Furthermore, NMR and CT techniques were used to observe the changes in the microstructure of BF concrete. The experimental results show that the BF contribution index Cf reaches the largest value when the BF content is around 3 kg/m3, approximately 2.7; in this case, the mechanical properties of BF concrete were also optimal, and the Cf was only 2.34 when the BF content was 7.5 kg/m3. NMR and CT test results show that there is a strong correlation between the BF morphological parameters and the distribution of pore structure in the concrete matrix. The overlapping contact of BF clusters led to the penetration of pores, which led the macro-pore proportion to increase dramatically. The increase in the macro-pore proportion is the main reason for the deterioration in concrete performance. In addition, these macro-pores may have adverse effects on the chloride ion permeability of BF concrete.The orientation, distribution, and contact point density of BF  in the concrete matrix play significant roles in the mechanical properties of BF concrete, but represent a weak point in current research. It is meaningful to study the morphological characteristics of BF in concrete. In this study, the transparent model test and joint blocking method were innovatively adopted to investigate the correlation of dosage with the BF morphological parameters and concrete mechanical properties. A focus on a BF dosage of 0–7.5 kg/m Fiber-reinforced concrete has been widely applied in civil infrastructure construction and underground engineering in recent years ,2. AmongAs a composite material, concrete’s mechanical performance is directly affected by its meso-structure ,10,11. IHowever, due to the opaque features of concrete, it is challenging to directly observe its inner meso-structure and the actual distribution of fibers; thus, it is necessary to adopt some advanced detection methods , such as2 solution, and the joint blocking method was used to analyze the morphological parameters  of BF. Compared with previous methods, the transparent model provides a new way to clarify BF dispersion characteristics. In addition, we combined this with NMR and CT technology to study the meso-structure of BF-reinforced concrete. The NMR technique was used to obtain the inner pore parameters of concrete with different BF contents; CT three-dimensional reconstruction technology was used to clarify the geometric characteristics of pores in the concrete. Through the comparative analysis of the transparent model, NMR, and CT test results, the mechanism of concrete performance deterioration was clarified. This is significant to further improve the gain effect of BF to concrete and optimize BF concrete’s mechanical properties in the future.At present, there are still barely suitable methods to study the dispersion form of BF (silicate composition) in concrete. To clarify the dispersion characteristics of BF and the influence mechanism of the morphological characteristics on the meso-structure, in this study, a transparent model test was adopted innovatively, which is frequently used in the field of geotechnical engineering. The distribution of BF in concrete was simulated by a transparent model, which was composed of quartz sand and CaBrIn this work, we used Conch brand P.O 42.5 Portland cement, the chemical composition of which is shown in 3, 1.5 kg/m3, 3 kg/m3, 4.5 kg/m3, 6 kg/m3, 7.5 kg/m3). The basic mix ratio of the concrete is shown in There were six different amounts of BF in the concrete , which was designed by the Changchun Academy of Mechanical Sciences . We followed the GB/T50081-2002  standard2 solution by using an Abbe refractometer, and adjust the refractive index of the CaBr2 solution to 1.4585. Step 4: Pour the CaBr2 solution into the quartz sand; after 1 min, pour the mixture into a PMMA box (100 × 100 × 100 mm). Step 5: After the lumen of the lighting system is adjusted to the most appropriate value, the front of the model can be photographed with an industrial camera to obtain the characteristic distribution image of BF in the transparent aggregate.The distribution of BF in concrete can be simulated by using a transparent model, which consists of a transparent aggregate and liquid medium with the same refractive index. The schematic flow of the transparent model test is shown in 2 relaxation spectrum. The T2 relaxation spectrum can reflect the characteristics of the liquid–solid interface, and the pore size determines the relaxation decay rate. Hence, the relaxation velocity directly reflects the pore structure information.Nuclear magnetic resonance (NMR) is a powerful tool to study the inner structure of opaque materials. It can detect porosity, pore distribution, permeability, etc. In nuclear magnetic physics, the relaxation spectrum is suitable to quantify the rate of signal decay. The peak of the monitored signal decays exponentially; it represents a TThe equipment selected for this test was the ZYB-Ⅱ vacuum pressure saturation device and the MesoME23-040V rock microscopic pore structure analysis system, developed by Suzhou Niumay Analytical Instrument Co., Ltd., Suzhou, China; the magnetic field intensity of the instrument was 0.5 T.The test was divided into five steps: (1) Sampling; (2) Vacuum water saturation; (3) Calibration; (4) Testing; (5) Data processing. Firstly, the concrete specimens with different BF contents were selected for coring and polishing, and then cylindrical specimens with 35 mm diameter and 50 mm height were produced. Because the NMR signal source is a hydrogen atom, it is necessary to fill the pores of the samples with water. After sufficient water saturation, we put the specimen into the instrument for testing. Then, the detected water signal was converted into pore information. In this way, pore information can be obtained. The flow of the nuclear magnetic resonance test is shown in Computed tomography (CT) technology was used to obtain the internal images of samples. However, there will be noise interference during the scanning, so preprocessing is needed to remove noise points in images to ensure quality. Furthermore, image cutting was necessary as the data of the original CT image were quite large, which is not conducive to processing and observation. The flow mainly included median filtering, 3D reconstruction, threshold adjustment, pore extraction, etc. Then, the internal characteristics of the BF and concrete mix were obtained. Avizo software  was used in this process, as shown in The HPC-225FB high-precision CT test system was used in this study; the working voltage of this system was 80 kV, the working current was 8 mA, the resolution was 10 μm, the number of scanning frames was 3 fps, and the pixel matrix was 1536 × 1930. The dimensions of the scanned BF concrete sample were consistent with the NMR sample .3, approximately 23.12 MPa, which is higher than 0 kg/m3 BF content concrete by approximately 9.4%. However, when the BF content increased to 7.5 kg/m3, the compressive strength of the concrete decreased by 13.2% compared to 3.0 kg/m3 BF content. The tensile strength reached the maximum value when the BF content was also 3.0 kg/m3, at approximately 1.9 MPa. The tensile strength of 3.0 kg/m3 BF content increased by 20.2% compared to 0 kg/m3 BF content, but the tensile strength of 7.5 kg/m3 BF content decreased by 13.7% compared to 3.0 kg/m3 BF content concrete. The flexural strength reached 2.81 MPa when BF content was 3.0 kg/m3, 6.1% higher than 0 kg/m3 BF content. When BF content increased to 7.5 kg/m3, the flexural strength decreased by 6.4% compared to 3 kg/m3 BF content.Curing time was 7 d: the compressive strength reached the maximum value when the BF content was 3.0 kg/m3, approximately 31 MPa, which was higher than 0 kg/m3 BF content concrete by around 9.9%. When the BF content increased to 7.5 kg/m3, the compressive strength of the concrete decreased by 10.3% compared to 4.5 kg/m3 BF content. The tensile strength reached the maximum value when the BF content was 3.0 kg/m3, around 2.42 MPa. The tensile strength of 3.0 kg/m3 BF content increased by 26.2% compared to 0 kg/m3 BF content, but the tensile strength of 7.5 kg/m3 BF content decreased by 22.2% compared to 3.0 kg/m3 BF content. The flexural strength reached 3.49 MPa when the BF content was 3.0 kg/m3, 8.1% higher than 0 kg/m3 BF content, and as the BF content increased to 7.5 kg/m3, the flexural strength decreased by 26.9% compared to 3 kg/m3 BF content.Curing time was 14 days: the compressive strength reached the maximum value when the BF content was 4.5 kg/m3, approximately 41.2 MPa, which was higher than 0 kg/m3 BF content concrete by around 31.3%. As the BF content increased to 7.5 kg/m3, the compressive strength of the concrete decreased by 14.6% compared to 3.0 kg/m3 BF content. The tensile strength reached the maximum value when the BF content was also 3.0 kg/m3, approximately 2.62 MPa. The tensile strength of 3.0 kg/m3 BF content increased by 16.6% compared to 0 kg/m3 BF content, but the tensile strength of 7.5 kg/m3 BF content decreased by 13.2% compared to 3.0 kg/m3 BF content. The peak flexural strength was 3.9 MPa when the BF content was 3.0 kg/m3, and the flexural strength of 3.0 kg/m3 BF content increased by 13% compared to 0 kg/m3 BF content. As the BF content increased to 7.5 kg/m3, the flexural strength decreased by 6.3% compared to the peak flexural strength. Curing time was 28 days: the compressive strength reached the maximum value when the BF content was 3.0 kg/m3. The mechanical properties of BF concrete will deteriorate when BF is used excessively; we hypothesize that the main reason for this phenomenon is the density of BF, which affects its dispersion form in a concrete aggregate, and then changes the microstructure of the concrete matrix. In the following, the test results of the transparent aggregate model are used to discuss the influence of content density on the morphological characteristics of BF concrete, and reveal the meso mechanism of the deterioration of BF concrete’s mechanical performance.It could be found that the best content range of BF in concrete is around 3.0–4.5 kg/mThe model image and binary images of the BF distribution pattern in the transparent model with different BF contents are shown in i is the image block number, i = 1, 2…, 9, and Pi is the set of pixels in the block image.In order to identify BF’s scattered morphological characteristics in the visual field, the joint blocking method was used, as shown in lH is three components in the HSV color system, if is the number of pixels with color value i of each component, and N is the total number of pixels in the local block of the image.The orientation and local color information of the visual field could be obtained by using the joint blocking method, and the HSV space was used for color feature extraction, as shown in jχ(i) and jψ(i) are local color histogram vectors of the image, i = 1, 2, …, m, θ is the average value of the χ2 distance (BF spacing) of all blocks, and the calculation formula is as follows:icosφirsinθ and isinφirsinθ represent the BF position vector in the transparent model.The BF distribution