Rainbow trout is one of the main freshwater fish species farmed in Romania, in a temperate continental climate. Blood biochemistry parameter values are well known for humans and other higher vertebrates, including some species of fish, and are fundamental for the assessment of physiological status. In this study, we examined the physicochemical parameters of water quality and the blood profile  of rainbow trout females reared in three farms, during the summer season, to monitor stress. These findings permit the progress of knowledge of blood parameters in fish and will encourage efforts to expand hematological and biochemical studies to assess the health of salmonids, which is essential for the sustainable development of aquaculture.Oncorhynchus mykiss) from three farms were investigated using different biomarkers during the summer season. In the given context, the main water physicochemical parameters were investigated and twelve biochemical parameters were measured from blood samples of rainbow trout reared in the Fiad, Șoimul de Jos, and Strâmba farms. We selected these farms because the genetic background of the rainbow trout is the same, with all studied specimens coming from the Fiad farm, which has an incubation station. Forty-five samples were collected monthly (May to August) throughout summer to observe the changes in the blood profile of rainbow trout. Principal component analysis showed a clear separation both among the studied farms and months. Furthermore, significant correlations (p < 0.05) between the majority of the biochemical parameters were found, indicating that the environmental parameters can influence several blood parameters at the same time. The present study provides several useful norms for assessing the welfare of rainbow trout, indicating that the relationships among different parameters are important factors in interpreting the blood biochemical profiles.Blood biochemistry parameters are valuable tools for monitoring fish health. Their baseline values are still undefined for a multitude of farmed fish species. In this study, changes in the blood profile of rainbow trout females ( Oncorhynchus mykiss, Walbaum, 1792, is the main target of Romanian aquaculture. The total aquaculture production in Romania in 2019 was 12,847 tons [Rainbow trout 847 tons . Knowled847 tons ,4. Harve847 tons . Environ847 tons . Differe847 tons ,11, or h847 tons .The initial physiological response of fish to stress involves the release of catecholamines and the activation of the hypothalamic–pituitary–interrenal axis (HPI) . The strSteroid hormones are involved in the regulation and progress of some physiological processes in vertebrates, such as embryonic development, sex differentiation, metabolism, immunological responses ,22, circMost studies on this subject focus on relations among intrinsic  and extrinsic factors . The correlations between these factors and rainbow trout welfare represent an important and current topic ,26,27,28In this study, we evaluated the fluctuations of the hematologic and hormonal profiles, as well as oxidative stress during the summer season, in three fish farms, when rainbow trout are subjected to technological stressors  and environmental stressors .All procedures involving animals were conducted following Romanian (Law 43/2014) and European legislation (EU Directive 63/2010). The study was approved by the Ethics Committee of the University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca (No. 148/2019).3.The study was carried out in three rainbow trout farms, in north-western Romania, in Bistrița-Năsăud county, namely Fiad (47°29′16.5″ N 24°22′59.4″ E), Strâmba (47°15′37.3″ N 24°40′10.2″ E) and Șoimul de Jos (47°09′55.8” N 24°50′03.2″ E) farms. These locations were selected because the rainbow trout genetic line found in all three farms are identical, with all studied individuals originating from Fiad farm, which has an incubation station. The feeding technique and the administered feed, Aller Gold , were the same during the entire experimental period, with the chemical composition as presented in 3) and nitrates (mg/dm3). Temperature, dissolved oxygen, pH, TDS, conductivity, resistivity, and salinity were monitored at sunrise, at 8 A.M., before feeding, at different points of the ponds , using the Hanna HI 9828/4-01 multiparameter. The methods used for determining ammonia (NH3), STAS 9800-2/71, nitrites (NO2), SR ISO 6777/1996, and nitrates (NO3), SR ISO 7890:1-1998, were applied by spectrometry. These parameters were monitored monthly, water samples being collected, transported, and analyzed at the Aquaculture Hygiene Laboratory of U.A.S.V.M. Cluj-Napoca, in less than four hours since collection, using a 0.5 L polypropylene container with a screw cap. The recorded physicochemical parameters of water were: temperature (°C), dissolved oxygen (mg/L), pH (pH/ORP meter), total dissolved solids (TDSs), conductivity (µS/cm), resistivity (Ω/cm), salinity (PSU), ammonia (mg/L), nitrites (mg/dmOncorhynchus mykiss) females were selected. Fish were fasted 24 h before sampling. The individuals were removed from the ponds and were immediately anesthetized with clove oil (eugenol 30 mg L−1) to reduce handling stress [Forty-five randomly selected fish (15 from each farm) were sampled every summer month  of 2019. During the experimental period, a total of 180 rainbow trout  count, hemoglobin (Hb), and hematocrit (Hct) from whole blood. The erythrocyte indices, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), and mean corpuscular hemoglobin concentration (MCHC) were calculated according to standard formulas . For anaSerum separator tubes (SSTs) were used for the chemical analysis of the blood. The samples were kept at 4 °C overnight, and the serum was obtained by centrifugation (15 min at 1000 xg). It was visually inspected to exclude hemolysis, which could influence results, and was stored at −80 °C until later analysis. The analyses were carried out using a HiPo MPP-96 Microplate Photometer spectrophotometer  at 340–620 nm wavelength intervals. Plasma cortisol was quantified by ELISA, using a commercial kit from DiaMetra S.r.l. , according to the manufacturer’s prospect. The minimum detectable concentration is 2.42 ng/mL at a 95% confidence limit.2), and progesterone (PROG) concentration in the serum were measured using the Fish Testosterone (T) Elisa, Fish 17β-estradiol (E2) Elisa, and Fish Progesterone (PROG) Elisa kits, according to manufacturer instructions. All Elisa kits were purchased from Aviva Systems Biology Co., Ltd. . The minimum detectable concentration was 0.1 ng/mL for T, 40 pg/mL for E2 and 0.5 ng/mL for PROG. The variation coefficients for intra- and inter-tests were <10%.Testosterone (T), 17β-estradiol  on the dataset of physiological parameters . The physiological response variables with the strongest associations with the first two PC axes were further analyzed utilizing analyses of variance , to check whether the location and/or month play important roles for the distribution of these parameters. We checked whether the data were normally distributed with the help of a Shapiro–Wilk test (shapiro.test function in RStudio). The following parameters were normally distributed: SOD and MCV. The values of GPx, cortisol, estradiol, testosterone, Ery, Hct, and MCH were not normally distributed. In the case of normally distributed data, we performed a two-way ANOVA followed by TukeyHSD post hoc test. For other data, we used the Kruskal–Wallis test followed by a pair-wise Wilcoxon post hoc test with Bonferroni correction. p < 0.05, and those p-values smaller than 0.001 were presented as p < 0.001. All analyses were performed in RStudio Version 1.2.5042 (2020). Next, as months tended to have a far stronger effect on the data, we performed correlations between monthly stress response variables and all other parameters . We considered strong positive or negative correlations when the coefficient was >0.5 or <−0.5, and moderate positive or negative correlations when the coefficient was >0.3 and <=0.5, <−0.3 and <=−0.5, respectively . Values p > 0.05).The results of the physicochemical water quality parameters measured in farms during the summer season are reported in Regarding water temperature (°C) a, we obsThe pH parameter c did not2  revealed that the first (PC1) and the second (PC2) components explained 45.97%  of the total variation . Usually2. The parameters that correlated most strongly negatively correlated with the first axis were: MCH and MCV. The strongest positive correlations with the PC2 axis were found in the parameters: GPx, SOD, Hct, and T. The parameter most strongly negatively correlated with the second axis was E2. These parameters were not particularly associated with any farm; however, there was a clearer association between them and months    . Higher Strâmba) a.p < 0.001)   . RBC cou= 0.117) b. p < 0.005) and DO , and strongly negatively correlated with MCV , temperature , conductivity , TDS  and NO2 . In June, the RBC count was strongly negatively correlated with MCV  and MCH . In July, the RBC count was strongly positively correlated with Hct , F , pH , conductivity , TDS , DO , and strongly negatively correlated with MCH , MCHC , NO2  and NO3 . In August, the RBC count was strongly negatively correlated with MCV  and MCH .In May, the RBC count was strongly positively correlated with resistivity   (p < 0.001), and strongly negatively correlated with RBC . In June, MCV values were strongly positively correlated with Hct  and MCH , and strongly negatively correlated with RBC . In July, MCV values were strongly positively correlated with Hct  and strongly negatively correlated with NO3 . In August, MCV values were strongly positively correlated with Hct  and strongly negatively correlated with RBC .The mean corpuscular volume (MCV) values varied significantly between months  . MCV val= 0.002) c. In Mayp < 0.001)  (p < 0.001) and MCV . In June, MCH values were strongly positively correlated with Hb , MCV , resistivity , DO , and strongly negatively correlated with RBC , pH , temperature , conductivity , TDS , salinity  and NO3 . In July, MCH values were strongly positively correlated with Hb  and MCHC , and strongly negatively correlated with RBC . In August, MCH values were strongly positively correlated with Hb , temperature , NO3 , and strongly negatively correlated with RBC  and DO .The mean corpuscular hemoglobin (MCH) values varied significantly between months  . MCH val< 0.001) d. In Mayp < 0.001) and months  , and Hct values were higher in Strâmba than those measured in Fiad (pStrâmba-Fiad = 0.01) (The hematocrit (Hct) values varied significantly between locations  . The Hct = 0.01) e. pMay–June < 0.001, pJune–July < 0.001, pJune–August < 0.001) (p < 0.001) and strongly negatively correlated with pH , salinity  and NH3 . In June, Hct values were strongly positively correlated with MCV . In July, Hct values were strongly positively correlated with RBC , MCV , T , F , pH , conductivity , TDS  and DO , and strongly negatively correlated with MCHC , NO2  and NO3 . In August, Hct values were strongly positively correlated with MCV  and strongly negatively correlated with MCHC .The Hct values in June were significantly higher than those recorded in the other three months (Wilcoxon rank sum test: < 0.001) f. In Mayp = 0.020) (Strâmba-Fiad = 0.047)  (p < 0.001), total dissolved solids (TDS) , and salinity , and strongly negatively correlated with resistivity . In August, GPx was strongly positively correlated with resistivity , NO2  and NH3 , and strongly negatively correlated with pH , conductivity , TDS  and salinity .Glutathione peroxidase (GPx) of rainbow trout varied significantly between farms  . The Str= 0.047) a. GPx vaarisons) b. In Junp < 0.001) (p = 0.641)   varied significantly between months  . The far= 0.641) . SOD valarisons) c. p < 0.001)  (p < 0.001) . June had higher F levels than August, but lower than May ; August had the lowest F levels compared to the other three months  (p < 0.001) and dissolved oxygen , and strongly negatively correlated with temperature , conductivity , TDS  and NO2 . In July, F was strongly positively correlated with Hct , RBC , T , pH , conductivity , TDS  and dissolved oxygen , and it was strongly negatively correlated with NO2 , NO3 , NH3  and temperature .F varied significantly between farms  . Șoimul < 0.001) a.. F val< 0.001) . F levelarisons) b. In May2 varied significantly between months   (p < 0.001)  (p < 0.001), resistivity , DO  and strongly negatively correlated with temperature , conductivity , TDS  and NO2 . In July, T was strongly positively correlated with Hct , F , conductivity , TDS  and DO , and strongly negatively correlated with NO2  and NO3 . The levels of E2 and T found circulating in the peripheral plasma of rainbow trout varied between 46.39 and 48.21 pg/mL for E2, and 0.44 and 0.61 ng/mL for T during the four months of the summer season.E< 0.001) . May hadarisons) c. T vari< 0.001) . Signifiarisons) d. In May2, MCH, MCV, GPx, SOD, Hct, and T.The values of the physicochemical parameters are within normal limits for rainbow trout aquaculture, and the PCA showed that the differentiating parameters between months and farms are: RBC, F, EThe results obtained in this study highlight the fluctuations of the hematological and hormonal profiles, and the oxidative stress in rainbow trout from three Romanian farms during the summer season. In addition, we have established the main physiological and environmental parameters that significantly differentiate analyzed farms and months, to offer a baseline necessary in the understanding of their physiological role in the immunological system of rainbow trout. This baseline is specific to a temperate-continental climate, with four seasons, such as in Romania .As other studies demonstrated, many biotic and abiotic factors can affect blood parameters in fish ,38,39. SUnder the conditions of adequate values of dissolved oxygen in the water and through the nitrification process, the low amount of ammonia found in the studied farms can be transformed into nitrites and nitrates. Nitrates are less toxic to fish over a long period throughout the entire nitrification process . Self-puChanges in the red blood component with manifestations of low/high RBC, Hct, and Hb values are a key feature of the clinical picture in different fish species. Erythrocyte indices help in diagnosing and treating anemia for each species, but the values of these indices are affected by age, sex, body mass, population density, environmental and species parameters .12/L [In our study, according to the PCA, the number of red blood cells (RBCs) varied significantly between farms. The analysis of the monthly variation of hematological parameters during the summer season showed that the number of RBCs was highest in May and June and lowest in July and August. As fish stockings, genetic background, feed, and age of fish were almost identical in the three farms, it is presumed that the differences in RBC count values occurred due to environmental parameters. Generally, the number of RBCs in the summer season is higher compared to that of other seasons, due to an intensified hematopoietic function caused by an increase in water temperature, a decrease in DO , an inte12/L . The mean corpuscular volume (MCV) and the mean corpuscular hemoglobin (MCH) values varied significantly between months. In a study of hematological parameters, Lone et al.  recordedThe hematocrit (Hct) values varied significantly between farms and months. Similar to RBC, the hematocrit is presumed to vary due to environmental parameters. As for the monthly variation, the Hct values in June were significantly higher than those recorded in the other three months. The reference limits of Hct are between 20% and 43% . In geneThe antioxidant defense system of the body, which contains enzymes such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx), is set to maintain the lowest potential levels of reactive oxygen species (ROS) in cells, and it is recognized as an essential component of an organism’s maintenance . PreviouIn the current study, Glutathione peroxidase (GPx) activity varied significantly between farms (Strâmba and Fiad), higher values of GPx being recorded in Strâmba farm compared to the other two farms. This difference was most likely the result of higher temperature values and lower DO, the thermal stress being more noticeable. Both SOD and GPx activity varied significantly between months. In May and July, the SOD and GPx values were lower than in June and August, being strongly correlated with conductivity, temperature, DO, total dissolved solids, and strongly negatively correlated with resistivity. The recorded results of SOD and GPx indicate that the rainbow trout metabolic compromise was higher in periods with higher temperatures and lower oxygen concentrations characteristic of summer months. Higher levels of DO may help to improve the oxidative parameters to cope with hyperoxia stress and to alleviate damage to tissue cell membranes. Ritola et al.  reportedO. mykiss after chemical exposure (49–110 nmol/L), handling and confinement (77–698 nmol/L), trapping .Blood cortisol (F) levels are initial indicators of acute and chronic stress in fish. The gills, intestines, and liver are important targets for F in fish. These organs reflect the two major actions of F in fish: regulation of hydromineral balance and energy metabolism. Other activities of F include reduced growth rate and suppression of reproductive and immune functions ,58. An i2) play an important role in hormonal control of reproductive functions. The main physiological function of steroid hormones in vertebrates is to regulate sexual development and reproduction. However, they have pleiotropic functions and beyond the “classical” function of the reproductive axis, they target several other physiological systems, including the immune system [2 varied significantly between months. Thus, the highest values were registered in May, observing a progressive decrease in the other months, the minimum being in August. T also varied significantly between months. As in the case of E2, significant differences were found between the values of T in May and those of the other months, when the lowest T values were recorded. These changes are considered normal, because rainbow trout spawning takes place in spring, beginning in March and ending in April in Romania, due to the temperate-continental climate with Mediterranean influences. In teleost fish, testosterone (T), 11-ketotestosterone (11KT), progesterone (PROG), and estradiol-17b  when lymphocytes were incubated in vitro with the steroids. They also observed that T synergizes with F to produce a greater inhibitory effect, suggesting that the two hormones act independently on different cells. The interactions between different hormones are often complex, but in many cases, changes in plasma hormone levels correspond to changes in the immune status and welfare of fish. A previous study has demonstrated that Estressor . In our  authors ,63,64,65Hematological and biochemical parameters are valuable tools for monitoring fish health. Their normal values are still undefined for the numerous farmed fish species. The results of this study reflect new aspects of blood parameters of rainbow trout in three farms and allow a better understanding of the influence of environmental conditions on the hematological and biochemical parameters in different locations. Clear monthly changes in the analyzed parameters were observed for all three salmonid farms studied, with the largest variation found in the case of Strâmba and Fiad farms. In future research, it would be necessary to increase the number of parameters investigated to expand the basic knowledge on the rainbow trout species and the stressors that may influence the growth and exploitation of this species.These results will encourage efforts to expand hematological and biochemical studies to screening programs to assess the health of salmonids in intensive aquaculture in Romania, to improve immunocompetence in farmed fish, which is essential for the sustainable development of aquaculture.


