The flow of polymer solution and heavy oil in porous media is critical for polymer flooding in heavy oil reservoirs because it significantly determines the polymer enhanced oil recovery (EOR) and polymer flooding efficiency in heavy oil reservoirs. In this paper, physical experiments and numerical simulations were both applied to investigate the flow of partially hydrolyzed polyacrylamide (HPAM) solution and heavy oil, and their effects on polymer flooding in heavy oil reservoirs. First, physical experiments determined the rheology of the polymer solution and heavy oil and their flow in porous media. Then, a new mathematical model was proposed, and an in-house three-dimensional (3D) two-phase polymer flooding simulator was designed considering the non-Newtonian flow. The designed simulator was validated by comparing its results with those obtained from commercial software and typical polymer flooding experiments. The developed simulator was further applied to investigate the non-Newtonian flow in polymer flooding. The experimental results demonstrated that the flow behavior index of the polymer solution is 0.3655, showing a shear thinning; and heavy oil is a type of Bingham fluid that overcomes a threshold pressure gradient (TPG) to flow in porous media. Furthermore, the validation of the designed simulator was confirmed to possess high accuracy and reliability. According to its simulation results, the decreases of 1.66% and 2.49% in oil recovery are caused by the difference between 0.18 and 1 in the polymer solution flow behavior indexes of the pure polymer flooding (PPF) and typical polymer flooding (TPF), respectively. Moreover, for heavy oil, considering a TPG of 20 times greater than its original value, the oil recoveries of PPF and TPF are reduced by 0.01% and 5.77%, respectively. Furthermore, the combined effect of shear thinning and a threshold pressure gradient results in a greater decrease in oil recovery, with 1.74% and 8.35% for PPF and TPF, respectively. Thus, the non-Newtonian flow has a hugely adverse impact on the performance of polymer flooding in heavy oil reservoirs. The worldwide demand for petroleum resources will continue to increase before new alternative energy sources are maturely developed and widely applied . With thInvestigating fluid flow in porous media can provide theoretical guides for improving the performance of polymer flooding and enhanced oil recovery (EOR) in heavy oil reservoirs . Many scIn addition, numerical reservoir simulation, a powerful approach, is a helpful complement to experiments in investigating the fluid flow in porous media and polymer flooding in heavy oil reservoirs. Compared with experiments, numerical reservoir simulation has advantages in efficiently predicting polymer flooding, as well as evaluating and analyzing factors controlling polymer flooding . With thIn this paper, physical experiments were conducted to study the rheology of the polymer solution and heavy oil, and their flow in porous media. The viscosities of a polymer solution with the same polymer concentration and an original heavy oil were measured at different shear rates to recognize their rheology, and the viscosities of polymer solutions with different polymer concentrations were measured to estimate the increasing viscosity capacity of the polymer. The micro-flow method was applied to measure the heavy oil TPGs, and a typical polymer flooding (TPF) experiment was carried out to analyze the heavy oil flow in porous media. In addition, a new mathematical model was proposed, and an in-house 3D two-phase simulator was designed with consideration of heavy oil TPG for polymer flooding in heavy oil reservoirs. The designed simulator was validated by comparing its simulation results with those from the ECLIPSE V2013.1 software  and the TPF experiments. Moreover, the designed simulator was applied to investigate the effects of non-Newtonian flow on production indicators, including a pressure difference, water cut, and oil recovery. These results can assist to improve the performance of polymer flooding in heavy oil reservoirs.The information of the materials including the polymer, brine, original heavy oil, and core samples are provided in One-hundred-and-ninety-nine milliliters of brine, whose ion component concentrations are shown in 1.096 g of polymer, whose properties are given in The stirrer speed of the JJ-1B stirrer was reduced to 100 rpm, and continued to run for 2 h.The JJ-1B stirrer was stopped, and the polymer solution was deoxidized, sealed and statically stored in a brown glass bottle for 12 h.Steps 1–4 were repeated, and 400 mL of polymer solution with a concentration of 5000 mg/L was obtained.Fifty milliliters of polymer solutions with a concentration of 5000 mg/L were diluted with 450, 200,116.67, 50 mL of brine, and 200 mL of polymer solution with a concentration of 5000 mg/L was diluted with 300 mL of brine.After dilution, all polymer solutions were stirred at 100 rpm for 0.5 h