (S3) and senescent (S4) stages, significantly higher fungal occurrences were recorded, up to 95%, including Rhizopus stolonifer, Aspergillus niger, Alternaria alternata, and Penicillium crysogenum. In the semi-bud stage of the flower, the fungal incidence in all significant fungi was lowered, except for Alternaria raphani and Aspergillus niger. Aspergillus terreus, Cladosporium cladosporioides, Fusarium fujikuroi, Curvularia lunata, Lasiodiplodia theobromae, Nigrospora oryzae. Penicillium lilacinum was found dominantly (88%), next, Nigrospora oryzae was found 86%, Aspergillus niger 86%, Fusarium fujikuroi 85%. Recorded at the senescent stage of the flower, compared to the blooming stage, it shows Penicillium lilacinum 73%, Aspergillus niger 65%, and Nigrospora oryzae 66%. Fusarium fujikuroi incidence is 50%. All fungal species appeared only from the Bloom Stage (S3) onward. S4 Stage (Senescent Flower) again showed peak fungal incidence across all listed species. The data highlights that fungal colonization begins primarily at the Bloom Stage (S3) and intensifies in the senescent stage.

Apple (Malus domestica) is a very important crop grown in Kazakhstan. Cytospora species are capable of causing destructive stem cankers on a wide range of woody plants, including apples, and can lead to twig and branch dieback. This study aimed to identify the Cytospora species responsible for canker disease of apple in Kazakhstan and to assess the susceptibility of major apple cultivars to these pathogens. Investigations were conducted in Almaty, Kazakhstan, during 2023 and 2024. Samples from symptomatic trees were collected, and nine Cytospora isolates were obtained from diseased apple trees. Multigene phylogenetic analysis based on combined sequence data of ITS, tef1-α, tub2, and LSU loci, together with morphological characteristics and pathogenicity assays, revealed two Cytospora species: C. leucostoma and C. sorbicola. The reactions of six apple cultivars (Gala, Golden Delicious, Red Delicious, Granny Smith, Fuji, and Jonaprince) to these species were evaluated, and statistically significant differences were found among cultivars (p < 0.05). The largest lesions occurred on Red Delicious and Fuji, indicating that these cultivars are the most susceptible. In contrast, lesion lengths on Jonaprince were significantly smaller than on all other cultivars, suggesting that Jonaprince is resistant to Cytospora species in Kazakhstan. This is the first report of C. leucostoma and C. sorbicola causing apple canker disease in Kazakhstan.

Cocoa (<i>Theobroma cacao</i> L.) is the main cash crop in Côte d'Ivoire and the primary source of income for producers. However, recent studies have highlighted the harmful effects of pod diseases on local production and the existence of a group of pathogens (<i>Lasiodiplodia </i>spp. and <i>Fusarium</i> spp.) that require better characterization in order to reorient control efforts. The objective of this study was to perform morphological and molecular characterization of isolates of <i>Lasiodiplodia</i> spp. and <i>Fusarium</i> spp. isolated from cocoa pods affected by rot. Samples of infected pods and soil from cocoa plantations were collected from seven locations in the Agneby-Tiassa region. After isolation on 11% agar culture medium, the isolates were purified on pea and carrot agar culture medium. Macroscopic and microscopic observations were made on these isolates. After DNA extraction, PCR tests with ITS4 and ITS5 primers were performed for molecular identification of the isolates. The PCR products obtained were transferred to a facility for sequencing. It should be noted that macroscopic and microscopic observations showed that <i>Lasiodiplodia</i> spp. isolates are light gray or gray-black in color, with thick-walled septate conidia. Fusarium spp. isolates are whitish in color, with oval microconidia and curved macroconidia. PCR test results showed that the isolates from the southeastern region of Côte d'Ivoire are <i>Lasiodiplodia theobromae</i> and <i>Fusarium solani</i>. Sequencing showed that these isolates were 100% similar to the species in the gene bank. This study will help to better target the fight against cocoa pod rot in Côte d'Ivoire by implementing sustainable control strategies.