position in the image is marked by the kernel function:iK can describe the imaging differences in the image; χ2 means can reflect the visual similarity of two kinds of color histograms, which provides an advantage for obtaining the BF’s morphological characteristics in the transparent model.The color histogram kernel function β was introduced to evaluate the BF dispersion in the transparent model. The expression was as follows:μ is the average number of fibers in the visual field, and β is the dispersion coefficient. Fibers dispersed well when β ∈ , and fibers dispersed poorly when β ∈ . β of BF is negatively correlated with the BF content. When the BF content is 1.5 kg/m3, β is 0.92, and the effect of BF dispersion is the best.The dispersion coefficient fC  was defined to evaluate the gain effect of BF on strength. The expression is as follows:Contribution index 3, C3 = 2.7. The contribution index was around 2.45 when the BF content was 6 kg/m3, and the contribution index was approximately 2.34 when the BF content was 7.5 kg/m3. It could be seen that Cf decreased gradually with increasing BF content. It can be inferred that when the BF content exceeds the optimal value, the excess part of BF agglomerates and does not exert the gain effect.The BF’s average contribution index with different BF contents is represented in 3, there are a few fibers overlapping in the visual field. For BF content of 4.5 kg/m3, some fiber clusters come into contact and overlap, and when the BF content is more than 4.5 kg/m3, there is a dramatic increase in fiber cluster overlapping, the trend of dispersion coefficient is shown in Based on the above analysis, excessive BF content will cause the fiber agglomerates to form clusters. When the BF content is more than 3.0 kg/mr < 0.01 μm), capillary pores (0.01 μm < r < 5 μm), and connected pores (r > 5 μm). As BF concrete increases, the percentage of gelation pores decreases gradually, and there is a significant increase when the BF content increases from 3 kg/m3 to 4.5 kg/m3. This indicates that BF content of 3–4.5 kg/m3 is the sensitive range for the gelation pores, and 1.5–4.5 kg/m3 is the sensitive range for the capillary pores. When the BF content ≤ 3 kg/m3, the content of connected pores is low, but when BF content ≥ 3 kg/m3, the content of connected pores increases dramatically with the increase in BF content.The results of the transparent model test show that the BF overlapping and agglomeration lead to a reduction in its contribution index on concrete. However, the main reason for the deterioration in concrete performance has not been determined. In order to identify the main reason for the deterioration in BF’s concrete mechanical performance, nuclear magnetic resonance technology was used to detect the characteristics of the inner structure of BF concrete. Because of the fiber clustering effect, there are only small gaps in between BF, which makes it challenging for the matrix to enter the fiber agglomerates, and there are crevices in the fiber agglomerates. These small gaps act as connectors to penetrate the primary pores in the concrete matrix, leading to changes in the pore structure in the concrete matrix. The total pore content increased with the increase in the BF content, and the total pore content significantly decreased with the increase in the curing time.3; the pore component difference caused by the BF content reached 32.6%. When the curing time reached 14 d, the main diameter pore component of the plain concrete sample was approximately 0.091%, but the main diameter pore component of concrete was approximately 0.117% when the BF content was 7.5 kg/m3; the pore component difference caused by BF content reached 28.5%. When the curing time reached 28 d, the main diameter pore component of the plain concrete sample was approximately 0.093%, but the main diameter pore component of concrete was approximately 0.113% when BF content was 7.5 kg/m3; the pore component difference caused by BF content reached 21.5%. The reason for this is as follows: with the increase in the curing times, the hydration reaction produces CSH, AFT, AFM, CH, etc. These materials will fill the pores in the concrete matrix, leading the total pore ratio to decrease gradually.When the curing time reached 7 d, the main diameter pore component of the plain concrete sample was approximately 0.095%, but the main diameter pore component of concrete was approximately 0.126% when the BF content was 7.5 kg/m3. When BF content > 3 kg/m3, the porosity increases substantially with the BF content increase. 3, the percentage of harmful pores increases dramatically.In previous studies, connected pores (r > 5 μm) were defined as the key factor affecting the mechanical properties of concrete ,21. Henc3, and they were the worst when the BF content was 7.5 kg/m3. Therefore, BF content 3 kg/m3 and BF content 7.5 kg/m3 were compared and analyzed. max ≤ 1 μm) of the BF concrete sample (3.0 kg/m3) is higher than that of the BF concrete sample (7.5 kg/m3). For pores of Feretmax > 1 μm, the BF concrete sample (BF content 3.0 kg/m3) is significantly lower than that of the BF concrete sample (BF content 7.5 kg/m3). This indicates that the proportion of small pores decreases with the BF content increasing from 3.0 kg/m3 to 7.5 kg/m3; conversely, the proportion of large pores gradually increases with the BF content increasing from 3.0 kg/m3 to 7.5 kg/m3.Reconstruction results of the pores in BF concrete are shown in 3, pores with roundness of 0.604–1 account for 86.7%. When the BF content is 3 kg/m3, the pores with roundness of 0.310–0.502 account for 89.2%. When the BF content is more than 4.5 kg/m3, the pores with roundness of 0.310–0.502 account for 91.7%. This means that most of the pores are round when the BF content is low and most of the pores are irregularly shaped when the BF content is high, as shown in 2 has a certain gap with that of the cement slurry, which may cause the distribution of BF to deviate. Nonetheless, it does not affect the trend of the results. CT or X-ray scanning are only suitable for studying the distribution morphology of fibers , which has a large gap with concrete in terms of density. Although the fluorescence tracer method has high accuracy, at present, the fluorescence tracking method is mainly used to detect the morphological form of synthetic fibers, which is easily labeled by a fluorescence tracer. For the silicate fiber, the effect of the fluorescence tracking method is still not ideal. Therefore, the detection method should be selected in a targeted manner for different types of fibers.Compared with X-ray and CT scanning, the transparent model method is more suitable for observing the distribution of silicate fibers, but it still has certain shortcomings. (1) The density of quartz is slightly lower than that of actual concrete aggregates. If converted according to the ratio of BF weight/model volume, there is a slight deviation between the quantity of BF added in the transparent soil model and the quantity of BF in the concrete sample. (2) For cementitious materials, the viscosity of the cement has a certain effect on the distribution of aggregates and fibers. However, in the transparent model test, the viscosity of the CaBr3), so that the size is close to the maximum particle size of concrete aggregates. If such specimens are tested, the test results will not be general. NMR has the same accuracy as the MIP method to study the pore structure of BF concrete. The NMR method can detect samples with a relatively large size, and the NMR sample preparation is simpler; therefore, the NMR method was adopted in this study, combined with CT reconstruction, and the geometric characteristics of pores in concrete with different BF contents were compared and analyzed.MIP is a powerful tool for studying pore parameters, such as pore radius, roar radius, and the pore–roar ratio of rock or concrete. This method has high accuracy, but the period for specimen preparation in the MIP test is too long, especially for the constant velocity mercury injection method. Typically, in order to shorten the test period, the specimen size must be as small as possible (around 1 cm(1)3. However, the mechanical properties of concrete will deteriorate when BF is used excessively. At 28 d, the compressive, tensile, and flexural strengths of concrete with a BF content of 7.5 kg/m3 were lower by 14.6%, 13.2%, and 6.3%, respectively, than when the BF content was 3.0 kg/m3.The mechanical test shows that BF concrete’s mechanical properties were the best when the BF content was around 3.0 kg/m(2)fC of BF was the largest when the BF content was 3 kg/m3, C3 = 2.7, but it was only 2.34 when the BF content was 7.5 kg/m3.The transparent model test and joint blocking method were used to explain the morphological characteristics and dispersion mechanism of BF in the concrete matrix. The results show that when the BF content exceeds the critical value, the excess part of BF agglomerates and does not display a gain effect. The contribution index (3)There is a strong correlation between BF’s morphological behavior and pore structure in the concrete matrix. The overlapping contact of BF clusters leads to the penetration of pores, which leads the macro-pore proportion to increase dramatically, and these macro-pores easily evolve into initial micro-cracks under loading. The increase in the macro-pore ratio is probably the main reason for the deterioration of concrete’s performance.In this paper, the correlation and the internal mechanism between BF content, BF morphological characteristics, and the mechanical properties of concrete were explored through experiments. The following conclusions were drawn. 2 solution was different from that of cement slurry, which may have caused the morphological properties of BF in the transparent model to differ from those in the concrete. Nonetheless, the trend of BF distribution in the transparent model with a change in BF content is the same as that in concrete. In future research, the transparent medium’s fluid viscosity should be also considered as an important factor.The viscosity of cementitious materials was also a significant factor affecting BF’s morphological features. In this study, the viscosity of the CaBr


