Aspergillus species consists of a group of opportunistic fungi that is virulent when the immunity of the host is compromised. Among the various species, Aspergillus fumigatus is the most prevalent species. However, the prevalence of fungal infections caused by non-fumigatus Aspergillus has been increasing. Polyenes, echinocandins and azoles are the three main classes of antifungal agents being used for the treatment of aspergillosis. Nevertheless, the incidence of resistance towards these three classes has been rising over the years among several Aspergillus spp. The side effects associated with these conventional antifungal agents have also limited their usage. This urges the need for the discovery of a safe and effective antifungal agent, which presents a major challenge in medicine today. Plants present a rich source of bioactive molecules which have been proven effective against a wide range of infections and conditions. Therefore, this present review intends to examine the current literature available regarding the efficacy and mechanism of action of plant extracts and their compounds against Aspergillus spp. In addition, novel drug delivery systems of plant extracts against Aspergillus spp. were also included in this review. Aspergillus, Candida, Cryptococcus, Pneumocystis jirovecii and endemic dimorphic fungi. In general, humans are relatively resistant to invasive fungal diseases. However, in immunocompromised patients and HIV-infected patients, the susceptibility toward opportunistic fungal infections is increased. Candida spp. are the most common isolates from immunocompromised patients, but Aspergillus or Cryptococcus may also be isolated from these patients [Fungal diseases were estimated to cause about 1.6 million deaths per year while more than 1 billion people were diagnosed with severe fungal diseases . Fungi cpatients . This capatients ,8.Aspergillus spp. is one of the most common causes of life-threatening fungal infection, especially in immunocompromised patients. Most infections involving Aspergillus spp. are caused by Aspergillus fumigatus; however, infections caused by non-fumigatus Aspergillus species, such as Aspergillus flavus, Aspergillus niger and Aspergillus terreus, have also been identified [entified ,10. Due entified .At present, three types of antifungal agents are used by clinicians for the treatment of fungal infections, namely polyenes, echinocandins and azoles. Resistance has been an issue due to the limited antifungal agents available while some fungi can be resistant to all three types of antifungal agents ,13. In aCandida spp. [Schistosoma mansoni [Aspergillus spp.Natural products play an important role in medical therapy. From January 1981 to September 2019, 1,881 new drugs were approved of which natural products constitute 4.6% while drugs derived from natural products constitute 18.9% . Many na mansoni . The useAspergillus infections are among the most common fungal diseases worldwide. There are several types of Aspergillus infections that have been diagnosed, which include but are not limited to allergic bronchopulmonary aspergillosis, chronic pulmonary aspergillosis, aspergillus spondylitis, invasive aspergillosis and cerebral aspergillosis. Aspergillus infections can infect both healthy and immunocompromised individuals, although infections are more common in the immunocompromised, and are often associated with a bad prognosis [Aspergillus spp., leading to COVID-19-associated pulmonary aspergillosis (CAPA) [Aspergillus infections [Aspergillus infections were estimated to be $1,254,833,662 in the United States in 2017 [Aspergillus infections  were also the second highest among all the other fungal diseases [rognosis . Invasivs (CAPA) . Patientfections ,7,8. Recfections . The tot in 2017 . The totdiseases .Aspergillus infection is caused by Aspergillus spp., which consists of more than 250 species, with fewer than 40 species being associated with infections in humans. A. fumigatus is the main species causing infection in humans and is the main contributor to death among infected patients. However, non-fumigatus Aspergillus species such as A. flavus, A. niger and A. terreus have also been isolated from patients with invasive aspergillosis [A. fumigatus remains the major pathogenic agent as compared to other species is unclear; however, it could be associated with its resilience in withstanding a wide range of environmental conditions in comparison to other species. In this context, A. fumigatus is able to tolerate a wide range of temperature and pH in which it can be isolated from decaying vegetation and soil with a temperature range of 12 °C to 65 °C and pH of 2.1 to 8.8 [A. fumigatus to survive the degradation of polysaccharides from plant cell walls and nitrogen sources acquired from the degradation of proteinaceous substrates [A. fumigatus to survive in the environment; however, its association with the enhanced pathogenic potential as compared to the other species will require further exploration and evidence.1 to 8.8 . In addibstrates ,27. ThesAspergillus infections, the three main classes of antifungal agents used are azoles, polyenes and echinocandins [Despite the discovery of several antifungal agents for the treatment of mycoses, their effectiveness is largely dependent on the ability to distinguish between the host and pathogenic cells which are both eukaryotic, consequently determining their specificity . For theocandins .Aspergillus spp. towards azoles has also been reported. A 5-year follow-up study revealed that 4.1% and 14.5% of A. fumigatus isolates were resistant to voriconazole and itraconazole, respectively. Among the voriconazole-resistance isolates, 95% have a cross-resistance with itraconazole. Regardless, the resistance level is highly variable depending on the type of azole drug used, the clinical settings, the patient’s background and the underlying infection [A. fumigatus from clinical and environmental samples across Asia revealed the range of 3% in China, Japan and Kuwait to 33% in China [A. fumigatus has also been associated with a higher mortality rate [A. fumigatus on the watch list in the CDC’s antibiotic resistance threats report, a category that includes uncommon germs that potentially spread and cause significant morbidity and mortality [Azoles, in particular triazoles, are the preferred agents for the treatment and prevention of invasive aspergillosis in most patients . Azoles nfection . In addiin China . Azole-rity rate ,39. In 2ortality .Aspergillus infections [Aspergillus infection, its use is limited due to the high incidence of nephrotoxicity [A. fumigatus clinical strains, which could greatly contribute to treatment failure [The next group of antifungal agents are the polyenes, in which amphotericin B is used in the treatment of fections . A well-fections . Howeverfections . Amphotetoxicity . In 2018 failure .fks1) which is involved in the synthesis of β-1,3-glucan, a major polysaccharide in the cell wall of the fungi [Aspergillus spp. Due to the limited oral bioavailability, echinocandins are administered only via intravenous infusion, thus are limited only to patients admitted to the hospital [A. fumigatus caused by the point mutation in the fks1 gene has been reported, causing treatment failure [The last group of antifungal agents used in the treatment and prophylaxis of aspergillosis are echinocandins . Echinoche fungi . This clhe fungi . Howeverhospital . This cohospital . In 2017 failure .Streptomyces nodosus [Glarea lozoyensis, Coleophoma empetri and Aspergillus nidulans, respectively [Interestingly, many of the current antifungals originated from natural sources. Amphotericin B, which was discovered in 1956 was derived from the bacteria  nodosus . On the ectively . Thus, tThe development of resistance to the currently utilized antifungal drugs makes it crucial to discover new classes of antifungals from natural products including plants . VariousPlants are a valuable source of a wide variety of biologically active chemical defence constituents, the secondary metabolites  . Numerou50) in cytotoxicity tests implies that a higher quantity of the extract would be needed to cause a toxic response. Extracts with LC50 > 0.1 mg/mL are regarded as having negligible cytotoxicity and the American National Cancer Institute (NCI) refer to an LC50 of ≤0.03 mg/mL to be toxic [Minimum inhibitory concentration (MIC) is defined as the lowest concentration of a substance that inhibits microbial growth . With rebe toxic ,66. Selebe toxic  and Peñabe toxic  as highlAspergillus activity of plant extracts is shown in Plants are used extensively as crude materials and as sources of pure compounds. Various techniques such as Soxhlet extraction, maceration, hydro-distillation, percolation, decoction, reflux extraction, pressurized liquid extraction, supercritical fluid extraction, ultrasound assisted extraction, microwave assisted extraction, pulsed electric field extraction, and enzyme assisted extraction have been employed to extract the bioactive compounds from plant materials ,69. The A. flavus and A. ochraceous [A. flavus and A. ochraceous. The lowest MIC value of 0.08 mg/mL was reported for Markhamia obtusifolia against A. flavus and Curtisia dentata against A. ochraceous after 24 and 48 h of incubation, with the latter, further yielded the highest total activity of 1583 mL/g against A. ochraceous [C. dentata whereas neophytadiene, palmitic acid and 4-(1E)-3-hydroxy-1-propenyl)-2-methoxyphenol were the major constituents of M. obtusifolia. Another study by Singh et al. [Acacia raddiana heartwood extract and fractions. Benzene fractions showed the greatest effectiveness against A. niger  whereas ethyl acetate fractions exhibited the highest effectiveness against A. flavus . β-sitosterol, palmitic acid and neophytadiene from other plant species have also been reported to be effective against Aspergillus spp. [Twenty-five South African medicinal plants selected on the basis of their availability were investigated against three phytopathogenic fungal strains including hraceous . The acehraceous . Gas chrh et al.  has alsolus spp. ,72,73.Milletia grandis revealed the lowest MIC value of 0.01 mg/mL against A. ochraceous after 24 and 48 h incubation period [M. grandis, the organic extracts of Warburgia salutaris, Bauhinia galpinii, Harpephyllum caffrum and Solanum panduriforme gave low MIC values of 0.01–0.10 mg/mL, and are thus good candidates for further investigation of their relevant antimicrobial compounds. The high phenolic contents of the plants may be responsible