For complex shaped materials, computational efficiency and accuracy of DEM models are usually opposing requirements. In the literature, DEM models of railway ballast often use very complex and computationally demanding particle shapes in combination with very simple contact laws. In contrast, this study suggests efficient DEM models for railway ballast using simple particle shapes together with a contact law including more physical effects. In previous works of the authors, shape descriptors, calculated in a shape analysis of two types of ballast, were used to construct simple particle shapes (clumps of three spheres). Using such a shape in DEM simulations of compression and direct shear tests, accurate results were achieved only when the contact law included additional physical effects e.g. edge breakage. A parametrisation strategy was developed for this contact law comparing DEM simulations with the measurements. Now, all the constructed simple particle shapes are parametrised allowing to study their suitability and relating their shape descriptors to those of railway ballast. The most suitable particle shapes consist of non-overlapping spheres, thus have a high interlocking potential, and have lowest sphericity and highest convexity values. In a micromechanical analysis of the four best performing shapes, three shapes show similar behaviour on the bulk and the micro-scale, while one shape differs clearly on the micro-scale. This analysis shows, which shapes can be expected to produce similar results in DEM simulations of other tests/load cases. The presented approach is a step towards both efficient and accurate DEM modelling of railway ballast. In the discrete element modelling of granular materials, particle shape and size representation plays an important role. Together with particle contact modelling and model calibration it determines the accuracy and reliability of the DEM model.Shape modelling is usually the crucial factor for computational efficiency. Therefore, from computational point of view it is interesting to ask: Is it possible to build an accurate DEM model using simple particle shapes  and combine them with a suitable contact law and a proper parametrisation? This study aims to contribute to answering this question using the example of railway ballast.As the single ballast stones have very complex shapes, including sharp edges and corners, particle shape modelling is frequently addressed in the literature on DEM modelling of railway ballast. Using 2D or 3D scanner data of ballast stones, rigid clumps of spheres are constructed in , 14, 19.The effect of particle shape is investigated in the literature intensively. In most studies, particle shape is varied (mostly with respect to certain shape descriptors) using the same contact law and parameters for all shapes and the simulated bulk behaviour is analysed. In the review paper , severalWhen a particular granular material is to be modelled via DEM, a comparison using different particle shapes should be accompanied by a parametrisation for each used shape. Such studies are rare in literature and application specific. An example is Coetzee , where cThe question how accurate particle shape of a given material must be modelled in DEM and how much of “shape inaccuracies” can be compensated by an individual parametrisation remains open. Although the different particle shapes in  could beThis work tries to contribute to answer the above question and continues previous works of the authors on DEM modelling of railway ballast. Figure A shape analysis with these two types of railway ballast was conducted, Suhr et al. , investiThe knowledge on ballast shape was used in  to systeContact modelling was addressed in , compariE and In , a paramThis work will use the knowledge gained from previous works to finish a complete analysis of DEM modelling of one type of railway ballast. 20 different simple particle shapes will be considered for the parametrisation to Kieselkalk ballast. While problems occurred for some particle shapes, 10 shapes are successfully parametrised to the experimental data. The shape descriptors of these shapes are compared to those of the ballast stones, to check for possible relations. A micromechanical analysis reveals that considering different particle shapes with their own set of parameters, the internal states of the packings can differ, although the bulk response is similar to the experimental one.This work is organised as follows: in Sect. e, flatness, f, sphericity, c, were evaluated. These shape descriptors are defined below in Eqs. , de, dee, flMoreover, three different curvature based angularity indices were compared. Only a newly introduced angularity index, the scaled Willmore energy, gave reasonable results in analytic tests, application to scanned data of sharp stones as well as artificially smoothed versions of the scanned stones. No difference could be seen between Calcite and Kieselkalk ballast with respect to this angularity index. As clumps of spheres will be used as DEM shapes, here the calculation of angularity makes no sense. Therefore, angularity will not be considered further.In , simple Obviously, the constructed clumps of spheres lack the angularity of real ballast stones. However, they are non-convex, which is an important property of ballast, and also allows for interlocking. Clumps of spheres can be arranged to obtain particles with higher or lower “surface roughness”, Soltanbeigi et al. . Roth anduced in . In addic and c and c and larger V showed a strong correlation with the porosity, chosen clump shapes and rejected ones could not be separated by volume. This is in contrast to the other shape descriptors, which do correlate strongly with the porosity, i.e. The shape descriptors of these chosen clumps (1–10 and 21–30) are plotted in Fig. s and in  neither  seen in . All choFor all DEM simulations in this work the software YADE  will be In this work, the CDM contact law will be used based on the experiences gained in , 23 withR is the current radius in the contact which is initialized with the equivalent radius R and R are derived in [R, increases in the contact law. In this formulation of the CDM law, the calculation of the tangential force remains unchanged from the Hertz–Mindlin law. In total the CDM law has four parameters: E, For two contacting spheres , the DEM software calculates the geometric overlap rived in  such thaure, see  for detaAs mentioned before, the simple particle shapes constructed in  will be Sample generation and pre-compaction is done using the method developed in  and willR. This procedure of sample generation and pre-compaction ensures that simulations using different sets of parameters and particle shapes all have the same mass and a similar initial porosity  have considerably lower values of mentclasspt{minimaThe parametrisation of the DEM models to the compression and direct shear tests is conducted using the method described in . In the est, see  for detaE, the parameter levels were chosen according to the measurements values of Kieselkalk ballast, see [As a first step of the parametrisation, compression tests are simulated using parameters belonging to a full Design of Experiments (DoE) plan. The levels of the parameters are given in in Table ast, see . For theAccording to the parametrisation strategy, outlined in , the simE and As discussed in , scatterFigur During the parametrisation process of shapes 21 to 29 severe problems occurred. As an example Fig. All remaining shapes 1–10 consist of clumps of non-overlapping spheres. For these shapes the parametrisation to Kieselkalk ballast was successful. The full details on the parametrisation results can be found in Appendix In Fig. In this subsection, the shape descriptors of the successfully parametrised particle shapes, 1–10, will be compared to those of the declined shapes and those of real ballast stones. Fig. On the contrary, such a distinction is possible with respect to sphericity and convexity. The sphericity values of shapes 1–10 lie between 0.7 and 0.77, while all declined shapes had higher sphericity values. The median value of the ballast stones’ sphericity is shortly below 0.75 and is thus in the range of values of shapes 1–10. The convexity values of shapes 1–10 lie between 1.23 and 1.35, while all declined shapes had lower convexity values. The values of shapes 1–10 are clearly higher than the median value of the ballast stones, but still lower than the maximal measured value of nearly 1.5. The declined shapes reach values below the ballast median value.The most dominant influence seen in the parametrisation is seen for the clump roughness angle. Shapes 1–10 are all non-overlapping clumps and thus have the highest potential for interlocking. Their clump roughness angles are all round 60Summarising, for the construction of simple particle shapes for DEM modelling of railway ballast the key influence factors are the clump roughness angle, sphericity and convexity, while clump surface area and volume also show some influence. Shapes with the highest clump roughness angles, lowest sphericity, highest convexity values and lowest surface area and volume gave the best results.c, see Fig. c decreases and It is interesting to relate the shape descriptors