Despite the impact of inappropriate prescribing on antibiotic resistance, data on surgical antibiotic prophylaxis in sub-Saharan Africa are limited. In this study, we evaluated antibiotic use and consumption in surgical prophylaxis in 4 hospitals located in 2 geographic regions of Sierra Leone.We used a prospective cohort design to collect data from surgical patients aged 18 years or older between February and October 2021. Data were analyzed using Stata version 16 software.Of the 753 surgical patients, 439 (58.3%) were females, and 723 (96%) had received at least 1 dose of antibiotics. Only 410 (54.4%) patients had indications for surgical antibiotic prophylaxis consistent with local guidelines. Factors associated with preoperative antibiotic prophylaxis were the type of surgery, wound class, and consistency of surgical antibiotic prophylaxis with local guidelines. Postoperatively, type of surgery, wound class, and consistency of antibiotic use with local guidelines were important factors associated with antibiotic use. Of the 2,482 doses administered, 1,410 (56.8%) were given postoperatively. Preoperative and intraoperative antibiotic use was reported in 645 (26%) and 427 (17.2%) cases, respectively. The most commonly used antibiotic was ceftriaxone 949 (38.2%) with a consumption of 41.6 defined daily doses (DDD) per 100 bed days. Overall, antibiotic consumption was 117.9 DDD per 100 bed days. The Access antibiotics had 72.7 DDD per 100 bed days (61.7%).We report a high rate of antibiotic consumption for surgical prophylaxis, most of which was not based on local guidelines. To address this growing threat, urgent action is needed to reduce irrational antibiotic prescribing for surgical prophylaxis. For the first time, we have used the World Health Organization (WHO)–defined daily dose (DDD) to estimate antibiotic consumption in surgical prophylaxis in 4 hospitals located in 2 geographic regions of Sierra Leone.There were a high rates of antibiotic use  and consumption (117.9 DDD per 100 bed days) in surgical antibiotic prophylaxis, which is consistent with local guidelines in only 410 (54.3%) surgeries.The most commonly used antibiotic was ceftriaxone 949 (38.2%) with a consumption of 41.6 DDD per 100 bed days.The Access antibiotics accounted for 72.7 DDD per 100 bed-days (61.7%), whereas consumption of the Watch antibiotics was 45.2 DDD per 100 bed-days (38.3%). No Reserve antibiotics were utilized.Factors associated with preoperative antibiotic prophylaxis were the type of surgery, wound class, and consistency of surgical antibiotic prophylaxis with local guidelines. Postoperatively, surgical wound type and consistency of surgical antibiotic prophylaxis with local guidelines were important factors associated with antibiotic use.Introduction However, the positive impact of antibiotics on health and other development sectors is threatened by the global increase in the levels of antimicrobial resistance (AMR) and the decline in the discovery of new antibiotics.The discovery of antibiotics heralded a major success in the treatment and prevention of infectious diseases. In 2019 alone, the global deaths attributable to AMR were 1.27 million, making AMR the leading infectious cause of death after coronavirus disease 2019 (COVID-19) and tuberculosis (TB).AMR is a growing public health problem because its global burden continues to increase and could reach an unprecedented level of 10 million deaths per year by 2050 if global efforts to control it are not mobilized. Consequently, the burden of AMR in LMICs is much higher than in high-income countries. This fact is reflected in the recent revelation that the West African subregion has the highest mortality burden due to bacterial AMR.Antibiotic consumption has increased in low- and middle-income countries (LMICs) due to improving economies and high burden of infectious diseases. In response to the numerous health and safety threats posed by AMR to populations, recent global efforts have been mobilized to prevent an increase in the burden of AMR. Despite these efforts, many gaps in the global AMR prevention and control remain, including the lack of routine surveillance data on antibiotic use and consumption in many countries in sub-Saharan Africa. At the national level, efforts are underway to reduce the AMR burden in Sierra Leone with support from the Fleming Fund. Even so, our previous studies have shown many challenges in the prevention and control of AMR in the country. Some of these challenges include a high burden of extended-spectrum β-lactase (ESBL)–producing gram-negative bacteria and gaps in hand hygiene implementation.AMR prolongs hospital stays, increases morbidity and mortality, diverts financial resources that could be used to improve health, and threatens efforts to combat infectious diseases. The use of antibiotics is essential to preventing surgicalsite infections. In many cases in low-income countries, however, the use of antibiotics for surgical prophylaxis is often inappropriate, unnecessary, and can exacerbate antibiotic resistance. Therefore, data on the use and consumption of antibiotics for surgical prophylaxis in medical institutions are crucial for formulating and implementing policies to address the problem of inappropriate use of antibiotics for surgical prophylaxis. Using this information, hospital-based antimicrobial stewardship programs are able to minimize or prevent the growing threat of AMR and can address the problem of limited access to essential antibiotics that are needed to treat infectious diseases affecting humans.The abuse and overuse of antibiotics, including surgical antibiotic prophylaxis, is one of the main reasons for the increased burden of AMR. and (4) to identify patterns of antibiotic consumption using the WHO Access, Watch, and Reserve (AWaRe) classification system.In this study, we assessed antibiotic use and consumption in surgical prophylaxis in 4 hospitals located in 2 geographic regions of Sierra Leone to inform antimicrobial stewardship interventions and monitor antibiotic consumption trends. The specific study objectives were (1) to determine the prevalence of antibiotic use for surgical prophylaxis, (2) to define the predictors of antibiotic use for surgical prophylaxis, (3) to evaluate antibiotics consumption using the 2021 Anatomical Therapeutic Chemical (ATC) and defined daily dose (DDD) method,In this study, we used a prospective cohort design to assess antibiotic use and consumption for surgical prophylaxis. Although MGH is the only government-owned hospital in its catchment area, Freetown has 8 hospitals that provide surgical services, with a total capacity of 890 beds. The study was conducted on 523 hospital beds (69%) in Freetown.We selected 4 hospitals for the study because they share similar characteristics with many secondary- and tertiary-care health facilities in Sierra Leone: Connaught Hospital (CH), Miliatary Hospital (MH), Makeni Government Hospital (MGH), and Lumley Government Hospital (LGH). LGH (42 beds) is a secondary-care facility, but the remaining 3 hospitals provide tertiary care. CH (300 beds), MH (181 beds), and LGH are in Freetown, the capital of Sierra Leone, with a population of 1 million, whereas MGH (207 beds) is located in a regional city, ∼170 km from the capital, with a catchment population of 606,544 or ∼8.6% of the Sierra Leonean population.With the exception of the national referral hospital (CH), which provides surgical services to the nonpregnant population, the remaining 3 hospitals provide surgical services to both pregnant and nonpregnant populations. All 4 hospitals are owned by the government of Sierra Leone. On average, each week, 20 adult patients undergo surgery at CH, 17 at MGH, 10 at MH, and 4 at LGH.The study included all adult (18 years or older) patients who received elective and emergency surgical services at 4 hospitals between March 2021 and October 2021 as part of a surgical-site infection surveillance program. Because the surveillance program excluded patients with amputation or other bone surgery, data on prophylactic surgical antibiotics in these procedures could not be included in the study. The Access antibiotics have a low resistance potential and should always be available to treat common bacterial infections. Because the Watch antibiotics are the highest-priority antimicrobials, they are key targets for monitoring the progress of hospital-based antimicrobial stewardship programs. The Reserve antibiotics are the last-line, used to treat multidrug-resistant infections and are the target of national AMS programs. When assessing antibiotic use in surgical prophylaxis, data were disaggregated by indication for surgical prophylaxis and related to the class of wound.Various standardized classification systems were employed to categorize antibiotic use or consumption patterns. The WHO AWaRe framework classifies antibiotics used for surgical prophylaxis as listed in the WHO Model List of Essential Medicine. Using the ATC system, antibiotics used for surgical prophylaxis were classified according to their anatomical, therapeutic and/or pharmacological, and chemical properties into all levels of classification at the subgroup level  and the chemical substance level . Antibiotic consumption was evaluated using the DDD per 100 bed days for each of the 4 hospitals. In assessing antibiotic consumption, aggregated surgical antibiotic prophylaxis data were estimated for each hospital.Antibiotic substances were further classified using the 2021 ATC classification system.P value <.05 was considered statistically significant.Trained nurses collected the data in the Epicollect software platform  from the patient files and patient and/or ward nurse interviews. After collection, the data were extracted into Microsoft Excel , cleaned, coded, and then transferred into Stata version 16 software  for analysis. Descriptive statistics such as frequencies and percentages were used to summarize demographic and clinical characteristics of study participants, as well as antibiotic consumption. The proportion of indications and the type or dose of surgical antibiotic prophylaxis consistent with the local antibiotic guidelines were determined using descriptive statistics. Binary logistic regression was employed to identify factors associated with antibiotic use. Results were adjusted for hospital variables and are presented for the pre- intra-, and postoperative periods. Firth correction was applied where observations were zero. A  To calculate DDD per 100 bed days, we divided consumption in DDD by bed days and multiplied by 100. A bed day was defined as an overnight stay in the hospital. We calculated the total DDD per 100 bed days in the sample and then determined the contributions to this total for the pre-, intra-, and postoperative periods, as well as for each antibiotic. We also calculated antibiotic consumption per 100 bed days for each hospital.The DDD values were calculated by converting the total amount of antibiotic dispensed into grams, which were then divided by the WHO-assigned DDD based on the 2022 version of the ATC/DDD index.Ethical approval was obtained from the Sierra Leone Ethics and Scientific Review Committee of the Ministry of Health and Sanitation, Government of Sierra Leone. Written informed consent was obtained from each participants.In total, 753 patients underwent surgery during the study period. Most had their surgeries at Connaught Hospital ; most patients were female ; and the median patient age was 30 years IQR, 10–87). The sociodemographic characteristics of patients in the 4 hospitals are summarized in Table 0–87. TheThe surgical procedures considered in this study were abdominal surgeries , caesarean sections , and nonabdominal surgeries . The wound classes were clean , clean contaminated , and contaminated . Although >90% of surgical wounds in LGH, MH, and MGH were clean, fewer than half of the surgical wounds at CH were clean .Of the 753 total surgeries, 723 (96%) had