Starches rich in amylose are promising functional ingredients for calory-reduced foods. In this research, a high-amylose Japonica rice starch (amylose content 33.3%) was esterified with octenyl succinic anhydride (OSA) to improve the functional properties. The OSA-modified derivatives were evaluated for structure and functional properties, with OSA-modified normal Japonica rice starch (amylose content 18.8%) used as control. Fourier transform infrared spectra confirmed the introduction of OSA groups to starch. OSA modification made little change to morphology and particle size of high-amylose starch, but decreased the relative crystallinity and pasting temperature and increased the pasting viscosity, swelling power, emulsifying stability, and resistant starch (RS) content. The changes of properties were related to the degree of substitution (DS). Typically, OSA-modified high-amylose starch at DS of 0.0285 shows polyhedral-shape granules, with a volume-average particle diameter of 8.87 μm, peak viscosity of 5730 cp, and RS content of 35.45%. OSA-modified high-amylose starch had greater peak viscosity and RS content and lower swelling power than OSA-modified normal starch of similar DS, but the two kinds of derivatives did not have a significant difference in emulsifying stability. The OSA-modified high-amylose Japonica rice starch could be used as an emulsifier, thickener, and fat replacer in food systems. Native starches often perform poorly in industrial applications and are tailored through physical or chemical modification to develop desirable functional properties, such as pasting properties, dispersity, and digestibility ,2. OctenThe properties of OSA-modified starch have been affected by many factors, including starch type, amylose content, degree of substitution (DS), etc. Wen et al.  found thOryza sativa L.) is one of the most important cereal crops, providing more than half of the world’s staple food. The Asian cultivated rice grown worldwide is classified into two major subspecies, Indica and Japonica. Indica rice is the major type of rice grown in the tropics and subtropics, for example, central and southern China, India, Pakistan, and Indonesia [Rice , a promising ingredient for calory-reduced food, was chemically modified with OSA. The structure and properties of the OSA-modified high-amylose Japonica rice starch were investigated and compared with OSA-modified normal Japonica rice starch (amylose content 18.8%). This research will be helpful for the design of modified starch with new and enhanced properties.High-amylose Japonica rice (variety Jiangtangdao1) was kindly supplied by the Shanghai Academy of Agricultural Sciences . Normal Japonica rice (varieties of huai5) was obtained from a local grain trader . Soybean oil (Golden Dragon Fish) was supplied by Yihai Kerry Group . Porcine pancreatic-amylase (P7545) and amyloglucosidase (A7095) were purchase from Sigma Chemical Co. . A glucose oxidase/peroxidase (GOPOD) kit was purchased from Megazyme International Ireland Ltd. . Other chemical reagents were of analytical grade unless otherwise stated.Starch was isolated from white rice according to the method of Wang et al. . AccordiStarch  was dispersed in water (140 mL) with stirring. The pH of the slurry was adjusted to 8.5 with 1 mol/L NaOH at 25 °C. OSA  was added to the slurry. The mixture was continuously stirred for reaction, with pH maintained at 8.5. After 6 h of reaction, the starch slurry was neutralized to pH 7.0 with 1 mol/L HCl and then centrifuged at 3000 r/min for 15 min . The residue was washed three times with water and once with acetone, and then dried at 40 °C for 24 h. The degree of substitution (DS) of OSA-modified starch was determined by titration following the method described by Song et al. .w/w) and pressed into transparent tablets under infrared light before measurement. Measurements were made in the region 4000–400 cm−1 at a resolution of 4 cm−1 with a total of 64 scans per spectrum.The Fourier transform infrared (FTIR) spectra of native and OSA-modified starch samples were collected on a Nicolet Nexus 670 FTIR spectrophotometer . Starch samples were mixed with dried KBr powder in a ratio of 1:100  at an accelerating voltage of 15 kV. Before examination, starch samples were sprinkled onto double-sided tape attached to a specimen holder and coated with gold in a vacuum evaporator.The particle size distributions of native and OSA-modified starch samples were measured with the laser light scattering method, by using a Malvern Mastersizer 3000 instrument . Distilled water was used as the suspension medium. Starch samples (100 mg) were dispersed in 5 mL of water and vortexed for 2 min . The dispersion was then swiftly transferred into the sample cell of the particle size analyzer.−1 and a scanning step of 0.033°. The relative crystallinity (%) was quantitatively estimated by using the Origin software  according to the method of Wang et al. [The crystalline structures of native and OSA-modified starch samples were evaluated using an Xpert PRO diffractometer , operating at 40 kW and 40 mA with a Cu Kα radiation source (λ = 0.154 nm). Starch samples were equilibrated to a moisture content of about 10% before the analysis. XRD patterns were acquired for a diffraction angle (2θ) range from 3° to 40° at a scanning speed of 10° ming et al. .The pasting properties of native and OSA-modified starch samples were measured using a Rapid Visco-analyzer  according to the method of Chen et al. . Starch g/g) using the following formula:Swelling power of native and OSA-modified starches were determined by heating starch–water slurries in a water bath at temperatures ranging from 50 to 80 °C in 10 °C intervals, according to the procedures of Wang and Copeland  with somSH is the height of the serum layer, and EH is the total height of the emulsion. A lower creaming index value indicates higher emulsion stability.The emulsifying stability of native and OSA-modified starches was measured according to the method of Lin et al.  with a sIn vitro digestibility of native and OSA-modified starches was analyzed according to the method of Ma et al. . BrieflyData are averages of duplicate observations and expressed as means ± standard deviations. An analysis of variance with a significance level of 5% was conducted, and Duncan’s test was applied to determine differences between means using the commercial statistical package .The DS of OSA-modified normal and high-amylose Japonica rice starches as a response to OSA addition are shown in −1 could be attributed to the vibration of O–H groups. The band at 2942 cm−1 was due to C–H stretching related with ring methane hydrogen atoms. The characteristic peak at 1643 cm−1 originated from the tightly bound water present in the starch. The peaks at 800–1200 cm−1 were due to stretching of the C–O bond.The FTIR spectra of normal and high-amylose Japonica rice starches with and without OSA modification are shown in −1 emerged after OSA modification. This peak was ascribed to the asymmetric stretching vibration of carboxylate RCOO– [−1 was observed in OSA-modified starches, which can be attributed to the characteristic C=O stretching vibration of an ester carbonyl group [−1 than OSA-HighAmyST-1. This result suggests that the increase of DS enhances the ester band absorption of OSA-modified high-amylose Japonica starch.For both normal and high-amylose starches, a new peak at 1571 cmte RCOO– . Anotheryl group . These rp > 0.05). These results suggest that particle size distribution of high-amylose starch was not significantly influenced by the OSA modification.The SEM images of normal and high-amylose Japonica rice starches with and without OSA modification are shown in p < 0.05). The RC of native high-amylose starch was 28.3%, and the derivatives at DS of 0.0177 and 0.0285 were significantly lower at 27.5% and 27.1%, respectively. This was consistent with previous research on OSA modification of indica rice starch and corn starch [XRD patterns of normal and high-amylose Japonica rice starches with and without OSA modification are shown in n starch ,29. Thesp < 0.05). This result was consistent with some previous research on other starches [p < 0.05) after OSA modification, which can be attributed to the weakening of the intermolecular hydrogen bond induced by the introduction of the bulky OSA group [p < 0.05). For OSA-modified high-amylose starch, pasting viscosities generally changed as DS increased. The incorporation of bulky OSA groups enhanced the overall pasting capacity of the starches, and the modified starches tended to paste more extensively [The pasting parameters of normal and high-amylose Japonica rice starches with and without OSA modification are shown in starches ,31,32. Zstarches  reportedSA group . SimilarSA group ,34. For ensively . Differeg/g, while that for native high-amylose starch increased from 2.13 to 7.47 g/g. The increase of amylose and long chains of amylopectin can strengthen the granular structure of starch and inhibit the swelling of granules during heating [p < 0.05). This may be ascribed to the weakening of the intermolecular hydrogen bond due to the introduction of OSA groups [p < 0.05). The two OSA-modified high-amylose starches with different DS had a slight difference in swelling power.The swelling powers of normal and high-amylose Japonica rice starches with and without OSA modification are shown in  heating ,35. AfteA groups . Like nap > 0.05). For normal starch, OSA modification improved the emulsifying stability, as revealed by the decreased creaming indexes of both freshly prepared and stored emulsions (p < 0.05). Compared with emulsions of native high-amylose starch, emulsion of OSA-HighAmyST-1 showed no significant difference in the creaming index of the freshly prepared emulsion (p > 0.05), but showed a slight decrease in the creaming index of the stored emulsion (p < 0.05), suggesting that the emulsifying stability of high-amylose starch was slightly improved after a low degree of OSA modification. The increase of DS enhanced the emulsifying stability of OSA-modified high-amylose starch. For both freshly prepared and stored emulsions, there were no significant differences between the OSA-HighAmyST-2-formulated emulsion and the OSA-NormalST-formulated emulsion in creaming indexes (p > 0.05). This result suggests OSA-modified high-amylose starch had an emulsifying stability comparable to OSA-modified normal starch of similar DS.The creaming indexes of normal and high-amylose Japonica rice starches with and without OSA modification are shown in p < 0.05). This result could be attributed to its higher amylose content and higher amounts of long-length chains (B2 and B3 chains). You et al. [p < 0.05). RS of OSA-modified high-amylose starch increased as DS increased (p < 0.05). Similar results were obtained by Remya et al. [p < 0.05). This result was in agreement with the difference of their native starches in RS content.The RDS, SDS, and RS contents of normal and high-amylose Japonica rice starches with and without OSA modification are shown in u et al.  suggestea et al. , who repOSA-modified high-amylose Japonica rice starch with different DS were prepared. The FTIR spectra confirmed the formation of ester linkage between OSA and starch. Compared with native high-amylose starch, the derivative did not show a significant change in morphology and particle size. OSA modification slightly decreased the relative crystallinity and pasting temperature of high-amylose starch, but increased the pasting viscosity, swelling power, emulsifying stability, and RS content. The changes of these properties were affected by DS. Compared with OSA-modified normal starch of similar DS, the OSA-modified high-amylose starch had a higher peak viscosity and RS content, lower swelling power, and comparable emulsifying stability. The modified high-amylose Japonica rice starch could potentially be employed as emulsifier, thickener, and fat replacer in the food industry, and the application needs further study. Due to the high content of RS, this starch derivative may have many potential health benefits, such as decreasing postprandial glycemic and insulinemic responses, lowering plasma triglyceride concentrations, improving whole-body insulin sensitivity, and reducing fat storage, which also deserve further research attention.