of shapes 1–10 to the parametrisation results shown in Fig. Two conclusions enhance the trust in the parametrisation method: first, for the considered similar shapes the obtained parameter sets are neighbouring. Second, their shape descriptors can be connected roughly to the identified parameters.In this subsection, it will be investigated, if micromechanical differences exist between the particle shapes, which give the best result in the parametrisation: shapes 5, 4, 7 and 3. They have a similar bulk behaviour: close to the experimental one. Remembering the previous section, shapes 4 and 5 have the same parameter set, the parameter set of shape 7 differs slightly, while the parameter set of shape 3 differs more. Both particle shape and the used parameters will influence the micromechanical behaviour. Please note that the coordination numbers of a packing strongly depends on the chosen particle shape representation. Due to the chosen simple particle shapes, the coordination numbers reported in this study are considerably higher than those reported in literature for DEM simulations of railway ballast.R, the coordination number and the percentage of sliding contacts. In the compression test, contact yielding plays an important role, which can be seen at the relatively high amount of yielding contacts. The behaviour of the four particle shapes is very similar: in the loading phase below 25 kN little till no yielding contacts occurs. With increasing load the percentage of yielding contacts rises up to almost 50% and drops sharply to zero in the unloading phase. The plotted mean values of R remain nearly constant during this loading cycle. During the loading cycle, contacts are lost and new ones are created, some of which yield. In total, the mean value of R remains mostly constant for the single shapes. The big differences seen between the shapes (between shape 3 and the remaining shapes) stem from the pre-compaction phase and the loading to the first load maximum (remember the parameter set of shape 3 differs more from the other shapes’ parameter sets).For the compression test, the specimens are compared during the second loading cycle. To characterise the internal state, the following quantities are plotted over the applied normal force in Fig. The coordination number increases slowly during the loading and decreases under unloading as expected. Clear differences between particle shapes can be seen, which remain nearly constant during the considered time.For the percentage of sliding contacts the following behaviour is seen. At the load minimum about 25% of the contacts slide. Under loading this percentage reduces sharply to roughly 5% and increases then slowly with increasing load to its maximum at about 50%. During unloading the percentage of sliding contact drops fast below 10% and increases then with reducing normal to about 30% at the load minimum. The behaviour of shapes 5, 4, and 7 is almost identical, while the one of shape 3 shows some deviations.For a micromechanical analysis of the direct shear test, the same quantities are plotted in Fig. R is nearly same for shapes 5, 4 and 7 and again much lower for shape 3, which can be related to the differing parameter set of this shape. For all shapes, a strong increase in the mean R is seen until 20 mm shear path followed by a moderate increase with further shearing. At shearing contacts, the geometric interlocking has to be overcome first. In some cases, this will be accompanied by yielding, probably with strong increases in R. Due to dilation the sample gets looser over the shear path and the increase in R slows down.The mean value of The coordination number reduces during shearing for all shapes. At the first half of the shear path this reduction is stronger than for the second half. As in the compression test, shape 3 shows the highest coordination number, followed by shape 4, 5, and 7. Compared to the compression test, the coordination numbers are lower, which can be explained by the fully unloading of the end of the compression test.Naturally, the percentage of sliding contacts is very high in the direct shear test. At the beginning of the test, values between 60 and 65% are seen which reduce to values between 40 and 50% after about 15 mm shear path and remain mostly constant during further shearing. The behaviour of the different shapes is very similar, with results for shape 3 slightly higher than for the other shapes.R: while shapes 5, 4 and 7 have very similar values, these are lower for shape 3. Thus, shapes 5, 4 and 7, who have similar parameter sets, show a similar bulk and micromechanical behaviour in the compression and direct shear test. This finding gives confidence that DEM models using these shapes for simulations of other tests/load cases would also produce similar results. However, this is less likely to be the case for shape 3, which has a similar bulk behaviour but differs on micromechanical scale, i.e. R. When different particle shapes are compared for the simulation of a certain material, a micromechanical analysis is advisable.Finally, the micromechanical behaviour of the four shapes 5, 4, 7 and 3 in the compression as well as in the direct shear test gives nearly identical results for the percentage of yielding contacts and the percentage of sliding contacts. Small differences between the shapes are seen in the coordination number for both tests. However, clear differences can be seen in the mean value of This work is the last part of a series of papers on DEM modelling of railway ballast. Two types of ballast named Calcite and Kieselkalk were tested for their bulk behaviour: in uniaxial compression tests Calcite showed a higher stiffness than Kieselkalk, while in a direct shear tests both types of ballast gave very similar results . To inveAll information from the experimental investigations were then used to build DEM models for both types of ballast. Aiming at computational efficiency, simple particle shapes (clumps of three spheres) were constructed in , which tE and The CDM law has four parameters, thus it is a huge task to parametrise DEM models for 20 particle shapes to the compression and direct shear tests. As a preliminary step, Suhr et al.  investigIn this study, the developed parametrisation method was applied to the 20 simple particle shapes to model Kieselkalk ballast. For some particle shapes, problems arose during parametrisation. Nevertheless, for particle shapes 1–10 the parametrisation was successful. The cost function used for the parametrisation was constructed such that values below 1 are considered acceptable. For shapes 1–10 the cost function values lie below 0.52 and thus all 10 shapes are considered acceptable for DEM simulation of Kieselkalk ballast.Shapes 1–10 consist of non-overlapping spheres and have the highest value of the clump roughness angle, indicating high potential for interlocking. From all constructed particle shapes, they have the lowest values of sphericity, the highest convexity index and mostly lower values of surface area and volume. In comparison to the shape descriptors of the ballast stones, the sphericity values of shapes 1–10 lie around the ballast stones’ median value. The convexity values of shapes 1–10 are clearly higher than the ballast median value, while they are still in the range of the ballast’s convexity values. Regarding flatness and elongation, some of shapes 1–10 are close the the ballasts median values, while others are farer away.In the parametrisation method, neighbouring parameter sets are obtained for the considered shapes, which are similar with respect to sphericity and convexity index and the clump roughness angle, which is nearly identical for shapes 1–10. Moreover, in DEM simulations using the different shapes, to some extent a decrease in the convexity index and an increase in sphericity of the shapes can be compensated by increasing the parameters R and percentage of sliding contacts. For the coordination number, small differences were seen between the shapes. Shape 3 also showed similar results regarding the percentage of yielding or sliding contacts and the coordination number. However, the mean values of R were considerably lower for this shape. This finding increases the confidence that DEM models using shapes 5, 4 or 7 for simulations of other tests/load cases would also produce similar results, as their bulk and micromechanical behaviour is similar. When different particle shapes are compared for the simulation of a certain material, a micromechanical analysis is advisable to detect cases like shape 3 .Considering the four shapes that gave the best results in the parametrisation, shapes 5, 4, 7 and 3, a micromechanical analysis was conducted. In the compression and the direct shear tests shapes 5, 4, and 7 showed nearly identical results regarding the percentage of yielding contacts, mean value of The presented results show that computationally efficient simple particle shapes combined with the CDM law are promising for DEM modelling of railway ballast. In contrast to high fidelity shape models, such simple particle shapes aim to capture key aspects of shape, such as interlocking potential. For a final assessment, validation tests with different load cases would be necessary.