Lysin motif (LysM) effectors contribute to the virulence of many pathogens, but the underlying mechanism of this type of effector remains poorly understood. Here, we identified a LysM-containing protein, named LtLysM2, in the plant opportunistic pathogen Lasiodiplodia theobromae. We demonstrated that LtLysM2 contributes to the virulence of the pathogen, is able to bind to chitin oligosaccharides, and can suppress chitin-triggered plant immune responses. Importantly, we showed that the grapevine (Vitis vinifera) protein VvSrc2, a membrane- and nuclear-localized protein homologous to the soybean (Glycine max) protein Src2 (Soybean genes regulated by cold 2), interacts with LtLysM2. Interestingly, the nuclear accumulation of VvSrc2 was elevated in the presence of LtLysM2. Ectopic expression of VvSrc2 in Nicotiana benthamiana enhanced resistance to L. theobromae. Additionally, the VvSrc2 protein interacted with the nuclear-localized RNA-binding protein VvUbp1, contributing to the programmed cell death evoked by VvUbp1. Our findings reveal a previously uncharacterized regulatory pathway in which VvSrc2 is upregulated after the recognition of LtLysM2 and serves as an intermediator to transduce extracellular signaling events into nuclear components during infection by L. theobromae through association with the downstream target VvUbp1 in the cell nucleus.

Background In coconut production, less than one third of the button nuts produced in an inflorescence eventually develop into mature nuts, as a result of fruit rot and premature nut fall diseases. Botryodiplodia theobromae is linked to the fruit rot and premature nut fall diseases of coconut, resulting in consequential yield loss of over 60%. Furthermore, the fungus poses a concern to plants, farmers, and plant health authorities because it may survive in plant materials as an endophyte, escaping the quarantine process. This study was carried out to analyse the cultural, morphological, and molecular characters of B. theobromae associated with these diseases in coconut fruits. Methods Eight isolates of B. theobromae used in this study were collected from diseased coconut fruit samples (showing signs of rot and freshly fallen nuts) in two major coconut producing areas in Nigeria. Pure culture of isolates were obtained using potato dextrose agar (PDA) medium. The culture medium and microscopy were used to also examine the cultural and morphological characters of isolates. For molecular characters of isolates, DNA was extracted from each isolate and amplified with a polymerase chain reaction (PCR) using a universal primer (ITS1F/4R) and three specific primers (Lt347-F/R, Bt2aF/bR, and EF1-728F/EF2-728R). Furthermore, PCR amplicons obtained from the ITS1F/4R primers were sequenced and analyzed using bioinformatics and phylogenetic methods. The genetic similarity and variation of isolates were also determined. Results The colony texture/color, ITS1/4 gene sequence information and phylogenetic analysis clustered the eight isolates of B. theobromae into five categories. The ITS1/4 sequence information using a blast search in the NCBI database, confirmed all eight fungal isolates as B. theobromae . Conclusion This study has provided a guide for proper categorization of B. theobromae , that is a prerequisite for early diagnosis and management of this pathogen in coconut producing areas.

Crown rot is a major postharvest disease affecting bananas (Musa spp.), resulting in substantial economic losses by compromising fruit quality and limiting marketability. In 2023, symptoms of crown rot were observed on banana fruits sourced from ten states across the northern and northeastern regions of Brazil. Pathogen identification was carried out using morphological characteristics and multigene phylogenetic analysis based on the internal transcribed spacer (ITS), translation elongation factor 1-alpha (tef1-α), β-tubulin (tub2), and RNA polymerase II subunit (rpb2). A total of 116 fungal isolates recovered from diseased fruit exhibited morphological traits consistent with the genus Lasiodiplodia. Phylogenetic reconstruction based on concatenated multilocus data revealed that the new isolates belonged to the species Lasiodiplodia brasiliensis, Lasiodiplodia hormozganensis, Lasiodiplodia iraniensis, Lasiodiplodia laeliocattleyae, Lasiodiplodia pseudotheobromae, and Lasiodiplodia theobromae. Pathogenicity tests confirmed that all Lasiodiplodia species/isolates experimentally induced crown rot symptoms on banana fruits of cultivars ‘Prata’, ‘Pacovan’, and ‘Terra’. Inoculated fungi were re-isolated from symptomatic tissues, which exhibited morphological characteristics identical to those of the original cultures, fulfilling Koch’s postulates. This study represents the first large-scale and comprehensive survey on Lasiodiplodia species associated with banana crown rot in Brazil, revealing that a complex of at least six phylogenetically distinct Lasiodiplodia species are involved in the disease etiology.Supplementary InformationThe online version contains supplementary material available at 10.1038/s41598-025-22739-z.

