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[This retracts the article DOI: 10.1007/s13205-021-02811-x.].
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Acrylamide, a Maillard reaction product, formed in fried food poses a serious concern to food safety due to its neurotoxic and carcinogenic nature. A "Green Approach" using L-Asparaginase enzyme from GRAS-status bacteria synergized with hydrocolloid protective coating could be effective in inhibiting acrylamide formation. To fill this void, the present study reports a new variant of type-II L-asparaginase (AsnLb) from Levilactobacillus brevis NKN55, a food-grade bacterium isolated using a unique metabolite profiling approach. The recombinant AsnLb enzyme was characterized to study acrylamide inhibition ability and showed excellent specificity towards L-asparagine (157.2 U/mg) with Km, Vmax of 0.833 mM, 4.12 mM/min respectively. Pretreatment of potato slices with AsnLb (60 IU/mL) followed by zein-pectin nanocomplex led to >70% reduction of acrylamide formation suggesting synergistic effect of this dual component system. The developed strategy can be employed as a sustainable treatment method by food industries for alleviating acrylamide formation and associated health hazard in fried foods.
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Acrilamida , Asparaginase , Coloides , Pectinas , Zeína , Asparaginase/química , Asparaginase/metabolismo , Acrilamida/química , Pectinas/química , Zeína/química , Coloides/química , Solanum tuberosum/química , CulináriaRESUMO
OBJECTIVE: The main purpose of the present study was to evaluate the therapeutic efficacy of Aloe vera-coated curcumin encapsulated nanoparticles in mitigating Alzheimer's disease progression in mice, by examining behavioural changes, biochemical markers, and histopathological alterations, thus elucidating its potential as a treatment strategy. METHODS: The green synthesis method was used to synthesise this nanoformulation, which was then characterised using a variety of techniques, including percentage encapsulation efficacy, UV-visible spectroscopy, DLS, FT-IR, FESEM, and EDX. Several in-vivo assessments, including behavioural evaluations, dose optimisation studies, oxidative stress marker estimation, and histological studies, were conducted to determine the potential therapeutic impact of nanoformulation on the Alzheimer-induced mice model. RESULTS: The synthesised nanoparticles show a mean diameter of 76.12 nm ±1.23, a PDI of 0.313 ± 0.02, a zeta potential of 6.27 ± 0.65 mV, and the percentage encapsulation efficiency between 90% and 95% indicating good stability of synthesised nanoformulation. With the help of Morris water maze, Y-maze, and novel object recognition assay, the learning capacity and memory were assessed, and the results show that the synthesised nanoformulation significantly decreased the transfer latency to reach baited arm or to the hidden platform within 7 days. CONCLUSION: The formulation demonstrated significant biochemical benefits and remarkable cognitive advantages, establishing it as a prospective therapeutic intervention option that is both safe and effective.
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Aloe , Doença de Alzheimer , Curcumina , Modelos Animais de Doenças , Nanopartículas , Animais , Curcumina/farmacologia , Curcumina/administração & dosagem , Curcumina/química , Aloe/química , Doença de Alzheimer/tratamento farmacológico , Camundongos , Nanopartículas/química , Masculino , Comportamento Animal/efeitos dos fármacos , Aprendizagem em Labirinto/efeitos dos fármacosRESUMO
Camels, known as the "Ship of the Desert," play a vital role in the ecosystems and economies of arid and semi-arid regions. They provide meat, milk, transportation, and other essential services, and their resilience to harsh environments makes them invaluable. Despite their similarities, camel breeds exhibit notable differences in size, color, and structure, with over 40 million camels worldwide. This number is projected to increase, underscoring their growing significance. Economically, camels are crucial for food production, tourism, and trade, with camel racing being particularly significant in Arab countries. Their unique physiological traits, such as low disease susceptibility and efficient water conservation, further enhance their value. Camel products, especially meat and milk, offer substantial nutritional and therapeutic benefits, contributing to their high demand. Genetic diversity studies have advanced our understanding of camels' adaptation to extreme environments. Functional genomics and whole-genome sequencing have identified genes responsible for these adaptations, aiding breeding programs and conservation efforts. High-throughput sequencing has revealed genetic markers linked to traits like milk production and disease resistance. The development of SNP chips has revolutionized genetic studies by providing a cost-effective alternative to whole-genome sequencing. These tools facilitate large-scale genotyping, essential for conserving genetic diversity and improving breeding strategies. To prevent the depletion of camel genetic diversity, it is crucial to streamline in situ and ex situ conservation efforts to maintain their ecological and economic value. A comprehensive approach to camel conservation and genetic preservation, involving advanced genomic technologies, reproductive biotechniques, and sustainable management practices, will ensure their continued contribution to human societies.
