Brazilian Journal of Microbiology最新文献

{"title":"Detection and characterization of a MERS-related coronavirus (Betacoronavirus cameli) in velvety free-tailed bats (Molossus molossus, Mammalia) captured in Southern Brazil.","authors":"Alexandre Sita, Micheli Filippi, Juliana Comerlato, Gabriela Espindola Birlem, Deivid de Souza da Silva, Larissa Mallmann, Vyctoria Malahyka de Abreu Góes Pereira, Meriane Demoliner, Juliana Schons Gularte, Alana Witt Hansen, Fernando Rosado Spilki, Matheus Nunes Weber","doi":"10.1007/s42770-026-02034-3","DOIUrl":"10.1007/s42770-026-02034-3","url":null,"abstract":"<p><p>Bat-borne coronaviruses are of increasing interest due to their potential for zoonotic spillover, particularly within the Betacoronavirus genus. Partial genomic sequences related to MERS-like CoV was identified in multiple hosts, including humans, camelids, and bats from the families Vespertilionidae and Molossidae. In this study, we investigated the presence of coronaviruses in 65 Molossus molossus bats captured in southern Brazil using nested RT-PCR and high-throughput sequencing (HTS). One individual (1.5%) tested positive in a rectal swab, and sequence analysis revealed the presence of a MERS-related coronavirus (Betacoronavirus cameli). Phylogenetic reconstruction based on five genomic contigs clustered the virus within the Merbecovirus subgenus, confirming its close relationship to MERS-CoV strains previously detected in bats and dromedary camels. M. molossus is a widely distributed, insectivorous bat species with synanthropic behavior, commonly roosting in urban areas and human dwellings. The detection of a MERSr-CoV in this species in southern Brazil expands the known host range of these viruses and suggests that members of the Molossidae family may serve as reservoirs. These findings underscore the importance of continuous genomic surveillance in Neotropical bats, particularly in the context of environmental changes that intensify contact between wildlife, humans, and domestic animals.</p>","PeriodicalId":9090,"journal":{"name":"Brazilian Journal of Microbiology","volume":"57 1","pages":""},"PeriodicalIF":2.1,"publicationDate":"2026-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13421570/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148618351","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Single spore propagation of native arbuscular mycorrhizal fungi associated with Khasi mandarin and their symbiotic efficacy in maize.","authors":"Vishal Kumar Mohan, Santa Ram Joshi","doi":"10.1007/s42770-026-02041-4","DOIUrl":"10.1007/s42770-026-02041-4","url":null,"abstract":"<p><p>Arbuscular Mycorrhizal Fungi (AMF) are important members of the soil microbiome that form symbiotic relationships with plant roots, improving nutrient uptake and stress resilience. In the present study, native AMF associated with the rhizosphere of Khasi Mandarin (Citrus reticulata Blanco) were isolated using the wet sieving and decanting method. Trap culture was used to increase the number of native AMF spores. Healthy spores were identified and used for single-spore inoculation to establish single-spore-derived cultures. Molecular identification of the AMF isolates was performed by amplification of the 18 S rDNA region. A controlled pot experiment was conducted to assess the effects of the obtained pure cultures on plant growth. Inoculated maize plants showed significant improvement in root and shoot biomass. This study focuses on evaluating the potential of native AMF isolates associated with Khasi mandarin and their use as cross-inoculants to develop biofertilizer that promote the growth of non-host plants.</p>","PeriodicalId":9090,"journal":{"name":"Brazilian Journal of Microbiology","volume":"57 1","pages":""},"PeriodicalIF":2.1,"publicationDate":"2026-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13421532/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148618273","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evaluation of genomic characteristics of Escherichia coli associated with bovine mastitis and the potential utility of alternative antimicrobial methods.","authors":"Jônathas Moreno Silva de Souza, Gabryel Bernardo Vieira de Lima, Daniela Dos Santos Cassimiro, Sheila da Silva, Thiago de Jesus Sousa, Danillo Sales Rosa, Renata de Faria Silva Souza, Fernando Antônio Gomes da Silva Júnior, Helinando Pequeno de Oliveira, Sílvia Cristina Osaki, Vasco Ariston de Carvalho Azevedo, Bertram Brenig, Flavia Figueira Aburjaile, Mateus Matiuzzi da Costa","doi":"10.1007/s42770-026-02040-5","DOIUrl":"10.1007/s42770-026-02040-5","url":null,"abstract":"<p><p>Mastitis is a disease that affects dairy cattle and can be caused by multidrug-resistant Escherichia coli, causing losses to the agricultural