The study uses solid-state fermentation to explore the co-production of industrial enzymes (amylases, cellulases, xylanases, and phytases). The experiments examine the key cultivation parameters, such as fermentation time (2 to 7 days), incubation temperature (25 to 35 °C), medium components, and the effects of inducers. Maximum enzyme activities were observed at 30 °C after 6 days of incubation, using paddy straw as a primary substrate. Supplementing paddy straw with rice bran significantly enhanced the activities of amylase, CMCase, FPase, and xylanase by 1.94x, 1.36x, 1.45x, and 2.14x, respectively using fungal strain Lasiodiplodia theobromae strain RBEE-GCS103. Further, enzyme production in enamel-coated metallic trays increased enzyme activities by 20-35% compared to production in 250 ml flasks. The present study highlights L. theobromae strain RBEE-GCS103 as a novel and potential co-producer for the industrially important enzyme utilizing underutilized agro-waste.

Lasiodiplodia theobromae, Pestalotiopsis clavispora and Fusarium spp. associated with pomegranate dieback and fruit rot in Tunisia

Dieback of Mango, primarily caused by Lasiodiplodia theobromae, poses a significant threat to mango production in Burkina Faso. This study evaluated the efficacy of biological and chemical treatments in controlling dieback and promoting tree recovery. Field trials were conducted in Koloko and Léo provinces across three mango orchards exhibiting varying disease incidences. Symptomatic tissues were collected for pathogen isolation and molecular confirmation via PCR, validating the presence of L. theobromae. Three biopesticide treatments (Plantsain, Fertisain, and their combination) were compared with a chemical reference (Azox + Manga Plus) and an untreated control, using a randomized complete block design with six replicates per treatment. Results indicated that Plantsain achieved the highest branch regeneration, with rates of 72.72% in Koloko and 60.49% in Léo, compared to 7.03% and 8.92% in untreated controls. Both disease incidence and severity decreased significantly with treatment, displaying a dose-dependent response. The combined Plantsain + Fertisain treatment at dose of 2 L/ha each also demonstrated high efficacy, comparable to the chemical Azox. These outcomes suggest that biopesticides not only significantly reduce the incidence and severity of the disease, the infection rate of L. theobromae, but can also promote the physiological regeneration of affected tissues. In conclusion, biological control strategies, taken individually or in combination, constitute sustainable and effective alternatives to chemical fungicides for the management of dieback in Burkina Faso.

Botryosphaeria canker and dieback is an important fungal disease caused by Botryosphaeriaceae spp. that affects the productivity of apple orchards in Chile and worldwide. In the field, studies on the management of this disease are focused on the protection provide by fungicides applied on pruning wounds. However, in Chile, information about the protection and efficacy of fungicides on apple trees against twig and branch pathogens is scarce. Therefore, in this study, we evaluated the efficacy of fungicides to control fungal trunk pathogens causing Botryosphaeria canker and dieback in apple trees using in vitro, greenhouse, and field trials. Isolates of Diplodia mutila, D. seriata, Neofusicoccum arbuti, and Lasiodiplodia theobromae obtained and characterized previously in Chile from apple trees with canker and dieback symptoms were used in this study. In vitro tests showed that benomyl, fluazinam, difenoconazole, and tebuconazole exhibited the lowest EC50 values, with means of 0.08, 0.09, 0.12, and 0.18 μg/ml, respectively. This study demonstrates that infection caused by D. mutila, D. seriata, L. theobromae, and N. arbuti can be significantly reduced using single-sprayed fungicides. The most effective in reducing infection on pruning wounds of apple trees by Botryosphaeriaceae were benomyl (66 to 76%), tebuconazole (47 to 68%), thiophanate-methyl (68 to 71%), boscalid + pyraclostrobin (47 to 63%), fluxapyroxad + pyraclostrobin (54 to 57%), and thiophanate-methyl + tetraconazole (63 to 74%). To our knowledge, this is the first study reporting the control of Botryosphaeriaceae causing canker and dieback in apple trees through the use of commercially available chemical fungicides, with different active ingredients and modes of action.