using the JJ-1B stirrer.The JJ-1B stirrer was stopped. To simulate the polymer degradation caused by a high shear rate in a near-wellbore region, all polymer solutions were sheared under 16,900 rpm for 35 s using a Waring 7012S blender  to perform pre-shearing.The Waring 7012S blender was stopped, and all polymer solutions were deoxidized, sealed, and statically stored in brown glass bottles for 12 h. Five-hundred, 250, 166.67, 500 and 100 mL of polymer solutions with concentrations of 500, 1000, 1500, 2000, and 2500 mg/L, respectively, were obtained.The polymer solution was prepared at an ambient temperature of 25 °C, and its procedure was as follows:−1 and the tests were performed on the Physica MCR301 advanced rotary rheometer at 25 °C.The rheological testing analyzed the polymer solution with a concentration of 2000 mg/L and heavy oil sample #1. The viscosity of the polymer solution with different concentrations was tested at a shear rate of 7.6 sThe experimental devices were connected according to The brine was used to displace the core sample at a flow rate of 0.05 mL/min until the volume of injected heavy oil sample reached four times the pore volume (PV) of the core sample, which had been saturated with the brine and placed for at least 24 h before the TPG measurement started.The heavy oil sample was used to displace the brine in the core sample at a flow rate of 0.05 mL/min after bypassing the oil column tube #1. The displacing flow rate was increased to 0.5 mL/min when the water cut at the outlet was lower than 2% until no water production. The core sample was saturated with bound brine and a heavy oil sample was obtained.The height of the oil column in the oil column tube #2 was lowered to about 5 cm, and the oil column in tube #1 was put into use. The height of the oil column in oil column tube #1 was raised to the same height as that in oil column tube #2. This condition was kept for 24 h.The heavy oil sample was used again to displace with a flow rate of 0.002 mL/min, and the height of the oil column in the oil column tube #1 increased gradually. The pressure gradient was the TPG of the heavy oil sample when the height of the oil column in oil column tube #2 began to rise.The flow chart in Notably, air was prevented from entering the measurement system to avoid its effect on the accuracy of the TPF measurement.The experimental devices were connected according to the flow chart, and the temperature of the thermotank was also set to 25 °C.The process of completely saturating a core sample with a heavy oil sample and bound brine was similar to that of the TPG measurement.Initial water flooding. After saturating, the brine was used to displace at a constant flow rate of 0.4 mL/min until the volume of the injected brine reached 1 PV.Polymer flooding. The polymer solution with a concentration of 5000 mg/L was used to displace at a constant flow rate of 0.4 mL/min until the volume of the injected brine reached 0.4 PV.Extended water flooding. The brine was used again to displace at a constant flow rate of 0.4 mL/min until the volume of the injected brine reached 2.6 PV.The flow charge of a typical polymer flooding (TPF) experiment is presented in Notably, avoiding air was also required during the TPF experiments.Only the oil and water phases were involved in the polymer flooding process and no mass exchange occurred between them.The mixture of water and polymer was ideal, and was presented only in the water phase.The fluids were compressible, and the rock was not only compressible but anisotropic.The flow was isothermal, and non-Newtonian flow was advised.The injection of polymer only affected the permeability of the water phase and did not affect the permeability of the oil phase.The effects of the capillary force and gravity were taken into account.The assumptions for the mathematical model included:Compared to water flooding, the polymer flooding process is more complicated and is accompanied by complex parameter changes. This is especially true for polymer flooding in heavy oil reservoirs. More parameter changes need to be considered and mainly include the viscosity changes of the water phase, polymer adsorption, the permeability reduction of the water phase, inaccessible pore volumes, and the TPG of heavy oil.3)−1; 3; 3; −1, 2. The viscosity of the water phase changes with the injection of the polymer solution and is affected by the mixing degree of the water and the injected polymer solution, polymer concentration, and a shear rate. First, the Todd-Longstaff mixing parameter was introduced to describe the mixing degree of the water and the injected polymer solution ,40. In tessed as ,42(2)μpsolution .