for their bioactivity. Furthermore, these plant species were generally non-toxic to Bovine dermis (LC50 = 0.01 to 0.68 mg/mL) and Vero cells (LC50 = 0.01 to 0.59 mg/mL), and W. salutaris showed the best safety margin with SI value of 34 [Among the ten selected South African medicinal plants, the organic extract of n period . In addiue of 34 .Zygophyllum coccineum, an edible halophytic plant [Z. coccineum demonstrated potent antifungal activity against A. fumigatus with MIC values of 15.63, 1.95, 3.90 and 31.25 µg/mL for its aqueous-ethanolic extract, chloroform fraction, ethyl acetate fraction and n-butanol fraction, respectively. In addition to the significant antifungal and antioxidant activity, the extract and fractions also demonstrated cytotoxic effects with IC50 values of 3.47, 3.19 and 2.27 µg/mL against MCF-7 (breast cancer), HCT-116 (colon cancer) and HepG2 (liver cancer) cell lines, respectively. The anticancer activity of the extract was also validated by in silico receptor-binding predicted energy levels and receptor-site docking feasibility of the major constituents found in the extract [Thirty-eight secondary metabolites of flavonoids, stilbene, phenolic acids, alkaloids and coumarin classes were identified by liquid chromatography-electrospray ionization-time-of-flight mass spectrometry (LC-ESI-TOF-MS) in the aqueous-ethanolic extract and subsequent fractions of ic plant . Most of extract .A. fumigatus [Tripleurospermum auriculatum demonstrated the greatest antifungal activity , followed by Centarea pseudosinaica , Plectranthus arabicus , Plectranthus asirensis , Centarea sinaica , Calendula tripterocarpa  and Koelpinia linearis . The high phenolic and flavonoid contents of the plants may be responsible for the protection against oxidative stress and paraneoplastic symptoms caused by cancer [50 up to 5000 mg/kg) and sub-chronic toxicity tests revealed that the plants have no toxicity and are considered safe for human use.Phytochemical screening and determination of antimicrobial activities of the desert plants growing in Saudi Arabia have reported notable antifungal activity against umigatus . Tripleuy cancer . FurtherA. niger and 5.00-27.00 mm against A. fumigatus [Foeniculum vulgare Mill. Apiaceae, Withania coagulans (Stocks) Dunal. Solanaceae, Camellia sinensis L. Kuntze Theaceae and Elettaria cardamomum (L.) Maton. Zingiberaceae showed the highest antifungal activity against the tested Aspergillus spp. . A statistically significant antifungal effect was also reported for polyherbal mixtures A, D and F against A. niger, A. fumigatus and Fusarium graminearum compared to fluconazole, which indicates their antifungal efficacy as fluconazole is not effective against these tested fungi [A combination of several medicinal herbs for extra therapeutic benefits is known as polyherbalism and it has been used in Ayurvedic, Chinese and Unani medicines since ancient times . Multiplumigatus . Recipe ed fungi . The preed fungi .Aspergillus spp.Essential oils (EOs) are mixtures of volatile compounds obtained from the plants by hydro-distillation, steam distillation, dry distillation, or mechanical cold pressing of plants . They arThymus decussatus were tested against several fungi including A. niger [T. decussatus showed strong antifungal activity against A. niger, with an MIC value of 0.49 μg/mL and inhibition zone of 47 mm (HD) and 48 mm (MAE). GC-MS analysis revealed the presence of oxygenated monoterpenes as the main components of both EOs with carvacrol as the main compound (94.40% for HD and 75.91% for MAE). The strong antimicrobial activity of T. decussatus may also be attributed to another main compound, p-cymene (3.61% for HD and 16.98% for MAE), in addition to carvacrol which has been reported to show antimicrobial activity in past studies. EOs from Asteriscus graveolens have been reported to show fungicidal activity against Alternaria sp. and Penicillium expansum in previous studies [A. graveolens EO also demonstrated a good antifungal effect against Candida albicans, A. niger, A. flavus and Fusarium oxysporum. The MIC results showed that A. graveolens EO strongly inhibited Aspergillus spp. used for testing, recording MICs values of 24.50 μg/mL against A. niger and 23.74 μg/mL against A. flavus. The higher amounts of α-thujone (17.92%) and carvacrol (14.14%) revealed by GC-MS may be responsible for the antifungal activity of the EO.EOs obtained by hydro-distillation (HD) and microwave-assisted extraction (MAE) methods from A. niger . Both HD studies  and a re studies  showed tMurraya paniculata showed moderate antifungal to strong antifungal activity against A. niger (MIC = 0.2 mg/mL), A. fumigatus (MIC = 0.1 mg/mL) and A. parasiticum (MIC = 0.2 mg/mL) [M. paniculata EO is due to the synergistic action among its components, as implied by higher MIC and MFC values of β-caryophyllene compared with the EO. Since the ratios of the lethal concentration to the minimum inhibitory concentration (MFC/MIC) were all less than or equal to 4, the EO oil and β-caryophyllene demonstrated fungicidal action instead of fungistatic action. From the time–kill curve study, the EO showed a good and rapid dose-dependent fungicidal effect within 4-8 hours for A. niger. Furthermore, the SI value of 3.06 was obtained when testing against tumorous cells of hepatocytes and fibroblasts. The toxicity of EOs against fungi may be attributed to terpenes and phenolic compounds, with several underlying mechanisms such as disruption of cell membranes, causing leakage of cellular material, inhibition of mitochondrial ATPase and the electron transport chain [EO from the fresh leaves of 2 mg/mL) . Thirteert chain .The antifungal activities of the EOs are not distinguished against all strains and their effective time of action is limited by their high volatility. Furthermore, their features such as hydrophobicity, instability and possible toxicity may compromise their use . EncapsuSatureja montana and Origanum virens were formulated and tested against A. flavus. Initially, after the incubation for 7 days, the colony forming units per gram (CFU/g) values of pure EOs were lower than the niosome-encapsulated EOs. However, the pure EOs lost their antifungal effect over time and the fungal growth increased with higher CFU/g values after incubated for 21 days, in contrast to the decreased fungal growth and lower CFU/g values for niosome-encapsulated EOs [Origanum vulgare and Thymus capitatus were reported to be active against various fungal strains including A. fumigatus and A. flavus, with MIC and MFC values two to four times lower compared to the pure EOs, indicating the improved antifungal activity of the EOs by nanoencapsulation. Encapsulation of Cuminum cyminum EOs by chitosan-caffeic acid nanogel considerably improved its performance with a lower MIC against A. flavus compared with free EO under sealed condition (350 ppm vs. 650 ppm) and non-sealed condition (800 ppm vs. >1000 ppm) [Aspergillus spp. are presented in Niosome-encapsulated EOs of ated EOs . Poly(ε-000 ppm) . Encapsu000 ppm) . The antThe bioavailability of plant bioactives is limited by poor solubility, poor stability due to gastric and colonic acidity, poor metabolism by gut microflora, poor absorption across the intestinal wall and active efflux mechanism as well as first-pass metabolic effects, failing clinical studies . Novel dIllicium verum EO with anethole as the major compound has reported MIC value of 0.7 μL/mL against aflatoxigenic strain A. flavus LHP-PV-1 as well as promising free radical scavenging activity and favourable safety profile with high LD50 value (11257.14 μL/kg) [I. verum EO-loaded chitosan nanoparticles  were lower than the EO in addition to the lower value of minimum aflatoxin B1 inhibitory concentration (MAIC) against A. flavus LHP-PV-1. Strong antifungal, antiaflatoxigenic and antioxidant activities of the encapsulated EO suggest its use as a plant-based preservative and shelf-life enhancer of food items, which must be confirmed in future long-term studies.Nanocarrier delivery has gained great attention to overcome the physicochemical and pharmacokinetic limitations of phytochemicals, enhance controlled release as well as improve the efficacy of bioactivities . Illiciu4 μL/kg) . EnhanceM. oleifera extract against C. albicans and C. glabrata were reduced from 125 and 250 μg/mL, respectively, to 62.5 μg/mL by the synthesized M. oleifera bismuth nanoparticles (NPs), indicating a synergistic effect [A. niger and A. flavus (250 μg/mL) were higher compared with the plant extract (62.5 μg/mL). Another study also demonstrated the poorer antifungal activity of M. oleifera copper NPs against A. niger and A. flavus (125 μg/mL) compared with the M. oleifera leaves extract (62.5 μg/mL) [Aspergillus spp.The use of plant extracts in the green synthesis of metallic nanoparticles may produce new nanomaterial with more potent and/or novel biological activities due to the synergistic effect . Using rc effect . However5 μg/mL) . Thus, tAspergillus spp. poses a life threat to society, and particularly immunocompromised patients, since many strains have shown resistance towards the existing antifungal agents. Medicinal plants provide new hope in the fight against Aspergillus infections. An increased action to identify and isolate new bioactive compounds that may potentially be used as a new treatment against Aspergillus infections is imperative. In this context, a large number of plant extracts and their constituents have shown potential as a therapeutic alternative. However, the implementation of plant extracts as a part of the therapeutic regimen is still challenging. Although in vitro studies showed promising results, in vivo studies should be conducted in order to establish a valid correlation with the in vitro data. Standardisation of the active compound in the plant materials is also necessary to ensure a reproducible and consistent antifungal activity in each batch of production. Due to problems associated with the bioavailability of plant extracts, studies on the novel delivery system of the plant extracts are highly warranted to enhance the therapeutic potential of these extracts. Additionally, interactions among different plant extracts, compounds, or antifungal agents should also be studied to maximise the antifungal efficacy while minimizing toxicity. This review summarizes the information on plant extracts with potent anti-Aspergillus effects that can aid in a faster and more efficient process in the development of new plant-derived antifungal agents against Aspergillus infections.