at least 1 dose of antibiotics in at least 1 period . Only 30 patients (4%) had surgery without any surgical antibiotic prophylaxis. In the preoperative period, 490 patients (65%) received antibiotics. Nearly all patients in MH and MGH  received antibiotics during the preoperative period. Also, one-third  received antibiotics during surgery . In the postoperative period, 692 patients (92%) received antibiotics. Although CH administered antibiotics to 77% of its patients, LGH placed all of its patients on antibiotics, and almost all patients received antibiotics at MH and MGH .As shown in Table P = .034) than nonabdominal surgeries. Compared to clean wounds, antibiotics were 6 times more likely to be used in clean-contaminated wounds  and 47 times more likely in contaminated wounds . Preoperative patients were >3-fold more likely to receive antibiotics when they are indicated in the local guidelines  postoperatively than nonabdominal surgeries.Using binary logistic regression analysis and adjusting for hospitals, the significant factors associated with preoperative surgical antibiotic prophylaxis were the type of surgery, wound class, and consistency of surgical antibiotic prophylaxis with local guidelines. Abdominal surgeries were nearly 3 times more likely to receive antibiotics for surgical prophylaxis,  were given postoperatively. Preoperative and intraoperative antibiotic use was reported in 645 cases (26%) and 427 cases (17.2%), respectively. Among the hospitals, MH had the highest rate of antibiotic use  followed by CH , LGH , and MGH .The most used antibiotic was ceftriaxone  with consumption of 41.6 DDD per 100 bed days. Cefuroxime and clarithromycin were each administered only once , whereas consumption for the Watch antibiotics was 45.2 DDD per 100 bed days (38.3%) . It was highest in the postoperative period at 59.9 DDD per 100 bed days. Its proportion ranged from 59.3% to 62.3% of total DDD per 100 bed days within the operative periods. Consumption rates for antibiotics in the Access category ranged from 47.2% in CH to 78.5% in MH. No antibiotics in the Reserve group were utilized. Other details on antibiotics consumption are shown in Figure  All 4 hospitals had a combined DDD of 117.9 per 100 bed days, which means that 1.179 DDD of antibiotic prophylaxis were administered for every procedure in these hospitals each day. The DDD reported in our study was higher than the DDD reported for surgical antibiotic prophylaxis in Nigeria (n = 17) and Indonesia (n = 30). The high rates of antibiotic use in our setting may be due to a lack of oversight of rational antibiotic prescribing. This rate may be reduced by a functional antibiotic stewardship system. In 2019, a national antibiotic consumption study in Sierra Leone reported a DDD of 19 per 1,000 inhabitants per day. Although a direct comparison with this study is somewhat inappropriate due to differences in methodology and study setting, national antibiotic consumption of 19 DDD was much lower than the 117.9 DDD reported in our study. The large differences between national and hospital-based data suggest that estimates of antibiotic consumption can provide better information if studies are performed at different levels.To the best of our knowledge, this is the first large-scale surgical antibiotic prophylaxis study in Sierra Leone. In this study, most patients received antibiotics for surgical prophylaxis, higher than the levels reported in Ghana, Gambia, and Turkey. One of the priority gaps identified in this study is the high rate of surgical antibiotics prophylaxis that does not follow local guidelines. In nearly half of the patients, indications for antibiotic prophylaxis were inconsistent with the local antibiotic guidelines. Similarly, the dose or type of antibiotics used for surgical prophylaxis was inconsistent with local guidelines in 75% of the surgeries. This finding may be explained by the fact that although all the hospitals in this study have drugs and therapeutics committees, the committees have broader responsibilities, including procurement and supply chain management, and they cannot effectively carry out AMS activities. In 2017, the first antimicrobial guideline in Sierra Leone was developed at CH. Although there were plans to scale-up the use of these guidelines to other hospitals, their implementation has not been supported by local AMS activities, such as dedicated leadership and a multidisciplinary approach, which has hindered their effective use. Therefore, hospital administrators and policy makers should take urgent action to develop robust strategies to address the inappropriate surgical antibiotics prophylaxis in Sierra Leone. Functional hospital-based AMS programs are a good platform to protect available antibiotics and prevent the development of resistance. Nonetheless, establishing an AMS program in a low-income country like Sierra Leone requires practical action and sustainable funding. Hence, support from the government and its partners is needed to set up and sustain AMS activities in hospitals.We identified gaps in our estimate of antibiotic consumption, which if not addressed urgently, could hinder the effort of the country in meeting the UN Sustainable Development Goals , especially for reducing AMR.Our study highlights significant differences in the pattern of antibiotic consumption in surgical prophylaxis, which is expected given the varying levels of service across these 4 hospitals. Paradoxically, however, the DDD of the national referral hospital that admitted more sick patients was lower than that of the other hospitals. This finding may be due to differences in human resources required for the rational use of antibiotics for surgical prophylaxis. A national oversight system for rational antibiotic prescribing in hospitals can address this variability in prescribing patterns. and with a study conducted in Nigeria, but it differs from a study conducted in Turkey and the WHO guidelines on surgical antibiotic prophylaxis. This fact adds to the background rationale that surgical antibiotic prophylaxis should follow local normative guidance.The most common antibiotics used for surgical prophylaxis in this study were ceftriaxone, metronidazole, and ampicillin. This finding is consistent with our previous study Most antibiotics used for surgical prophylaxis in this study were in the Access group and none were in the Reserve group, probably due to the unavailability of drugs in this group or perhaps due to their high cost. However, this finding shows good prescribing practice because the WHO guidelines recommend that at least 60% of total antibiotic consumption in any particular setting should be in the Access group. In making this assertion, it is important to note that the AWaRe classification of antibiotics provides an overall national target and is not specifically designed to classify antibiotic use patterns in individual hospitals. Owing to their higher resistance potential, Watch antibiotics should be a key target for AMS interventions, especially in hospitals where ceftriaxone  is the most common antibiotic used for surgical prophylaxis.Antibiotics in the Access category are widely available and affordable and are used as first- or second-line treatment.Our study had both strengths and limitations. A large sample size of patients who received surgical services from 4 different hospitals in 2 geographic regions of Sierra Leone was included in this study. Although not nationally representative, this evidence provides insight into the wider use of antibiotics for surgical prophylaxis and can be used to promote and implement AMS programs. We used local unpublished antibiotic guidelines developed in 2017 to determine the appropriateness of indications, doses, and types of antibiotics for surgical prophylaxis. Because these guidelines were not developed based on local evidence, it may not represent the true picture of surgical antibiotic prophylaxis in this setting. Also, the study population was largely unbalanced, which may have affected the representation of pregnant women. We did not assess surgical antibiotic prophylaxis in private hospitals, which can be done in future research.In conclusion, we report a high rate of antibiotic consumption for surgical prophylaxis, most of which was not based on antibiotic guidelines. To address this growing threat to global health security, key stakeholders in the AMR and infection prevention and control response should take immediate steps to reduce irrational antibiotic prescribing for surgical prophylaxis by establishing local normative guidelines and training healthcare workers to follow the surgical antibiotic prophylaxis guidelines.


S. aureus (MRSA) therapy after 3 days of hospitalization. We conducted interviews at 10 Veterans’ Affairs medical centers (VAMCs) during the preimplementation period (N = 16 antibiotic stewards) and postimplementation (N = 13 antibiotic stewards) ~12 months after program initiation. Preimplementation interviews focused on current stewardship programs, whereas postimplementation interviews addressed the implementation process and corresponding challenges. We also directly asked about the impact of COVID-19 on stewardship activities at each facility. Interviews were transcribed and analyzed using consensus-based inductive and deductive coding. Codes were iteratively combined into barrier and facilitator groupings. Barriers identified in the preimplementation interviews included challenges with staffing, the difficulties of changing prescribing culture, and academic affiliates . Facilitators included intellectual support , facility support, individual strengths of antibiotic stewards , and resources such as VA policies mandating stewardship. By the postimplementation phase, all sites reported a high volume of COVID-19 cases. Additional demands were placed on infectious disease providers who comprise the antibiotic stewardship teams, which complicated the implementation of SSTOP. Many barriers and facilitators mentioned were similar to those identified during preimplementation interviews. Staffing problems and specific providers not “getting it [stewardship activities]” continued, whereas facilitators centered around strong institutional support. Specific pandemic-related barriers included slow down or stoppage of stewardship activities including curbing of regular MRSA screening practices, halting weekly stewardship rounds, and delaying stewardship committee planning. Pandemic-specific staffing problems occurred due to the need for “all hands on deck” and challenges with staff working from home, as well as being pulled in multiple directions, . Furthermore, an increase in antibiotic use was also reported at sites during COVID-19 surges. Our findings indicate that SSTOP implementation met with barriers at most times; however, pandemic-specific barriers were particularly powerful. Sites with strong staffing resources were better equipped to deal with these challenges. Understanding how the program evolves with subsequent COVID-19 surges will be important to support the broad implementation of SSTOP.Effective stewardship strategies such as an “antibiotic timeout” to encourage prescriber reflection on the use of broad-spectrum antibiotics are critical to reduce the threat of multidrug-resistant organisms. We sought to understand the facilitators and barriers of the implementation of the Antibiotic Self-Stewardship Timeout Program (SSTOP), which used a template note integrated into the electronic health record system to guide decision making regarding anti- methicillin-resistant Funding: NoDisclosures: None