(3)γ˙eqDuring the flow of polymer solution through the rock, some polymer molecules will inevitably be adsorbed on the inner surfaces of pores in the rock . Here, tThe adsorbed polymer molecules will cause a permeability reduction of the water phase . Here, twhich is :(5)Rk=13 and 3.Polymer cannot enter some small pores when the polymer solution flows through the rock, which results in an inaccessible pore volume . Here, tHeavy oil needs to overcome a TPG to flow , and the2, 3, 2, and The flow of the water phase was in accordance with Darcy’s law, and its flow equation was :(7)νw→=3, and Because the heavy oil flow through the rock no longer follows Darcy’s law, the oil phase flow equation had to be corrected with consideration of the TPG of heavy oil :(8)νo→=for water:for polymer:for oil:where 3/m3, respectively. 3/(s·m3), respectively. The source term is negative, and the sink term is positive. 3.According to the conservation of mass, the continuity equations of all components in ground standard conditions can be obtained by combing the flow equations:Although the flow and continuity equations described the basic flow characteristics, the interrelationship between some physical quantities in the equations needed to be additionally described by the auxiliary equation and the equations of state.The auxiliary equations included:The equations of fluids, rock and rock-fluids included:The above equations provided a general description of polymer flooding in heavy oil reservoirs and might have multiple solutions. Therefore, definite conditions were given to obtain the unique solution for this specific study. The definite conditions included the initial and boundary conditions.3.The initial conditions included the distribution of initial pressure, saturation and polymer concentration:The boundary conditions included the outer and inner boundaries. The outer boundary was a closed boundary with no-flow, which was:for water:for polymer:for oil:where x direction between the grid blocks 2·m, in which x, y and z directions in m, respectively; and x direction. −1.3/s, respectively. Production is negative, and injection is positive. Although the definite conditions were given, it was very difficult to solve the above partial differential equations by analytical methods , especiaEquations (26)–(28) can form a system of nonlinear equations. To solve it and ensure stability during the calculation process, the full implicit method was applied. For the solution of each specific time step, the Newton-Raphson method, an iterative method, was applied. More details can be seen in Chen’s research ,51. FinaR2) reaches 0.9881. Their relationship expression can be written as:−1, respectively. The flow behavior index of the polymer solution is 0.3655, less than 1, showing a typical shear thinning. From the plot of the original heavy oil, the relationship between the shear stress and shear rate is linear, but the line with the R2 of up to 0.9998 does not pass through the origin of the coordinate axes. Their relationship expression can be given as:−1, respectively. It illustrates that the heavy oil has a typical Bingham fluid rheological feature [The rheological charts of the polymer solution with a concentration of 2000 mg/L and the original heavy oil are presented in  feature ,53.R2 of up to 0.996 can be expressed as follows:−1 is 29.57 mPa·s, which is about one-sixth that at 5.6 s−1. The main reason for the shear thinning of the polymer solution is that the entanglements between the polymer molecules are destroyed at a higher shear rate, resulting in a decrease in hydrodynamic radius and a reduction in the viscosity of the polymer solution [The viscosities of the polymer solution with a concentration of 2000 mg/L and the original heavy oil at different shear rates are indicated in solution ,55. The solution . This exR2 of up to 0.9943 is:Different from the relationship between polymer solution viscosity and shear rate, the viscosity of the polymer solution significantly increases with an increase in polymer concentration. The viscosity of the polymer solution with a concentration of 2500 mg/L is 278 mPa·s, which is about 34 times greater than that with a concentration of 500 mg/L. The main reason for this finding is that the longer molecular chains and more entanglements are in the polymer solution with a higher polymer concentration, resulting in a larger hydrodynamic radius, which increases the viscosity of the polymer solution .R2 of 0.9688 is:The measured TPG of heavy oil is given in It can be also seen from The simulation results of Case 1 run by the ECLIPSE V2013.1 software were used to compare Case 1 run by our designed simulator to validate the designed simulator without consideration of TPG because it is a recognized commercial numerical reservoir simulator, and its simulation results are authoritative . The maiDifferent from the validation without considering TPG, the validation with considering TPG cannot be conducted by comparing the simulation results of the designed simulator with those of the ECLIPSE V2013.1 software because no widely accepted commercial numerical reservoir simulation software including ECLIPSE V2013.1 software considers TPG ,35. To vThe flow behavior index characterizes the extent to which the fluid deviates from