Congenital absence of the inferior vena cava (IVC) triggers collateral vessel growth to drain the peripheries and abdominal organs. This causes venous stasis and increases the risk of deep vein thrombosis (DVT) and pulmonary embolism. Typically, patients with absent IVCs present before 30 years of age, with bilateral DVT symptoms triggered by intense exercise. The abnormality can remain undetected as computed tomography imaging is not usually performed. Due to the increased risk of clotting, these patients should be on life-long anticoagulation. Raising clinical awareness of this condition, to ensure appropriate investigations and treatment, is important. Abnormally developed inferior vena cavae (IVCs) affect ~4% of the population, including variants such as agenesis, hypoplasia, duplication and interruption , 2. SuchThe agenesis or absence of an IVC is rare, prevalence estimated 0.0005–1% . Causes An absent IVC necessitates collateral vessel formation to drain the abdominal organs and peripheries . These iA 24-year-old man presented to hospital with leg and hip pain, commencing during high-intensity interval training, an activity he regularly undertook. The pain originated in his right leg, spreading to the left and both hips. Additionally, he reported 1 day history of dyspnoea worse on exertion and associated dizziness. He denied symptoms of cough, fever, haemoptysis or pleuritic chest pain. He reported intentional weight loss of 3.5 stone over 1 year.His past medical history comprised of asthma. He took no regular medications and didn’t smoke. There was no reported family history. He works as a financial manager.Clinical observations were heart rate 103, respiratory rate 20, blood pressure 126/69 and oxygen saturations 100% on room air. Heart sounds were normal and chest was clear. There was no palpable lymphadenopathy. His right calf was swollen compared with the left and tender on palpation. Coordination, power and sensation of both legs were intact.An electrocardiogram (ECG) revealed normal sinus rhythm and no cardiovascular strain . A chestA computed tomography (CT) pulmonary angiogram and thorax–abdomen–pelvis showed extensive bilateral PEs  and 3. TA thrombosis screen including; paroxysmal nocturnal haemoglobinuria, thrombophila screen, anti-nuclear antibody, antiphospholipid antibodies, beta-2-glycoprotein level, anticardiolipin levels, Hep B&C, human immunodeficiency virus screen and JAK-2 mutation screening was negative.The gentleman was initially administered treatment-dose tinzaparin pending vascular MDT. Mechanical thrombectomy was considered but deemed not in his best interest due to the high risk of venous re-occlusion in the presence of persisting venous outflow obstruction. Lifelong anticoagulation was recommended. Given his haemodynamic stability throughout admission thrombolysis was deemed unnecessary. Anticoagulation with rivaroxaban caused rectal bleeding and he was switched to apixaban, which has been well tolerated. He remains asymptomatic at 1-year post-diagnosis, under follow-up from the haematology team.The IVC is made up of four parts, the hepatic, supra-renal, renal and infra-renal veins . The phrAn absent IVC could be due to perinatal thrombosis . One casWhile hypoplasia of an infra-renal IVC is a rare but well-documented anomaly, the CT in this case demonstrates a normal infra-renal IVC with a hypoplastic suprarenal IVC , 7. ThisTo our knowledge, there are three reported cases of PE with associated IVC abnormalities . The mecThere were 62 documented cases of abnormal IVC presenting with DVT as of 2010 . IntenseCurrent NICE guidelines do not require a CT for patients with DVT unless there is concern of cancer . Without


Fusarium graminearum causing head scab (HS) or head blight (HB) disease in wheat is one of the nasty fungi reported to cause significant grain quality and yield loss. Biological control using endophytic bacteria has emerged as a prospective option for containing fungal diseases in an environmentally benevolent, durable, and sustainable manner. In this regard, 112 endophytic bacilli were isolated from the anthesis stage (Zadok’s growth stage 65) from five different wheat genotypes with an aim to identify prospective antagonistic strains against F. graminearum. The molecular identity of the strains was confirmed by matching 16S rRNA sequences of bacterial strains with the gene sequences of type strains available in the National Center for Biotechnology Information database and reported 38 different species of Bacillus in all the five wheat cultivars. Further, it has been observed that only fourteen strains  displayed in-vitro antagonistic effect against Fusarium graminearum fungus. Furthermore, the three endophytic Bacillus strains showing the strongest antagonistic effect  under in-vitro antagonistic assay were selected for field experiments. In a two-year consecutive field study, a combination of three strains (B. clarus NOK09 + B. subtilis NOK33 + B. amyloliquefaciens NOK109) displayed a remarkable reduction in HS disease index by 81.47% and 77.85%, respectively. Polymerase chain reaction assay detected three genes  involved in antibiotic biosynthesis pathways. Additional attributes such as potassium solubilization, siderophore release, and hydrolytic enzyme  synthesis have been observed in these strains. Overall, the present study was successful in profiling endophytic bacilli and selecting the combination of effective antagonistic endophytic Bacillus strains that could be the best alternative for the sustainable and ecological sound management of HS disease in wheat under field conditions.  Fusariumgraminearum causing head scab (HS) or head blight (HB) disease in wheat is one of the nasty fungi reported to cause significant grain quality and yield loss. The major outbreaks of HS disease in wheat from major wheat-growing countries of the world, including Australia, Asia, Canada, Europe, and South America, have been well documented  and incubated at 37 ± 2 °C temperature for two days. An aliquot (100 µL) of remaining water after the last wash was inoculated on NA Petri plates to inspect the occurrence of different bacterial colonies for seven days. Different microscopy and biochemical tests  were also conducted in three replicates for preliminary confirmation of the Bacillus spp. [Healthy ear head samples were collected from wheat genotypes  to identify their nearest match. Molecular Evolutionary Genetics Analysis (MEGA) version 11 [Bacillus sp. available in National Center for Biotechnology Information  database. The gene sequences were deposited in the NCBI databank to get the gene accession numbers  produced by  Neumann . Amplifirsion 11  tool was numbers . The aliF. graminearum. Briefly, a 5 mm diameter segment of fungal mass of highly virulent F. graminearum NFG1 isolate was positioned in the mid-point of Petri plates amended with PDA: NA (1:1). Each strain (~5 × 108 cfu ml−1) was streaked in a straight line closer to the border of Petri plate. The inoculated plates were incubated at 30 ± 2 °C. F. graminearum inoculated Petri plate without endophytic strain, and neutral bacterial strain (Bacillus pumilus NOK68) served as a control. Data on the growth of F. graminearum was recorded in mm till the control plate (without endophytic strain) was completely filled. F. graminearum growth inhibition (%) by each endophytic strain was computed by the formula quoted by Sharma et al. [Dual-culture plate assay was used to confirm the antagonistic capabilities of endophytic bacterial strains against a et al. . The assBacillus strains that reflected maximum antagonistic action towards F. graminearum under in-vitro conditions were chosen for testing their bio-control potentialities against HS disease in susceptible wheat genotype (PBW 343) at Crop Protection Experiment Area, ICAR-Indian Institute of Wheat and Barley Research, Karnal, Haryana, India. Under each field treatment, wheat seeds were sown in six lines (each line of 3 m long) with 22.5 cm line spacing. All the experiments were planned in a randomized complete block design with five replicates per treatment. Different treatments comprised of antagonistic endophytic bacteria (~107 cfu ml−1) and without endophytic bacteria were used to inoculate 150 wheat ear heads  at anthesis time (Zadoks growth stage 65). The bacterial suspension (2 mL per wheat ear head) in each plot was applied by a compressed air sprayer in the late afternoon (at 4:00PM). The control treatment (T8) was only sprayed with highly virulent F. graminearum isolate (NFG1). An additional control treatment (T9) comprised of fungicide  sprayed alone. Field assessment of HS incidence and HS severity was made at the late milk stage. Disease data was recorded on 40 ear heads per replicate (200 ear heads treatment −1) as per the disease rating described by Stack and McMullen [Three McMullen . DiseaseChrome azurol (CAS) agar assay  was carrBacillus strains. The polymerase chain reaction temperature profiles and primers information given in Three endophytic strains  displaying strong antifungal activities (>70%) were tested for the presence of different AMP genes in The field experiments performed to check the antagonistic effects of selected bacteria were arranged in complete randomized block design (CRD) with three replicates. A one-way analysis of variance (ANOVA) was performed to test the significance of each treatment. A post hoc comparison of mean values was carried out by performing Duncan’s multiple range test (DMRT).viz., DBW187, HD2967, PBW343, HD3086, and Agra local) represent the Bacillus genus. The genotypes DBW187, HD2967, PBW343, HD3086, and Agra local contain 24, 27, 17, 26, and 18 Bacillus strains of endophytic origin, respectively. The bacterial strains belonged to 28 different species  and reveal a high degree (100%) of sequence resemblance with the type strain sequences in available in EZ Biocloud e-server . It has been observed that the wheat genotype DWB 187 contained the maximum number of distinct Bacillus species  displayed antagonist action towards highly virulent F. graminearum NFG1 isolate (F. graminearum NFG1 (F. graminearum growth (p < 0. 