sector. To address this, research into new substances with antimicrobial activity is crucial. Thus, the objective of this study was to evaluate the efficacy of extracts of Commiphora leptophloeos, Amburana cearensis, Agave sisalana, and highly soluble polypyrrole (Hs-PPy) against E. coli isolates. Eight E. coli isolates from bovine mastitis and a standard strain were used. The presence of efflux pumps was investigated by ethidium bromide extrusion. Sequencing was performed on the Illumina HiSeq 2500 platform using a 2 × 150 bp library with quality assessed via FastQC and assembly completed using Unicyler. Genome annotation and the identification of resistance and virulence genes were carried out using PROKKA and PanViTa, respectively. Species confirmation as E. coli was achieved via Jspecies and pyANI public genomes from NCBI. Finally, antimicrobial activity was determined by broth microdilution. Among the natural products tested against the isolates only A. cearensis showed antibacterial activity, with a concentration of 12,500 µg/mL, whereas Hs-PPy showed activity at 250 µg/mL. The isolates tested positive for the presence of efflux pump genes, but only 3 isolates expressed the mechanism in vitro. The natural products were not effective in eliminating the bacteria. Hs-PPy was the only compound that demonstrated bactericidal activity against the bacteria, proving to be an alternative for bactericidal action, even in the presence of genes that inhibit its action against a wide variety of compounds.</p>","PeriodicalId":9090,"journal":{"name":"Brazilian Journal of Microbiology","volume":"57 1","pages":""},"PeriodicalIF":2.1,"publicationDate":"2026-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13421616/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148618265","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Isolation and characterization of Lactococcus lactis from Anabas testudineus (Bloch, 1792) as a potential probiotic for sustainable Aquaculture.","authors":"Nitul Ali, Priyam Sarmah, Dandadhar Sarma","doi":"10.1007/s42770-026-02037-0","DOIUrl":"10.1007/s42770-026-02037-0","url":null,"abstract":"<p><p>The present investigation provides the first report of Lactococcus lactis isolated from the intestinal tract of Anabas testudineus (Bloch, 1792), emphasizing its probiotic attributes and biosafety for aquaculture applications. From 100 bacterial isolates, three strains (GUNA, GUN-1, GUN-2) were identified as L. lactis via 16 S rRNA sequencing. These isolates displayed pronounced antagonism against Aeromonas hydrophila and A. jandaei, along with marked tolerance to gastrointestinal stressors, including low pH (71-80% survival at pH 2.0-3.0), bile salts (69-78% at 0.3-0.5% oxgall), and phenol (66-91% at 0.2-0.4%). Functional traits such as auto-aggregation (21-35%), co-aggregation (15-24%), and hydrophobicity (18-29%) indicated strong adhesion potential. All isolates were non-hemolytic, gelatinase-negative, and devoid of biogenic amine production, confirming their safety. Antibiotic profiling revealed resistance to streptomycin, tetracycline, and ciprofloxacin, but sensitivity to ampicillin. In vivo assays with Labeo rohita and Cirrhinus mrigala confirmed 100% survival without adverse effects. Collectively, these findings highlight L. lactis from A. testudineus as a safe, potent probiotic and a promising bio-alternative to antibiotics in aquaculture.</p>","PeriodicalId":9090,"journal":{"name":"Brazilian Journal of Microbiology","volume":"57 1","pages":""},"PeriodicalIF":2.1,"publicationDate":"2026-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13421708/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148618316","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Decoding the genome of Acinetobacter baumannii from a urinary tract infection case: A comprehensive analysis of resistance, virulence, and genetic mobility attributes.","authors":"A S Smiline Girija, Kannika Parameshwari Kannan","doi":"10.1007/s42770-026-02036-1","DOIUrl":"10.1007/s42770-026-02036-1","url":null,"abstract":"<p><p>Acinetobacter baumannii is an opportunistic multidrug-resistant (MDR) pathogen frequently associated with healthcare-associated infections, including urinary tract infections in critically ill patients. In this study, a carbapenem-resistant A. baumannii (CRAB) isolate, SGK_8A, obtained from an ICU patient, was subjected to whole-genome sequencing (WGS) to explore its genomic features. Species identification and antimicrobial susceptibility testing using the VITEK^® 2 system confirmed resistance to multiple antibiotics, while susceptibility to tigecycline and colistin was retained. WGS was performed using the Illumina NovaSeq 6000 platform, followed by de novo assembly and functional annotation using