Newtonian fluid  and can The comparison results of pressure difference, water cut, LPL flow diversion ratio, and oil recovery of Cases 6–9, the production indicator reductions of Cases 7–9 vs. Case 6, and 3D remaining oil saturation distributions after initial water flooding, polymer flooding and extended water flooding of Cases 6 and 9 are illustrated in In summary, the polymer solutions shear thinning has a disadvantageous impact on the polymer flooding including the PPF and TPF in the heavy oil reservoirs, and the effect of polymer solution shear thinning on the PPF is less than that on the TPF after comparing the simulation results of these specific cases.In order to investigate the effect of heavy oil TPG on the four main production indicators—pressure difference, water cut, LPL flow diversion ratio and oil recovery of polymer flooding including both the PPF and TPF, Cases 10–15 were carried out, where the parameters of Cases 10–12 were the same as Case 3 except that their heavy oil TPGs were 5, 10 and 20 times that of Case 3, respectively. The parameters of Cases 13–15 were the same as Case 6 except that their heavy oil TPGs were 5, 10 and 20 times that of Case 6, respectively.The comparison results of pressure difference, water cut, LPL flow diversion ratio and oil recovery of Cases 6 and 13–15; the production indicator reductions of Cases 13–15 vs. Case 6, and 3D remaining oil saturation distributions after initial water flooding, polymer flooding and extended water flooding of Cases 6 and 13–15 are demonstrated in In summary, the heavy oil TPG has a detrimental impact on the polymer flooding including the PPF and TPF in the heavy oil reservoirs, and the effect of heavy oil TPG on the PPF is much less than that on the TPF based on comparisons of the simulation results of these specific cases. Additionally, the effect of heavy oil TPG on the PPF is also less than that of polymer solution shear thinning on the PPF, but the effect of heavy oil TPG on the TPF is more than that of polymer solution shear thinning on the TPF as noted in the comparison of simulation results in this paper.Cases 16 and 17 were conducted to analyze the combined effect of TPG and shear thinning on the four main production indicators including pressure difference, water cut, LPL flow diversion ratio and oil recovery of polymer flooding including both the PPF and TPF, where the parameters of Cases 16 were the same as Case 3 except for the polymer solution flow behavior indexes of 0.18 and the fact that its heavy oil TPG was 20 times that of Case 3, and the parameters of Cases 17 were the same as Case 6 with the exception of the polymer solution flow behavior indexes of 0.18 with a heavy oil TPG 20 times greater than that of Case 3.The comparison results of pressure difference, water cut, LPL flow diversion ratio and oil recovery of Cases 6 and 17, the production indicator reductions of Case 17 vs. Case 6, and 3D remaining oil saturation distributions after initial water flooding, and polymer flooding and extended water flooding of Cases 6 and 17 are presented in In summary, the combined effects of both polymer solution shear thinning and heavy oil are adverse on the polymer flooding, including the PPF and TPF in the heavy oil reservoirs, and the combined effect is greater than the single effect by comparing the simulation results of these specific cases.R2 of 0.9881; its flow behavior index is 0.3655, and its viscosity decreases as shear rate increases, showing a typical shear thinning performance. The shear stress and shear rate of the original heavy oil is linear with the R2 of up to 0.9998, however, the fitting line does not pass through the origin of the coordinate axes; the viscosity of the original heavy oil is infinite at the low shear rate and remains at an approximate but steady 202.7 mPa·s as the shear rate increases, presenting Bingham fluid characteristics. The heavy oil needs to overcome the TPG to flow, and its TPG increases with a reduction in rock permeability and an increase in heavy oil viscosity. Moreover, the heavy oil TPG and the mobility have an excellent power-law relation as expressed by the R2 of 0.9688, and the heavy oil TPG increases with a decrease in mobility. Furthermore, the new 3D two-phase polymer flooding simulator considering the non-Newtonian flow was designed and validated with high accuracy and reliability by comparing its simulation results with those performed by the commercial software, as well as with the results obtained from the TPF experiments. Additionally, the effect of non-Newtonian flow on the production indicators was studied using the deigned simulator. According to the simulation results, the oil recoveries of the PPF and TPF, with the polymer solution flow behavior indexes of 0.18, are 1.66% and 2.49% lower than those of the PPF and TPF, with the polymer solution flow behavior index of 1, respectively, demonstrating that the polymer solution shear thinning has a disadvantageous effect on