05), among which B. clarus NOK09 (77.3% F. graminearum growth inhibition), B. subtilis NOK33 (71.9% F. graminearum growth inhibition), and B. amyloliquefaciens NOK109 (79.4% F. graminearum growth inhibition) displayed more than a 70 % inhibitory effect on fungal growth  formation . It has Bacillus strains  were displayed in ituD), bacillomycin (bmyC), and surfactin (srfA) gene produced a single specific amplicon of 647, 395, and 201 bp, respectively, in NOK9, NOK33, and NOK109 strains.PCR amplification of AMP genes in three endophytic Bacillus strains  which showed the greatest bio-control activity (>70%) towards F. graminearum NFG1 under in-vitro conditions were chosen to appraise their bio-control potential to contain HS disease in wheat under field conditions and observations were presented in F. graminearum NFG1 isolate) in both the years (p < 0.05) relative to the untreated endophyte control (T8)  . Maximumrol (T8) . In bothF. graminearum against fungicides are mounting and ultimately resulted in the loss of their field efficacy [Bacillus spp. allied to the anthesis stage of wheat for the eco-friendly and sustainable management of HS under natural field conditions. In the current research investigation, the diversity of cultivable endophytic Bacillus strains allied with healthy wheat ear heads of five different wheat cultivars  has been explored. It is important to mention here that healthy wheat ears were selected for the exploration of Bacillus endophytes because of the fact that the healthy plant parts harbor a diverse and distinct type of microbial population of endophytic bacteria than unhealthy plants [Bacillus strains displaying excellent antagonistic activity in different crops have been reported by various workers [F. graminearum infection [Bacillus species from wheat genotypes and explore their antifungal activity for the field management of HS in wheat. Head scab (HS) is acknowledged as one of the prime diseases of wheat because of its global presence in all wheat-growing countries and is ranked as the fourth biggest threat to successful quality wheat production . Due to efficacy ,50. Furty plants ,52. Early plants . Marag ey plants  reported workers ,55,56,57nfection . Moreovenfection , there iBacillus strains were recovered from wheat ear heads of five different cultivars. These results are in harmony with earlier published literature, where a culture-dependent approach was employed to isolate the bacterial endophytes from different regions of the plant [B. aerophilus, B. albus, B. atrophaeus, B. australimaris, B. badius, B. cabrialesii, B. cereus, B. clarus, B. coahuilensis, B. dafuensis, B. ferrooxidans, B. fungorum, B. glycinifermentans, B. haikouensis, B. halotolerans, B. haynesii, B. mojavensis, B. mycoides, B. nakamurai, B. paralicheniformis, B. paramycoides, B. pseudomycoides, B. rugosus, B. safensis, B. siamensis, B. stratosphericus, B. swezeyi, B. tequilensis, B. velezensis, B. wiedmannii, B. yapensis, B. zanthoxyli, and B. zhangzhouensis with wheat ear head for the first time.A series of published literature indicated that investigation of the diversity of the plant microbiome is one of the potential approaches to identify novel and effective microorganisms as a fungal antagonist and plant growth promoter ,57,58. Bhe plant ,60,61. IBacillus strains prior to field validations [Bacillus strains of antagonistic nature against F. graminearum. Based on the 16S rRNA sequence analysis performed in the present study, the antagonistic bacteria recovered from wheat ear heads were identified as B. clarus NOK09, B. mojavensis NOK16, B. subtilis NOK33, B. rugosus NOK47, B. mojavensis NOK52, B. clarus NOK59, B. coahuilensis NOK72, Bacillus cabrialesii NOK78, B. cabrialesii NOK82, B. rugosus NOK85, B. amyloliquefaciens NOK89, B. australimaris NOK95, B. pumilus NOK103, and B. amyloliquefaciens NOK109. These research findings agree with earlier reports, where B. amyloliquefaciens, B. pumilus and B. subtilis strains have been described as potential antagonists [A large body of published reports on bacterial endophytes has focused on the plausible applications of isolated strains in managing agriculturally important diseases and established dual-culture assays as one of the golden standards for assessing the antagonistic effectiveness of isolated idations ,63. Simiagonists ,65,66.Bacillus strains  showing maximum bio-control activity against F. graminearum reflected strong character for potassium solubilization and siderophore production. These results are in harmony with earlier reports, where the presence of endophytic bacterial strains showing strong attributes of potassium solubilization and siderophore production were observed [B. cereus, B. amyloliquefaciens, and B. subtilis subsp. subtilis from pearl millet host and further suggesting their function as a growth promoter, nutrient mobilizer, and disease defender. Here, it is important to mention that siderophore is a small, low molecular weight  iron-chelating agent which binds to the available iron making it unavailable for the phytopathogens. It also supports the plant by making iron easily available for various biological processes operating inside the plant system. Numerous recent studies reported siderophore production as the most common trait in bacterial endophytes associated with plants [Bacillus strains were also found to be positive for the production of chitinase, protease, lipase, amylase, and pectinase enzymes. Additionally, it appears that these hydrolytic enzymes could be responsible for the remarkable inhibition of F. graminearum. Here, it is worth mentioning that with the help of these extracellular hydrolytic enzymes, endophytes can penetrate the plant tissue and play a vital role in endophytic colonization and establishment inside the host plant [In the current study, three endophytic observed ,68,69. Kobserved  also rech plants ,71,72. Ah plants . In the st plant ,75. F. graminearum had three diverse antibiotic biosynthesis genes  that are linked with the production of the antibiotics iturin, bacillomycin, and surfactin. The obtained results are in conformity with other workers that revealed the prevalence and linkages of numerous antifungal peptide genes in diverse types of Bacillus strains [Bacillus species contain diverse types of AMP genes that have a role in the biosynthesis of antibiotics with specific modes of action [itu gene is essential in enhancing fungal cell membrane permeability [srf gene is linked with biofilm synthesis [bmy associated with the degradation and alterations of the cell wall and cell membrane of fungal hypha [PCR detection of AMP genes indicated that three antagonistic strains showing maximum bio-control activity against  strains ,78,79,80f action . For inseability . On the ynthesis  and bmy al hypha ,83.B. clarus NOK09, B. subtilis NOK33, and B. amyloliquefaciens NOK109) reflecting >70% mycelial growth inhibition against F. graminearum under in-vitro conditions in single or in combination was performed to test the hypothesis. Here, it is pertinent to mention that microbial consortia function in a strong and structured network and offers additional protection for the host to survive better than individual microorganisms under adverse conditions [Paenibacillus sp. strain B2 and Arhtrobacter spp. strain AA was found more effective than a single-strain inoculum for promoting resistance against Septoria tritici blotch disease under drought stress [B. subtilis B2, B. thuringiensis B10, and Enterobacter cloacae B16) also showed noteworthy bio-control effects against Sclerotinia stem rot disease in tomatoes [Bacillus subtilis RC 218 and Brevibacillus sp. RC 263) under semi-controlled field conditions resulted in a significant reduction in the HS severity level in wheat [Bacillus strains of endophytic origin along with individual antagonists. The results emanating from the present investigation clearly evidenced the effectiveness of the combination of three distinct strains (B. clarus NOK09 + B. subtilis NOK33 and B. amyloliquefaciens NOK109) followed by the consortia of two strains in suppressing HS infection in wheat. However, positive and noteworthy antagonistic effects were achieved with all the test strains when used singly or in combination under in-vivo bio-control experiments. The reason that a single strain may be less effective can be associated with poor competitiveness against native microflora and fluctuating environmental conditions [Bacillus sp. MN54 + Trichoderma sp. MN6) was applied. Besides this, the results obtained in the current study with respect to the bio-control efficacy agree with earlier published literature, where a significant level of disease protection has been documented by spraying microbial consortia [B. clarus NOK09 + B. subtilis NOK33 and B. amyloliquefaciens NOK109 when compared with the antagonistic effect of individual strain treatments on HS disease containment. Similar types of observation regarding the effectiveness of endophytic bacterial consortia in controlling the purple blotch disease and enhancing the growth and yield of shallots have been demonstrated by Resti et al. [P. fluorescens Pf1 and B. subtilis (EPCO16 and EPC5) in controlling the Fusarium wilt incidence in chilli by 17–30%. However, in the current study, better percent disease suppression in terms of HS incidence (62.52 to 63.86%) and severity (59.14–61.18%) was obtained when the combination of B. clarus NOK09 + B. subtilis NOK33 and B. amyloliquefaciens NOK109 spray were inoculated on the wheat plants. A large number of bacterial endophytes from diverse types of plants have been identified and evaluated for bio-control activities for several decades to control fungal diseases in India ,84,85. Hnditions ,87. For t stress . Similartomatoes . On parain wheat . These snditions . Here, inditions , who notonsortia . In addii et al. . In a pai et al.  also docBacillus of entophytic nature, where only ten species displayed antagonism towards F. graminearum. Based on field results, it can be proposed that the combination of three endophytic Bacillus mixes (B. clarus NOK09 + B. subtilis NOK33 + B. amyloliquefaciens NOK109) in place of a single strain could be an important approach for the fieldmanagement of HS disease of wheat in an eco-friendly and sustainable manner.In conclusion, it can be inferred that wheat genotypes at the anthesis stage harbor 28 distinct species of