Passive muscle stiffness of the RF and VL and knee flexion ROM did not significantly change at any measurement time compared with before dermal suction (p > 0.05). Our preliminary results suggest that dermal suction improves hip extension ROM immediately after dermal suction of the quadriceps femoris, followed by a return to the pre-prescription level 30 min after. However, the effect was prolonged for 120 min and disappeared before 24 h.This study aimed to investigate the acute and prolonged effects of dermal suction on joint range of motion (ROM) and passive muscle stiffness. Eight-minute dermal suction was prescribed for the quadriceps femoris in 15 participants. Hip extension ROM, knee flexion ROM, and passive muscle stiffness of the rectus femoris (RF) and vastus lateralis (VL) were measured before and immediately, 30 min, 60 min, 120 min, 24 h, and 48 h after dermal suction. Passive muscle stiffness was measured using shear wave elastography. Hip extension ROM significantly increased immediately ( Flexibility, a component of physical fitness, is believed to be an important factor in sports performance and injury prevention. Dermal suction is one of several methods for improving flexibility. Dermal suction treatment is called cupping therapy in traditional Chinese medicine. In cupping therapy, a vacuum is applied to a targeted skin area . SeveralIn the aforementioned studies, joint ROM was used as an index of joint flexibility. However, all structures around the joint, such as the skin, muscles, and tendons, contribute to joint flexibility . TherefoThe prolonged effects of dermal suction intervention are important for strength and conditioning coaches or physical therapists who perform dermal suction in clinical practice on joints or muscles. Stretching ,12 and fWe recruited participants among students and researchers at the investigator’s university and received details of the experiment at the time of recruitment. A total of 30 people received the information, of which 15 gave their consent to participate in this study. Participants comprised seven men and eight women who were recreationally active. They did not participate in competitive athletic events, and regular resistance training did not exceed 3 times per week. We requested the participants refrain from strenuous exercise from 24 h before the examination. No participants complained of any injuries specific to the leg. We explained the purpose, content, methods, and risks of the study orally and in writing before the examination. We obtained written informed consent from all the participants. The local ethics committee approved this study .We designed this single-arm study to investigate the acute and prolonged effects of dermal suction on joint ROM and passive stiffness of muscle. In this study, we targeted the right leg in all participants. We measured joint ROM of the hip and knee and passive muscle stiffness of the rectus femoris (RF) and vastus lateralis (VL) before (PRE) and immediately (POST), 30 min, 60 min, 120 min, 24 h, and 48 h after 8 min of dermal suction, which we targeted to the right quadriceps femoris.We measured hip extension and knee flexion ROMs as indices of joint flexibility. We measured these ROMs using the modified Thomas test . The modWe photographed all tests from the sagittal plane  and analyzed using open source software . We marked three anatomical landmarks  to obtain the hip and knee angles. We defined hip extension ROM as the angle formed between a line of the lateral femoral epicondyle and greater trochanter and a horizontal reference line. In addition, we defined knee flexion ROM as the angle formed between a line with respect to the greater trochanter and the lateral femoral epicondyle and a line with respect to the lateral malleolus and lateral femoral epicondyle. We performed each ROM measurement twice and utilized the average of both values for analysis.After measuring the joint ROM, we measured the passive muscle stiffness of the RF and VL. During the measurements, the participants lay in a supine position with their knees fully extended and were instructed to relax all their muscles. We used an ultrasound shear wave elastography scanner  with a linear array transducer  to measure the passive stiffness of muscle. We used shear wave velocity (SWV) (m/s) as an index of muscle stiffness. We placed the probe at the midpoint of the femur  for RF and VL. We measured the SWV twice for each muscle. The measurements of the SWV were performed by a researcher with more than 10 years of experimental experience with ultrasonography. We calculated the SWV over the region of interest, which we manually set for RF and VL. We used ultrasound shear wave elastography scanner built-in software (Q-box) to analyze SWV. We averaged the values of three images and utilized the average value of each measurement for the analysis.In dermal suction, we referred to Enomoto et al. (2021), who examined the acute effects of dermal suction on passive joint and muscle stiffness . We usedPrior to the examination, we conducted a pilot study using five participants to assess reliability. We twice measured hip extension and knee flexion ROM and passive muscle stiffness of the RF and VL for each participant, and then we calculated intraclass correlation coefficients (ICC) and 95% confidence intervals (CIs).α > 0.05) from the least-significant ones with a backward stepwise deletion procedure to arrive at a minimum adequate model [p < 0.05.We performed linear mixed models using RStudio statistical software version 1.4.1106  . We addete model . We evalThe age and physical characteristics of the participants are presented in 2 = 7.777, df = 1, p = 0.032), 60 min , and 120 min  after treatment compared with PRE. However, hip extension ROM at 30 min , 24 h , and 48 h  after treatment revealed no significant differences compared with PRE.The hip extension ROM for each measurement is shown in 2 = 2.621, df = 1, p = 0.633; 30 min after treatment: χ2 = 0.638, df = 1, p = 1.000; 60 min after treatment: χ2 = 3.343, df = 1, p = 0.405; 120 min after treatment: χ2 = 0.558, df = 1, p = 1.000; 24 h after treatment: χ2 = 0.220, df = 1, p = 1.000; and 48 h after treatment: χ2 = 0.297, df = 1, p = 1.000).The knee flexion ROM for each measurement is depicted in 2 = 2.230, df = 1, p = 0.812; 30 min after treatment: χ2 = 0.919, df = 1, p = 1.000; 60 min after treatment: χ2 = 1.816, df = 1, p = 1.000; 120 min after treatment: χ2 = 4.398, df = 1, p = 0.216; 24 h after treatment: χ2 = 0.093, df = 1, p = 1.000; and 48 h after treatment: χ2 = 0.360, df = 1, p = 1.000).2 = 4.319, df = 1, p = 0.226; 30 min after treatment: χ2 = 3.609, df = 1, p = 0.345; 60 min after treatment: χ2 = 4.581, df = 1, p = 0.194; 120 min after treatment: χ2 = 0.103, df = 1, p = 1.000; 24 h after treatment: χ2 = 0.082, df = 1, p = 1.000; and 48 h after treatment: χ2 = 0.589, df = 1, p = 1.000).The SWV of the VL for each treatment is shown in In this study, we compared joint ROM and passive muscle stiffness before and immediately, 30 min, 60 min, 120 min, 24 h, and 48 h after 8 min of dermal suction and obtained the following main findings. Dermal suction increased hip extension ROM immediately, 60 min, and 120 min after dermal suction compared with baseline; however, it was not significantly different at 30 min, 24 h, and 48 h after dermal suction. On the other hand, passive muscle stiffness of both the RF and VL and knee flexion ROM did not change significantly at any measurement time compared with the pre-treatment level. The results revealed that hip extension ROM increased significantly immediately, 60 min, and 120 min after dermal suction but not at 30 min, 24 h, and 48 h after dermal suction . Some poAs mentioned above, hip extension ROM increased significantly immediately, 60 min, and 120 min after dermal suction, with changes ranging from 1.37° to 2.14° when compared with those before dermal suction . ChiacchIn contrast to hip extension ROM, knee flexion ROM showed no significant changes at any measurement time after dermal suction compared with that before dermal suction . It is d3; triceps surae, 819 cm3) [In our study, passive muscle stiffness of both the RF and VL showed no significant changes at any measurement time after dermal suction compared with that before dermal suction . These r819 cm3) . In addi819 cm3) ). These 819 cm3) . This st819 cm3) . On the 819 cm3) . Accordi819 cm3) : 20.7° (819 cm3)  19.1° (S819 cm3)  24.3° (S819 cm3) ; hence, 819 cm3) .This study had some limitations. First, this study was a single-arm study with no control group, making it difficult to determine whether the improvements in hip extension ROM were due to the dermal suction treatment, the effects of natural history, or a placebo effect . Future This preliminary study provides information regarding acute and prolonged changes in passive muscle stiffness and joint ROM after dermal suction. Our results suggest that hip extension ROM improves immediately after dermal suction of the quadriceps femoris, followed by a return to baseline 30 min later. However, the effect was prolonged for 120 min and disappeared before 24 h. Moreover, our results also suggest that dermal suction has limited acute and prolonged effects on knee flexion ROM and passive muscle stiffness of the RF and VL. These results provide preliminary findings to design better future studies on dermal suction.