In this Editorial, we report on the trend of manuscript submissions during the peak of COVID-19 pandemic in 2020. The total monthly new manuscript submissions from 2018 to 2020 were compared to examine the overall trends in submission and publication activities.Apart from that, we also report on the changes in the journal’s administration and the external recognition received by the journal. The emergence of COVID-19 as a global pandemic in early 2020 and the impose of movement control order by the government has affected many aspects of both personal and professional lives of people around the world.Malaysian Journal of Medical Sciences (MJMS)’ electronic manuscript submission system : 0.394CiteScore: 2.0H-index: 25The Professional Evaluation Panel of the Majlis Penerbitan Ilmiah Malaysia (MAPIM)-Kementerian Pengajian Tinggi (KPT) Award 2019 has chosen a manuscript published in the first volume of MJMS 2019 to win the Best Journal Manuscript Award  International Index. This event is held annually but due to COVID-19 pandemic, it has been postponed to 30 March 2021 and was conducted via Cisco Webex and Facebook Live MAPIM. The details of the winning manuscript were as follows:


Pearl millet is an underutilized and drought-resistant crop that is mainly used for animal feed and fodder. Starch (70%) is the main constituent of the pearl millet grain; this starch may be a good substitute for major sources of starch such as corn, rice, potatoes, etc. Starch was isolated from pearl millet grains and modified with different physical treatments (heat-moisture (HMT), microwave (MT), and sonication treatment (ST)). The amylose content and swelling capacity of the starches decreased after HMT and MT, while the reverse was observed for ST. Transition temperatures (onset (To), peak of gelatinization (Tp), and conclusion (Tc)) of the starches ranged from 62.92–76.16 °C, 67.95–81.05 °C, and 73.78–84.50 °C, respectively. After modification , an increase in the transition temperatures was observed. Peak-viscosity of the native starch was observed to be 995 mPa.s., which was higher than the starch modified with HMT and MT. Rheological characteristics (storage modulus (G′) and loss modulus (G′′)) of the native and modified starches differed from 1039 to 1730 Pa and 83 to 94 Pa; the largest value was found for starch treated with ST and HMT. SEM showed cracks and holes on granule surfaces after HMT as well as MT starch granules. Films were prepared using both native and modified starches. The modification of the starches with different treatments had a significant impact on the moisture, transmittance, and solubility of films. The findings of this study will provide a better understanding of the functional properties of pearl millet starch for its possible utilization in film formation. Poaceae, and is a native African millet that is cultivated in the arid and semiarid tropics of Africa and Asia. The worldwide production of millet is 31,019,370 tonnes, and India is top producer of millet  followed by Niger  and Nigeria  [Pearl millet is a drought-resistant crop that belongs to the grass family, the Native starch had some drawbacks such as the lesser stability of the starch paste when given high shear, acidic, or freezing conditions, which affects the quality of food products ,3. As suMicrowave treatment (MT) is one of the starch modification techniques which can be used to change the functional characteristics of starches . MicrowaThe preparation of films and coatings based on polysaccharides, such as starch, has shown great growth in recent years due to the interesting mechanical and barrier properties observed in films prepared from these materials . Dai et Pearl millet cultivar (HC-20) was collected from Chaudhary Charan Singh Haryana Agricultural University, Hisar (HR), India. The chemicals used were of analytical grade.2S2O5, 18–20 h) and ground in a grinder . To remove the protein and fibrous matter, the slurry was passed through nylon cloth (100 mesh) and centrifuged . To dry the starch, a universal oven  was used .Sandhu and Singh’s  method oPearl millet starch was treated with HMT using the method of Collado . The moiFor MT, starch moisture content was adjusted to 25% with distilled water and mixed properly. Vacuum bags were used to hold the starch to be used for MT, and the bags were vacuumed, pressed to ensure uniform thickness (3.5 mm), and kept for 24 h at 4–6 °C before microwave treatment .ST was applied to starch by following the method of Majeed et al. . A starcThe amylose content of the starches was evaluated using the method of Williams et al. . The swew/w) was added to an aluminium pan, and an empty pan was used as reference. Pans were hermetically sealed and heated from 40 to 150 °C at the rate of 5 °C/min. Thermal transitions of the starch samples were defined as To (onset), Tp (peak of gelatinization), and Tc (conclusion), and ∆Hgel referred to the enthalpy of gelatinization. Enthalpies were calculated on the basis of dry starch. These were calculated automatically. The gelatinization temperature range (R) and peak height index (PHI) were calculated as 2(Tp-To) and ∆H/(Tp-To), respectively.Thermal characteristics of the starches were analysed using a DSC . For measurement, a starch to water ratio 1:3 . The starch suspension (8%) was equilibrated at 50 °C for 1 min. and heated at 6 °C/min to 95 °C. After holding at 95 °C (2.7 min), the suspension was cooled to 50 °C at 6 °C/min and held at 50 °C for 2 min. Peak viscosity (PV), breakdown (BV), trough (TV), setback (SV), final viscosity (FV), and pasting temperature (PT) were obtained from the pasting graph.Rheological characteristics of samples were calculated with rheometer (MCR-52). A parallel plate system was used (4 cm diameter) and a gap size of 1000 µm was maintained throughout the study. Starch paste (10%) was prepared by heating the starch suspension at 85 °C in a water bath for three minutes. Before loading the sample on the rheometer, the sample was cooled to room temperature. A frequency sweep test was performed from 0.1–100 rad/s at 25 °C. The storage modulus (G′), loss modulus (G′′), and damping factor (tanδ) were evaluated using the graph.XRD pattern of starches were analyzed using the diffractometer . Diffractograms were evaluated at 25 °C over a 2θ range of 4–40 with a step size of 0.02 and a sampling interval of 10 s.Morphological characteristics were evaluated using SEM . A starch suspension (1%) was prepared using ethanol and the sample was loaded onto an aluminium stub and was coated with gold–palladium (60:40). During analysis, an acceleration potential of 10 kV and a magnification power 1500× were used.PSD of the starch was analyzed using a laser diffraction particle size analyzer . Sonication (3 min) was applied to the starch sample prior to estimation. The refractive indexes were 1.33 and 1.53 for distilled water and starch during analysis.w/w) was added after cooling the starch slurry to room temperature and stirred at 150 rpm for 20 min. After passing the starch slurry through a muslin cloth, solutions were cast on Teflon coated baking trays and dried in the oven  at 50 °C for 16 h. After cooling, the films were peeled off and stored at 25 °C/53% relative humidity for 48 h for further measurements.Starch films were prepared by following the method of da Rosa Zavareze et al.  with somThe method of Galus et al.  was opteTriplicate observations were submitted to one-way analysis of variance (ANOVA) using Minitab Statistical Software version 15 .Amylose content (AC) plays a key role in the characteristics and applications of starch. Different methods are applied for the calculation of AC, but the iodine binding spectrophotometry method is a basic method for the determination of AC . In thisThe light transmittance of starches ranged between 13.53 to 30.52%. The largest value was evaluated for HMT starch. Light transmittance was measured for the storage period of 130 h at 4 °C . With anThe thermal analysis data for the native and modified starches are tabulated in p < 0.05) modification impacts were found on the pasting characteristics of starches , and some starch granules are shown to have some pore like structures on the granule surface . After HThe particle size distributions of the starches are tabulated in Properties of native and modified starch films are tabulated in p < 0.05) variation in the characteristics of the modified starches were observed based on the type of treatment provided to the starch. After modification, SP decreased for starch treated with HMT and MT, while solubility power increased after modification. Modified starches showed higher transition temperature compared to native starch. The pasting and rheological properties were also altered after treatment. The values of PV and G′ decreased after HMT and MT while reverse was observed for starch treated with ST. The morphological properties of the starch treated with ST showed the least effect on the granule surface, while starch granules treated with HMT had the highest effect. Films prepared using modified starches showed less moisture and higher solubility and opacity content compared to films prepared using native starch. The findings suggest that physical modifications can improve the stability and gel texture of starches, which will improve the applications of starches.In present study, pearl millet starch was modified using HMT, MT, and ST. Significant (


This study revealed the underlying mechanisms involved in the puffing process of dried cassava starch gel by exploring the development of the puffed structure of gel upon sand-frying, chiefly focused on the changes in the multi-scale structure and the physicochemical properties of starch. The results suggested that the sand-frying-induced puffing proceeded very fast, completed in about twenty seconds, which could be described as a two-phase pattern including the warming up (0~6 s) and puffing (7~18 s) stages. In the first stage, no significant changes occurred to the structure or appearance of the starch gel. In the second stage, the cells in the gel network structure were expanded until burst, which brought about a decrease in moisture content, bulk density, and hardness, as well as the increase in porosity and crispness when the surface temperature of gel reached glass transition temperature of 125.28 °C. Upon sand-frying puffing, the crystalline melting and molecular degradation of starch happened simultaneously, of which the latter mainly occurred in the first stage. Along with the increase of puffing time, the thermal stability, peak viscosity, and final viscosity of starch gradually decreased, while the water solubility index increased. Knowing the underlying mechanisms of this process might help manufacturers produce a better quality of starch-based puffed products. As is well known, starch is a versatile ingredient widely used in various foods as a thickener, stabilizer, or a gelling agent. Starch is also the most important nutrient, which provides the major energy for the human body . More imBesides the puffing regime, starch possessing the appropriate structure is usually applied to develop high-quality starch-based puffed snacks. For instance, Joshi et al. selected the rice varieties  to produce superior puffed products, which was credited by their having the proper ratio of amylose to amylopectin . Dias etA clear understanding of the underlying mechanisms involved in the puffing process is needed for obtaining a better quality of puffed products . CurrentCassava starch was purchased from Guangxi Honghao Starch Development Co., Ltd. . Its proximate composition was measured by AACC methods, namely starch (76-11), amylose (61-03), crude protein (46-13), crude lipid (30-25), moisture (44-15A) and ash (08-17) . The resCassava starch (40 