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This study explored the genetic diversity and evolutionary history of riverine and swamp buffaloes in India, utilizing complete mitochondrial genome sequences. Through comprehensive sampling across varied agro-climatic zones, including 91 riverine buffaloes from 12 breeds and 6 non-descript populations, along with 16 swamp buffaloes of the Luit breed, this study employed next-generation sequencing techniques to map the mitogenomic landscape of these subspecies. Sequence alignments were performed with the buffalo mitochondrial reference genome to identify mitochondrial DNA (mtDNA) variations and distinct maternal haplogroups among Indian buffaloes. The results uncovered the existence of 212 variable sites in riverine buffaloes, yielding 67 haplotypes with high haplotype diversity (0.991), and in swamp buffaloes, 194 variable sites resulting in 12 haplotypes, displaying haplotype diversity of 0.950. Phylogenetic analyses elucidated the genetic relationships between Indian buffaloes and the recognized global haplogroups, categorizing Indian swamp buffaloes predominantly into the SA haplogroup. Intriguingly, the haplogroup SB2b was observed for the first time in swamp buffaloes. Conversely, riverine buffaloes conformed to established sub-haplogroups RB1, RB2, and RB3, underscoring the notion of Northwestern India as a pivotal domestication site for riverine buffaloes. The study supports the hypothesis of independent domestication events for riverine and swamp buffaloes, highlighting the critical role of genetic analysis in unraveling the complex evolutionary pathways of domestic animals. This investigation contributes to the global understanding of buffalo mitogenome diversity, offering insights into this important livestock species' domestication and dispersal patterns.
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Búfalos , Variação Genética , Genoma Mitocondrial , Haplótipos , Filogenia , Animais , Búfalos/genética , Índia , DNA Mitocondrial/genética , Feminino , Herança MaternaRESUMO
Camels play a crucial socio-economic role in sustaining the livelihoods of millions in arid and semi-arid regions. They possess remarkable physiological attributes which enable them to thrive in extreme environments, and provide a source of meat, milk and transportation. With their unique traits, camels embody an irreplaceable source of untapped genomic knowledge. This study introduces Axiom-MaruPri, a medium-density SNP chip meticulously designed and validated for both Camelus bactrianus and Camelus dromedarius. Comprising of 182,122 SNP markers, derived from the re-sequenced data of nine Indian dromedary breeds and the double-humped Bactrian camel, this SNP chip offers 34,894 markers that display polymorphism in both species. It achieves an estimated inter-marker distance of 14 Kb, significantly enhancing the coverage of the camel genome. The medium-density chip has been successfully genotyped using 480 camel samples, achieving an impressive 99 % call rate, with 96 % of the 182,122 SNPs being highly reliable for genotyping. Phylogenetic analysis and Discriminant Analysis of Principal Components yield clear distinctions between Bactrian camels and dromedaries. Moreover, the discriminant functions substantially enhance the classification of dromedary camels into different breeds. The clustering of various camel breeds reveals an apparent correlation between geographical and genetic distances. The results affirm the efficacy of this SNP array, demonstrating high genotyping precision and clear differentiation between Bactrian and dromedary camels. With an enhanced genome coverage, accuracy and economic efficiency the Axiom_MaruPri SNP chip is poised to advance genomic breeding research in camels. It holds the potential to serve as an invaluable genetic resource for investigating population structure, genome-wide association studies and implementing genomic selection in domesticated camelid species.