We investigated the roles of different concentrations of chemical synthetic spherical silver nanoparticles (AgNPs) in protecting pepper seedlings of the Mecca region, which were naturally and artificially infected by the pepper mild mottle virus (PMMoV). The virus shows many infection symptoms, including pepper leaf deformation with filiform leaves and severe mosaic symptoms. Our study focused on the antiviral activity of different concentrations of spherical nanoparticles in controlling PMMoV infecting pepper seedlings. PMMoV identification was confirmed via DAS-ELISA using the following antiserum: PMMoV, cucumber mosaic virus (CMV), tobacco mosaic virus (TMV), tomato mosaic virus (ToMV), potato virus Y (PVY), and tomato spotted wilt virus (TSWV). The presence of PMMoV was confirmed using electron microscopy and reverse transcription polymerase chain reaction (RT-PCR). We evaluated the effects of exogenously applied different concentrations of AgNPs on CMV infection rate, infection severity, virus concentration, and the concentrations of photosynthetic pigments chlorophyll a, chlorophyll b, carotenoid content, phenolic compounds, and protein components in virus-infected plant cells that were treated with three different concentration of nanoparticles  compared to the positive and negative control. Among the vegetable crops of great interest and importance is the pepper crop of the Mecca region, which is grown commercially. Recently, pepper plants have been used to evaluate the effects of antimicrobials and viruses  and to sSeveral viruses infect pepper plants, including PMMoV, CMV, ToMV TMV, PVY, and TSWV ,9,10,11.The use of safer and more environmentally friendly nanoparticle synthesis methods, including biological methods using many living organisms such as primitive nuclei (from bacteria and actinomyces) or eukaryotes (fungi and plants), is gaining worldwide attention ,16. SyntIn our study, we hypothesized that inhibition of viral infection in infected pepper plant cells by synthetic silver nanoparticles (AgNPs) by chemical reduction methods would be the safest way to curb the viral invasion of plant cells . This moSilver nanoparticles synthesized by the chemical reduction method had a size average of 36.32 nm and played a significant role in the defense mechanism of pepper plants against viral infection. When pepper leaves infected with PMMoV symptoms were treated with three different concentrations of AgNPs in our study, they could limit viral replication inside the leaves and reduce the appearance of infection symptoms on virus-infected plant leaves compared to that in the control ,24.Samples of pepper leaves that showed clear symptoms of PMMoV were collected from the different pepper crops in the Mecca region. These symptoms included transparency of the veins (vein clearing) accompanied by severe mosaic formation, reduction in the blade of the leaves (filiform leaves), and small size with distortions of the leaves deformation see a. The saThe presence of PMMoV was confirmed via RT-PCR using a primer specific to the virus RNA-dependent RNA polymerase (RdRp) gene amplification. The authors of ,28 reporMicroscopic examination of very thin sections of infected and healthy pepper leaves using transmission electron microscopy (TEM) showed a random distribution of the globular virus particles with a laminated aggregate of crystalline particles inside the infected cells see c,d,e. ThA surface plasmon resonance (SPR) device at a wavelength of 450 nm, measured using an ultraviolet spectrophotometer, was used to corroborate the configuration of the AgNPs ,33,34. TAs shown in The treatment of seedlings with AgNPs 24 h after a viral infection led to no symptoms of viral infection in plants. We also found that treatment with AgNPs during viral infection led to a great decrease in the appearance and severity of infection symptoms in the virus-infected leaves compared to that in the seedlings treated 72 h after viral inoculation, which did not affect viral infection or disease severity.B. licheniformis, showed excellent in vitro antiviral activity against the bean yellow mosaic virus. The significance of the particular antiviral activity is highlighted, given the significant yield reduction in fava bean crops resulting from bean yellow mosaic virus infections in many African countries [This may be due to the inability of the AgNPs to activate induced systemic resistance (ISR) of the plant against PMMoV infection. Moderate reductions in all symptoms were observed when the virus-infected plants were treated with AgNPs simultaneously during the infection in group 2. This result was an agreement with that of , which rountries ,38,39,40In general, the virus has a great ability to affect the breaking down of light pigments inside infected plant cells or indirectly influence the ability of the light pigments to participate in the photosynthesis process, which leads to the emergence of an external infection display such as mosaic symptoms or mottling, and therefore when pepper seedlings were treated with silver nanoparticles after infection with the virus for 24 hours it led to an increase in the content of light pigments in the treated cells, and thus a disappearance in the external symptoms of infection compared to other treatments. Additionally, increasing the concentration of silver nanoparticles led to an increase in the accumulation of the photopigments inside chloroplasts in infected cells, compared to that by treatment with low concentration of nanoparticles during the pre-viral replication phases. In infected pepper leaves inoculated with PMMoV, the total phenolic content was lower than that in healthy leaves. However, infected leaves treated with three concentrations of AgNPs showed a significant increase in phenolic content compared with untreated infected leaves shown in . These rInfected pepper leaves recorded a high accumulation rate of total soluble protein compared to that in healthy leaves, but infected leaves treated with 200, 300, and 400 µg/L of AgNPs 24 h after PMMoV inoculation were recorded as having inferior build-up of total soluble protein compared to other treatments and the healthy and infected controls. On the other hand, in this study we recorded the lowest decrease of total soluble protein when infected pepper plants were treated with 400 µg/L AgNPs 24 h after inoculation compared with other concentrations of AgNPs shown in .In infected pepper leaves inoculated with PMMoV, the total phenolic content was lower than that in healthy leaves. However, leaves treated with AgNPs showed a significant increase in phenolic content at all treatment concentrations. All plants treated with AgNPs showed a higher build-up of phenolic content compared to that of infected leaves. Infected pepper leaves recorded a high accumulation of total soluble proteins compared to that of healthy leaves, but infected leaves treated with 400 µg/L AgNPs 24 h after PMMoV inoculation were recorded as having an inferior build-up of total soluble proteins compared to 200 µg/L of AgNPs and the control. These results are consistent with those of , which mCapsicum annuum L leaves collected from different crops in the Mecca region generally exhibited symptoms of typical PMMoV infection.Samples of infected pepper Chenopodium giganteum leaves to purify the virus and record the shape of local legions surrounded with little halo edges that would appear on infected leaves; the virus was then multiplied using artificial mechanical transmission on healthy sweet pepper cv.