g) was suspended in water (50 mL), and the resultant slurry was poured into a stainless tray (21.0 × 16.5 × 2.50 cm). The slurry was spread uniformly on the bottom of the tray by gently shaking it, and then the tray was horizontally floated over a boiling water bath for 15 s. Subsequently, the tray was immersed in boiling water for approximately 30 s. After that, the tray was immediately removed and then immersed in a tap water bath for approximately 15 s to cool down. As a result, a starch gel formed on the bottom of the tray . The gelTo puff the dried starch discs, fine sand (300 g) was loaded in an iron pan and heated to 200 ± 10 °C with the aid of an induction cooker . The temperature of the sand bed was measured using an AS842A infrared thermometer . Then starch discs were fed into a hot sand bed and quickly blended with a stainless steel spatula to achieve uniform frying. A video of puffed starch gel preparation was provided as bp), true density (p), porosity (ε), hardness, crispness, and color. The volume of the starch disc was determined by a fine sand replacement method, and the puffing ratio was defined as the ratio of disc volume before and after puffing [bp, g/mL) was obtained by dividing the disc’s mass by its volume. The true density (p) was determined by a toluene displacement method as described previously by Joshi et al. [p−bp)/p × 100%. The hardness and crispness were determined by a puncture test method with the aid of a TA.XT plus texture analyzer  equipped with a spherical probe (P/0.5s) [The puffing performance of the dried starch discs was evaluated in terms of puffing ratio, moisture content, bulk density  . A trigg(P/0.5s) ,29. The 2, approximately 50 mg) was first put into a dilatometer and then evacuated.The cross-sectional images of starch discs were recorded by scanning electron microscopy (SEM)  at an accelerating voltage of 10 kV. The pore size distribution was determined with a mercury porosimeter  with pressure varied from 0.1 to 61,000 psia. Before measurement, the sample (1 cmemp), peak time (PT), peak viscosity (PV), trough viscosity (TV), final viscosity (FV), breakdown (BD), and setback (SB) were recorded [To determine the physicochemical properties of starch, the discs were firstly ground and passed through a 200-mesh sieve. The water solubility index (WSI) was measured according to the method described by Kaisangsri et al. . The pasrecorded .Tg) of the starch disc was evaluated under tensile mode using a Q800 Dynamic Mechanical Analyser . The samples were heated from 30 °C to 200 °C with a heating rate of 5 °C/min. The oscillating frequency was set at 1 Hz. In this study, the temperature corresponding to the peak value of tan δ curve was taken as Tg.The thermal stability was analyzed by a thermogravimetric analyzer  under nitrogen by heating the samples from 30 °C to 600 °C at a rate of 10 °C/min. The glass transition temperature (θ)–40°(2θ) at a scanning rate of 2°/min. The MDI-Jade 6.0 software  was utilized to calculate the relative crystallinity. For FT-IR, the obtained spectrum was processed by deconvolution using a PeakFit software  [The ordered starch structure in the discs was analyzed by an X’Pert3 Powder X-ray diffractometer  and a Spectrum100 FT-IR spectrometer . The diffractometer was operated at 40 kV and 40 mA across the diffraction angle range of 4° .g for 10 min, and the resulting supernatant was filtered through a 0.22 μm PTFE membrane. An aliquot of the filtrate (100 μL) was applied onto a high-pressure size exclusion chromatography system , coupled with a multi-angle laser light scattering detector  and a refractive index detector . A set of chromatographic columns connected in series, i.e., Ohpak SB-805 HQ, Ohpak SB-804 HQ, and Ohpak SB-803 HQ , were used to separate. DMSO containing 5 mM LiBr was used as the eluent, and the flow rate was controlled at 0.3 mL/min. Astra 5.3.4.20 software  was used to analyze the collected data from light scattering. The weight-average molecular weight (wM) and number-average molecular weight (nM) were calculated using the second-order berry method.The molecular weight of the starch was determined according to a previous method with some modifications . The staw/v) and 5 μL of isoamylase (1000 U/μL) were added to 2.5 mL of a gelatinized sample and stirred for 24 h at 37 °C. Then, the mixture was heated for 5 min to deactivate the enzymes. After placing 600 μL of the mixture in a centrifuge tube, it was dried by blowing nitrogen at room temperature. Finally, the dried sample was dissolved in 600 μL of mobile phase, centrifuged at 12,000× g for 5 min, and 20 μL of supernatant was analyzed by HPAEC-PAD. The ICS5000 system  equipped with a 4 × 50 mm CarboPac PA1 guard column and a 4 × 250 mm Dionex™ CarboPac™ PA10 analytical column was used. The sample was eluted with a mixture of 0.2 mol/L NaOH and 0.2 mol/L NaAC. The flow rate was set at 0.5 mL/min. PeakNet  was used to analyze the experimental data.The chain length of amylopectin molecules was analyzed with high-performance anion-exchange chromatography coupled to a pulsed amperometric detector. Briefly, fractionated amylopectin (5.0 mg) was suspended in 5 mL of double-distilled water and heated in a boiling water bath for 60 min with an intermittent vortex. The sodium acetate buffer , NaN3  by SPSS statistical software . A comparison of means was carried out using Duncan’s test. A p > 0.05). The puffing ratio, porosity, whiteness (L*), total color difference (ΔE*), and crispness of the gel were progressively increased from beginning at 7 s (puffing stage), whereas the moisture content, redness (a*), yellowness (b*), and hardness were decreased step by step. Finally, the physical properties of the gel were not altered fundamentally. At the heating stage, the surface temperature of the gel did not reach its glass transition temperature (Tg)  , so the The cross-sectional microstructure of gels at different puffing moments are pre-sented in It can be seen from p < 0.05) within the heating stage. However, in the puffing stage, the WSI of cassava starch increased slowly. This might be caused by the large degree of degradation of cassava starch in the heating stage. Nevertheless, cassava starch was only degraded slightly during the puffing stage [The water solubility index represents the number of small molecules dissolved in water . As illung stage . These rng stage .p > 0.05) in all samples. The peak time ranged from 4.16 to 4.53 min. The final viscosity, peak viscosity, trough viscosity, setback, and breakdown ranged from 3256 to 4730 Cp, from 2639 to 5655 Cp, from 1910 to 3226 Cp, from 1346 to 1818 Cp, and from 729 to 2429 Cp, respectively. It is worth noting that both peak viscosity and final viscosity were markedly reduced (p < 0.05) in the sand-frying puffing process, which mainly occurred in the puffing stage of the starch gel. It has been reported that both peak viscosity and final viscosity of grain  starch in the gun puffing process were reduced [The pasting properties can shed light on the alterations in viscosity of starch suspension when heated, and reflect the changes of starch structure. The pasting curves  and its  reduced . Lai and reduced . Hidalgo reduced . The decThe thermal stability of cassava starch under different puffing moments was evaluated, and relevant results, including thermogravimetric (TG) and corresponding derivative TG (DTG) curves, are presented in θ = 5.6° and 2θ = 17° at the puffing time of 0 s. This was a typical B-type crystallinity pattern. With an increase of puffing time, the diffraction peaks at 2θ = 17° would disappear, and the intensity of diffraction peaks at 2θ = 5.6° was reduced. These results suggested that puffing might be closely related to the diffraction peak of 2θ = 17°. In addition, the long-range crystalline structure of cassava starch was transformed into an amorphous structure in the sand-frying puffing process [The XRD spectra of cassava starch under various puffing times were presented in  process . This po process ,42. In 2 process . Similar process . In this−1 was obtained as presented in −1 represented molecular order and crystallinity of starch. 1020 cm−1 represented the amorphous or disorder phase of starch. The ratio of 1047/1020 cm−1 can be used to quantify the molecular order degree of starch [−1 was gradually reduced as the puffing time increased, indicating that the molecular order degree of cassava starch decreased during the sand-frying puffing process, which primarily occurred in the heating stage. A similar result has been reported by Kuo et al. [−1 was reduced in the steam cooking or extrusion puffing process, thus influencing the molecular order degree of rice starch. These results indicate that the low molecular order degree of starch might be conducive to puffing.f starch . As showo et al. . They diwM and nM are summarized in wM and nM were decreased with the increase of puffing time. Liu et al. also found that the degradation of the molecular weight of corn starch occurred during the puffing process [The SEC–MALLS–DRI chromatograms and graphs of molecular weight versus elution time of cassava starch under various puffing times were obtained, as illustrated in  process . In this1 (DP 13–24), chain B2 (DP 25–36), and chain B3 (DP ≥ 37) [2, chain B3, and average chain length (CL) decreased from 11.1% to 8.2%, from 6.9% to 2.6%, and from 18.4 to 16.6, respectively. However, there was an increase in the relative proportion of chain A and chain B1 from 32.3% to 35.9%, and from 49.7% to 53.3%, which proved that cassava amylopectin was degraded in the sand-frying puffing process, thus reducing the molecular weight of cassava starch. Li et al. also reported the analogous findings. They discovered that the relative proportion of the A chains (21.1%) of potato amylopectin remained unchanged when it was treated with moisture-heat treatment , but that the relative proportion of B1 chains increased from 49.1% to 49.6%, and those of the B2 and B3 chains were reduced from 17.8% to 17.6%, and from 12.0% to 11.6%, respectively [The chain length distribution of cassava amylopectin at different puffing times were presented in DP ≥ 37) . The resectively . Due to In this study, the underlying mechanisms involved in the puffing process of dried cassava starch gel were revealed. The results indicated that the puffing of starch gel was a complex process, and the whole process was mainly divided into two stages, namely the heating stage and the puffing stage. In the heating stage (0~6 s), starch gel experienced no significant changes in its appearance and structure. In the puffing stage (7~18 s), the cells in the gel network structure expanded until they burst, which caused changes in the physical properties of gel when the surface temperature of the gel reached glass transition temperature. The destruction of the crystalline structure and the short-range molecular order structure of cassava starch and molecular degradation occurred simultaneously in the above two stages, with molecular degradation mainly occurring in the first stage. Meanwhile, the thermal stability, peak viscosity, and final viscosity of the starch gradually decreased, while the water solubility index increased along with the enhancement of puffing time. Although this work only figured out the underlying mechanisms for the puffing process of cassava starch gel, the physics of the process were quite similar to other starch gels. Therefore, this work might help manufacturers produce a better quality of starch-based puffed products. However, it must be noted that the present study only involved the puffing of starch gel related to the time scale, whereas the puffing of starch gel concerning moisture and temperature scales has not been conducted. This should be addressed in subsequent investigations.