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Camelus , Análise de Sequência com Séries de Oligonucleotídeos , Polimorfismo de Nucleotídeo Único , Animais , Camelus/genética , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Filogenia , Domesticação , Cruzamento/métodos , Genótipo , Técnicas de Genotipagem/métodosRESUMO
The current study attempts to investigate the differences in gene expression in longissimus thoracis muscles between sheep breeds acclimated to diverse environments. Changthangi sheep inhabits the cold arid plateau of Ladakh, at an altitude above 3000 m with prevalence of rarefied atmosphere. Muzzafarnagri sheep, on the other hand is found in the sub-tropical hot and humid plains at an altitude of about 250 m. Comparative transcriptomics was used to provide a molecular perspective of the differential adaptation of the two breeds. RNA sequencing data was generated from four biological replicates of the longissimus thoracis muscles from both breeds. The common genes expressed in both breeds were involved in muscle contraction and muscle fibre organization. The most significant pathways enriched in Changthangi muscles were glycogen metabolism, reduction of cytosolic Ca++ levels and NFE2L2 regulating anti-oxidant, while those in Muzzafarnagri were extracellular matrix organization and collagen formation. The hub genes identified in Changthangi were involved in hematopoiesis and HIF signaling pathway, suggesting the molecular acclimatization of Changthangi to the high altitude cold desert of Ladakh. The nodal genes discovered in Muzzafarnagri sheep were associated with the extracellular matrix which accentuates its significance in the development, growth and repair of muscles. The observed transcriptomic differences underscore the morphological and adaptive disparity between the two breeds. The candidate genes and pathways identified in this study will form the basis for future research on adaptation to high altitude and body size in small ruminants.
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Ladakh, one of the highest inhabited regions globally, hosts the unique Changthangi goat, renowned for producing Pashmina, the world's most luxurious natural fiber. In comparison, the fiber derived from Changthangi sheep is considered next only to Pashmina. This research endeavors to compare the skin transcriptome profiles of Changthangi goats and Changthangi sheep, aiming to discern the molecular determinants behind the recognition of Changthangi goats as the source of Pashmina. Drawing upon previously conducted studies, a collective of 225 genes correlated with fiber characteristics were extracted from the differentially expressed genes noticed between the two species (p-value of ≤ 0.05 and a log2 fold change of ≥ 1.5). These genes were analyzed using DAVID software to understand their biological functions and to identify enriched KEGG and Reactome pathways. The protein-protein interaction networks were constructed using Cytoscape, cytoHubba, and STRING to focus on key genes and infer their biological significance. Comparative transcriptome analysis revealed significantly higher expression of genes involved in signaling pathways like Wnt, MAPK, PI3K-Akt, Hedgehog, associated with fiber development and quality in Changthangi goats. These pathways play crucial roles in hair follicle (HF) formation, maintenance of epidermal stem cells, and fiber characteristics. Findings also highlight the enrichment of cell adhesion molecules and ECM-receptor interaction, emphasizing their roles in HF structure, growth, and signaling. This investigation offers an in-depth understanding of the molecular intricacies governing Pashmina production in Changthangi goats, providing valuable insights into their unique genetic makeup and underlying mechanisms influencing the exceptional quality of Pashmina fibers.
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Perfilação da Expressão Gênica , Cabras , Pele , Transcriptoma , Animais , Cabras/genética , Cabras/metabolismo , Pele/metabolismo , Ovinos/genética , Ovinos/metabolismo , Mapas de Interação de Proteínas/genética , Transdução de Sinais/genética , Lã/metabolismo , Fibra de LãRESUMO
BACKGROUND: Quantitative real-time PCR (qPCR) is a highly reliable method for validating gene expression data in molecular studies due to its sensitivity, specificity, and efficiency. To ensure accurate qPCR results, it's essential to normalize the expression data using stable reference genes. METHODS: This study aimed to identify suitable reference genes for qPCR studies in goats by evaluating 18 candidate reference genes (ACTB, BACH1, B2M, GAPDH, HMBS, HPRT1, PGK1, PPIA, PPIB, RPLP0, RPL19, RPS9, RPS15, RPS28, SDHA, TBP, UXT, and YWHAZ) in 10 different caprine tissues (heart, intestine, kidney, liver, lung, muscle, rumen, skin, spleen, and testis). An integrated tool called RefFinder, which incorporates various algorithms like NormFinder, GeNorm, BestKeeper, and ΔCt, was used to assess the stability of expression among these genes. RESULTS: After thorough analysis, ACTB, PPIB, and B2M emerged as the most stable reference genes, while RPL19, RPS15, and RPS9 were found to be the least stable. The suitability of the selected internal control genes was further validated through target gene analysis, confirming their efficacy in ensuring accurate gene expression profiling in goats. CONCLUSION: The study determined that the geometric average of ACTB, PPIB, and B2M creates an appropriate normalization factor for gene expression studies in goat tissues.