(Sirtaki) seedlings. Following the incubation period for viral growth and propagation, the symptoms of the infection started appearing and were recorded [The presence of pepper mild mottle virus (PMMoV) was confirmed in infected pepper leaf samples collected from different crops in the Mecca region via DAS-ELISA, as explained by ref.  using anrecorded .Artificial mechanical transmission was used to transfer the virus artificially from infected pepper leaves to healthy sweet pepper cv.(Sirtaki) seedlings using an abrasive substance to allow the virus to enter the cell without dying.Infected plant juice was extracted using a phosphate-buffer solution (pH 7) to maintain the stability of the virus during the extraction. Then, the infected seedlings were sprayed with water, and it was during the incubation period of the virus (inside the cells) that the symptoms of the infection started appearing on the newly grown seedlings, which was after approximately 3 weeks .Ultra-precise sections of the mesophyll tissue of the infected leaves were examined using an electron microscope (JOEL-JEA100 CX.) to detect the presence of inclusion bodies inside the infected cells, as well as the intracellular changes caused by the virus, compared to the healthy sections. Tissues were cut, fixed, and stained according to standard procedures described in ref. .The presence of the virus in all infected and healthy plant samples was tested via DAS-ELISA using antibodies specific to PMMoV  at a wavAccording to the methods described in ref. , specifi3) was added as a starting material to synthesized spherical silver nanoparticles after heating at 60 °C with vigorous stirring to 20 mL (8.0 mM trisodium citrate C6H5O7Na3 solution) reducing agent and surfactant pre-heated at 60 °C. The mixture was stirred well for 20 min, then the mixture was cooled to room temperature with continuous stirring [A quantity of 80 mL of silver nitrate  with 0.15405 nm Cu Kα radiation. The size and morphology of the spherical AgNPs produced were studied by TEM using the methods of ,35,48,49The sweet pepper seedlings cv. (Sirtaki) were planted for germination on wet filter paper at room temperature for 2–3 d, and pepper seedlings were transferred to sterilized containers filled with soil and stored in a greenhouse. After 3 weeks of growth, plants of similar sizes were selected and divided into three groups, and each group was divided into eight treatments: T1: , T2: , T3: , T4: , T5: (infected), T6: (NPs-200 µg/L and infected), T7: (NPs-300 µg/L and infected), T8: (NPs-400 µg/L and infected). Three replicates were allocated for each treatment, and the three groups were divided as follows: the first group was treated with nanoparticles at the time of viral inoculation, the second group was treated with nanoparticles 72 h before the inoculation, and the third group was treated with nanoparticles 24 h after the inoculation .The leaves of healthy and virus-infected pepper plants were treated with AgNPs. After three weeks of treatment, the virus concentration, infection rate, and infection capacity were recorded based on the scale recommended by ,50:DiseaThe photosynthetic pigments  were evaluated as described by  in the t662) − (0.99 × E644) = mg/g (F.wt.)Chl a = (9.784 × E644) − (4.56 × E662) = mg/g (F.wt.)Chl b = (21.426 × E440.5) − 0.268 (chla + chlb) = mg/g (F.wt.)Car=  using a pyrogallol standard curve, according to .Total soluble protein content was evaluated in the leaves of infected plants treated with AgNPs and compared with that in the control, according to .The SPSS program was used to perform all statistical analyses of the tables under study, with all numerical results subjected to investigation of disagreement (ANOVA).Chenopodium giganteum. PMMoV RNA-dependent RNA polymerase (RdRp) gene was discovered in infected leaves by RT-PCR when fragments of an expected size of 830 bp were amplified. A study of the ultra-structure showed the presence of laminated aggregate of crystalline particles and viral particles associated with abnormalities in the shape and composition of both the nucleus and chloroplasts. The TEM images confirmed the production of AgNPs at nanoscale; most of them were mono-dispersed with spherical shapes and found to be an average 36.32 nm, confirmed using TEM images. Healthy plants in the experiment were not affected by all concentrations of AgNPs. In infected pepper leaves inoculated with PMMoV, the total phenolic content was lower than that in healthy leaves. However, leaves treated with AgNPs showed a significant increase in phenolic content at all treatment concentrations. All plants treated with AgNPs showed a higher build-up of phenolic content compared to that of infected leaves. Infected pepper leaves recorded a high accumulation of total soluble proteins compared to that of healthy leaves, but infected leaves treated with 400 µg/L of AgNPs 24 h after PMMoV inoculation were recorded as having an inferior build-up of total soluble proteins compared to 200 µg/L of AgNPs and the control.In our study, symptoms included transparency of the veins (vein clearing) accompanied by severe mosaic formation, reduction in the blade of the leaves (filiform appearance), and small size with distortions of the leaves (deformation), which are those of typical infection with PMMoV observed in pepper leaves. The samples that tested positive in the ELISA served as the virus source in our study. The virus was biologically purified after isolation using a single local lesion technique; the local legions were surrounded with little halo edges on


Prosopis juliflora (Pj) as an energy feedstock. Proximate, ultimate and biochemical characterization of Pj pods were conducted, and thermal behaviour and calorific values were determined. Cellulose and hemicellulose were isolated and characterized by reliable methods. The overall characterization has revealed the Pj pods as a potential feedstock for bioenergy. The collected Pj pods contain (% w/w) moisture 7.89 ± 0.002, volatile matter 87.67 ± 0.002, ash 0.21 ± 0.002, fixed carbon 4.23 ± 0.002 with a calorific value of 17.62 kg/MJ. The CHNS content was (w/w %) carbon 41.77, nitrogen 3.58, sulfur 26.3 and hydrogen 6.55. The biochemical composition analysis yields (% w/w) on a dry basis; cellulose 26.6 ± 0.18, hemicellulose 30.86 ± 0.27, lignin 4.71 ± 0.12, protein 11.63 ± 0.12 and starch 1.1 ± 0.06 and extractives 30.56 ± 0.008. The isolated cellulose and hemicellulose were analyzed and confirmed by CP/MAS &1H NMR, FTIR, TG-DSC, SEM, XRD, and TGA. The present results revealed that the tested biomass, Prosopis juliflora, could be used as a feedstock in biorefinery for bioenergy.The production of renewable and sustainable biofuels using inevitable wastes is a promising alternative to the alarming depletion of fossil fuels. Significantly, the sustainable biorefinery of lignocellulosic waste, as an alternative fuel source, is a prognosticating approach to tackle many agricultural/forestry residues and offers a circular economy as well as environmental benefits. But, the heterogeneity of lignocellulosic biomass is one of the major bottlenecks in lignocellulosic biorefinery. Thus the characterization of lignocellulosic biomass is essential to understanding the feedstock's nature, composition and suitability for biofuel production. The present study taps evergreen spiny non-edible pods of  Energy demand is anticipated to increase by around 50% over the next 20 years, with predictions of 778 EJ by 