This study was to determine whether coronary artery bypass grafting without cardiopulmonary bypass  could reduce early postoperative mortality and major complications compared with conventional coronary artery bypass grafting with cardiopulmonary bypass  by experienced surgeons.From January 2016 to June 2020, isolated CABG was performed in 1200 patients  in Wuhan Union Hospital. The propensity score matching was used to adjust for differences in baseline characteristics between the ONCABG and OPCABG groups. After 1:1 matching, 404 pairs for each group were selected to compare outcomes within 30 days after surgery. All the operations were completed by experienced surgeons that had completed more than 500 on-pump and 200 off-pump CABG, respectively.P < 0.001) and a higher rate of incomplete revascularization  than the ONCAB group. There was no significant difference in early postoperative mortality between ONCAB and OPCAB groups . However, patients in the OPCAB group had a lower risk of postoperative stroke , new-onset renal insufficiency , respiratory failure , reoperation for bleeding , and required less ventilator assistance time  and intensive care unit (ICU) time .After propensity matching, the two groups were comparable in terms of preoperative characteristics. The OPCAB group had less vein graft (2.5 ± 1.0 vs. 2.7 ± 0.9; In our study, patients undergoing OPCAB had fewer postoperative complications and a faster recovery. It is a feasible and safe surgical approach to achieve revascularization when performed by experienced surgeons. Coronary artery bypass grafting (CABG) reduces mortality and improves quality of life in patients with extensive coronary artery disease. Conventional coronary artery bypass grafting is performed on cardiopulmonary bypass (CPB) and remains the standard intervention for coronary artery disease requiring surgery for coronary revascularization . The tecThis was a retrospective study conducted in our Medical Center that included 1200 patients underwent isolated CABG from January 2016 to June 2020. Of these, 429 received ONCAB and 771 received OPCAB. The selection of patients undergoing either on-pump or off-pump CABG is at the discretion of the surgeon prior to surgery. Patients with conversion from off-pump to on-pump surgery were included into OPCAB group. All surgeons operating on patients had extensive experience in both procedures. The Medical Ethics Committee of Tongji Medical College, Huazhong University of Science and Technology approved the ethics of this study (IORG No. IORG0003571) and patient informed consent was waived.Both surgical approaches used a standard median sternum incision to expose heart. For the OPCAB procedure, several standard coronary artery stabilizers and cardiac positioning techniques were used. For the ONABG, standard cardiopulmonary bypass techniques were used and myocardial protection was performed by intermittent antegrade cold blood cardioplegia. In both surgical approaches, the grafts were derived from the left internal mammary artery (LIMA), great saphenous vein or radial artery. The conventional anastomosis is LIMA with left anterior descending (LAD) artery, great saphenous vein and/or radial artery anastomosis with other target vessels. Except for differences in surgical procedures, anesthesia and in-hospital management for patients were similar.The choice of technique (ONCAB or OPCAB) was made by the operating surgeon. The demographic variables were included age and sex. Also recorded were the concomitant medical diseases including hypertension, diabetes, peripheral arterial disease, carotid artery stenosis, renal artery stenosis, stroke, chronic renal insufficiency, renal-replacement therapy, chronic obstructive pulmonary disease (COPD). Cardiac variables included myocardial infarction, percutaneous coronary intervention (PCI), atrial fibrillation, intra-aortic balloon pump (IABP), cardiotonic drugs use, left ventricular ejection fraction (LVEF), left ventricular end-diastolic diameter (LVEDD), and number of diseased vessels. Variables associated with revascularization included number of grafts, distal arterial and venous anastomosis, and incomplete revascularization rate. The primary postoperative outcome included death, stroke, non-fatal myocardial infarction and new renal failure requiring dialysis. Other postoperative outcome included new-onset atrial fibrillation, low cardiac output syndrome, IABP use, new renal insufficiency, respiratory failure or infection, reoperation for bleeding and sternum infection. Hospital stay time, postoperative intensive care unit (ICU) time and ventilator assistance time were also collected to assess clinical efficacy. Definitions of these variables are provided in the 2 tests or Fisher's exact test. Normally distributed continuous variables are expressed as mean ± SD, while non-normally distributed continuous variables are shown as median or interquartile range. Normally distributed data were analyzed by Student's t-test, while non-normally distributed data were analyzed by Mann-Whitney U test. P-value <0.05 was considered statistically significant. IBM SPSS  was used to analyze data.All clinical data in this study were obtained from the electronic medical record system of our hospital. The propensity score matching method has been described above. We report categoric variables as counts with percentages and analyzed by χP < 0.001) but slightly lower left ventricular ejection fraction  than the ONCAB group. The proportions of males (P < 0.001), peripheral arterial disease (P = 0.039), stroke (P = 0.018), atrial fibrillation (P = 0.038) and urgent surgery (P = 0.013) were higher in the OPCAB group. There was no significant difference in the proportions of hypertension, diabetes, hyperlipidemia, chronic obstructive pulmonary disease (COPD), and renal insufficiency between the two groups (P > 0.05) (p = 0.502). The discriminative power of propensity score matching quantified by measuring the recipient operating characteristic area was found to be moderate , indicating a good balance of preoperative risk between the two groups. After adjustment for propensity score, 404 pairs for each group were selected and the preoperative characteristics of patients in two groups were comparable . In unadjusted studies, the OPCAB group had a higher average age  . Then, wmparable .P < 0.001) and fewer vein grafts (P < 0.001) compared with ONCAB group. Patients in OPCAB group had a higher rate of incomplete revascularization (P = 0.042) (P = 0.066) were comparable between the two groups. However, the OPCAB group still had fewer vein grafts  and higher rates of incomplete revascularization   . After p= 0.049) .P = 0.008). However, the incidence of postoperative mortality (P = 0.209), myocardial infarction (P = 0.488) and renal failure requiring dialysis (P = 0.242) were similar between the two groups . As with unmatched results, no significant difference existed in the postoperative mortality (P = 1.000), myocardial infarction (P = 0.787) and renal failure requiring dialysis (P = 0.590) between the two groups , new renal insufficiency (P = 0.011), respiratory failure (P = 0.037) reoperation for bleeding (P = 0.042) and sternum Infection (P = 0.045) compared with ONCAB. Besides, OPCAB reduced postoperative ventilator assistance time (P < 0.001) and ICU stay time (P < 0.001) significantly. There was no significant difference in rate of IABP use (P = 0.803) and low cardiac output syndrome (P = 0.562) between two groups , respiratory failure (P = 0.001) and reoperation for bleeding (P = 0.012). Postoperative ventilator assistance time (P < 0.001) and ICU stay time (P < 0.001) were shorter in the OPCAB group. No significant difference existed in the postoperative new-onset atrial fibrillation (P = 0.220), IABP use (P = 0.331), low cardiac output syndrome (P = 0.112) and sternum infection (P = 0.499) between the two groups , but it was lower than that of several large prospective multicenter clinical studies comparing ONCAB and OPCAB such as the ROOBY trial (17.8%) and GOPCABE trial (34%)  , 38. Dueal (34%) , 40. Howal (34%) , 42. BesY trial 1.8% and Gal (34%) . HoweverY trial 1.8% and Gal (34%) . As one There are several limitations in this research. Firstly, the study was retrospective, which may have confounding factors though propensity matching has been used to reduce bias. Secondly, the study was a single-center study, the general applicability of the results was worth discussing. In addition, the study lacked a comparison of long-term survival data, which is also important for evaluating the efficacy of surgery. Finally, although propensity matching can correct for selection bias, covariate imbalance and potential confounding, it has the disadvantage that a part of individuals end up not matching and were excluded from the analysis, resulting in a loss of both generalizability and precision .In conclusion, our study showed that with well-developed technique and experienced operator, OPCAB can reduce postoperative complications within 30 days. It is a feasible and safe surgical approach to achieve revascularization when performed by experienced surgeons.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by the Medical Ethics Committee of Tongji Medical College, Huazhong University of Science and Technology approved the Ethics of this study (IORG No. IORG0003571). Written informed consent for participation was not required for this study in accordance with the national legislation and the institutional requirements.CW, YJ, SC, and XC designed the research. CW, YJ, and YS conducted the research. RT and QW collected the data. CW and DW analyzed the data. CW and SC wrote the paper. XJ and ND done data analysis and language proofreading. SC had primary responsibility for final content. All authors read and approved the final manuscript.This work was supported by the National Natural Science Foundation of China (Grant No. 81873489).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Brevundimonas albigilva TT17 was isolated from human skin. This strain can grow on Triton X-100. Here, we present the whole-genome sequences of this strain and B. albigilva KEME 9005-016T. This result demonstrates the first information on the genome sequences of the species of B. albigilva. Brevundimonas belonging to the family Caulobacteraceae comprises Gram-negative, nonfermenting, and aerobic bacilli  medium agar  containihttps://www.bioinformatics.babraham.ac.uk/projects/fastqc/)  . The reaectively . The defectively .T. The assembly report provided by NCBI (ASM2357356v1) showed that TT17 is a type category of B. albigilva. The pairwise BLAST comparison between the open reading frames (ORFs) in the two genomes using RAST serve showed overall very high similarities. Still, some clusters with lower similarities and nonmatches are scattered as found in the TT17 genome  and SRX15388290 (MinIon seq) for strain TT17 and SRX14829765 (MinIon seq) for KEME 9005-016T.The assembled genome sequences of