In recent years, diseases of tea plants (Camellia sinensis) caused by Lasiodiplodia theobromae have significantly impacted the quality and yield of tea leaves in China and India. However, the pathogenic mechanisms of L. theobromae in tea plants remain unclear. In this study, the relative expression levels of ethylene receptor 2 (ETR2, historically referred to as ethylene response 2) mRNA was analyzed at different hours post inoculation at the first, second, and third leaves on the tea shoot inoculated with L. theobromae hyphae. The results revealed spatiotemporal differences in CsETR2 expression, suggesting its involvement in disease resistance. Subcellular localization analysis indicated that CsETR2 was localized in the endoplasmic reticulum of leaf cells in Nicotiana benthamiana. Through transient overexpression, antisense oligonucleotide (AsODN) silencing in tea leaves, and transgenic N. benthamiana assays, it was confirmed that CsETR2 positively regulated resistance in N. benthamiana. 3,3'-diaminobenzidine/nitroblue tetrazolium staining and quantitative analysis of reactive oxygen species (ROS) scavenger genes demonstrated that CsETR2 mediated disease resistance through ROS-mediated mechanism. As a key component in ethylene (ET) signal transduction, the expression of CsETR2 was positively correlated with genes in the ET transduction pathways under overexpression or silencing conditions. β-glucuronidase and dual-luciferase assays revealed that PC-5p-1042181_5 inhibited CsETR2 mRNA expression by targeting its mRNA, thereby reducing disease resistance. Further experiments with transient overexpression, AsODN silencing, and transgenic N. benthamiana validated the negative regulatory role of PC-5p-1042181_5. This study uncovers a novel disease resistance mechanism involving ROS-ET interference mediated by PC-5p-1042181_5-CsETR2 module, offering new insights for breeding disease-resistant tea cultivars.

Bleeding disease in tangerine plants is caused by the fungus Botryodiplodia theobromae. This study aimed to determine the effect of Cyperus rotundus L. extract on the growth of B. theobromae mycelia and spores in vitro, and to determine the best concentration of C. rotundus extract in inhibiting the growth of the fungus. The study was conducted using a Completely Randomized Design (CRD) consisting of 7 treatments, namely C. rotundus extract concentrations of 0.0% (control), 0.1%, 0.2%, 0.3%, 0.4%, and 0.5%, as well as a fungicide treatment containing the active ingredient carbendazim 0.1%. The observed variable was fungal growth measured by colony diameter. The results showed that the treatment of sedge rhizome extract was able to suppress the development of B. theobromae fungal colonies. This nutmeg rhizome extract has the potential to be developed as a botanical pesticide to control B. theobromae fungus, and a concentration of 0.5% nutmeg rhizome extract has the highest inhibitory power against the growth of B. theobromae fungus, namely 56%.

Bleeding disease in tangerine plants is caused by the fungus Botryodiplodia theobromae. This study aimed to determine the effect of Cyperus rotundus L. extract on the growth of B. theobromae mycelia and spores in vitro, and to determine the best concentration of C. rotundus extract in inhibiting the growth of the fungus. The study was conducted using a Completely Randomized Design (CRD) consisting of 7 treatments, namely C. rotundus extract concentrations of 0.0% (control), 0.1%, 0.2%, 0.3%, 0.4%, and 0.5%, as well as a fungicide treatment containing the active ingredient carbendazim 0.1%. The observed variable was fungal growth measured by colony diameter. The results showed that the treatment of sedge rhizome extract was able to suppress the development of B. theobromae fungal colonies. This nutmeg rhizome extract has the potential to be developed as a botanical pesticide to control B. theobromae fungus, and a concentration of 0.5% nutmeg rhizome extract has the highest inhibitory power against the growth of B. theobromae fungus, namely 56%.

The study was conducted to evaluate the morphological characteristics and determine the cause of the blight disease of two varieties of lotus, Hong and Bach Diep, in My Duc commune and Chuong My ward - Hanoi. The research method included evaluating morphological characteristics according to UPOV standards (2023), isolating the fungus from diseased lotus samples according to the method of Burgess & cs. (2008), testing artificial infection in pots and mason jars. The results showed that both varieties have pink flowers, showy flowers fragrant, suitable for planting as ornamental plants and cultivation in eco-tourism areas. The Hong lotus variety has single-petal flowers, while the Bach Diep lotus variety has double-petal flowers. The fungus Lasiodiplodia theobromae was identified as the cause of the blight disease on two lotus varieties in My Duc commune and Chuong My ward. Both lotus varieties in the experiment were infected at a very severe level (level 9), with clear diseased leaves, petiole and the plants blighted after 10-14 days. The research results provide a scientific basis for proposing effective disease prevention and control measures. This is an important premise to protect production, preserve the environment and develop lotus growing areas in an ecological and sustainable direction.