20352. But the exhaustion of fossil fuels and the subsequent increase in greenhouse gas emissions have become urgent problems3. These harmful greenhouse gases also harm human health because they cause cancer4. Thus finding an alternative energy source that can satisfy energy needs also be environmentally benign is vital4. In contrast to other environmentally friendly energy sources, including wind, solar, geothermal, marine, and hydropower, biomass may directly produce fuel and chemicals5.The foundation of modern civilization, energy is essential to most human activities. Modernizing our lives and cities would not have been possible without energy6. The energy from the lignocellulosic biomass is greatly interesting since it is abundantly available, technically feasible and economically viable7. Several technologies, including gasification, combustion, pyrolysis, enzymatic hydrolysis and fermentation, are employed to turn biomass into fuel and platform chemicals3. Thermochemical processes often demand a large amount of energy, including a solvent or catalyst. In contrast, the biochemical approach has a protracted time and has less effective at breaking down the resistant biomass3. Various lignocellulosic biomass like sugarcane bagasse8, berry waste2, jatropha seeds9, sunflower straw10, sorghum stem11, rice straw12, waste peach pulp13, wheat straw14, corn stover15 and so forth have been studied for the effective production of biofuels.Lignocellulosic biomass comes in various wastes, including woody, aquatic detritus, farm manure and byproducts, and agricultural wastes16. Cellulose is the primary polymer that contains long chains of cellobiose units, a renewable resource that is safe for the environment, inexpensive, non-toxic, biodegradable, and biocompatible17. Hemicellulose comprises monosaccharides such as mannose, galactose, arabinose, glucose, rhamnose, xylose and uranic acid18. Lignin is a high molecular weight, complex structure containing cross-linked phenolic monomers that provide cell wall rigidity, impermeability and resistance against microbial attack19.The lignocellulosic feedstock contains majorly cellulose 40–50%, hemicellulose 25–35% and lignin 15–20%4. Nevertheless, there is a constant demand for environment-friendly disposal or refinery of various lignocellulosic biomass20. From the standpoint of waste valorization, the fractionation of this lignocellulosic residue into cellulose, hemicellulose, and lignin is the primary step for producing a variety of products with a higher economic value, such as biofuel, cosmetics, biodegradable films, paper, and plastic materials, while minimising the associated environmental impacts19.The lignocellulosic biomass usage in biorefineries for biofuels offers reduced greenhouse gas emissions, zero waste generation and significant sustainability in the circular bioeconomy21. By recycling trash using pyrolysis to create biochar, energy, and value-added goods, agricultural and animal waste disposal can lessen its negative environmental effects22. Biochar production from lignocellulose is simple, reliable, technically feasible and commercially viable23. Using biochar as a feed for animal growth reduces methane formation and enhances anaerobic digestion23.One of the most promising technologies for converting lignocellulosic biomass into high-value products like biochar, syngas, and bio-oil without oxygen is pyrolysisProsopis juliflora pods, for its suitability as bioenergy feedstock. The Prosopis juliflora (Pj) plant is a drought-tolerant, evergreen tree with drooping branches of the Leguminosae family as an energy stock24. The native of the Pj pods to arid and semi-arid regions of the world. Pj plants have been colonized in many countries, such as North America, Africa and Asia. Pj plants have flexible branches, a green–brown, a twisted stem, and curving multi-seeded pods with a hardened pericarp and flattened25. Pj plants can survive in a very harsh and poor soil environment, control erosion, improve soil fertility, and reduce soil salinity in drylands. Due to invasive properties, Pj plants grow as much as 9 m in length and 50 cm in diameter. The fruit is a non-dehiscent curved pod of 10–15 cm in length, 0.8–1.0 cm in width, 2–4 mm thick, and contains a hard endocarp, soft and heavy mesocarp and woody endocarp with many seeds24. The pods generated by the Prosopis species are legume pods containing high levels of carbohydrates, protein, and starch used as animal feed for the past decades. Prosopis species are made up of exocarps, fibrous endocarps and fleshy mesocarps. The nutritional value of a plant might vary depending on the phenological stage, species, plant portion, world region, and growing climatic conditions26.The present study aims to exploit one such lignocellulosic biomass, 27 as biopesticides28, gums for antioxidants29, extracts for antiplasmodial activity30, seed powder for the removal of Pb (II)31 and all parts of the plant for antibacterial activity32. Pj plants have higher carbon and less hydrogen content, leading to the best carbon-effective ratio for biorefinery. Pj pods were used as a raw material in a few fermentations to produce cellulolytic enzymes33, xylanase34, oxytetracycline35, ethanol36, bio-oil37 and biohydrogen38 and biochar production under pyrolysis39. To the best of our knowledge, the structural characterization of Pj pods was not yet reported.Pj plant has been investigated for several applications, leaf extract to treat sore eyes, sore throat, diarrhoea, and open wounds40. The physicochemical properties of biomass help design the biomass conversion process, including feeding, conversion, sorting of intermediate products, and collection of products. Biomass physical, chemical and thermal properties are related to the gasification process, and the knowledge of biomass composition is also essential for solid waste management41. Therefore, the present study deals with the proximate, ultimate, biochemical and thermal properties of the Prosopis juliflora (Pj) pods to aid the selection of Pj pods as a feedstock for bioenergy and value-added products.The systemic characterization of biomass is essential for biofuel production35.Pj pods  were collected from Pudukkottai, Tamil Nadu, India, as per the ASTM International standards—ASTM E1757-1. The plant biomass was identified by botanist Dr Sam Aldrin, SASTRA Deemed University, India. The raw pods were collected from the same region for the entire study to prevent changes due to different nutritional content. The collected pods were washed completely with tap water and dried under sunlight. The dried pods were ground to achieve a 0.4–0.1 mm particle size using an ASTM sieve40. Essential biofuel quality can be determined by proximate and ultimate analysis42.Proximate analysis of Pj pod powder was conducted to estimate the moisture, volatile matter, ash content and fixed carbon40.The conventional oven-dry method (NREL) was used to estimate the moisture content according to ASTM E1756-01. Briefly, 10 g of Pj pods powder was weighed in the crucible and heated in a hot air oven at 105 °C for 4 h. Pj pods powder was removed from the crucible and allowed to cool in a desiccator. The moisture content of Pj pods was calculated using Eq.  content was estimated as the difference in weight of moisture, volatile matter and ash content from the initial biomass. The fixed carbon content of Pj pods was calculated using Eq. 40.3\docu40.The elemental composition of Pj pod powder, including carbon, nitrogen, oxygen, hydrogen