We report a case of a mycotic abdominal aortic aneurysm caused by invasive group B streptococcus. Given the anatomical suitability with healthy segments of aortoiliac vessels, in situ repair was performed. A cryopreserved femoral vein graft was chosen because of risks of graft reinfection and negated the need for bilateral femoral vein harvest. The patient remained clinically well and the graft patent with no concerns at 6 months of follow-up. A review of literature on group B Streptococcus aortitis was performed. Mycotic aneurysms account for less than 2% of surgically repaired aortic aneurysms but are nevertheless associated with marked morbidity and a mortality rate of 15%-50%.Salmonella and Staphylococcus, and less commonly Klebsiella, Campylobacter, Escherichia coli, group A Streptococcus, Enterococcus, Treponema pallidum, Mycobacterium tuberculosis, or fungi (particularly Candida or Aspergillus).The predominant causative organisms include 9/L and C-reactive protein was mildly elevated at 11.5 mg/L. All other laboratory workups were otherwise unremarkable. She had an abdominal/pelvis computed tomography (CT) scan that showed nonspecific fat stranding around a mildly dilated aorta maximally measuring 3.0 × 2.7 cm . Blood cultures remained negative after 5 days. Infectious diseases was consulted, and the patient was switched to ceftriaxone for 6 weeks, followed by oral amoxicillin for a year thereafter. The patient was discharged 10 days postoperatively. At 6 months of follow-up, she remained clinically well and CT imaging demonstrated resolution of surrounding stranding without any signs of pseudoaneurysm or deformities.The patient remained stable and her pain completely resolved after the procedure. The surgical tissue culture surprisingly grew GBS (GBS is a gram-positive, facultative anaerobic bacteria often implicated in skin and soft-tissue infections, bacteremia without a focus, endocarditis, osteomyelitis, neonatal meningitis, urinary tract infections, and pneumonia.,Before the advent of antibiotics, the most common cause of mycotic AAAs was infectious endocarditis. More common nowadays is hematogenous or lymphatic bacterial spread in individuals with risk factors such as arterial damage from iatrogenic manipulation, immunodeficiencies, and intravascular drug use.Surgery is an essential component in the management of mycotic AAAs and may include ex situ or in situ repair. Potential disadvantages to ex situ repair include concerns of graft patency, as well as aortic stump rupture caused by residual tissue infection.When opting for in situ repair, there are several graft options. Prosthetics such as Dacron or polytetrafluoroethylene may be used, and despite impregnating with rifampin, the risk of graft infection must be taken into consideration. Other means of reducing infection risk may be employed including soft-tissue coverage with a pedicle of uninfected tissue such as the omentum.More recently, cryopreserved venous allografts have shown promise for their low rates of reinfection and reintervention. In a study of 23 suprainguinal arterial reconstructions using cryopreserved venous allografts—5 of which were mycotic AAAs—graft reinfection occurred in only one patient and two required reintervention for anastomotic dilatation.,,15In our review of the literature, we did not find any previous cases of a cryopreserved venous allograft used in the repair of a mycotic AAA caused by GBS. Among the nine other reported cases, one used an iliac arterial homograftMycotic AAAs caused by GBS infection are rare with generally favorable surgical outcomes in previous reports. Cryopreserved femoral vein allograft may be an ideal choice for in situ repair to mitigate concerns of graft reinfection while negating the morbidity of femoral vein harvests. The long-term durability of cryopreserved femoral vein for aortic reconstruction remains to be determined.


Clostridium thermocellum, can have cost-effective cellulosic biomass utilizations, such as consolidated bioprocessing, simultaneous biological enzyme production and saccharification. However, these processes require a longer cultivation term of approximately 1 week. We demonstrate that constituents of the C. thermocellum membrane vesicle fraction significantly promoted the growth rate of C. thermocellum. Similarly, cell-free Bacillus subtilis broth was able to increase C. thermocellum growth rate, while several B. subtilis single-gene deletion mutants, e.g., yxeJ, yxeH, ahpC, yxdK, iolF, decreased the growth stimulation ability. Metabolome analysis revealed signal compounds for cell–cell communication in the C. thermocellum membrane vesicle fraction  and B. subtilis broth . These findings suggest that the constituents in membrane vesicles from C. thermocellum and B. subtilis could promote C. thermocellum growth, leading to improved efficiency of cellulosic biomass utilization.The cultivation of the cellulolytic bacterium,  Clostridium thermocellum (Acetivibrio thermocellus)  hydrogenase maturase, lactate dehydrogenase, pyruvate-formate lyase, Pfl-activating enzyme, phosphotransacetylase, and acetate kinase genes, which eliminated formate, acetate, and lactate production, and reduced H2 production, presented a titer of 2.2% ethanol from 6% Avicel cellulose [C. thermocellum I-1-B produced 2.4% ethanol from 8% cellulose [Thermoanaerobacterium saccharolyticum produced 3.8% ethanol from 9.2% Avicel cellulose in 146 h [C. thermocellum can be simplified consolidated bioprocessing (CBP). This is a promising strategy because it eliminates the need to add lignocellulose-degrading enzymes that significantly increase the cost of biofuel production [ellulose . A straiellulose . The ethoduction ,12.C. thermocellum, form carbohydrate-active enzyme (CAZyme) complexes that are termed cellulosomes [C. thermocellum preferentially utilizes cellooligosaccharide, and glucose tends to accumulate in the culture broth [C. thermocellum from 10% cellulose or 12% alkali pretreated rice straw by approximately 7.7% over 10 days [Some cellulolytic bacteria, including ulosomes ,14,15,16re broth . Supplem 10 days . This teC. thermocellum produces extracellular membrane vesicles (MVs) that are released into the broth [Bacillus subtilis, B. anthracis, Streptomyces coelicolor, Listeria monocytogenes, Staphylococcus aureus, Streptococcus mutans, S. pneumoniae, and Clostridium perfringens [Ruminococcus spp., a cellulolytic bacterium that resides in the ovine rumen. DNA molecules ranging in size from <20 to 49 kb, and from 23 to 90 kb are attached to MVs from Ruminococcus sp. YE73 and Ruminococcus albus AR67, respectively. Thus, MVs can function as vectors for horizontal gene transfer to confer cellulolytic activity, as documented in the mutant strain Ruminococcus sp. YE71 [Bacteroides fragilis and B. thetaiotaomicron are equipped with hydrolytic enzymes and are important in polysaccharide degradation [Fibrobacter succinogenes are enriched with CAZymes, and intact MVs are able to degrade a broad range of hemicelluloses and pectin [C. thermocellum may utilize MVs to deliver cellulosomes, which enhance the cellulolytic activity of C. thermocellum [We previously reported that he broth . MVs arehe broth ,21. MVs fringens ,26,27,28sp. YE71 . MVs froradation ,31. MVs d pectin . We havemocellum .Pseudomonas aeruginosa, a hydrophobic cell–cell communication signal termed Pseudomonas quinolone signal is released from the bacteria via MVs [Alteromonas and Halomonas as the sole carbon source, indicating that MVs should be considered in the marine food web and may have important roles in the carbon flux of the ocean [Mycobacterium tuberculosis, the causative agent of tuberculosis, increased MV production in response to iron restriction has been observed [M. tuberculosis.MVs contain various compounds that include DNA and RNA. These cargos are delivered to neighboring cells. MVs have several important functions related to cell–cell interactions. In  via MVs ,34. MVs he ocean . In Mycoobserved . These MC. thermocellum and B. subtilis can promote C. thermocellum growth. Metabolome analysis was also performed to identify the candidate compounds with the growth stimulation.In this study, we demonstrated that the MV fractions collected from C. thermocellum DSM 1313  culture was inoculated in 5 mL of CTFUD medium 2SO4, 1.5 g/L KH2PO4, 130 mg/L CaCl2 2H2O, 500 mg/L L-cysteine-HCl, 11.56 g/L 3-morpholinopropanesulfonic acid, 2.6 g/L MgCl2 6H2O, 1 mg/L FeSO4 7H2O, 4.5 g/L Bacto yeast extract, 1 mg/L resazurin, pH 7.0) containing 0.5% cellobiose  with 16 × 125 mm Hungate tubes , and cultured at 60 °C under anaerobic conditions with nitrogen gas [One hundred microliters of ogen gas .B. subtilis KAO/NAIST chromosomal deletion mutants [B. subtilis . B. subtilis strains were aerobically cultured in Luria Bertani broth at 37 °C. mutants  and BKE  mutants  were obtC. thermocellum and B. subtilis culture was centrifuged at 10,000× g for 2 min at 4 °C, and the supernatant was filtered through a 0.22-μm syringe filter to remove cells. The filtrate was centrifuged at 179,000× g for 1 h at 4 °C and the pellet was washed twice with 2 mL of sterile phosphate-buffered saline (PBS). The pellet was resuspended in PBS and used as the MV fraction. The MV fraction was kept on ice before use.Five milliliters of MVs were visualized using transmission electron microscopy. Six microliter aliquots of the MV fraction was added to 300-mesh carbon and formvar-coated copper grids and incubated for 1 min. After removing the extra solution with filter paper, each specimen was stained with 2% phosphotungstic acid. The sample was observed with a JEM-1011 microscope  at an accelerating voltage of 80 kV.C. thermocellum DSM 1313 culture was inoculated in 5 mL of CTFUD medium containing 0.5% cellobiose with the supplementation of the collected MV fraction. C. thermocellum was cultured at 60 °C under anaerobic conditions with nitrogen gas. The C. thermocellum growth was evaluated with optical density of the broth at 600 nm.One hundred microliters of C. thermocellum MV fraction was treated with 10 mg/L surfactin, and the filtrate obtained after ultrafiltration with Vivaspin 2-100 K  was used to obtain the constituents in MVs. Cell-free supernatants of B. subtilis trpC2 and trpC2 yxeJ broth were prepared by centrifugation and filtration with a 0.22-μm syringe filter. These specimens were homogenized with zirconia beads in 75% methanol, and the supernatants were collected after centrifugation at 15,000× g rpm for 10 min. The supernatants were applied to a MonoSpin C18 column  and were filtered through a 0.22-μm syringe filter.The m/z: 80–1200; full scan resolution, 70,000; and MS/MS scan resolution, 17,500. The obtained data were analyzed with PowerGetBatch and MFSearcher [LC-MS analysis was performed on an Ultimate 3000 rapid separation LC (RSLC) and the Q Exactive system . Ultimate 3000 RSLC analysis was performed with the following parameters: column, InertSustain AQ-C18 (GL Science); column temperature, 40 °C; injection volume, 2 µL; solvent flow rate, 200 µL/min. The eluting solution was 0.1% formic acid containing 2% acetonitrile. The Q Exactive system had the following parameters: measurement time, 3–30 min; ionization method, electrospray ionization; measurement mass range, C. thermocellum and T. saccharolyticum strains produced 3.8% ethanol from cellulose for 6 days [C. thermocellum cultivation with BGL supplementation for 10 days reportedly produced 76.7 g/L glucose from alkali pretreated rice straw [C. thermocellum is an important factor in improving the efficiency of CBP and BSES. In this study, we collected MVs from C. thermocellum broth  promoted C. thermocellum growth by accelerating the growth rate  of myo-inositol cysteine to cysteine [Among these 23 genes, the functions of several genes have been experimentally evaluated. The paragine . The ioleactions ,47,48. Winositol . YydF issubtilis . Kim et t growth . PQQ, wht growth . PQQ hast growth ,55,56. Wis broth . More th protein ,58. ahpCeroxides . Utilizaeroxides . A rangeeroxides . YxdK iseroxides . Co-cult ability . The yxecysteine .C. thermocellum MVs and analyzed them using LC-MS/MS. Among the 534 detected peaks, the intensities of seven peaks were significantly higher in the fraction where MVs had been disrupted by surfactin compared to MVs not disrupted using surfactin . These include cis-11-methyl-2-dodecenoic acid from g others . DSFs arg others ,68,69,70l medium . Dean etoduction .B. subtilis trpC2 and trpC2 yxeJ (B. subtilis trpC2 compared to that of trpC2 yxeJ (B. subtilis trpC2 broth were estimated by MS/MS analysis (B. subtilis trpC2 broth. Nicotinamide reportedly enhances growth of both Gram-negative and Gram-positive bacteria, such as M. avium, Propionibacterium acnes, S. aureus, and B. macerans [Vibrio cholerae O1 [Pseudomonas and Aeromonas strains has been reported [Agrobacterium. 6-Paradol was reported to have significant anti-adhesive activity against S. aureus [Subsequently, we compared the metabolites in the broth of pC2 yxeJ . Among tpC2 yxeJ . The stranalysis . Diversemacerans ,74,75,76lerae O1 . The utireported ,79. Nopa. aureus .C. thermocellum. Additionally, the MV constituents with growth stimulation were described by LC-MS/MS analysis. These findings suggest that the constituents in membrane vesicles could promote C. thermocellum growth, leading to improved efficiency of cellulosic biomass utilization.In this study, we demonstrated that constituents in membrane vesicles significantly promoted the growth rate of