Mycoplasma hyopneumoniae, the causative agent of swine respiratory disease, demonstrates differences in virulence. However, factors associated with this variation remain unknown. We herein evaluated the association between differences in virulence and genotypes as well as phenotype . Strains 168 L, RM48, XLW-2, and J show low virulence and strains 232, 7448, 7422, 168, NJ, and LH show high virulence, as determined through animal challenge experiments, complemented with in vitro tracheal mucosa infection tests. These 10 strains with known virulence were then subjected to classification via multilocus sequence typing (MLST) with three housekeeping genes, P146-based genotyping, and multilocus variable-number tandem-repeat analysis (MLVA) of 13 loci. MLST and P146-based genotyping identified 168, 168 L, NJ, and RM48 as the same type and clustered them in a single branch. MLVA assigned a different sequence type to each strain. Simpson’s index of diversity indicates a higher discriminatory ability for MLVA. However, no statistically significant correlation was found between genotypes and virulence. Furthermore, we investigated the correlation between virulence and biofilm formation ability. The strains showing high virulence demonstrate strong biofilm formation ability, while attenuated strains show low biofilm formation ability. Pearson correlation analysis revealed a significant positive correlation between biofilm formation ability and virulence. To conclude, there was no association between virulence and our genotyping data, but virulence was found to be significantly associated with the biofilm formation ability of M. hyopneumoniae.The online version contains supplementary material available at 10.1186/s13567-022-01109-x. Mycoplasma hyopneumoniae strains on pigs are variable. However, little remains known about the reason or elements responsible for such differences. In this study, our aim was to identify factors associated with variations in M. hyopneumoniae virulence.The incidence of mycoplasma pneumonia of swine (MPS) has witnessed an increase, resulting in significant economic losses to the pig industry across the world . The patM. hyopneumoniae strains. Pig challenge experiments are the gold standard method for virulence assessment. The virulence characteristics of M. hyopneumoniae strains can be evaluated using in vitro models, such as swine tracheal mucosa. Some studies have explored the association between M. hyopneumoniae virulence and genotypes. Genotyping plays a vital role in epidemiological investigations and in bacterial identification and differentiation. Genotyping techniques are usually easy to perform, require minimal bacterial load, and results are quickly generated ,,43], 744M. hyopneumoniae shows the ability to form biofilms [M. hyopneumoniae. Thus, we suggest that differences in the virulence of strains can be correlated with their biofilm formation ability. Future studies should assess a higher number of strains to validate the presence of a correlation between biofilm formation ability and virulence. Moreover, we need to enhance our understanding of virulence genes that may be related to biofilm formation ability of various strains.Biofilms represent one of the main virulence traits associated with the pathogenesis of diverse pathogens , 44. A wbiofilms . Herein Additional file 1. Serological tests ofM. hyopneumoniaeIgG antibodies for detection ofM. hyopneumoniaeantibody-negative pigs.Additional file 2. The presence ofM. hyopneumoniaein lung tissues (qPCR) and clinical observations of pigs post-infection.Additional file 3. Correction of virulence and results of the in vitro tracheal infection.Additional file 4. Results of DNA sequencing based on MLST, P146 gene sequencing and MLVA.Additional file 5. Correlation of virulence and biofilm.


Functional ingredients for human health have recently become the focus of research. One such potentially versatile therapeutic component is fucose-containing sulfated polysaccharides (FCSPs), referred to as fucoidans. The exploitation of marine brown algae provides a rich source of FCSPs because of their role as a structural component of the cell wall. Fucoidans are characterized by a sulfated fucose backbone. However, the structural characterization of FCSPs is impeded by their structural diversity, molecular weight, and complexity. The extraction and purification conditions significantly influence the yield and structural alterations. Inflammation is the preliminary response to potentially injurious inducements, and it is of the utmost importance for modulation in the proper direction. Improper manipulation and/or continuous stimuli could have detrimental effects in the long run. The web of immune responses mediated through multiple modulatory/cell signaling components can be addressed through functional ingredients, benefiting patients with no side effects. In this review, we attempted to address the involvement of FCSPs in the stimulation/downregulation of immune response cell signaling. The structural complexity and its foremost influential factor, extraction techniques, have also attracted attention, with concise details on the structural implications of bioactivity. Marine natural products are widely researched and have produced various promising bioactive components that can be useful functional elements with multiple health benefits . The fieThe survival of multicellular organisms largely depends on their capacity to fight infections and recover from injuries. Therefore, the immune system is fundamental. A diverse array of cytokines, signaling proteins, lipid molecules, and transcription factors are involved in immune functions . ComplexThus, this review explores FCSPs from marine algae, referring to their structural diversity and extraction techniques accompanied by impurities in isolating standard fucoidan. Furthermore, we reviewed the effects of fucoidans from marine algae on anti-inflammatory and immunomodulatory cell signaling pathways. The influence of the structure on bioactivity was briefly addressed. This review focused on seaweed-extracted fucoidan and its activity. However, fucoidan extraction is not limited to seaweeds from marine resources, and the potential of fucoidans extends beyond the bioactivities addressed herein. Fucoidans have multiple biological benefits that are beyond the scope of this article and are worth addressing in another project.FCSPs are referred to as fucoidans . FucoidaFucus vesiculosus was presented by Patankar et al. (1993) [Ascophyllum nodosum suggesting a revised structure for fucoidan [2-O-sulfation and 2,3-O-desulfation. Crude fucoidan was extracted using the method described by Mabeau et al. (1990), and acid hydrolysis was conducted using the process described by Colliec et al. (1994) [F. serratus consists of a backbone with alternating 3- and 4-linked α-fucopyranose residues [F. serratus, performing air and vacuum drying, and then milling the algal biomass at room temperature with a 4:2:1 MeOH:CHCl3:H2O mixture to eliminate the color compounds in the structure. Then, the material was mechanically stirred with CaCl2, and the crude polysaccharide was obtained through centrifugation, combined, dialyzed, and lyophilized to form a mixture consisting of 32.8% fucose, 24.4% glucose, 18.9% SO3Na, 2.9% xylose, 2.6% galactose, and 1.8% mannose. Column fractionation was performed on the above sample containing DEAE-Sephacel (Pharmacia) in Cl− form, with gradient concentrations of NaCl. Ion-exchange chromatography produced four different fractions with considerable yields [Sargassum stenophyllum was reported to have two types of galactofucans, which resembled the fucosylated chondroitin sulfates from sea cucumbers [2, and further precipitated with cetylpyridinium chloride to obtain a higher yield of fucoidan fractions. CaCl2 was used to dissolve the pyridinium salts and reprecipitation was performed using ethanol. Moreover, insoluble and soluble fractions were obtained via the centrifugation of ethanol-insoluble material dissolved in water. Another study focused on the order Fucales using S. polycystum and revealed a novel sulfated galactofucan. This polymer had a backbone of 3-linked α-L-fucopyranose and 4-linked sulfation. Furthermore, the backbone contained random branches of 2-linked α-D-galactopyranose, which were sulfated at position 4.Although FCSPs are diverse, their structure has been used to classify them depending on their order to ease the pace of study. The order Fucales has been widely discussed. The backbone structure of FCSP from . 