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Perfilação da Expressão Gênica , Cabras , Masculino , Animais , Cabras/genética , Cabras/metabolismo , Perfilação da Expressão Gênica/métodos , Algoritmos , Coração , Reação em Cadeia da Polimerase em Tempo Real/métodos , Padrões de ReferênciaRESUMO
Quantitative PCR (qPCR) is a widely-used technique for quantifying the expression of target genes across various tissues, as well as under different pathological and physiological conditions. One of the challenges associated with this method is the need to identify optimal reference genes (RGs) that maintain consistent expression levels under diverse experimental settings, thereby ensuring accurate biological interpretation. In this study, we conducted a thorough analysis of 18 candidate RGs (ACTB, BACH1, B2M, GAPDH, HMBS, HPRT1, PGK1, PPIA, PPIB, RPLP0, RPL19, RPS9, RPS15, RPS28, SDHA, TBP, UXT, and YWHAZ) across 10 ovine tissues (muscle, skin, kidney, liver, intestine, rumen, lung, testis, heart, and spleen) obtained from five individual sheep. We aimed to identify genes with stable expression across these tissues. A literature-based survey helped us shortlist candidate genes representing various functional classes from multiple livestock species. We employed four algorithms: geNorm, NormFinder, BestKeeper, and Delta Ct (ΔCt), to rank these genes based on their stability. A consistent trend in the rankings was observed across these different algorithms. RefFinder was then used for a comprehensive ranking, integrating the outputs from the various methods. ACTB, PPIB, BACH1, and B2M emerged as the most stable RGs, while RPS9, RPS15, and PGK1 displayed variable expression. We validated our findings through qPCR analysis of four target genes (ACTN2, CRYAB, DLK1, and TRIM54) in the skin samples from two different sheep breeds. Based on these results, we recommend ACTB, PPIB, BACH1, and B2M as reliable internal control genes for qPCR experiments involving diverse ovine tissues.
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Algoritmos , Gliceraldeído-3-Fosfato Desidrogenases , Masculino , Animais , Ovinos/genética , Coração , Reação em Cadeia da Polimerase em Tempo Real/métodos , Testículo , Perfilação da Expressão Gênica/métodos , Padrões de ReferênciaRESUMO
Human skull has always been used for victim identification in forensic odontology. The gender-dimorphic bone of the skull is the mandible. The gonial angle has frequently been investigated for gender estimation with variable results and requires further exploration. We aim to compare the efficacy of gonial angle estimation by ancient methods of lateral cephalometric tracing compared with more recent digital analysis methods for gender estimation in the Indian population. Lateral cephalograms of 191 (96 M and 95F) cases above the age of 17 years were retrieved. Cephalometric analysis of gonial angle on radiographs was done using both manual cephalometric tracing method and digitally using Adobe Photoshop software. The results were subjected to statistical analysis for evaluation. The mean gonial angle was higher in females (125.05; 123.77 and 125.28) than in males (122.583; 121.715 and 122.008) using both manual and digital methods. On applying the logistical regression analysis (LRA), the digital method showed the highest gender estimation accuracy of 60.7% followed by Burstone's analysis (57.1%) and manual conventional analysis (56.5%). Burstone's analysis (57.9%) correctly identified increased females, whereas digital analysis (62.5%) and manual conventional analysis (59.4%) accurately recognised increased males. The present study showed a higher gender estimation accuracy using digital methods as compared to manual methods, but it still lacks the credibility to be used as a sole factor for predicting the gender of an individual. Hence, a cumulative factor must be taken into consideration for gender identification which would provide more promising results.