Fig (Ficus carica) orchards in the Salento peninsula (southeastern Apulia region, Italy) are increasingly affected by decline syndromes whose etiology remains poorly resolved. In this paper, we provide a first characterization of a complex disease outbreak, integrating field surveys, fungal isolation, molecular phylogenetics, and pathogenicity assays. Symptomatic trees displayed chlorosis, defoliation, cankers, vascular discoloration, and wilting, frequently associated with bark beetle galleries. Mycological analyses revealed a diverse assemblage of fungi, dominated by Botryosphaeriaceae (including Neofusicoccum algeriense, and Lasiodiplodia theobromae), the Fusarium solani species complex (notably Neocosmospora perseae), and Ceratocystis ficicola. While C. ficicola was isolated with lower frequency, its recovery from adult beetles—including Cryphalus dilutus—supports a role in insect-mediated dissemination in addition to soilborne infection. Pathogenicity tests demonstrated that N. algeriense and N. perseae, together with C. ficicola, caused severe vascular lesions and wilting, confirming their contribution to fig decline. By contrast, other Fusarioid strains showed no pathogenicity, consistent with their role as latent or stress-associated pathogens. This study provides the first evidence that N. algeriense and N. perseae act as pathogenic agents on fig, highlights their interaction with C. ficicola within a multifactorial decline syndrome, and identifies dual epidemiological pathways involving both soil/root infection and insect-facilitated dissemination via beetles such as C. dilutus. These findings redefine fig decline in the Salento peninsula (southern Italy) as a multifactorial disease rather than a single-pathogen outbreak, with significant implications for diagnosis, epidemiology, and integrated management strategies.

The study was conducted to isolate and select the fungal isolates causing stem end rot of pomelo fruit and characterized by strong growth. Pomelos showing signs of stem end rot or stem wilt were collected. Sixteen fungal isolates were isolated from fruit samples collected from two districts Binh Dai and Mo Cay Bac of the Ben Tre province, Vietnam. The experiment with 16 fungal isolates was completely randomized, with 3 replicates, each replicate being a Petri dish. Nine fast growing fungal isolates were selected, with the diameter of fungal colonies ranging from 8.60 to 9.00 cm, 48 h after inoculation. These nine isolates when inoculated to Pomelo fruits caused pomelo fruit rot symptoms and their pathogenicity was confirmed following Koch’s postulates. The range of infection diameter on pomelo fruits was 0.85-3.00 cm. The three fungal isolates causing the earliest and the most severe pomelo fruit rot were identified as Lasiodiplodia pseudotheobromae PL-M01-A4-B and PL-B01-A7-B, and L. theobromae PL-M01-A6-B by sequencing the ITS region, with 100% similarity. Keywords: Fruit rot, Lasiodiplodia pseudotheobromae, Lasiodiplodia theobromae, pomelo, pathogenic factor, stem end rot.

Lasiodiplodia theobromae and Lasiodiplodia brasiliensis associated with dieback and canker in cupuaçu trees (Theobroma grandiflorum) in Brazil

Four Botryosphaeria spp. isolated from apple with dieback symptoms, Diplodia mutila, Diplodia seriata, Neofusicoccum arbuti, and Lasiodiplodia theobromae, were subjected to different conditions to induce sporulation, then freeze-dried and stored in glass vials and ampoules at a temperature of 4 ° C using two protective media (skimmed milk powder in water at 20% and a mixture of skimmed milk powder at 20% plus 5% inositol). Viability was assessed after storage periods of 1, 90, 180, and 365 days. Low-nutrient growth conditions on 2% water agar supplemented with pine needles, incubated under UV light (λ = 350 nm) and at 24 ° C, effectively stimulated sporulation of all four Botryosphaeriaceae species. The survival rate of the isolates was similar. Overall viability showed slight but significant differences depending on the type of protective medium and storage container used for the freeze-dried cultures (p < 0.001). Among the tested media, the highest viability was maintained in vacuum-sealed glass ampoules using either a medium containing 20% skimmed milk powder with 5% inositol or 20% skimmed milk powder alone.