1993) , who profucoidan . The pre93 , who . (1994) . Bilan eresidues . β-Xylose yields . Sargassucumbers . They weChorda filum was assessed for its fucoidan fractions [2-O-acetylated random α-(1-3)-linked fucose residues have been reported. Similar to fucoidan isolated from the previously mentioned species, fucoidan fractions have been obtained via ion-exchange chromatography using DEAE Sephadex A-25. Structural analysis of fucoidans is prevalent for the order Fucales but has also been reported for the order Laminariales. Wang et al. (2010) obtained three fractions of low-molecular-weight fucogalactan from Laminaria japonica [L. japonica using water and further purified through ethanol precipitation. Usoltseva et al. (2019) assessed purified sulfated fucans from L. longipes and Saccharina cichorioides  [S. cichorioides, the sampling material can be reduced to a practical size of 1 mm; ethanol, acetone, and chloroform were added to increase the efficiency of the extraction process. Fat was removed from the sample, followed by the addition of 0.1 N HCl (2 L) at room temperature. Ultrafiltration was performed to concentrate the sample to up to 20% of the initial volume, and the sample was washed four times with 96% ethanol for the further precipitation of polysaccharides. Subsequently, 96% ethanol was used to wash the pellet, and air-dried samples were used in further experiments. A DEAE-cellulose column was used for the fractionation of fucoidan [Cladosiphon okamuranus from the order Chordariales showed a linear backbone of α-(1-3)-fucopyranose, half of which was sulfate substituted at the C4 position. Some fucopyranoses were O-acetylated [The brown seaweed ractions . Its cheariales) . Accordifucoidan . Other aetylated . Backbonetylated .F. vesiculosus compared with other periods. They also reported that the fucose content was lower in the sterile phase than in the fertile phase [L. japonica showed increased levels of fucose content during the reproductive phase [It is evident that the structural composition of fucoidans varies among different brown seaweed species. We have elaborated on multiple structures discussed in previous literature depending on seaweed order, but neither of those supports a consistent basic structure of fucoidan. This proves that the structural traits of FCSPs cannot be predicted or categorized due to their complexities among each other. Though the chemical composition of fucoidans is complex, the basic substituents are fucose and sulfate. Moreover, the fucoidans are consisted of other monosaccharides, uronic acid, and protein. Other than these the structural features will be determined by age, geographic region, and the harvesting season. A previous study reported that the fucose content increased during the sporulation period in le phase . Moreovele phase . Anotherve phase .The nature of the chemical components, method of extraction, and interfering substances influence the extraction . Most poThe source of extraction of the sulfated polysaccharides is marine brown algae rather than marine invertebrates. The molecular weight, sulfation percentage, point of sulfation, and monosaccharide composition are vital to the bioactivity of fucoidan. Therefore, purification should be performed under mild conditions to avoid structural shifts . The appMacrocystis pyrifera, in which hydrolysis results in uronic acid [Fucoidan extraction using dilute acetic acid has a history of over 100 years, with written scientific records. In 1913, Kylin termed these compounds fucoidan . Currentnic acid . The remnic acid ,13. In anic acid ,37.Purification of high-quality fucoidan requires further steps assisted by column chromatography. One such method is anion-exchange chromatography. The sulfate ester groups linked to the polymer backbone provide anionic charges even at low pH values, which can be utilized during this chromatographic technique. The resin used in this method is quaternary ammonium, which forms strong coulombic interactions with the loaded sample. Elution is performed with the application of a NaCl gradient, where its concentration increases at the later stages of the column purification mobile system. The elution of lower sulfate-containing fractions occurs first, whereas the high sulfate fractions elute last ,38. The F. vesiculosus [S. muticum [F. evanescens was extracted using the method described by Hmelkov et al. (2017) [Jasso et al. (2011) implemented microwave-assisted extraction (MAE) with optimum reaction conditions to extract sulfated polysaccharides from iculosus . They suiculosus ,42. This muticum . Fucoida. (2017) . SimilarS. horneri using AMG, Celluclast, Viscozyme, and Alcalase. The chemical properties of each isolate were evaluated for their anti-inflammatory potential [The efficient extraction of seaweed polysaccharides is hindered by the complexity and rigidity of the cell wall. This has led to the use of carbohydrates to degrade the cell wall, thus releasing polysaccharides and other bioactive components. Each carbohydrase must be implemented at its optimal pH and temperature to obtain higher yields with promising bioactive properties . Sanjeewotential . The extotential .F. vesiculosus and A. nodosum which are Atlantic representatives of Phaeophyceae and exhibit similar chemical composition and biological properties [Availability, suitable extraction methods, algal sources, yield, and purity should be considered for mass production. A suitable procedure for commercial fucoidan production was published in 1952 . Recentloperties . In addiInflammation can arise in any tissue in response to multiple conditions, such as infectious, toxic, or autoimmune injury. Inflammation does not reflect a single, straightforward mechanism but is a complex set of axes that interact to overcome detrimental situations. However, if the situation is not properly handled through this multifaceted mechanism, it can lead to persistent tissue damage via the recruitment of cellular members of the immune system. The neutral status of inflammation does not arise passively and exhibits proper maintenance of health requirements by ample positive actions where it does not entail activation of inflammation. Modulation of inflammation is an alternative approach, although pathogenic events are unknown . HoweverDiscussions regarding the health benefits of food beyond basic nutrition have increased public awareness in recent years. The health-promoting effects of foods and the implementation of these bulk sources as functional foods have become a widely researched topic. Thus, significant developments in the natural products of nutraceuticals, pharmaceuticals, and functional foods have opened up broader topics in the research community . The potU. pinnatifida intervened in obesity-associated inflammation-related metabolic syndrome [U. pinnatifida downregulated the above inflammatory factors, resulting in the inhibition of adipogenesis. Early reports explained that TNF-α regulates both MCP-1 and PAI-1 [S. horneri was used to isolate low-molecular-weight fucoidan. The FTIR and NMR results collectively signify the structural similarity to commercial fucoidan. The chemical composition of the low-molecular-weight (~45 kDa) fractions (SHC4–6) reveals the structure–activity relationship bearing 43.15 ± 0.36% polysaccharide and 29.92 ± 0.29% sulfate levels. Polyphenol and protein interferences were negligible, and fucose was as high as 36.50 ± 1.45%. The authors discussed the involvement of the nuclear factor kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) pathways in the downregulation of inflammation caused by fine dust.Kim et al. 2012) reported that fucoidan isolated from syndrome . Inflammnd PAI-1 . The anand PAI-1 . The hig012 reporS. horneri [2, and pro-inflammatory cytokines in RAW 264.7 cells via downregulating the NF-κB signaling cascade [Regulation of the immune response gene expression is critical for the maintenance of immune homeostasis. One of the most significant effector pathways involved in inflammation is NF-κB, which is a transcriptional regulator composed of Rel-protein homo- or heterodimers . This re horneri . Among t cascade . Researc cascade . Pozhari cascade .Another pathway that has been subjected to broader discussion involves MAPKs. This protein structure is well-conserved from unicellular eukaryotes to multicellular organisms. Specifically, this structure includes three proteins, i.e., extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38, which sequentially phosphorylate to initiate the signaling cascade. MAPKs are not only involved in the regulation of inflammation but also play an important role in cell survival, proliferation, differentiation, and death. Huang et al. published a detailed review of each MAPK component .S. swartzii with a high sulfate content against LPS-stimulated inflammatory responses in RAW 264.7 macrophages [S. swartzii was 33.99 ± 0.17%. Although the authors did not propose a detailed structure of the isolated fucoidan, the chemical composition showed a higher sulfate content and fucose monosugar. Therefore, the active fraction may have influenced the results. According to this report, canonical p65 nuclear translocation was evident using immunofluorescence techniques. TLRs, which are located in the cytoplasmic membranes of macrophages, share a toll-interleukin receptor domain. These signals are cascaded from MyD88, IL-1 receptor-associated kinases (IRAKs) to NF-κB [Padina commersonii [P. commersonii significantly inhibited their expression, altering downstream signal transduction. Another study on fucoidan derived from the invasive species S. horneri reported the inhibition of LPS-induced inflammation via the NF-κB and MAPK pathways [2, and pro-inflammatory cytokines (TNF-α and IL-6) decreased [Jayawardena et al. (2020) analyzed fucoidan derived from rophages . They reto NF-κB . Sanjeewmersonii . TLR2/4 pathways . The phoecreased ,55.S. cristaefolium, in addition to reducing NF-κB activation, dramatically reduced the production of iNOS by decreasing the phosphorylation of p38, ERK, and JNK in LPS-stimulated RAW 264.7 cells [S. horneri, a brown seaweed, had anti-inflammatory properties [MAPKs are key players in inflammatory processes, such as upstream NF-κB signaling molecules . Wu et a.7 cells . Anotheroperties . These e2 via the arachidonic acid pathway. Thus, the modulation of COX-2 is as important as that of iNOS in inflammatory pathogenesis [NO Is used as a primary evaluation cross-point in inflammation studies. It is a vital end product and is regulated by iNOS. Mammalian cells are capable of producing three forms of nitric oxide synthase (NOS) depending on the cell type: iNOS, eNOS, and nNOS, where l-arginine acts as the primary substrate. NO production exceeds regular levels in the presence of immunological reactions; thus, it is involved in pathogenesis . The resogenesis . Inflammogenesis . Cytokinogenesis . TherefoS. japonica against LPS-induced inflammation in macrophages [The Janus kinase (JAK)-signal transducer of activators of transcription (STAT) pathway is another vital