the polymer flooding in the heavy oil reservoirs, and its effect on the PPF was less than that on the TPF. For the heavy oil, considering a TPG of 20 times greater than its original value, the oil recoveries of PPF and TPF are reduced by 0.01% and 5.77%, respectively, illustrating that the heavy oil TPG has a negative effect on the polymer flooding in the heavy oil reservoirs, and its effect on the PPF is slight, less than that on the TPF. However, a better polymer EOR after the polymer flooding of TPF with the greater heavy oil TPG is obtained, strongly proving the importance of polymer flooding for heavy oil reservoirs. In addition, the effect of heavy oil TPG on the PPF is also less than that of polymer solution shear thinning on the PPF, while the effect of heavy oil TPG on the TPF is more than that of polymer solution shear thinning on the TPF. The oil recoveries of the PPF and TPF, with the polymer solution flow behavior indexes of 0.18 and the heavy oil TPGs 20 times the original heavy oil TPG, are 1.74% and 8.35% lower than those of the PPF and TPF, without shear thinning and heavy oil TPF, respectively. To minimize the effects of non-Newtonian flow, including polymer solution shear thinning and heavy oil TPG, on polymer flooding in heavy oil reservoirs, some methods including applying shear resistant polymer and heavy oil TPG reduction methods have been proposed [In this study, physical experiments and numerical simulations were combined with the aim of investigating the effect of non-Newtonian flow on polymer flooding in heavy oil reservoirs and giving theoretical and technical guidance. The physical experiment results showed that the flow of both the polymer solution with a concentration of 2000 mg/L and original heavy oil exhibit the non-Newtonian flow characteristics. The shear stress and shear rate of the polymer solution with a concentration of 2000 mg/L indicates a good power law relationship with the proposed ,62. In tproposed ,64 based


Assessments of Indoor Environment Quality (IEQ) present a very significant challenge when analyses are undertaken mainly in buildings that include a particularly sensitive and vulnerable population, such as elderly people. In order to maintain an indoor environment that is adequate for occupants, it is necessary to comply with a set of requirements  regarding concentrations of airborne pollutants and hygrothermal comfort conditions. This paper studies IEQ in compartments in 3 buildings in two cities in central Portugal, Viseu and Covilhã, which hold elderly care centers. The following environmental parameters were continuously recorded: air temperature, relative humidity, concentration of carbon dioxide, formaldehyde, and total volatile organic compounds and ventilation rates. An analysis of the obtained results was performed, taking recommended guidelines and threshold values into account, thus making it possible to evaluate the IEQ conditions and hygrothermal comfort in the selected indoor spaces. On the basis of the conclusions reached and the observed problems of hygrothermal comfort and indoor pollutants in the indoor spaces, a number of recommendations are proposed, specifically in terms of climate control, ventilation, and maintenance, in order to obtain an overall improvement of IEQ. In developed countries, people spend more than 90% of their time indoors. Indoor environmental conditions are strongly related to health, well-being, and overall performance . With peAlthough IEQ plays an important role in all types of buildings, particular consideration should be paid to Elderly Care Centers (ECC). The perception of the elderly as a risk group with regard to inadequate indoor environments, due mainly to the presence of pollutants in the indoor air, is essentially due to the fact that the elderly population’s immune defenses are reduced, and many of them suffer from multiple chronic diseases . Every bWith regard to the age pyramid in Portugal, approximately 19% of the population is aged 65 or over. The Aging Index, i.e., the relationship between the number of people over 65 years of age and the number under the age of 14, has shown an increasing trend in recent decades, mainly due to the decrease in the birth rate ,8. In shStudies in ECC are rare, perhaps because the premise that in these places, problems associated with the Indoor Air Quality (IAQ) are less important, due to the relatively low occupancy density. The most common studies in this field are related to comfort analyses . HoweverMore research is needed to better describe the IAQ where the elderly people are passing most of their time. This research is intended to contribute to that effort by providing information on elderly care premises in Portugal.Tint e RH), with minimum values of air change rates (ACH) and with maximum values of concentrations of indoor air pollutants. Portuguese regulations provide parameters for thermal considerations in buildings and IEQ, with the exception of RH, for which