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Mandíbula , Crânio , Masculino , Feminino , Humanos , Adolescente , Crânio/diagnóstico por imagem , Cefalometria/métodos , Software , Povo AsiáticoRESUMO
The peculiarity of Indian cattle lies in milk quality, resistance to diseases and stressors as well as adaptability. The investigation addressed selection signatures in Gir and Tharparkar cattle, belonging to arid ecotypes of India. Double digest restriction-site associated DNA sequencing (ddRAD-seq) yielded nearly 26 million high-quality reads from unrelated seven Gir and seven Tharparkar cows. In all, 19,127 high-quality SNPs were processed for selection signature analysis. An approach involving within-population composite likelihood ratio (CLR) statistics and between-population FST statistics was used to capture selection signatures within and between the breeds, respectively. A total of 191 selection signatures were addressed using CLR and FST approaches. Selection signatures overlapping 86 and 73 genes were detected as Gir- and Tharparkar-specific, respectively. Notably, genes related to production (CACNA1D, GHRHR), reproduction (ESR1, RBMS3), immunity (NOSTRIN, IL12B) and adaptation (ADAM22, ASL) were annotated to selection signatures. Gene pathway analysis revealed genes in insulin/IGF pathway for milk production, gonadotropin releasing hormone pathway for reproduction, Wnt signalling pathway and chemokine and cytokine signalling pathway for adaptation. This is the first study where selection signatures are identified using ddRAD-seq in indicine cattle breeds. The study shall help in conservation and leveraging genetic improvements in Gir and Tharparkar cattle.
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Genoma , Polimorfismo de Nucleotídeo Único , Feminino , Bovinos/genética , Animais , Polimorfismo de Nucleotídeo Único/genética , Fenótipo , Índia , ReproduçãoRESUMO
Introduction: The use of platelet-rich plasma (PRP) has gained popularity in orthodontics in the past decade. PRP is an autologous concentration of platelets rich in growth factors and is hypothesized to play a role in orthodontic tooth movement (OTM) due to its bone remodeling capacity. Objective: The aim of this study was to determine the effect of PRP on the rate of tooth movement in humans. Materials and Methods: Fourteen patients requiring bilateral extraction of upper first premolars were included in the study. PRP (×4) was prepared and injected in the alveolar mucosa, distal to canine on experimental sides. Canine retraction was initiated on the same day of PRP injection using NiTi closed coil springs on both sides. The amount of canine retraction was measured and compared between both sides at the 4th, 8th, 12th, and 16th weeks by measuring the distance between lateral incisor and canine on the cast. Results: Results showed mean displacement of 1.355 mm, 1.232 mm, 1.191 mm, and 1.085 mm on experimental side and 1.265 mm, 1.126 mm, 1.031 mm, and 0.879 mm on control side at the 4th, 8th, 12th, and 16th weeks, respectively. Although increased OTM (13.85% or 1.1 times faster) was observed on experimental side at all time intervals compared to the control group, the result was not statistically significant. Conclusion: 4× concentration of PRP does not accelerate OTM significantly.
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This study explored the maternal genetic diversity in the pig genetic resources of India by analyzing a mitochondrial D-loop fragment and comparing it with the corresponding sequences of previously published studies involving domestic pigs and wild boars. Sequencing of 103 samples representing different domestic pig populations revealed existence of 32 maternal haplotypes. The indices of haplotype and nucleotide diversity in Indian domestic pigs were 0.9421 and 0.015, respectively. Median-Joining network revealed that Indian pigs belong to Clade A and show conformity to 6 haplogroups reported worldwide (D1a, D1a1, D1a2, D1e, D1h and D3a). Among these, D1e and D1a2 were shared with Asian wild boars too. Interestingly, haplotype sharing was evident between Indian pigs and samples from other countries representing Africa, Asia, Europe and Oceania. This study substantiates India's contribution as a possible pig domestication center and highlights the importance of the Indian subcontinent in dispersal of the species to other continents. Additionally, genetic evidence suggested the influence of trading routes and historical interactions in shaping pig genetic exchange. Overall, this investigation provides valuable insights into the genetic diversity, historical migration, and domestication of Indian domestic pigs, contributing to the broader understanding of global pig genetic resources and their evolutionary history.