American Journal of Preventive Cardiology. We cannot express how grateful we are to all of you who have given the Journal the opportunity to publish your work, as well as our dedicated editorial board and associate editors, the leadership of the American Society for Preventive Cardiology (ASPC), and our publisher Elsevier. You all believed in the promise and potential of the Journal from the beginning, and without your support and effort, the Journal’s productivity to this point would have not been possible. The acceptance and indexing of the Journal into Pub Med Central earlier this year, barely a year after its inception, is testament to its importance in publishing the best work in preventive cardiology. The Journal has received to date well over 200 submissions, including more than 125 original contributions and more than 30 state-of-the-art reviews, with an average time to first decision of 2.5 weeks.We are proud to announce with this issue that we have now completed two years of publication of the Journal again brings more content to advance the prevention field forward. The issue begins with unique ASPC practice statement on enrollment of women and diverse populations in cardiovascular clinical trials and a commentary on disparities in COVID-19 and implications for preventive cardiology. Some original research articles of note include one on age and sex disparities in hypertension control in the Multiethnic Study of Atherosclerosis, how thoracic aortic calcification may improve risk prediction and help inform statin eligibility, a simulation analysis of cost effectiveness of genetic testing in familial hypercholesterolemia, as well as one on the interplay of sleep duration and inflammation on cardiovascular mortality. Other original contributions also nicely round out the issue as well as a state-of-the-art review on a paradox of atrial fibrillation involving hypertension and stroke.This 8th issue of the Journal, and wish you all a safe and successful New Year. With the COVID-19 pandemic hopefully coming in better control, we hope to see many of you in the near future!As we close in on the Holidays, we again thank all of you who have contributed to the success of the


Compounds with lower dynamic stiffness are a better solution from the tyre/road noise point of view. The article presents the constructed test stand for the evaluation of dynamic stiffness both in in situ and in laboratory conditions. As a result of the tests, it was found that poroelastic pavements have a much lower dynamic stiffness (from 138.3 to 143.0 dB re. 1 N/m) compared to the asphalt concrete pavement (150.3 dB re. 1 N/m). In the group of poroelastic pavements, lower dynamic stiffness is characteristic for pavements with a binder course of porous asphalt. The results of the research are a contribution to further work on the influence of the dynamic stiffness of the pavements on the tyre/road noise level. The conducted measurements and analysis of the results prove the usefulness of the proposed test stand for determining the dynamic stiffness of bituminous mixtures in laboratory and field conditions. This is confirmed by the coherence between the force and acceleration signals at the level of at least 0.96—which indicates a very good validation of the test results with a random error lower than ±5% with 90% confidence level. Solutions with a poroelastic wearing course are a special type of road pavement. Such a surface is characterized by the fact that the upper layer of the surface is made of a mineral-rubber-asphalt mixture. Such a mixture contains more than 20% air void content and about 20% rubber granules. A poroelastic pavement has a positive effect on the reduction in the tyre/road noise level. This is due to better sound absorption because of high void content, and lower stiffness as a result of the use of rubber granulate to partially replace mineral aggregate. Research on poroelastic pavement was carried out as part of the SEPOR project (TECHMATSTRATEG1/347040/17/NCBR/2018) [Differences in sound levels in the frequency range above 500 Hz are partly due to the different grain size of the aggregate and different void content. Considering the same maximum aggregate grain size of PSMA8 and PAC8 mixtures and similar void content, it can be assumed that the differences in sound levels result from varying levels of stiffness of these layers. Contrary to the assessment of the load capacity of road surfaces, in the case of tyre/road noise, pavements with lower stiffness are the better solution. In the literature, despite general records on the impact of the stiffness of the upper pavement layers on the tyre/road noise level, there are no clear findings in this regard.Results of testing of the rolling noise of car tyres confirm the need to develop a test stand that could be used to reliably determine both the dynamic stiffness of bituminous mixtures in laboratory conditions and road pavements in field conditions.The first configuration of the test stand was presented in . This paThe dynamic stiffness of asphalt mixtures is usually determined under laboratory conditions. Such tests can be carried out with the use of universal testing machines or other dedicated machines and devices. Usually, they are also equipped with thermal chambers that enable the testing of samples under various temperature conditions.a two-point bending test on trapezoidal specimens or prismatic specimens ,a three-point bending test on prismatic specimens or a four-point bending test on prismatic specimens (3PB-PR and 4PB-PR tests),an indirect tensile test on cylindrical specimens ;a direct uniaxial test, in which cylindrical specimens are subjected to direct tension-compression (DTC-CY test) or only direct tension (DT-CY test). In the latter case, when the test consists only of direct tension, the specimen may also have a prismatic shape (DT-PR test).The detailed method of preparing bituminous samples and performing stiffness measurements is presented in standard DIN EN 12697-26:2012 . In accoThe research methods specified in standard  were useA slightly different methodology for determining the dynamic modulus for bitumen samples in laboratory conditions was proposed in the report of the American Transportation Research Board  and stanIn a very similar way, when using the repetitive sinusoidal dynamic compressive-axial load, the dynamic modulus of bituminous samples under laboratory conditions was also tested by Walubita et al.  and ZhanIn laboratory conditions, attempts were also made to test the dynamic stiffness of samples from bituminous mixtures in the same way as was developed for materials used under floating floors in dwellings. The detailed method of specimen preparation and the procedure for testing the dynamic stiffness of these types of material are included in standard EN 29052-1 . The assThe stiffness measurements carried out in laboratory conditions are closely related to the geometry and dimensions of the tested samples and do not take into account the influence of the stiffness of the lower structural layers of the pavement on the tested stiffness. The method of preparing/levelling the surface of the samples (or the lack of their preparation) may also significantly affect the obtained results. Therefore, more and more attempts are being made to test dynamic stiffness directly on road surfaces.Field research is most often based on impulse excitation realized with impact/modal hammers and the recording of motion response with one or more accelerometers (or other types of transducers) bound or bolted to the wearing course of the pavement. An example of this type of research is described in studies ,18. To dPulse excitation to test the stiffness of asphalt pavements was also used in the experimental device “The PiScan Probe” . In addiUnfortunately, despite the fact that the impact excitation generated with the use of modal hammers are quick, uncomplicated and convenient in the implementation of various types of field measurements , they do not provide such a high Signal-to-Noise ratio as the excitations generated by the use of modal shakers/exciters . Impact Most probably, it is the significant non-linearity  and the use of impulse excitation to test the stiffness of the road surface that should explain such large differences between the determined (3.9–17.7 GPa) and the expected (2.5–3.2 GPa according to the IKRAM Group Sdn. Bhd. Standard) values of elastic modulus and a difference of up to 56% between what was expected and what was determined with the use of the “The PiScan Probe” to assess the thickness of the asphalt layer .That is why a reliable determination of asphalt pavement stiffness in in situ conditions should be based on modal exciters rather than on impulse excitation.A good starting point for developing a methodology for this type of research could be laboratory methods using portable modal exciters to determine various types of measures of stiffness of the tested material. Unfortunately, the stiffness of the asphalt pavement cannot be determined in situ in accordance with the guidelines of standard . The proA much better solution when it comes to the possibility of conducting field measurements aimed at reliable determination of the dynamic parameters of asphalt mixtures, equated with their stiffness, is the application of the guidelines used in modal analysis and dynamic diagnostics of buildings, machines, and various types of vehicles, which are included in ISO 7626-2:1990 . The staThe solution to enable testing the driving-point dynamic stiffness of asphalt mixtures at the field site in a manner similar to that described in  was pres, than the loads occurring in the case of materials used for floating floors (0.4–4.0 kPa)—for which the dedicated dynamic stiffness test procedure has been developed [Taking into account the maximum grain size of aggregate in asphalt mixtures used in typical road surfaces up to 11–14 mm), the dimensions of the circular plate were considered insufficient for mediating in the transfer of dynamic forces to the tested samples adopted in . Moreove mm, the eveloped . The obtAt the stage of modernisation of the test stand for dynamic stiffness determination, an assumption was made about the possibility of carrying out measurements with the Resonant and Non-resonant Methods in laboratory conditions (on samples of any shape and dimension) and with the Non-resonant Method in field conditions implemented