pathway associated with diverse cytokines, interferons, growth factors, and related molecules. Receptor-associated JAKs are activated by ligand binding to cognate receptors. Successive STAT activation conveys rapid signaling from the cell surface to the nucleus, allowing subsequent translocation of STATs to the nucleus, where they modulate target gene expression . Ye et arophages . This reF. vesiculosus [F. vesiculosus [F. vesiculosus were investigated in spleen DCs [U. pinnatifida. RAW 264.7 macrophages expressed chemokines (RANTES and MIP-1α), whereas murine splenocytes produced IL-6 other than chemokines [F. vesiculosus [Immunomodulation is the navigation of the immune defense system via medication. Immune response enhancement has been reported for many polysaccharides obtained from natural resources . Yang eticulosus . This priculosus , providileen DCs , and theemokines . Do et aiculosus . This reC. okamuranus accumulated more in the liver of rats than in their blood serum. Furthermore, the transport mechanism of fucoidan was identified, and active transport through the cell membrane was suggested as the most probable mechanism [F. vesiculosus exhibited superior skin penetration by topical application in rats and that a 100 mg/kg dose showed a prolonged half-life compared to that of intravenous injection [S. japonica has been reported to reduce inflammation, oxidative stress, hepatorenal injuries, and DNA damage via the downregulation of pro-inflammatory cytokines such as IL-1β, IL-6, and TNF-α in streptozotocin-induced diabetes mellitus in rats [S. hemiphyllum significantly reduced the TNF-α levels [F. vesiculosus did not have a significant effect in this in vivo model [F. vesiculosus had a significant downregulatory effect on IL-10, IL-1β, and IL-1α in a dextran sulfate sodium-induced mouse model of acute colitis [Chnoospora minima, Ecklonia cava, and S. horneri decreases the production of COX-2 and NO in LPS-induced zebrafish embryos [U. pinnatifida showed significantly reduced tissue infiltration with inflammatory cells and bone and cartilage destruction in a rheumatoid-arthritis mouse model [U. pinnatifida and Turbinaria ornate downregulated arthritis in rats induced by Freund’s adjuvant by decreasing the levels of PGE2, TNF-α, and IL-6 in blood plasma [T. decurrens significantly reduced the expression of IL-1 β and COX-2 in a formalin-induced edema mouse model. AlKahtane et al. (2020) performed an anti-inflammatory study of L. japonica fucoidan against microcystin-LR-induced liver damage and found a significant reduction in the levels of IL-6, TNF-α, and IL-1β in the serum of mice [S. fluitans against carbon tetrachloride-induced liver injury in rats. The results of this study confirmed a significant reduction in the levels of IL- 1β and TNF-α [F. vesiculosus on an LPS- and Porphyromonas gingivalis-injected periodontitis mouse model, which acted by downregulating IFN-γ [C. okamuranus fucoidan by decreasing the pro-inflammatory cytokine levels, such as TNF-α, IL-1β, and IL-6, and downregulating NF-κB signaling [The anti-inflammatory and immunomodulatory activities of fucoidan are solidified not only in vitro but also in vivo. Previous studies have revealed that oral administration of these polysaccharides results in low levels in the plasma and high levels in particular organs; for example, fucoidan isolated from echanism ,78,79. Tnjection . The ant in rats . A simil in rats . Furtherα levels . Matsumovo model . However colitis . In anti embryos ,86. Low-se model . Fucoidad plasma ,89. Fuco of mice . Chale-Dnd TNF-α . In adding IFN-γ . Furtherignaling .U. pinnatifida significantly increased the levels of natural killer cells, neutrophils, and pro-inflammatory cytokines in the body and downregulated apoptosis. F. vesiculosus fucoidan also elevated DC maturation, Th1 immune responses, and cytotoxic T-cell activation, and increased the production of antibodies after exposure to antigens [F. evanescens, L. cichorioides, and L. japonica has been shown to activate the immune system. Makarenkova et al. (2012) demonstrated that the interaction between fucoidan and TLRs increases the production of cytokines and chemokines. According to these results, fucoidan binds to TLRs and enhances NF-κB signaling; this results in the production of cytokines and inflammatory responses [U. pinnatifida. Furthermore, this treatment elevated the production of pro-inflammatory cytokines in a herpes simplex virus (HSV-1)-infected mouse model [F. vesiculosus, U. pinnatifida, A. nodosum, and M. pyrifera, revealing significant upregulation of IL-8, IL-6, and TNF-α production. M. pyrifera showed superior activity among these seaweeds [U. pinnatifida increased IFN-γ production in a UVB-induced mouse model. However, this study reported that it did not increase cytokine production but reduced edema and leukocyte migration to the skin [The anti-inflammatory activity of fucoidan has gained much attention, and studies have shown its immunomodulatory potential. Some studies have revealed that low-molecular-weight fucoidan has anti-inflammatory activity and high-molecular-weight fucoidan exhibits immunomodulatory activity . Howeverantigens . Furtheresponses . Hayashise model . Jang etseaweeds . Fucoidathe skin .®. This was a combination of M. pyrifera (10% w/w), F. vesiculosus (85% w/w), L. japonica (5% w/w), zinc, vitamin B6, and manganese. The results showed an overall reduction in symptoms such as stiffness and pain during physical activities. Furthermore, a 1000 mg/day dose showed prominent results [C. okamuranus on the activation of natural killer cells in cancer survivors. The results suggested that 3 g of fucoidan was a safe dose for patients, and recurrence of tumors significantly decreased [In addition to these in vitro and in vivo evaluations, clinical trials have been reported by Myers et al. (2016). According to the results of this study, patients  with osteoarthritis were treated with fucoidan. In brief, five participants were treated with 100 mg/day and seven participants were treated with 1000 mg/day of Maritech results . Nagaminecreased .U. pinnatifida was orally supplemented to BALB/c mice, and its effects on viral replication in the HSV-1-infected in vivo model were assessed. Oral supplementation of fucoidan was determined to protect the mice against viral infection based on the survival rate and lesion scores. Fucoidan administration enhanced cytotoxic T lymphocyte activity. In vitro assays confirmed improvements in phagocytic activity and B-cell blastogenesis upon treatment with fucoidan. In conclusion, this study suggested that the effect of orally administered fucoidan largely depends on gut immunity in animals. Tabarsa et al. (2020) reported the immune-boosting properties of fucoidan from Nizamuddinia zanardinii [Hayashi et al. (2008) assessed the extended scope of fucoidan overstimulation of both innate and adaptive immune defense functions, revealing the potential of fucoidan to inhibit viral replication . Fucoidanardinii . This acUndaria pinnatifida and F. vesiculosus by the Australian manufacturer Marinova as “Generally Recognized as Safe (GRAS)”. The European Commission approved fucoidan extracts from the same species as a novel food [Owing to their low toxicity and great oral bioavailability, fucoidan and fucoidan-derived products have recently been commercialized in different sectors, including cosmetics, dietary supplements, and animal-feeding supplements . Fucoidavel food ; respectvel food .U. pinnatifida on letrozole, tamoxifen, and hormone-treated breast cancer patients. This study showed no significant difference in the risk of clinically significant interactions and adverse effects of fucoidan treatment [® injection) along with fucoidan powder bis in die (twice a day in Latin); the administration of Taiwan oligo fucoidan® effectively increased the disease control rate of mCRC patients [Despite the use of fucoidan and fucoidan-derived products as food and dietary supplements, there are several examples of the combination of fucoidan with clinical, chemical, and chemotherapeutic agents as combination therapy. In Taiwan, a human clinical study of lung cancer patients showed that treatment with cisplatin and fucoidan efficiently increased the survival rate of patients . Studiesreatment . A prosppatients . The respatients .Brown algae are an approachable and substantial source of sulfated polysaccharides. Brown algal fucoidans have attracted attention as active ingredients for a wide array of medicinal applications. Some brown algal species are cultivated on a large scale and frequently consumed as food or food additives where occasional functional food and drug development interests are exhibited. This review article explores the structural diversity of fucoidans, extraction methodologies, and the associated impurities. Algal fucoidans have complicated heterogeneous structures. Despite the few reports on the structural elucidation of selected species, refined structures are not yet clear. Studies have been carried out with purified fucoidan, and most published research has been conducted with relatively crude fucoidan preparation, hindering the elucidation of the structure–activity relationship. Defined fucoidans and their conformational investigations could lead to a better understanding of this matter to fill the gaps. Chemical modifications in fucoidans, such as sulfation at predetermined positions, could considerably improve their respective bioactivities. Deeper studies on fucoidan structure will possibly facilitate the development and utilization of brown algal resources. Among the potent biological activities of fucoidans, this review delves into immunomodulatory and anti-inflammatory effects. Even though most research requires successful human trials, marine algal fucoidans conclusively provide therapeutic interventions to improve immunity and prevent immune disorders through cellular signaling. However, the number of clinical studies on this topic is limited, impeding knowledge of fucoidan pharmacokinetics. Consequently, this review aims to facilitate the awareness of brown algal fucoidans as a viable source of research material and a sustainable, utilizable material for industrial applications.