and sulfur, was determinedCHNS analysis was carried out using CHNS Elemental analyser model Elemental Vario EL III Germany, analysed at CECRI Karaikkudi, India.43.X-ray fluorescence spectrophotometry was used to estimate the mineral composition of the Pj pods. Briefly, the Pj pods powder was mixed with boric acid as a binder under high pressure (22 tons) to make a pellet. The pellet results were recorded in X-ray fluorescence (XRF) spectrometer S8 Tiger, Bruker AXS, Germany, analysed at SASTRA Deemed University, Thanjavur, India, using a 4-kW rhodium anode X-ray tube44. The acid bleach method was used to isolate and estimate the Pj pods' cellulose. Briefly, 5 g of Pj pods powder was loaded in a Soxhlet apparatus with water and ethanol and reflexed for 8 h to eliminate the wax. The dewaxed Pj pods powder was bleached using 1.5% sodium chlorite for 2 h at 70 °C at pH 3.5. The bleaching procedure was repeated until a consistent white colour was obtained, indicating pure cellulose. The obtained cellulose was filtered, dried and weighed45.Pulping and bleaching are two crucial steps to extracting cellulose effectively from lignocellulosic biomass45.The filtrate from the acid bleaching step was treated with 1 M NaOH at 65 °C for 2 h, neutralized with 6 M HCl at pH 5.5 and precipitated using 46.Acid hydrolysis was used for the isolation of lignin. Briefly, 1 g of Pj pod powder was hydrolyzed with 72% sulfuric acid, autoclaved for 2 h at 121 °C and filtered off. The filtrate was considered acid-soluble lignin, and the residue was acid-insoluble lignin. Further, the acid-soluble lignin was estimated using a UV–Visible spectrophotometer and Eqs.  & 5)46.46.5)46.4UV (abs), average absorbance for sample; ε, absorptivity (specific wavelength); ODW sample, sample weight in mg; path length, the path length of UV–Vis cell in cm.44.Ethanol and benzene extract fat, wax, oil, gums, sugars and other pigmenting components from LCB. In a Soxhlet apparatus, 2.5 g of dried Pj pods powder was extracted with 150 mL of ethanol at 70 °C and reflexed for 8 h. The extracted Pj pods powder was dried in a hot air oven at 105 °C for 2 h. The weight difference between moisture-free pods and extractive complimentary pods powder is represented as extractives47. Briefly, 0.5 g of Pj pods powder was mixed with 10 mL of concentrated sulfuric acid and 5 g of catalyst mixture . The mixture was taken in a digestion tube, loaded into the digester, and heated to 400 °C until the mixture turned light green, indicating the complete digestion process. The digested sample was distilled in a kelplus distylemba distillation system with a hose connected (Model classic dx). One side of the hose was connected with a conical flask and loaded with 20 mL of 4% boric acid and mixed indicator . The other side of the hose was loaded with 1 M NaOH and distilled. The digested sample was heated by passing the vapour constantly, and the released ammonia was absorbed into a conical flask. With the absorption of ammonia, the pinkish colour turned to green colour. The collected distillate was titrated against 0.02 N of sulfuric acid until the sample colour was changed from green to pink. Micro Kjeldahl total nitrogen was estimated from the titer values. Crude protein was calculated using Eq. The anthrone method was used to estimate the starch. Briefly, the 0.1 g Pj pods powder was mixed with 25 mL 80% of hot ethanol and 5 mL of distilled water. The mixture was stirred thoroughly, centrifuged, and decanted off. The residue was again centrifuged with 30 mL of 80% hot ethanol. Two extracts were combined under pressure in a boiling water bath. The alcoholic extract was mixed with 5 mL of distilled water and 6.5 mL of perchloric acid. The mixture was stirred for 30 min at room temperature and centrifuged. The pellet was suspended in 5 mL of distilled water; the perchloric acid extraction was repeated and centrifuged. Two extracts were combined, diluted to 100 mL and filtered.49.Further, 5 mL of the filtrate was diluted to 100 mL with distilled water and used for starch analysis. The starch solution, 1 mL, was diluted with 1 mL of water, and 10 mL of anthrone reagent was added. The mixture was boiled in a boiling water bath for 12 min, cooled, and the amount of glucose was determined by a spectrophotometer. Starch was estimated from the glucose, assuming the 0.9 g starch yields 1 g glucose. Anthrone reagent was prepared as 10 mg of anthrone in 10 mL of concentrated sulfuric acidThe calorific value of Pj pods powder was determined with a Bomb calorimeter . In a bomb, 0.46 g of moisture-free Pj pods powder was taken in a crucible, kept inside the bomb, and filled with 30 bar pressure of oxygen to combust the Pj pods powder.50of model SDT Q-600 to identify the thermal stability of Pj pods, cellulose and hemicellulose. The thermal analysis was carried out under a nitrogen atmosphere of 0.5 lb/in2 with a 100 mL/min flow rate. The different stages of degradation were observed when the components started heating from 37 to 1000 °C.TGA was performed in a Thermal analyzer40 at SASTRA Deemed University. The wavelength was measured in transmission mode from 400 to 4000 cm−1 by the ATR  method51. Methanol was used to clean the ATR crystal before use. Later, the crystal was scooped with samples until they covered the surface area of the crystal and pressed with a mechanical anvil.The functional group of Pj pods, cellulose and hemicellulose were determined by Fourier Transform Infra-red Spectroscopy Spectrum one, Perkin Elmer USA13C spectrum of cellulose and hemicellulose was studied by Bruker Avance HD 500 MHz spectrophotometer from IITM, SAIF, CSAIF, Chennai, India44. The following condition was analyzed: resonance frequency at 125.76 MHZ; proton 90° pulse sequence was 4.30 µs; contact pulse 3300 µs, recycle delay was 5 s; acquisition time 0.02 s; 1024 scan per spectrum were used.CP/MAS 44.The SEM imaging of dried pods, acid-treated pods, cellulose and hemicellulose, was studied by Vega 2 TE-SCAN microscope with a speeding voltage of 10.0 kV. The magnification of the isolated cellulose was focused on up to 500X50 at Bishop Heber College, Trichy, India.The isolated cellulose crystallinity index was analysedusingPANalyticalXpert343.X-ray Fluorescence Spectroscopy analysis was conducted to estimate the chemical elements in the Pj pods powder. The mineral composition was studied by Bruker SPECTRA plus for S8 TIGER X-ray Fluorescence Spectroscopy52. Table 53 and decreases the yield of biomass combustion. Moisture content is represented as the quantity of water present per unit of Pj pods powder, and the low moisture content yields more combustion. High moisture content will decrease the yield of biomass combustion54. Pj pods showed a higher volatile matter content of 87.67 ± 0.002% w/w, which could be a valuable source for pyrolysis. The fuel reactivity has also been influenced by volatile matter55. The high volatile content of biomass has a propensity to easily burn and degrade during the thermochemical reaction, ideally yielding a higher proportion of bio-oil during the pyrolysis56.The proximate and ultimate analyses were used to determine the composition of Pj pods40. The amount of total minerals in biomass indicates the ash content or total ash63. The higher and lower heating values, the amount of solid material to release, dust particles in combustion gases, boiler fouling, and corrosion are all strongly correlated with the ash content64. In addition, a high amount of ash content acts as a heat