directly on various types of road surfaces.In the test stand, described in detail by the authors in , the inp2) than that adopted in the test stand described in [The impedance head is not directly attached to the tested structure. It is usually screwed to the dedicated hexagonal mounting bases with a diameter of over a dozen mm, which are bound to the tested structure. Taking into account the typical size of the aggregate grains used in bituminous mixtures and taking into account the operating ranges of the TIRA TV 51144IN inertial exciter  as well as the PCB 2888D01 impedance head, it was considered a reasonable compromise to transfer the loads from the exciter to the tested samples or the road surface through a steel circular plate with a diameter of 30 mm, which provides over 4.5 times the load transfer surface . The test stand with mounted stainless steel and polyamide stingers is shown in As a result of the research, it was found that in the case of asphalt mixtures, a much better solution is to use an atypical stinger made of polyamide  or made of stainless steel. Therefore, in further testing, whether carried out on bituminous samples in laboratory conditions or directly on the road surface, a stinger made of rigid polyamide was used.In order to increase the mobility of the test stand in laboratory conditions, a massive cast iron measuring/scribing plate  was mounted on a custom designed steel support structure equipped with a set of three swivel wheels, which allows researchers to move the stand freely. Additionally, a three-point mechanism for precise screw lifting (for transport/displacement) and lowering was mounted on the steel supporting structure to secure it in a horizontal position on the laboratory floor for the duration of measurements . The supporting structure of the measuring base is shown in Other modifications were also made to the test stand. Instead of the 32-channel SCADAS Recorder data acquisition system from SIEMENS, a 16-channel SCADAS Mobile measurement system  was used . Additionally, a control measurement was introduced based on the CMOS Multi-Function Analogue Laser Sensor—IL series produced by Keyence International, with an IL-600 sensor head (measuring range from 200 to 1000 mm) and the IL-1000 type amplifier. The laser displacement measurement system is shown in To power the data acquisition system and the amplifier and exciter in the field, a Honda EU20i single-phase portable generator with a nominal power of 1.6 kW (max 2.0 kW), IP23 protection rating, and weight of approx. 21 kg was used.The study was divided into two stages. In the first stage (stage No. 1), measurements of dynamic stiffness were carried out directly on selected road surfaces (in situ). In the second stage (stage No. 2), the dynamic stiffness was determined in laboratory conditions on samples taken at the in-situ test sites.In situ measurements (stage No. 1) were carried out in Poland on the experimental section of the road near Kartoszyno (on 17 March 2021) and on pavements located in the Bialystok University of Technology campus (on 30 April 2021). In the case of the experimental section in Kartoszyno, four test sites were selected on pavements with a poroelastic wearing course made of PSMA5 and PSMA8 with SMA11 and PA11 as binder courses. At the Bialystok University of Technology campus, research was carried out in one location with a typical asphalt concrete (AC) surface on a concrete base course.In each of the adopted test sites, three measurements were taken at points separated by at least 30 cm from one another. At each test site, samples were taken for further testing in laboratory conditions. The mobile test stand and exemplary test sites are shown in In the second stage of the study (stage No. 2), measurements of dynamic stiffness were carried out in the laboratory of the Bialystok University of Technology on samples taken from road pavements where in situ measurements were carried out (from five test sites). The samples drilled from the pavement were made to the standardized dimensions  . The thiIn For each of the samples, three measurements of dynamic stiffness were carried out with the use of identical data acquisition settings and the signal generated by the exciter—as in the case of in situ measurements.To maximize the possibilities of the test stand, measurements of dynamic stiffness were carried out in a frequency bandwidth that is twice as wide 2048 Hz) as in previous tests 048 Hz as, with thIn accordance with the recommendations of ISO 7626-2  the timeAnalysis of preliminary test results show that the coherence of the recorded signals for both in situ tests and lab samples of the pavement are close to 1.0 . With 500 averages and such a high level of coherence of the recorded signals, the random error should be significantly lower than ±5% .The ISO 7626-2  and PN-EWith the applied static load from the exciter at the level of about 10 kg and the diameter of the load plate of 30 mm, the sample load at the driving-point was 138.8 kPa.The registration and analysis of the recorded data was carried out using the LMS Test.Lab Spectral Testing software .In the following graphs , the exeTo facilitate the visual comparative analysis of the obtained dynamic stiffness, the following diagrams have compiled averaged values from individual measurement series—independently for in situ tests  and laboTo visualize the differences in the dynamic stiffness obtained as a result of field tests carried out directly on asphalt surfaces and tests carried out in laboratory conditions, all averaged values were summarised in one graph :The dynamic stiffness for selected frequencies  is also summarised in tabular form . AdditioThe measurements and analysis of the recorded signals show that in each case there is a very strong coherence between force and acceleration signals . In the most unfavourable case (measurement of the dynamic stiffness on a poroelastic pavement with a wearing course of the PSMA8 with PA11 as binder course), the coherence does not fall below the level of 0.96. According to the guidelines of ISO 7626-2 , with thAnalysing the results of dynamic stiffness measurements carried out directly on the tested poroelastic pavements, it was found that the pavement made of the PSMA5 with the PA11 as binder course was characterized by the lowest stiffness. Dynamic stiffness for selected frequencies in the range of 100–1000 Hz is from 137.1 to 139.6 dB re. 1 N/m. Slightly higher stiffness was obtained in the case of the poroelastic pavement PSMA5 with SMA11 as binder course (139.6–141.3 dB re. 1 N/m). Much higher averaged dynamic stiffness was obtained for the poroelastic pavement made of PSMA8 with PA11 as binder course (140.7–143.0 dB re. 1 N/m). The highest dynamic stiffness in the case of poroelastic pavements was obtained for the PSMA8 with SMA11 as binder course (142.1–143.6 dB re. 1 N/m). A pavement with a wearing course of asphalt concrete (AC) on a concrete base course is characterized by a much higher dynamic stiffness than poroelastic pavements and is in the range of 149.9–151.5 dB re. 1 N/m.The data summarised in In the dynamic stiffness tests under laboratory conditions on samples taken from road surfaces, the same trend of changes was found as in the field tests. The lowest dynamic stiffness is characteristic of the PSMA5 with PA11 as binder course (139.9.3–140.5 dB re. 1 N/m). Slightly higher values were obtained for the PSMA5 with SMA11 as binder course (140.0–141.1 dB re. 1 N/m). In the case of PSMA8 with SMA11 and PA11 as binder courses, quite similar levels of average dynamic stiffness were obtained (139.7–142.6 dB re. 1 N/m).Similarly, as in the case of dynamic stiffness tests of the road surface, also in the case of laboratory tests, the highest values were obtained for samples of asphalt concrete (147.1–155.6 dB re. 1 N/m).The values of standard deviations for the results of measurements carried out in laboratory conditions on samples of poroelastic layers did not exceed the value of 1.2 dB re. 1 N/m. For asphalt concrete (AC) samples on a cement concrete base course, the standard deviation was 0.6 dB re. 1 N/m for frequencies of 100 and 300 Hz. The significant scatter at higher frequencies (4.5 dB re. 1 N/m at 500 Hz) requires further analysis.When comparing the averaged dynamic stiffness, it can be stated that there is a good agreement between the values determined in the laboratory tests and the results obtained in situ on road surfaces .There were, however, some differences in the values of dynamic stiffness in the range of certain frequencies. They were caused by different test conditions. In the laboratory, the tests were carried out with the use of a rigid cast iron surface plate, and under in situ conditions, a flexible substructure and subsoil were under the binder course of the pavement.The determined results confirm the lower stiffness of the PSMA5 (with the maximum grain size of 5 mm) compared to the PSMA8 (with the maximum grain size of 8 mm). In the case of the PSMA5, a more favourable effect of the PA11 as binder course on the dynamic stiffness of the pavement was also noted. In both field and laboratory tests, a significantly higher dynamic stiffness of the asphalt concrete pavement was found.Based on the findings, it can be concluded that the dynamic stiffness influences the differences in the levels of tyre/road noise between poroelastic surfaces and asphalt concrete surfaces see . Such a As part of the research, a similar trend was found in the distribution of dynamic stiffness of the tested pavements and layers, both in the case of field measurements and tests in laboratory conditions.The lowest dynamic stiffness is characteristic of the PSMA5 on the porous asphalt PA11 as binder course . Slightly higher stiffness was established for the PSMA5 on the SMA11 as binder course (140.5 and 140.6 dB re. 1 N/m). Even higher values of dynamic stiffness were found for the PSMA8 with PA11 and SMA11 as binder courses (141.6 and 142.0 dB re. 1 N/m and 141.5 and 143.0 dB re. 1 N/m). The highest dynamic stiffness in the group of tested mixtures was determined for asphalt concrete (AC) on a cement concrete as base course (150.1 and 150.3 dB re. 1 N/m).Slight differences in the values of dynamic stiffness in the range of certain frequencies in the in situ and laboratory conditions result from different test conditions. In the laboratory, the tests were carried out with the use of a measuring base (in the form of a rigid cast iron surface plate), and under in situ conditions, a flexible substructure and subsoil were under the binder course of the pavement.At the same time, satisfactory values of standard deviations within individual measurement series were obtained. Slightly larger dispersions of the values of dynamic stiffness in the vicinity of the mean values were noted in the case of in situ tests.The findings concerning the dynamic stiffness of poroelastic and asphalt concrete layers made as part of the study confirm its influence on the level of rolling noise. The constructed test stand enables the testing of mineral-asphalt mixtures (including poroelastic mixtures) in laboratory conditions before deciding to use them on a specific road in real field conditions. However, it does not allow for the assessment of the pavement load capacity due to the assumed range of the applied loads. It is necessary to continue the research with the use of the proposed test stand and tire/road noise measurements to develop the relationship between the stiffness and the level of generated sounds.The conducted measurements and analysis of the results prove the usefulness of the proposed test stand for determining the dynamic stiffness of bituminous mixtures in laboratory and field conditions. This is confirmed by the coherence between the force and acceleration signals at the level of at least 0.96—which indicates a very good validation of the test results with a random error lower than ±5% with 90% confidence level.