Dermatophagoides pteronyssinus (Der p) asthma and/or allergic rhinitis (AR). Forty-four patients received subcutaneous Der p-AIT for 2 years, and eleven patients received only symptomatic treatment. The patients needed to finish their questionnaires at each visit. Serum and salivary Der p-specific IgE, IgG4, and IgE-blocking factors (IgE-BFs) were measured at 0, 4, 12, and 24 months during AIT. A correlation between them was also evaluated. Subcutaneous Der p-specific AIT improved the clinical symptoms of children with asthma and/or AR. The Der p-specific IgE-BF significantly increased at 4, 12, and 24 months after AIT treatment. Serum and salivary Der p-specific IgG4 significantly increased with the time of the AIT treatment, and significant correlations between them at different time points were observed (p < 0.05). Furthermore, there were significant correlations (R = 0.31–0.62) between the serum Der p-specific IgE-BF and Der p-specific IgG4 at the baseline, 4, 12, and 24 months after the AIT (p < 0.01). The salivary Der p-specific IgG4 levels also demonstrated a certain correlation with the Der p-specific IgE-BF. Der p-specific AIT is an effective treatment for children with asthma and/or AR. Its effect was associated with increased serum and salivary-specific IgG4 levels, as well as an increased IgE-BF. Non-invasive salivary-specific IgG4 may be useful for monitoring the efficacy of AIT in children.At present, there are no effective, non-invasive, and objective indicators to evaluate the efficacy of pediatric house dust mite (HDM)-specific allergen immunotherapy (AIT). A prospective, observational study was performed in children with  Dermatophagoides pteronyssinus (Der p) and Dermatophagoides farinae (Der f) [Along with economic development and social progress, the prevalence of allergic disease has increased significantly. In China, house dust mites (HDMs) were the most prevalent allergens in patients with asthma and/or allergic rhinitis (AR) . Further (Der f) . Current (Der f) .Der p-specific IgA antibodies are dominant in the serum and saliva of non-allergic children, while Der p-specific IgE and IgG4 antibodies predominate in allergic children, suggesting the specific IgA has a key role in the healthy immune response to mucosal allergens [Der p-IgG4 and Der p-IgA levels in AR patients were significantly lower than those in the healthy controls, while Der p-IgE levels were significantly higher [Der p-IgG4 and Der p-IgA in the maintenance phase compared with the up-dosing phase, whereas Der p-IgE only increased in the up-dosing phase, further indicating a protective role of IgG4 or IgA against allergies [Sublingual HDM-AIT could induce symptom improvement and immunomodulation . A threellergens . Salivary higher . Moreovellergies .Der p-AIT, and their correlation became stronger over the treatment time. Furthermore, the IgE-blocking antibodies induced by the subcutaneous Der p-AIT are mainly allergen-specific IgG4, not IgA [Der p AIT in children with asthma and/or AR, observing whether salivary IgG4 levels would be an alternative immunological marker of AIT treatment.Due to its competitive ability to bind allergens with IgE, IgG4 is considered an IgE-blocking antibody and a marker of immune tolerance . This Ig not IgA . TherefoDer p and had specific serum IgE against Der p. All the included patients were followed up to assess AIT within 24 months of recruitment. This study was approved by the Ethic Review Board of Children’s Hospital, Zhejiang University School of Medicine (2016-IRBAL-056). Informed consent was obtained for all the subjects who were under 18 from a parent and/or legal guardian.In this prospective observational study, children with asthma and/or AR were enrolled. Mild-to-moderate asthma and AR were diagnosed according to the GINA guidelines and ARIA guidelines, respectively. All the patients were skin prick test positive for Der p Alutard SQ vaccine . The treatment protocol followed the recommended up-dosing schedule of 16 weeks before reaching a maintenance dose of 100,000 Alutard SQ, with a duration of 24 months for the AIT. The patients were observed in the outpatient for at least half an hour after each administration in order to detect and treat any immediate adverse reaction.The standardized HDM-specific subcutaneous immunotherapy was performed at the clinic. The patients received subcutaneous injections of the aluminum-formulated Patients were requested to assess their symptoms and record the medication use over the past week at each visit during the whole course of treatment. The symptom and mediation use were scored based on WAO recommendations and our previous study ,11. PatiThe patients were also asked to fill out the rhinoconjunctivitis quality of life questionnaire (RQLQ) and asthma quality of life questionnaire (AQLQ) in the past week at each visit. The RQLQ is composed of 7 domains, including activity limitation, sleep, non-nose/eye symptoms, practical problems, nose symptoms, eye symptoms, and emotional function, with 28 questions; and the AQLQ included 31 questions involving activity limitations, symptoms, emotional function, and exposure to environmental stimuli, based on previous studies ,12. Theig for 10 min) of blood samples and were then stored at −20 °C until the assays. Unstimulated saliva samples were collected from the oral cavity using standard Salivette devices  according to the manufacturer’s instructions. The swab was placed in the mouth for 5 min and then reinserted in the Salivette. Samples were immediately placed on ice and centrifuged at 3000× g for 10 min. The supernatants were collected and stored at −20 °C until further analysis.Serum and saliva samples were obtained from all the children at 4-time points . Serum samples were collected after centrifugation  and was calibrated against ImmunoCAP-specific IgE. The Der p-specific IgG4 levels were determined by a sandwich ELISA. In brief, the microplate was coated with 10 μg/mL of a Der p extract  protein and then blocked with 2% casein and subsequently incubated with diluted standards, controls, and serum samples. Mouse monoclonal antibodies to human IgG4  were used at 1/10,000 for detection and revealed by HRP-labeled goat anti-mouse IgG  and then followed by Tetramethylbenzidine visualizing. The assays were further calibrated against ImmunoCAP-specific IgG4  by measuring the specific IgG4 concentration in mgA/L of Der p IgG4 in the ELISA standard stock solutions by ImmunoCAP. The lower limit of quantification (LoQ) of serum Der p IgG4 was 3.33 μg/L, and the LoQ of salivary Der p IgG4 was 0.67. Serum samples needed to be diluted at least 5-fold due to the different matrix effects, whereas there was no need to dilute saliva samples.Serum and salivary IgE and IgG4 against the us study . Der p-sDer p-specific IgE-BF was measured using the ADVIA system  based on previous studies [The  studies ,12. In gt-test or Mann–Whitney U test. Scatter plots were used to evaluate the correlations between the two variables. All the statistical analyses and graphics were carried out with R software packages (R version 4.0.3). p < 0.05 was considered statistically significant.The continuous variables were expressed as mean and standard deviation or percentile values and compared using the Student’s Der p-specific IgE and IgG4 levels, and salivary Der p-specific IgG4 levels between the two groups. Although asthma symptom scores from the AIT patients were higher than in the control group, other questionnaire data, including the rhinitis symptom score, ocular symptom score, VAS symptom score, medicine score, and rhinitis and asthma quality of life scores, showed no significant differences between them.p < 0.05), which display a continuous downward trend , suggesting a cross-reaction between them , further indicating their extensive cross-reactivity.The reatment . Howeverreatment . Althougc IgE-BF . AdditioDer p-specific IgG4 was detected in all the patients, including the AIT group and control group. After the AIT treatment, all the patients had detectable Der p-specific IgG4 at 4, 12, and 24 months. Furthermore, serum Der p-specific IgG4 significantly increased with the time of the AIT treatment , 12 , and 24 months  of the AIT  between the serum Der p-specific IgE-BF and Der p-specific IgG4 at the baseline , 4 , 12 , and 24  months after the AIT (Der p-specific IgG4 levels also demonstrated a certain correlation with the Der p-specific IgE-BF (Der p-specific IgG4 and serum-specific IgE-BF before treatment (p = 0.025, As the AIT treatment continued, the serum  the AIT . It is nc IgE-BF . Given treatment A. After Der p-specific AIT improved the clinical symptoms of children with asthma and/or AR. Its efficacy was closely associated with increased serum and salivary Der p-specific IgG4 levels, consistent with an increased IgE-BF. Most importantly, salivary Der p-specific IgG4 levels showed the same trend as serum Der p-specific IgG4 levels with the treatment time, which can contribute to assessing the efficacy of Der p-specific AIT in children.The present study demonstrated that Der p-specific AIT treatment. Furthermore, this displays a continuous downward trend with the treatment time, indicating that AIT had a better long-term clinical efficacy for children with asthma and/or AR. In addition to the long-term efficacy for children with AR, single-allergen subcutaneous immunotherapy was also beneficial for AR caused by multiple allergens, including HDMs [A large number of studies have confirmed that AIT is effective in patients with allergic diseases ,13,14,15ing HDMs . These fing HDMs ,18.Der p-specific IgG4 increased after 4 months and were closely associated with the symptom scores at 12 months of AIT [Der p-specific IgG4 significantly increased with the time of the AIT treatment, closely associating with improved clinical symptom scores during the two-year treatment. Serum-specific IgG4 levels showed to be a time- and dose-dependent increase in the serum inhibitory activity for both the IgE-BF and IgE-facilitated allergen binding during subcutaneous AIT, suggesting serum IgG4 possibly as a useful alternative marker of the clinical response to AIT [In general, IgG4 is considered to be a benign, non-pathogenic antibody. Food-specific IgG4 cannot fully illustrate food allergy or intolerance, but it is likely a physiological response of the immune system to food component exposure . IgG4 mas of AIT . Our pres of AIT . The pree to AIT .Der p-specific IgG4 levels were detected only in a small number of patients compared with the serum IgG4 levels at the baseline. There was no doubt that saliva Der p-specific