sink during pyrolysis or combustion that will reduce the fuel energy process65 and biomass with a greater ash content generates more char leftovers during pyrolysis56. Pj pods with a low ash content of 0.21 ± 0.002% w/w, comprising phosphorous, silica, aluminium, sodium, iron, calcium, and potassium, could be better feedstock for fuel energy production. However, ash is one example of a solid fraction that has the potential to be used in agriculture because of its high nitrogen, phosphorous, and potassium content2.Ash is composed of mineral and inorganic matter of biomass and affects the combustion rate of biomass40. The high amount of fixed carbon represents a high energy level. In most gasifiers, the conversion of FC into gases dictates the pace of gasification and its yield, making fixed carbon a significant parameter for gasification study66. Knowing fixed carbon's shape and hardness can aid choose the right combustion equipment since they reveal a fuel's caking qualities67. The high volatile matter will increase biochar yield in thermochemical conversion processes. Pj pods have a fixed carbon content of 4.23 ± 0.002% w/w, producing low char during combustion.Fixed carbon (FC) is the amount of leftover Pj pods powder after releasing volatile matter, moisture, and ash68, low greenhouse emissions42 and thermochemical conversion processes such as pyrolysis, torrefaction and gasification2 and that can produce streams of liquid, solid, and gaseous products. Higher carbon content and a lower oxygen content must result in a higher heating value since the carbon content of biomass is directly correlated with the heating value, and the oxygen content is inversely correlated with the heating value69. The amount of carbon and oxygen are directly correlated with the heating value. Pj pods have a high amount of carbon, meaning that the more significant percentage of carbon, the higher the heating value of the fuel.The elemental composition of Pj pods was determined by ultimate analysis Table . The low64. One of the primary gases emitted during the thermal reactions of biomass is a nitrogen oxide (NOx). Therefore, these emissions need to be controlled as they affect human health. (NOx) consist of seven oxides that are considered the most emissions in an air70. When burning fuel, NOx emissions can be reduced in seven major methods by employing a catalytic combustion system, reduced air preheat, low NOx burner to lower the peak temperature, air staging and natural gas burning. Similarly, the ozone-depleting NO2 can also be treated in a similar way70.Additionally, a more significant oxygen content decreased the heating value of Pj pods, and biomass nitrogen content is crucial for evaluating nitrogen oxide (NOx) emissions, an air pollutant71.As determined by XRF, the micronutrients Zn, Mn, P, Cu, Ca, Mg, Na, K, and Fe were present in the whole Pj pods Table . The mic50. Lignin is the major obstacle to biomass conversion as it inhibits microorganisms' growth72.Chemical analysis data for biomass is crucial information when examining the reaction properties of solid fuels. The term "holocellulose" refers to the total cellulose, which is made up of cellulose and hemicelluloses, and extractives constitute the significant portion of Pj pods powder, 88% with less protein, lignin and starch Table . During 72. Pj pods constitute 57.76 ± 0.14% w/w holocellulose less than the Pj bark, 77%50.Sodium chlorite-based bleaching is an effective method to isolate cellulose from biomass. Alkaline treatment for the isolation of hemicellulose was a widely reported method, as it effectively cleaves the ester bond between lignin and hemicellulose47 and Pj seeds as 4.5%54.The amount of protein in Pj pods was estimated by the micro Kjeldahl method. Pj pods have shown a good amount of protein, 11.63 ± 0.12% w/w, an excellent alternative and low-cost animal feed. Further, the exact protein composition has to explore to assess the nutritional value, net protein utilization and digestibility. The protein content of Pj pods from Ecuador was reported as 10.1% (dry basis)73, and the high amount of extractives needs further evaluation.With the high amount of hollocellulose and less lignin content of 4.71 ± 0.12% w/w, Pj pods could be an effective source for energy conversion through biochemical approaches. The negligible amount of starch 1.23 ± 0.06% w/w was determined, similar to starch obtained from waxy maize starch 1.2 ± 0.9%74. The higher HHV also indicates the Pj pods significance as a bioenergy source. The HHV value was calculated by using Eq. . Therefore, lignin does not exhibit a straightforward weight loss derivate peak−1 corresponding to the axial angular bending of the O–H bond in hydroxyl groups in α cellulose, showing how the hydroxyl group is broadly distributed throughout the cellulose and hemicellulose structures85. Spectral peaks at 1635 cm−1 were assigned carbonyl stretching C–O for the acetyl group in hemicellulose and the aldehyde group in lignin. The C–O–C bond deformation of hemicellulose and cellulose was assigned to 1422 cm−1. The spectral peak at 1056 cm−1 can be assigned to the C–O–C group in Pj pods, and the overlap of stretching vibrations of glycosidic bonds in glucomannan and xylan85. A spectral peak of 606 cm−1 can be assigned to β glycosidic linkages within sugar units , with potentially smaller aspect ratios, giving a large surface area for chemical pretreatment Fig. A. Due to75. Removing amorphous components causes the cellulose index to rise, resulting in the more significant intensity peak associated with bleached fibres99. In Pj pods, cellulose, a well-defined crystalline peak, was observed at 2θ = 10, 22.85 and 38.27 . There were three typical peaks for Pj pods cellulose at 2θ angles of 2θ = 10º, 22.85º and 38.27º regions related to cellulose type I . There is a direct correlation between the degree of crystallinity and the hardness of the samples. The sample becomes more difficult as the crystallinity index risesProsopis juliflora (Pj) pods were characterised using FTIR, SEM, XRD, NMR, HHV and TGA. Lignin, cellulose, and hemicellulose were isolated from Pj pods and evaluated. Pj pods have shown a low high carbon content of 41.77% w/w, a high calorific value of 17.073 MJ/kg, 57.76 ± 0.14% w/w of holocellulose and ash content of 0.21 ± 0.002% w/w. All the parameters were in good agreement with the reported literature. The composition and functional properties of holocellulose, and lignin may differ based on the structure and species of the biomass. However, only marginal differences can be observed in individual biomass based on seasonal and geographical variation. The significant amounts of holocellulose, 57.76 ± 0.14% w/w and the whole carbon content encourage high-value applications of Pj pods in biomass biorefinery. Minor proportions of lignin make it suitable for the fermentative production of biofuels. The present results indicated the significance of the valorization of Prosopis juliflora (Pj) pods. However, further in-depth investigations are required to efficiently utilize the same in environmental remediation, biomass-biorefinery and biofuel technologies. Recently our team has reported the hybrid hydrolysis and fermentation of Prosopis juliflora (Pj) pods for hydrogen production38. The scaleup of hydrogen production is in progress. Further, the combined thermochemical and biochemical routes for refining Prosopis juliflora (Pj) pods for bioenergy and other value-added products could contribute to the circular economy.