Unicycler, RAST, PATRIC, CARD, VFDB, and antiSMASH. The draft genome consisted of 3,939,487 bp with a GC content of 38.95% and an N50 value of 77,016 bp. Genome annotation identified 281 functional subsystems associated with cellular metabolism, stress response, and defense mechanisms. Phylogenetic analysis indicated close relatedness to reference strains ATCC 17,978, SDF, and AB30. Resistance gene analysis detected several β-lactamase genes, including bla_OXA- 23, bla_OXA- 51, and bla_OXA- 144, along with multiple additional antimicrobial resistance determinants. The genome also contained genes associated with adhesion, biofilm formation, and other virulence-related functions. Prophage analysis identified five phage regions, including a high-coverage contig that may represent a mobile genetic element (MGEs). Additionally, biosynthetic gene clusters and simple sequence repeats were identified within the genome. Overall, these findings provide a genomic overview of antimicrobial resistance determinants, virulence-associated genes, and MGEs present in the CRAB isolate SGK_8A. The results contribute to the growing genomic data on A. baumannii and may support future comparative and functional studies.</p>","PeriodicalId":9090,"journal":{"name":"Brazilian Journal of Microbiology","volume":"57 1","pages":""},"PeriodicalIF":2.1,"publicationDate":"2026-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13421585/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148618285","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Impact of VP2 mutations on viral fitness in canine parvovirus.","authors":"Tamiris Silva Lopes, Brenda Picoli Gheno, Ahmed Abd El Wahed, Uwe Truyen, André Felipe Streck","doi":"10.1007/s42770-026-02035-2","DOIUrl":"10.1007/s42770-026-02035-2","url":null,"abstract":"<p><p>Canine parvovirus type 2 (CPV-2) is a highly contagious pathogen responsible for severe gastroenteritis in carnivores, particularly in dogs. Continuous antigenic evolution has resulted in the emergence of CPV-2a, CPV-2b, and CPV-2c variants, defined by amino acid substitutions in the VP2 capsid protein. In this study, genetically altered CPV-2 viruses carrying common VP2 mutations (S297A, V300G, D305Y, Y324I, N426D, N426E, and T440A) were generated to evaluate their effects on viral behavior in vitro. Mutations were individually introduced into an infectious backbone (strain 447), classified as CPV-2a based on VP2 sequence analysis, and compared with the ancestral CPV-2 backbone (strain 265). Viral dynamics were assessed using quantitative PCR and immunofluorescence assays to examine extracellular viral DNA and intracellular infection, enabling discrimination between replication and virion release. Distinct profiles were observed among mutants, particularly for N426D, N426E, and T440A, and the impact of each substitution was influenced by the genetic background. Polyclonal antibodies induced by commercial CPV-2 vaccines neutralized both the CPV-2 control and the CPV-2 N426E mutant, indicating cross-reactivity. These findings demonstrate that VP2 evolution must be interpreted through functional approaches rather than solely by variant classification and provide experimentally based information relevant for molecular surveillance and vaccination strategies.</p>","PeriodicalId":9090,"journal":{"name":"Brazilian Journal of Microbiology","volume":"57 1","pages":""},"PeriodicalIF":2.1,"publicationDate":"2026-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13407408/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148590359","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Prebiotics and probiotics in animal nutrition: implications for food safety, sustainability, and circular production systems.","authors":"João Victor Dos Anjos Almeida, Julia Memrava Cabrera, Mauro de Medeiros Oliveira, Ricardo Pinheiro de Souza Oliveira, Alessandro M Varani","doi":"10.1007/s42770-026-02033-4","DOIUrl":"10.1007/s42770-026-02033-4","url":null,"abstract":"<p><p>The pursuit of sustainable and efficient animal production systems has intensified global interest in microbiota-targeted nutritional strategies. This review synthesizes current research on prebiotics and probiotics, selectively utilized microbial substrates and beneficial live microorganisms, respectively, that confer health benefits, across major livestock and aquaculture sectors, including poultry, swine, ruminants, fish, and shrimp. It examines the fundamental role of host-specific gut microbiomes in regulating health and productivity and elucidates how prebiotics, alone or in synergistic formulations with probiotics, modulate these microbial communities. Mechanisms of action involve enhanced nutrient digestibility, improved intestinal morphology and barrier integrity, selective