IgG4 levels significantly increased after AIT treatment, in agreement with the trend of improved clinical symptoms. Furthermore, significant positive correlations were observed between serum- and salivary-specific IgG4. The positive correlation between the serum and salivary-specific IgG4 levels to allergens further indicated that IgG4 could be passively transferred rather than locally secreted [Notably, in the present study, salivary secreted . Differesecreted , while osecreted . Althougsecreted , our finThe present study had some weaknesses that should be considered. First, because of the difficulty in performing a randomized controlled trial in pediatric populations, we did not carry out a double-blind, randomized, placebo-controlled trial, possibly leading to selection bias. Second, all the patients or their parents were asked to record medications and symptoms at every visit; due to a longer follow-up interval, this could have interfered with the patient’s description of their symptoms. Third, the sample size was relatively small due to the difficulty in collecting pediatric patients and obtaining blood samples. Especially for the control patients who did not receive AIT, it was difficult to obtain blood samples at adequate follow-up intervals.Der p-specific AIT improves the clinical symptoms of children with asthma and/or AR. The efficacy of AIT was closely associated with increased serum and salivary-specific IgG4 levels, consistent with an increased IgE-BF. Furthermore, salivary Der p-specific IgG4 levels showed the same trend as the serum Der p-specific IgG4 levels with the treatment time. Therefore, salivary-specific IgG4 measurements may be useful for monitoring its efficacy on children with asthma and/or AR during allergen-specific AIT.We can conclude that


Rhizoctonia cerealis is the causal agent of sharp eyespot, a devastating disease of cereal crops including wheat. Several metalloproteases have been implicated in pathogenic virulence, but little is known about whole-genome metalloproteases in R. cerealis. In this study, a total of 116 metalloproteases-encoding genes were identified and characterized from the R. cerealis Rc207 genome. The gene expression profiles and phylogenetic relationship of 11 MEP36/fungalysin metalloproteases were examined during the fungal infection to wheat, and function of an upregulated secretory MEP36 named RcFL1 was validated. Of 11 MEP36 family metalloproteases, ten, except RcFL5, were predicted to be secreted proteins and nine encoding genes, but not RcFL5 and RcFL2, were expressed during the R. cerealis infection process. Phylogenetic analysis suggested that MEP36 metalloproteases in R. cerealis were closely related to those of Rhizoctonia solani but were remote to those of Bipolaris sorokiniana, Fusarium graminearum, F. pseudograminearum, and Pyricularia oryzae. Expression of RcFL1 was significantly upregulated during the infection process and induced plant cell death in wheat to promote the virulence of the pathogen. The MEP36 domain was necessary for the activities of RcFL1. Furthermore, RcFL1 could repress the expression of wheat genes coding for the chitin elicitor receptor kinase TaCERK1 and chitinases. These results suggest that this MEP36 metalloprotease RcFL1 may function as a virulence factor of R. cerealis through inhibiting host chitin-triggered immunity and chitinases. This study provides insights on pathogenic mechanisms of R. cerealis. RcFL1 likely is an important gene resource for improving resistance of wheat to R. cerealis through host-induced gene silencing strategy.  Triticum aestivum L.) is one of the most important staple crops worldwide [Rhizoctonia cerealis van der Hoeven [R. cerealis can also infect other important economical crops and bio-energy plans, including barley (Hordeum vulgare), rye , sugar beet (Beta vulgaris), cotton (Gossypium spp.), potato (Solanum tuberosum), and turfgrasses [R. cerealis and study mechanisms underlying the fungus–host interactions. Thus, we have recently completed the genome sequencing and assembling/annotation of the wheat sharp eyespot pathogen R. cerealis strain Rc207  to establish databases for further study on pathogen–host plant interactions and the fungal virulence factors. Bread wheat  named FoMEP1, which acts in concert with the serine protease FoSep1 to cleave chitinases produced by tomato. The double deletion mutant of FoMep1 and FoSep1 can reduce virulence of F. oxysporum on tomato [Colletotrichum graminicola has been demonstrated to have the role in the degradation of chitinases like other fungi fungalysins, and to enhance virulence of this pathogenic fungus causing the maize anthracnose disease [Pyricularia oryzae (Magnaporthe oryzae), functions as an effector and is recognized by the rice cytoplasmic resistance protein Pi-ta [R. cerealis, MEP43 metalloprotease RcMEP1 was shown to be required for the pathogen virulence [Phytophthora infestans were reported to be potential pathogenicity factors in P. infestans [R. cerealis.Metalloproteases (MEPs) are multiple families of hydrolytic enzymes; the majority is zinc-dependent due to two zinc-binding histidine residues in a HEXXH motif ,19,20. An tomato . The M36 disease . Avr-Pitin Pi-ta . In R. cirulence . Moreovenfestans . HoweverR. cerealis Rc207 genome, and examined their transcriptional profiles in time-course during the fungal infection to wheat. Since almost among 11 genes coding MEP36 metalloproteases  in the R. cerealis genome, nine genes, but not RcFL5 and RcFL2, were expressed and ten proteins, except RcFL5, were predicted as secreted proteins, we analyzed the sequences of MEP36 metalloproteases and their genes’ structures, phylogenetic relationship, and transcriptional profiles during R. cerealis infection to wheat. Notably, we investigated the functional roles of the MEP36 metalloprotease RcFL1 and its MEP36 domain in the fungal virulence to wheat, and the results suggested that RcFL1 acted as a virulence factor through inducing plant-cell death and inhibiting the expression of chitinases (TaChit3 and TaChitIV) and chitin elicitor receptor kinase-encoding gene (TaCERK1) in the treated wheat. In this study, we identified a total of 116 metalloprotease-encoding genes from the R. cerealis Rc207. The results showed that the Rc207 genome contained a total of 116 genes encoding metalloproteases, belonging to 24 families  proteins, which may play significant roles in pathogen infection  was used to group all metalloprotease sequences into ortholog groups (OGs). These 116 metalloproteases from R. cerealis Rc207 were clustered in 65 OGs by OrthoFinder , pattern 2 , pattern 3 , and pattern 4 , containing 6, 7, 8, and 9 introns, respectively. Furthermore, significant differences in size (28–875 bp) between the exons were observed. These results suggested that these MEP36 metalloprotease genes had variable and complex gene structures  residues and a Fungalysin/Thermolysin Propeptide Motif (FTP) with a length of 32–50 aa residues . Thus, e peptide . Using D peptide . The pre peptide . Multipls 49.70% .R. cerealis (RcFLs) and those from other plant–pathogens, we constructed a phylogenetic tree based on the multiple amino acid sequence alignment on the 11 RcFLs metalloproteases and 91 MEP36 metalloproteases from other fungal plant–pathogens, namely Bipolaris sorokiniana (ND90Pr), Fusarium graminearum (PH-1), Fusarium pseudograminearum (CS3096), Pyricularia oryzae , and Rhizoctonia solani   .R. cerealis Rc207 with seven MEP36 metalloproteases from R. solani AG-8 , nine MEP36 metalloproteases from R. solani AG-1 IB , three MEP36 metalloproteases from R. solani AG-1 IA , eleven MEP36 metalloproteases from R. solani AG-3 Rhs1AP , and eleven MEP36 metalloprotease from R. solani 123E . Group III (Green) contained RcFL1, RcFL2, RcFL3, RcFL5, RcFL7, three MEP36 metalloproteases from R. solani AG-8 , three MEP36 metalloproteases from R. solani AG-1 IB , an MEP36 metalloprotease from R. solani AG-3 Rhs1AP (EUC61642.1), and a MEP36 metalloprotease from R. solani 123E (KEP51322.1). None of the R. cerealis RcFLs proteins were clustered in Group II (blue), IV (yellow), and V (red). The resulting phylogenetic tree divided these MEP36 metalloproteases into five groups . The GroR. cerealis infection to wheat, we performed RNA-sequencing to investigate transcript dynamics of the metalloprotease genes in the R. cerealis Rc207 strain at five infection time-points  to wheat and in culture (vitro) mycelia. The transcriptomes showed that 82, 82, 98, 116, and 116 genes were expressed at 18, 36, 72, 96, and 240 hai, suggesting there was a temporal sequence leading to full expression of all genes  at 18, 36, 72, 96, and 240 hai, respectively  were significantly up-regulated during R. cerealis infection process  compared to in vitro fungal mycelia (RcFL1 (Rc_11192.1) were markedly upregulated during the R. cerealis infection process  compared to in vitro fungal mycelia  can induce the cell death of wheat or N. benthamiana leaves. To obtain purified RcFL1 and RcFL1-MEP36 proteins, the full RcFL1 or the RcFL1-MEP36 peptide-encoding sequences were separately sub-cloned in fusion to the His-TF tag of the pCOLD Trigger Factor (TF) vector, generating two recombinant protein vectors pHis-TF-RcFL1 and pHis-TF-RcFL1-MEP36. After induction by 0.5 mM isopropyl-β-d-thiogalactoside at 16 °C, the resulting pHisTF-RcFL1 and pHis-TF-RcFL1-MEP36 as well as the control His-TF proteins were highly expressed in E. coli DE3, respectively. After purification using Ni+ resin and elution with imidazole buffer, these proteins were examined through SDS-PAGE , the His-TF-RcFL1 (expressing the full RcFL1 protein) and His-TF-RcFL1-MEP36 (expressing the M36 domain of RcFL1) were able to trigger more obvious necrosis and plant cell death on the infiltrated leaves of susceptible wheat cv. Wenmai 6  areas in these wheat leaves were assessed. As a result, the larger disease lesion sizes were present in His-TF-RcFL1 and His-TF-RcFL1-MEP36 infiltrated leaves compared to those infiltrated with the His-TF control , the control His-TF, RcFL1, or RcFL1-MEP36 proteins were individually infiltrated into leaves of a  control A. The st control B. These TaCERK1 (TraesCS6D01G403200.1), a wheat chitin elicitor receptor kinase CERK1, and two chitinase-coding genes (TaChit3 and TaChitIV) in the susceptible wheat cv. Wenmai 6 leaves infiltrated with His-TF-RcFL1 or the His-TF control. The results showed that the expression levels of TaCERK1, TaChit3 and TaChitIV were significantly decreased by the RcFL1 protein in the His-TF-RcFL1 infiltrated wheat leaves compared to His-TF infiltrated wheat leaves and healthy wheat leaves (TaCERK1 and chitinases (TaChit3 and TaChitIV) in wheat.To explore if RcFL1 functional role is related to the chitin elicitor pathway or chitinases in wheat, RT-qPCR was deployed to examine transcript levels of t leaves , suggestR. cerealis Rc207. They were divided into 24 protein families, including 34 putative secretory metalloproteases with a predicted signal peptide. In the plant pathogen P. infestans, 91 metalloproteases were identified and divided over 21 protein families, including 