European standardization provides recommendations.In order to maintain an indoor environment with acceptable air quality and levels of thermal comfort, it is necessary to comply with the reference levels for temperature and relative humidity , formaldehyde (CH2O), and total volatile organic compounds (TVOC)), as well as hygrothermal comfort conditions  in three ECC in the cities of Viseu and Covilhã, Portugal; see In order to contribute to the deepening the knowledge on this subject, this article presents the results of measurements of indoor pollutants  ECC 1: 7 rooms and 1 living room; (2) ECC 2: 5 rooms and 3 living room.A physical characterization of the two buildings and some of the compartments studied is presented in 2/resident. In the case of ECC 2, this ratio is grossly disregarded.According to the regulations in Portugal , the minInformation about the interior cladding of the studied compartments, windows, and corresponding indoor and outdoor solar protection is presented in Portuguese thermal regulations  indicateA physical characterization of the studied building (ECC 3) and compartments is presented in Information about the interior cladding of the studied compartments, windows, and corresponding indoor and outdoor solar protection is presented in The protective thresholds for the considered physico-chemical pollutants are set out in 2 as a tracer gas (in post-occupancy periods), as described in ASTM E741  in the living rooms. In view of the available equipment, only one point of analysis of the various parameters in each compartment was considered .Two trials were conducted: the first in winter, i.e., between December 2015 and January 2016, and the second in spring, i.e., between March and April 2016.Outdoor temperature measurements were obtained from a meteorological station at the University of Beira Interior (UBI), located at approximately 680 m altitude. Regarding the location chosen to place instruments inside the compartments and to assess the ACH, the criteria already presented were followed.The measurements of the pollutants were performed continuously; each reading was measured over a minimum period of 5 min . Due to Ni is the number of measurement points in zone i and Ai is the area of zone i (m2).The minimum number of analysis points for the various indoor air quality parameters to be measured was calculated by applying the following expression to the total area of the compartment, rounding up to the unit :(1)Ni = µ) refers to the arithmetic mean.In all presented results (cities of Viseu and Covilhã), the calculated mean  Threshold of CO2 protection exceeded (1625 ppm for a mean of 8 h); (3) Threshold of TVOC protection exceeded (0.52 ppm for a mean of 8 h); (4) Threshold of CH2O protection exceeded (0.08 ppm for a mean of 8 h).In the analysis of the results, according to Tint and RHint, this “excessive” percentage is considered to be reasonable in assessments of indoor environments, since existing regulations are relatively recent, and the buildings have envelope and HVAC systems that are mostly unprepared for the new, very high standards. Moreover, if the percentage defined is too small , implementing climate control in these spaces would become quite expensive.As for In the compartments of ECC 1, in both trials, approximately constant temperatures and relative humidity were recorded; they were within the recommended values . This may be because the building in question is recent, with improved systems relative to the ECC 2. An example of this is the existence of double-glazed windows with aluminum frames, without thermal brakes, in all of the compartments, as well as the existence of thermal insulation in the envelope of the building.2 and TVOC), recorded in the compartments of ECC 1 during the second trial.2 which is above the protection threshold. From 2 remained below the regulatory limits. The high values may therefore be due to the high density of occupancy of the living room, reduced ACH, or even to possible combustion processes, since the kitchen and the dining hall are located in the vicinity of the living room; (2) TVOC concentrations were not of concern in any of the analyzed compartments. The standard deviation values of the TVOC of the rooms were high, and we may conclude that the most influential parameters (type of activities and ACH) vary quite a bit during occupancy.According to the results presented in RH values; (4) The rooms had excessive percentages of time where the RH values registered were higher than the recommended maximum limit.The thermal comfort of rooms 1 and 2 and living rooms 1 and 2 of ECC 2 was assessed using the adaptive model proposed by LNEC , as showTint and Tmp .Considering the use of active air conditioning , the results revealed the existence of a thermally comfortable environment in practically all compartments, with the temperature data within the established “comfortable” guideline values.2 and TVOC) recorded in the compartments of ECC 2 during