stimulation of beneficial bacteria (e.g., Lactobacillus, Bifidobacterium), suppression of pathogens (e.g., Escherichia coli, Salmonella, Vibrio), and potentiation of innate and humoral immune responses. Consistent evidence demonstrates that microbiota-targeted interventions improve key zootechnical indicators, including weight gain, feed conversion ratio, and carcass quality, while enhancing animal welfare and resilience to environmental and physiological stressors. A pivotal finding is their ability to reduce reliance on antimicrobial agents, addressing antibiotic resistance and aligning with consumer demand for ethical and natural production systems. Furthermore, this review highlights the valorisation of agro-industrial by-products (e.g., fruit pomace, cereal brans) and novel substrates such as Hermetia illucens (black soldier fly) as functional, cost-effective sources that embed animal nutrition within a circular bioeconomy framework. By integrating multidisciplinary findings from terrestrial and aquatic species, this review concludes that microbiota-based functional nutrition is indispensable for advancing next-generation sustainable animal production, uniting productivity, environmental stewardship, and animal welfare while supporting safe, ethical, and climate-resilient food systems worldwide.</p>","PeriodicalId":9090,"journal":{"name":"Brazilian Journal of Microbiology","volume":"57 1","pages":""},"PeriodicalIF":2.1,"publicationDate":"2026-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13400549/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148577067","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Pangenomic and genomic plasticity analyses of the genus Rickettsia.","authors":"Paula Cristina de Magalhães, Andrei Giacchetto Felice, Siomar de Castro Soares","doi":"10.1007/s42770-026-02030-7","DOIUrl":"10.1007/s42770-026-02030-7","url":null,"abstract":"<p><p>The Rickettsia genus comprises obligate intracellular bacteria transmitted by arthropods and responsible for clinically relevant zoonoses, rickettsioses, such as spotted fever and typhus. The difficulty of cultivating these bacteria in vitro reinforces the importance of in silico approaches, such as pangenomic and genomic plasticity analyses. This study analyzed 165 genomes from 31 Rickettsia species available in the REFSEQ (NCBI) database. Tools such as Orthofinder, ANIclustermap, Gegenees, and Mauve were used to classify genes into core, shared, and singletons, assess genomic similarity, and identify structural rearrangements. The results indicate that the genus has an open pangenome (α = 0,842), suggesting high genetic variability and adaptive and expansion potential. Species such as R. typhi exhibited a nearly closed pangenome (α = 0,999), with high genomic conservation, whereas R. rhipicephali showed an open pangenome (α = 0,876), reflecting greater plasticity and intraspecies diversity. Functional categorization of genes revealed that the core genome is associated with vital functions, while singletons include genes related to genetic mobility, indicating possible acquisition through horizontal transfer. Synteny analysis demonstrated high gene conservation in R. typhi and extensive structural reorganization in R. rhipicephali. Statistical correlation reinforced the stability of the core genome regardless of pangenome expansion and revealed an inverse relationship between the number of singletons and the value of α. The findings demonstrate the existence of contrasting evolutionary trajectories within the Rickettsia genus, with conserved, specialized species coexisting alongside genetically dynamic species that are more adaptable to different niches. Thus, this study expands the understanding of clonality, genomic plasticity, and functional diversity within the genus, providing support for future investigations into virulence factors, vaccine targets, and bacterial evolution.</p>","PeriodicalId":9090,"journal":{"name":"Brazilian Journal of Microbiology","volume":"57 1","pages":""},"PeriodicalIF":2.1,"publicationDate":"2026-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13400510/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148576972","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Upstream endotoxin removal in recombinant protein purification: an optimised pre-cell disruption strategy for reducing endotoxin contamination of inclusion bodies in Escherichia coli.","authors":"Shahin Hadadian, Mina Sepahi, Mohaddeseh Sheikh Abbasi, Samira Komijani","doi":"10.1007/s42770-026-02015-6","DOIUrl":"10.1007/s42770-026-02015-6","url":null,"abstract":"<p><p>Endotoxin contamination remains a critical challenge in recombinant protein production using Gram-negative bacteria, particularly for intracellularly