25 metalloproteases with a predicted signal peptide or signal anchor [R. cerealis genome, the MEP1, MEP14, MEP28, MEP35, MEP36, and MEP64 families contain 3, 3, 9, 8, 10, and 1 secretory metalloproteases, respectively. Previously, only 5 MEP35 proteins were identified from next-generation genome sequences of R. cerealis Rc207. These results suggest that the advanced whole-genome assembly of the R. cerealis Rc207 is high-quality.Metalloproteinases, in which zinc is an essential metal ion for the catalytic activity, have been identified with the pathogenicity and virulence of pathogenic fungi ,24,25,28l anchor ,27,29,30R. cerealis. With the exception of RcFL5, 10 remaining RcFLs all contained a signal peptide with a length of 19–23 aa residues. Nine encoding genes, but not RcFL5 and RcFL2, were expressed during R. cerealis infection process to wheat. Such facts may imply that RcFL1-RcFL11, but not RcFL5 and RcFL2, may play roles in the fungal infection. Phylogenetic analysis indicated that RcFL4, RcFL6, RcFL8, RcFL9, RcFL10, and RcFL11 from R. cerealis Rc207 with 41 MEP36 metalloproteases from R. solani AG-8, R. solani AG-1 IB, R. solani AG-1 IA, R. solani AG-3 Rhs1AP, and R. solani 123E were clustered into the group I, while RcFL1, RcFL2, RcFL3, RcFL5, RcFL7, with three MEP36 metalloproteases from R. solani AG-8, three MEP36 metalloproteases from R. solani AG-1 IB, a MEP36 metalloproteases from R. solani AG-3 Rhs1AP, and a MEP36 metalloprotease from R. solani 123E were clustered into the Group III. None of the R. cerealis MEP36 proteins were clustered with other MEP36 metalloproteases of plant pathogens B. sorokiniana (ND90Pr), F. graminearum (PH-1), F. pseudograminearum (CS3096), and P. oryzae. These data suggest that MEP36 metalloproteases in R. cerealis were more closely-related to those of R. solani but were remote to those of B. sorokiniana (ND90Pr), F. graminearum (PH-1), F. pseudograminearum (CS3096), and P. oryzae .The current analysis showed that ten (90.91%) of 11 MEP36 metalloproteases are predicted to be secreted proteins. MEP36 family metalloproteases all harbor a HEXXH motif, in which two histidine residues function as the first and the second zinc ligand. Several MEP36 metalloproteases have been shown to play important roles in pathogenicity/virulence of other pathogens ,21,22. HR. cerealis are able to induce plant cell-death and may act as a virulence factor of R. cerealis [RcFL1 was markedly upregulated during the R. cerealis infection process  compared to the fungal grown inculture. Furthermore, the functional analyses demonstrated that the secretory R. cerealis MEP36 metalloprotease RcFL1 could induce plant cell-death and necrotic lesions, and promoted the fungal virulence. Moreover, our analyses suggest that the MEP36 domain is necessary for the function of the metalloprotease RcFL1 in the virulence of R. cerealis.Necrotrophic fungal pathogens utilize an array of effectors to induce plant cell death which may facilitate the growth of the necrotrophic pathogens ,30. Our cerealis . Here, gRhizoctonia isolates [Verticillium dahlia, is required for full virulence of the pathogen through preventing host chitin-triggered immunity [F. verticilloides could completely cleave the classs IV chitinase ChitA from maize [R. cerealis MEP36 metalloprotease RcFL1 could repress the expression of chitinases (TaChit3 and TaChitIV) and gene coding for the chitin elicitor receptor kinase TaCERK1 in host wheat. The data imply that the MEP36 metalloprotease RcFL1 functions as a virulence factor of R. cerealis possibly through preventing the host chitin-triggering immunity and reduced transcript accumulation of chitinases. It is very interesting to determine experimentally the detailed role of RcFL1 in the loss of this MAMP system in future. Taken together, the fungal-pathogen MEP36 metalloprotease RcFL1 can promote plant cell death and turn down host defense measure to help R. cerealis infect wheat.Chitin is a key component of the cell walls of pathogenic fungi, such as isolates . Plants isolates ,23,31,32isolates ,18,22. Fom maize . InteresR. cerealis Rc207 at wheat tillering stage [R. cerealis Rc207 and in vitro mycelia were sampled for deep RAN-Sequencing. Sharp eyespot-susceptible wheat cultivar Wenmai 6 plants were grown in a greenhouse under a 13-h light (~22 °C)/11-h dark (~10 °C) regime. Using the toothpick inoculated method, the base sheath of wenmai 6 plant was inoculated with ng stage . The sheN. benthamiana plants were grown under standard glasshouse conditions at 25 °C with a 12-h light and 12-h dark regime.R. cerealis Rc207, a highly aggressive strain collected in Northern China, was isolated in Shandong [The necrotrophic fungus All the primers and the sequences in the study are listed in −10 was used to identify members of the candidate metalloprotease gene family from the R. cerealis Rc207 genome sequence. Signal peptide was used to predict using SignalP v4.0 [https://www.lynnon.com/download/ (accessed on 29 June 2022)). The expression profiles of metalloprotease family genes were drawn by the TBtools software [R. cerealis and 91 MEP36 proteins from other plant-pathogenic fungi , and R. solani  downloaded from the NCBI database (File S1).The function of metalloproteases was annotated with the Hmmer software  based onalP v4.0 , and putsoftware . A neighsoftware  based onR. cerealis Rc207 in vitro mycelia and R. cerealis-inoculated sheaths and/or stems, or RcFL1-treated leaves of the susceptible wheat cv. wenmai 6 plants were extracted using the TRIzol reagent  [According to the manufacturer’s instruction, total RNAs from CA, USA) . ReverseR. cerealis infection process in wheat, fungal, or wheat RNAs were extracted for examining transcripts of the RcFL1, TaChit3, TaChitIV, and TaCERK1 genes. Following the procedure described in Dong et al. [RcFL1) of R. cerealis or the target genes  of wheat were calculated using the 2−ΔΔCT method [Actin gene (TaActin) and the R. cerealis Actin gene (RcActin) were used as the internal reference, respectively. Three independent biological replications were evaluated. The g et al. , RT-qPCRT method , where wRcFL1 and its partial sequence comprising the MEP36 domain were separately subcloned in fusion to the His-TF tag of the pCOLD TF vector, generating the expression vectors pHis-TF-RcFL1 and pHis-TF-RcFL1-MEP36. The DNAs of the resulting pHis-TF-RcFL1 and pHis-TF-RcFL1-MEP36 fusion constructs as well as pCOLD TF vector were each transformed into competent cells of E. coli BL21 (DE3), respectively. After these, E. coli BL21 cells were cultured in LB medium and induced with 0.5 mM isopropyl-β-d-thiogalactoside at 16 °C at 100 rpm overnight. The recombinant proteins were purified with Ni-NTA resin and eluted [The full encoding sequence of d eluted . FinallyN. benthamiana [Cell death-inducing activities of the RcFL1 and RcFL1-MEP36 proteins of the heterologous expressed proteins  were assayed by infiltrating samples into detached leaves from fully expanded secondary of two-month-old wheat plants or four-week-old thamiana ,38. R. cerealis Rc207. These leaves were placed in Petri dishes containing filter paper saturated with sterile distilled water and kept under a 16-h day/8-h night regime at 25 °C. Leaf lesions were calculated by length×width and photographed at 3 days post inoculation (dpi) with R. cerealis Rc207 [Following protein infiltration for six hours, leaves were inoculated in the same location with 50 µL mycelial suspension of is Rc207 ,38. Six R. cerealis Rc207 and were divided over 24 protein families, including 34 secretory metalloproteases with a predicted signal peptide. All the 116 metalloproteases genes were expressed at the late stages, 96, and 240 hai, of the fungal infection to wheat. Of 11 MEP36/fungalysin family metalloproteases, 10 MEP36 were predicted to be secreted proteins and nine encoding genes were expressed during R. cerealis infection process to wheat. Phylogenetic analysis suggested that these MEP36 metalloproteases in R. cerealis were closely-related to those of R. solani but were remote to those of B. sorokiniana, F. graminearum, F. pseudograminearum, and P. oryzae. Interestingly, an MEP36 metalloprotease gene RcFL1 displayed a significantly upregulated expression during the infection process, and RcFL1 has been verified to function as a virulence factor of R. cerealis possibly through inhibiting host chitin-triggered immunity and chitinases. Thus, RcFL1 likely is a promising gene resource for improving resistance of wheat and other crop plants to R. cerealis through host-induced gene silencing strategy. This is the first systematic inventory of whole-genome metalloproteases in R. cerealis.In this study, a total of 116 metalloproteases were identified from the whole-genome sequences of the wheat sharp eyespot


Both Bacillus sphaericus and the Streptococcus aureus bacterial strains were evaluated for their ability to precipitate calcium carbonate (CaCO3). These strains were obtained from the Department of Biotechnology at GLA University in Mathura. This research aims to solve the issue of augmenting the tension and compression strengths of concrete by investigating possible solutions for environmentally friendly concrete. The sterile cultures of the microorganisms were mixed with water, which was one of the components of the concrete mixture, along with the nutrients in the appropriate proportions. After that, the blocks were molded, and then pond-cured for 7, 28, 56, 90, 120, 180, 270, and 365 days, respectively, before being evaluated for compressibility and tensile strength. An investigation into the effect that bacteria have on compression strength was carried out, and the outcomes of the tests showed that bacterial concrete specimens exhibited an increase in mechanical strength. When compared to regular concrete, the results showed a maximum increase of 16 percent in compressive strength and a maximum increase of 12 percent in split tensile strength. This study also found that both bacterial concrete containing 106, 107, and 108 cfu/ml concentrations made from Bacillus sphaericus and Streptococcus aureus bacteria gave better results than normal concrete. Both cluster analysis (CA) and regression analysis (RA) were utilized in this research project in order to measure and analyze mechanical strength.The focus of this research is to isolating and identifying bacteria that produce calcite precipitate, as well as determining whether or not these bacteria are suitable for incorporation into concrete in order to enhance the material's strength and make the environment protection better. In order to survive the high “potential of hydrogen” of concrete, microbes that are going to be added to concrete need to be able to withstand alkali, and they also need to be able to develop endospores so that they can survive the mechanical forces that are going to be put on the concrete while it is being mixed. In order to precipitate CaCO Cement is the sole component that is made, whereas the rest are naturally occurring and sourced from the area [2 emissions. Because of its adaptability, concrete is employed in the