the first trial showed that: (1) Room 1 and living room 2 had high mean CO2 values. Nevertheless, none of the compartments analyzed presented mean TVOC values of concern; (2) With the exception of living room 1, all the compartments analyzed had a maximum mean value of CO2 which was above the protection threshold.The statistical analysis of the indoor pollutants .With respect to the values presented, and taking the measurement period into account (≈ 5 min), we may conclude that the triple room presented elevated levels of formaldehyde. No potential source was identified; therefore, we speculate that this may be due to the wall coverings or other materials used. These values require further investigation.The hygrothermal comfort conditions are shown in It should be noted that in the ECC 3, the measurement in the living room was not performed simultaneously with the other two compartments, which is why this compartment is not shown in An analysis of nd trial took place in the spring of 2015, continuously recording concentrations of CO2 and TVOC only at one point, with a 2-min time interval and a “Maximum measurement period” between 1 and 8 days. The 22 and TVOC. The initial points of the CO2 decay method are shown; they were used to determine the ACH through the decay technique. Points with maximum levels of TVOC are also shown.2 and TVOC occurred predominantly between 10:00 and 11:00 a.m., followed by a marked decay of CO2. This conjugation of behavior may be the result of cleaning actions, followed by opening windows.From 2 in the living room, ECC 1 has an IEQ which is within the regulatory threshold values and standards; (2) In ECC 2, the rooms have high percentages of time with temperatures below regulation standards, and reveal worrying RH values ; (3) In ECC 2, the rooms have excessive periods with temperatures below regulation, and reveal RH values that raise some concerns ; (4) With the exception of living room 1, all rooms in ECC 2 had maximum mean values of CO2 which were above the protection threshold; (5) In assessing the thermal comfort by applying the adaptive comfort model proposed by LNEC and regarding ECC 2, the rooms are comfortable overall; (6) The ACH were found to be insufficient, with values generally below the required levels for all of the analyzed compartments.From the results obtained in both ECC in Viseu, the following conclusions may be drawn: (1) With the exception of the concentration of COFrom the results obtained in the ECC in Covilhã, the following conclusions stand out: (1) The RH values in the compartments do not raise concern; (2) The triple room has an excessive percentage of time with below regulation temperatures (perhaps due to opening windows during cleaning actions); (3) The living room has very low ACH values; (4) The triple room has high levels of formaldehyde.The previous conclusions imply the need for action concerning the heating, maintenance, and ventilation conditions, so that the spaces operate within the appropriate conditions of comfort and air quality.


Understanding the area-specific resource use of inpatient psychiatric care is essential for the efficient use of the public assistance system. This study aimed to assess the geographical variation in psychiatric admissions and to identify the prefecture-level determinants of psychiatric admissions among recipients of public assistance in Japan.We identified all recipients of public assistance who were hospitalized in a psychiatric ward in May 2014, 2015, or 2016 using the Fact-finding Survey on Medical Assistance. The age- and sex-standardized number of psychiatric admissions was calculated for each of the 47 prefectures, using direct and indirect standardization methods.R2 = 28% and R2 = 23%, respectively). The sensitivity analyses, using medical cost as the outcome and data from different survey years and subgroups, showed similar findings.A total of 46,559 psychiatric inpatients were identified in May 2016. The number of psychiatric admissions per 100,000 population was 36.6. We found a 7.1-fold difference between the prefectures with the highest (Nagasaki) and lowest (Nagano) numbers of admissions. The method of decomposing explained variance in the multiple regression model showed that the number of psychiatric beds per 100,000 population and the number of recipients of public assistance per 1,000 population were the most important determinants of the number of psychiatric admissions (We identified a large geographical variation in the number and total medical cost of psychiatric admissions among recipients of public assistance. Our findings should encourage policy makers to assess the rationale for this variation and consider strategies for reducing it. The inpatient cost for mental disorders accounted for 15% of the total medical cost in recipients of medical assistance, whereas the proportion was much smaller for the entire population (4%).5,6 The national government suggests that approximately 20% of patients