expressed, positively charged proteins prone to lipopolysaccharide (LPS) binding. This study introduces an efficient upstream strategy for LPS removal before the cell disruption, aimed at reducing the endotoxin contamination load in the downstream purification process. A recombinant cationic protein (named as 4mer-S3-DP) was expressed in Escherichia coli (E. coli) BL21 (DE3), and biomass was treated with varying concentrations and incubation times of Tris-HCl and EDTA, as defined by a central composite design (CCD) of response surface methodology (RSM). Endotoxin levels in inclusion bodies (IBs) were quantified using a chromogenic Limulus Amebocyte Lysate (LAL) assay, and statistical modelling was performed to optimize treatment conditions. The final quadratic model demonstrated high predictive accuracy, and response surface analysis revealed that both insufficient and excessive treatment conditions could increase endotoxin contamination. Optimal conditions, consisting of 70 mM Tris-HCl for 30 min and 60 mM EDTA for 5 min, achieved a 2.7-log reduction in endotoxin levels (99.8%) without compromising protein integrity, as confirmed by SDS-PAGE analysis. Experimental validation of the optimum criteria predicted by this model showed no significant difference between predicted and observed endotoxin levels of IBs. This upstream intervention as a simple and effective endotoxin removal approach before the cell lysis could be adaptable to other recombinant proteins expressed in Gram-negative bacterial.</p>","PeriodicalId":9090,"journal":{"name":"Brazilian Journal of Microbiology","volume":"57 1","pages":""},"PeriodicalIF":2.1,"publicationDate":"2026-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13396096/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148577130","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Association of D-limonene and nisin: the action on vegetative cells and spores of Alicyclobacillus spp. in processed orange juice.","authors":"Jean Lopes da Silva, Emilly Brito Ferreira, Laines Cassiano Sumera, Maria Fernanda Giugiolli, Evandro Bona, Danielle Lazarin Bidóia, Fabrícia Gimenes, Jane Martha Graton Mikcha, Beatriz Cervejeira Bolanho Barros, Benício Alves de Abreu Filho","doi":"10.1007/s42770-026-02024-5","DOIUrl":"10.1007/s42770-026-02024-5","url":null,"abstract":"<p><p>Contamination by Alicyclobacillus spp. is a challenge in the juice and acidic beverage industry, since it can cause spoilage and, consequently, economic losses. Natural preservatives are interesting strategies as an alternative search for microbial growth control in food, in particular the use of D-limonene (DL) and nisin (Nis). This study aimed to evaluate the antibacterial activity of the DL combination and Nis against five strains of Alicyclobacillus spp. A. acidoterrestris (CBMAI 0244^T); A. herbarius (CBMAI 0246^T); A. acidiphilus (CBMAI 0247^T); A. hesperidum (CBMAI 0298^T) and A. sendaiensis (KCTC 3843^T). The interaction among the compounds was initially determined in culture medium by the checkerboard method, and the efficacy was evaluated by disk diffusion tests, Minimum Inhibitory Concentration (MIC), and Minimum Bactericidal Concentration (MBC). Subsequently, the combination was evaluated in orange juice to determine the Minimum Sporicidal Concentration (MSC). The checkerboard results against A. acidoterrestris revealed synergism, with a Fractional Inhibitory Concentration Index (FICI) of 0.07. The combination (DL + Nis) demonstrated larger inhibition halos (24-26 mm) compared to DL (10-11 mm) and Nis (20-24 mm) alone. The MIC values of the combination were reduced, ranging from 0.12 to 3.90 µg/mL which means it was bactericidal for all strains. In reconstituted orange juice, the combination was effective, with a MSC of 0.48 µg/mL against A. acidoterrestris spores. Electron microscopy revealed significant morphological damage and signs of cell lysis in treatments with the combination, suggesting a loss of membrane integrity consistent with the observed synergistic effect. Additionally, DL showed antioxidant activity of 0.23 µmol Trolox/100 g (ABTS), 91.67 µmol Trolox/100 g (FRAP), and electronic nose (E-nose) analysis demonstrated that the combination did not significantly alter the volatile profile of the juice. Therefore, the combination of D-limonene and nisin represents a natural alternative with potential for the control of Alicyclobacillus spp. in commercial orange juice.</p>","PeriodicalId":9090,"journal":{"name":"Brazilian Journal of Microbiology","volume":"57 1","pages":""},"PeriodicalIF":2.1,"publicationDate":"2026-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13396087/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148577025","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}