construction of a wide variety of structures, including bridges, large buildings, off-shore constructions, airports, sidewalks, railroad beds, and deep foundations, despite the fact that it is fragile and has a low resistance to stress [3) calcium carbonate has been more popular in the field of building engineering. It is seen as a potentially fruitful novel method for extending the useful life of cement-based buildings [3 calcium carbonate precipitation that results from metabolic activity of various microbe species, such as sulfate-reducing microbes, ureolytic microbes, nitrate reducing microbes, and oxidation of organic microbes, is what allows this method to self-heal the gaps in the concrete and the inevitable microcracks that will form in the concrete. For instance, ureolytic microbes are responsible for the production of an enzyme known as urease, which decomposes urea into carbonate ions [3 in a given environment. This enzyme catalyzes the hydrolysis of urea into carbon dioxide and ammonia, which ultimately results in a rise in both the pH and the amount of carbonate in the surrounding/environment in which the microbes are found. The incorporation of microbial species into concrete has increased its strength and endurance, providing additional benefits in the form of environmentally friendly and cost-effective alternatives. Since bacteria thrive at alkaline pH, the concrete's resistance to alkali assault, chemical attacks, freeze-thaw strike, and drying shrinkage is significantly improved [3) as a precipitate; however, for the selected bacteria to provide the greatest advantages, they must be alkaliphilic and thermophilic. Bacillus sphaericus, which is an alkaliphile and precipitates (CaCO3) calcium carbonate with a higher density, and Streptococcus aureus, which is also an alkaliphile and increases the compressive strength of concrete, are both beneficial microorganisms [3, which filled the gaps and improved the texture of the concrete as a result, was thought to be responsible for the increase in compression strength that was observed. Additionally, “Ryu et al.” came to the conclusion that the highest increase in compression strength was observed at a microbial cell concentration of 30 × 105 cfu/mL, and also that the compressive strength declined once this value was exceeded [6 cells per milliliter of solution [Concrete is thought of as a homogeneous substance since it is created by combining cement, coarse and fine aggregate, and water in a certain ratio. Concrete is a porous material and is sensitive to various assaults such as chloride, COas pores , 2. Sincas pores –5. Cementhe area . The mano stress –9. A greuildings –12. The ate ions , 14. Accimproved –17. In aimproved –20. The improved , 22. An rganisms , 24. Forexceeded . In a sisolution . Predictsolution , 28.In order to find a solution to the problem of increasing the tensile and compression strengths of concrete, the purpose of this study is to investigate potential options. Microorganisms grown in sterilized cultures were combined with water, which was one of the components of the concrete mixture, along with the nutrients in the right amounts. After that, the blocks were cast, and then they were pond-cured for 7, 28, 56, 90, 120, 180, 270, and 365 days, respectively, before being assessed for compressibility and tensile strength. In order to assess and investigate the nature of mechanical strength, this research study made use of both cluster analysis (CA) and regression analysis (RA). After a thorough study of old papers, the researcher found that not much research has been done with Bacillus sphaericus and Streptococcus aureus bacteria and also found that till date no researcher has done a thorough study of the results through cluster analysis for Bacillus sphaericus and Streptococcus aureus bacterial concrete. The researcher used three different concentrations, namely 106, 107, and 108 cfu/ml, of Bacillus sphaericus and Streptococcus aureus bacteria to determine the relationship between bacterial concrete and normal concrete.This paper shows the introduction of the study in OPC (ordinary Portland cement) of the grade 43 was utilized in this experimental endeavor. In accordance with IS 4031-1996 , this OP3 crystals on concrete suggests that the presence of the microbes had no negative implication on the hydration of reaction, which forms a dense CaCO3 crystal in liquid medium [Both Bacillus sphaericus and Streptococcus aureus were obtained from the Department of Biotechnology at GLA University in Mathura. The fact that the microorganisms Bacillus sphaericus and Streptococcus aureus are able to survive in the highly alkaline surroundings of concrete during the formation of CaCOd medium , 61. Ured medium , 62.The gathering of particular experimental data that contributes to a better understanding of microorganism concrete and the features it possesses is the primary aim of the current experimental research (durability and strength). In the current experimental inquiry, tests on the behavior of hardened and fresh characteristics of ordinary concrete grade and normal grade concrete without and with the inclusion of microbes have been carried out. These studies were carried out as part of the experimental study. Concrete in its hardened state is subjected to the appropriate laboratory tests, which enable the concrete strength, such as compression strength and tension strength. Research process methodology is shown in   Phase 1: growth of bacteria and culturing technique of bacterial concrete  Phase 2: to study the compression strength and split tensile strength of bacterial concrete  Phase 3: regression analysis and cluster analysisThe primary purpose of this experimental inquiry is to investigate the strength of normal concrete and bacterial concrete. The current work may be broken down into three distinct stages:3. This medium includes baking soda, ammonium chloride, urea, and CaCl2, and it was dissolved in distilled water [Both Bacillus sphaericus and Streptococcus aureus were cultivated in a medium that was designed for the improved generation of CaCOed water . In ordeed water , 41. Dured water  to deter3 of concrete is 400 kg. Casting concrete into cubes of size 100 millimeters by 100 millimeters by 100 millimeters is carried out to determine the concrete's compressive strength, and casting concrete into cylinders of size 100 millimeters in diameter and 200 millimeters in length is carried out to determine the concrete's split tensile strength. Mix design is shown in Concrete of the grade M30 was mixed in accordance with the standards set out in IS 10262-1982 . For an The evaluation of the effect that the incorporation of microorganisms into the mixture has on the compression strength and stiffness of cement concrete blocks is another factor that is taken into consideration –67. The In order to evaluate the tension strength of each mixture, three cylinders measuring 100 mm by 200 mm were casted for every combination. In order to create these cylinders, a mold made of cast iron and steel was utilized. Casting and curing times for each mix percentage were as follows: seven days, twenty-eight days, fifty-six days, ninety days, one hundred eighty days, two hundred seventy days, and three hundred sixty-five days. In accordance with the Indian Standard code IS 516–1959 , a splitThe findings of the experiments were evaluated using regression analysis , which lσ) [R2) [The results of an experiment are depicted in σ)  but alsoσ) [R2)  that werR2 (0.8813), indicates that the regression line as well as the statistics of compression strength and split tensile strength values has a solid connection with one another. As can be seen from equation (The following value for the regression coefficient, Multivariate statistical techniques are typically utilized for the categorization, analysis, and interpreting of big data sets. These approaches are also employed for the decrease of a dimension of complicated datasets with a minimal loss of original data . ClusterThrough cluster analysis, it was found that the compression strength of bacterial concrete SP1, SA 1, SP 2, and SA 2 increases in the same way, and similarly, compression strength of SP3 and SA3 increases in the same group. Also, the compression strength of normal concrete sets itself apart from bacterial concrete. Cluster analysis results for compression strength are displayed in Through cluster analysis, it was found that the tension strength of bacterial concrete SP 3, SA3, and SA1 increases in the same way, and tensile strength of SP 1, SA2, and SP 2 increases in the same group. Also, the split tensile strength of normal concrete sets itself apart from bacterial concrete. Cluster analysis results for split tensile strength are displayed in The study of this research work has shown that the mechanical strength of concrete can be increased by using Bacillus sphaericus and Streptococcus aureus bacteria for bio-concrete.7 cfu/ml produced better results for mechanical strength.Using both bacteria in concrete with different concentrations showed that microbial concrete with a concentration of 106, 107, and 108 cfu/ml concentrations made from Bacillus sphaericus and Streptococcus aureus bacteria gave better results than normal concrete.This study also found that both bacterial concrete containing 10Value of the regression coefficient indicates that the regression line as well as the statistics of compression strength and split tensile strength values has a solid connection with one another.Through cluster analysis, it was found that the compression strength of bacterial concrete SP1 and SA1 increases almost equally, similarly, the compression strength of bacterial concrete SP2 and SA2 increases almost equally, and both the compressive strength of both SP3 and SA increases almost equally in a similar group. And in the cluster analysis for split tensile strength, three groups were formed, in which in the first group the tensile strength of SP3, SA3, and SA1 increases almost equally, and in the second group the tensile strength of SP1, SA2, and SP2 is mutual, which grows almost evenly, and in the third group comes the normal concrete.Research organizations from all over the worldwide have been interested in the employment of microbes for the purpose of increasing the longevity of construction materials as a consequence of the encouraging findings obtained so far. Our grasp of the opportunities and constraints presented by biotechnological processes on construction materials may unquestionably benefit from the work that has been carried out by a number of research groups that have concentrated their attention on a variety of materials. Studies are still being conducted on the preservation of nutrients and microbial products since these factors have an effect on the survival, development, and creation of biofilms. “Bacillus sphaericus and Streptococcus aureus bacterial concrete” appears to become the most viable technique for generating crack-resistant concrete in the coming days, according to the findings of this research as well as the prior research that has been conducted.