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Domesticação , Sus scrofa , Suínos/genética , Animais , Sus scrofa/genética , Índia , Mitocôndrias/genética , Haplótipos/genética , Filogenia , Variação Genética/genética , DNA Mitocondrial/genéticaRESUMO
Backyard poultry farming contributes to food security, nutrition, and the regular income of rural farmers in India. Their products have a niche market here and fetch higher prices than those of commercial poultry. Improved varieties are being developed to overcome the slow growth, late sexual maturity, and low production of indigenous breeds, while retaining their positive attributes. A comprehensive study was conducted to analyze the functional attributes of meat from the Jabalpur color (JBC), a colored, improved dual-purpose synthetic line, developed by Nanaji Deshmukh Veterinary Science University, Jabalpur, India. The birds were managed in a deep litter system under a backyard type of housing (night shelter and free range). Primal meat cuts (breast and thigh) of the male birds (n = 20/group) were evaluated at the age of marketing. The corresponding attributes were compared with the results obtained for commercial Cobb (400) broilers. The protein concentration of JBC breast (25.65 ± 0.39 g/100 g of tissue) and thigh (19.04 ± 0.23 g/100 g of tissue) meat was superior (p ≤ 0.05) to that of Cobb broilers. Established assays (in vitro) identified a better (p ≤ 0.05) antioxidation capacity in the JBC meat. High-performance liquid chromatography confirmed a considerable quantity of functional biomolecules (carnosine, anserine, and creatine) in the JBC breast and thigh meat extracts. The average carnosine concentration (mg/g of tissue) was 2.66 ± 0.09 and 1.11 ± 0.04 in the JBC breast and thigh meat, respectively. The mRNA expression was quantified by qRT-PCR for the carnosine-related genes: ß-alanine transporter (SLC36A1), carnosine-synthesizing enzyme (CARNS1), and carnosine-degrading enzyme (CNDP2); this explained the comparable carnosine in the JBC and Cobb meat. Meat extracts from both genetic groups (JBC and Cobb) had high anti-glycation potential. Higher protein content and antioxidant capacity, along with the bioactive dipeptides in the JBC meat, herald exciting research opportunities for its use in improving the traditional backyard poultry farming system.
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To contribute to the knowledge of maternal genetic diversity in domestic donkeys, this study investigated the mitochondrial DNA variations and analyzed the genetic structure in Indian donkeys based on 31 mitogenome sequences representing four breeds/populations (Agra, Halari, Kachchhi and Spiti). A total of 27 haplotypes with a haplotype diversity value of 0.989 were evident in the donkey genetic resources of India. The genetic differentiation between the investigated populations was evaluated using population pairwise FST values, which showed maximum differentiation between Kachchhi and Halari donkeys. The Neighbor-Joining (NJ) tree based on the whole mitogenome sequence and the Median-Joining (MJ) network for partial D-loop fragment showed clear demarcation of Indian donkeys into Nubian and Somali clades, substantiating African maternal origin of Indian domestic donkeys. The topology of the MJ network excluded the Asian wild asses as the possible progenitors of Indian donkeys. Halari and Agra donkeys showed conformity exclusively to the Nubian lineage of the African wild asses. However, representation of both the Nubian and Somali lineages was observed in Kachchhi and Spiti donkeys. Comprehensive analysis carried out by retrieving D-loop sequences from different countries representing Asia, Africa, Europe and South America revealed existence of shared haplotypes across geographically isolated regions of the globe. This observation is indicative of utility of donkeys as pack animals across inter-continental trading routes during development of human civilizations. Our results represent a valuable contribution to maternal genetic diversity of Indian donkeys and provide insights into the worldwide spread of the species following initial domestication in Africa.
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DNA Mitocondrial , Equidae , Animais , Humanos , Equidae/genética , Filogenia , DNA Mitocondrial/genética , África , Domesticação , Haplótipos , Variação GenéticaRESUMO
RNA sequencing-based expression profiles from pectoralis major muscles of black meat (Kadaknath) and white meat (broiler) chicken were compared to identify differentially expressed genes. A total of 156 genes with log2 fold change ≥ ± 2.0 showed higher expression in Kadaknath and 68 genes were expressed at a lower level in comparison to broiler. Significantly enriched biological functions of up-regulated genes in Kadaknath were skeletal muscle cell differentiation, regulation of response to reactive oxygen, positive regulation of fat cell differentiation and melanosome. Significant ontology terms up-regulated in broiler included DNA replication origin binding, G-protein coupled receptor signaling pathway and chemokine activity. Highly inter-connected differentially expressed genes in Kadaknath (ATFs, C/EPDs) were observed to be important regulators of cellular adaptive functions, while in broiler, the hub genes were involved in cell cycle progression and DNA replication. The study is an attempt to get an insight into the transcript diversity of pectoralis major muscles of Kadaknath and broiler chicken. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-023-03682-0.