hospitalized at psychiatric wards should be discharged from the hospitals when the conditions  for community-based care are given.6 To reduce potentially avoidable long-term stays in psychiatric wards, local governments have implemented various programs for promoting the discharge from the hospital to the community among recipients with mental disorders.6 Understanding the area-specific resource use for inpatient psychiatric care is essential for the reduction of potentially avoidable long-term stays and can thus contribute to the appropriate use of the public assistance system through enhancement of independence rather than institutionalization. Local governments will assess the rationale for the number of psychiatric admissions in their own prefectures in comparison to that of Japan as a whole. Moreover, the national government will need to judge whether a geographical variation in the number of psychiatric admissions appears to be justified based on the aims of the public assistance system.In 2015, there were approximately 2 million recipients of public assistance in Japan,To date, there is no information on the area-specific number of psychiatric admissions among recipients of public assistance. We aimed at assessing the geographical variation in the number of psychiatric admissions and at identifying the prefecture-level determinants of psychiatric admissions among recipients of public assistance in Japan.5 The FSMA is conducted every year by the Ministry of Health, Labour and Welfare in Japan. It collects all electronically issued medical claims reviewed every June  among all recipients of medical assistance. Since 2014, data of the FSMA include parameters, such as prefectural identification numbers based on the location of the residence, patient identification numbers, diagnostic codes, procedural codes, length of the hospital stay, and total medical cost per month. We obtained anonymous patient-level data from the FSMA conducted in 2014, 2015, and 2016.We conducted an ecological study using data from the Fact-finding Survey on Medical Assistance (FSMA). A detailed description of the FSMA has been reported elsewhere.3The Japanese local government system is composed of 47 prefectural governments. There is a large heterogeneity in the prefecture-level potential determinants of resource use for inpatient psychiatric care . For exaAll claims of patients hospitalized at a psychiatric ward for at least 1 day during May of the study year  were identified. To avoid duplicates, we included only claims for psychiatric care provided in May. For patients who were hospitalized at more than two hospitals in May, we included one claim, issued from the hospital with the longer stay.7 Aggregated data from the Population Census in 2015 were used as the standard population.4 We selected the unit of population as 100,000 population for the direct standardization method. In the analyses of the number of psychiatric admissions, we used the standardized claim ratio (SCR), which is analogous to the standardized mortality ratio and enables the comparison of the number of psychiatric admissions in a specific prefecture to that in Japan as a whole (reference standard).8,9 The empirical Bayes estimates for the SCRs were obtained using the Poisson-Gamma model along with the moment estimator10; the 95% equal-tail credible intervals (CrIs) for the SCRs were obtained using posterior distribution.11 In the analyses of the total medical cost, we used the multiplicative effect (ME), which also enables the comparison of the total medical costs in a specific prefecture to that in Japan as a whole (reference standard). We obtained the estimates for the MEs using the Poisson model along with the natural logarithm of the number of the population as an offset term; the 95% confidence intervals (CIs) for the MEs were obtained using the robust standard error.12In the main analysis, we used data from the FSMA, 2016. First, we calculated the age- and sex-standardized number and total medical cost of psychiatric admissions for each of the 47 prefectures, using direct and indirect standardization methods.13–15: (1) the number of recipients of public assistance per 1,000 population in 2015,3,4 (2) the number of treated patients with mental disorders per 1,000 population in 2014,16 (3) the number of visits for psychiatric nursing care at home per 100,000 population in 2014,4,17 (4) the number of psychiatric beds per 100,000 population in 2016,18 (5) the average length of stay in psychiatric beds in 2016,18 (6) the number of beds in institutions covered by long-term care insurance per 1,000 population aged ≥65 years in 2015,4,19 (7) the percentage of depopulated municipalities in 2014,20 (8) the percentage of population living alone in 2015,21 and (9) unemployment rate.20,22 Multicollinearity was assessed by examining Pearson correlation coefficients between all pairs of the independent variables, tolerance, and variance inflation factor (VIF) values. As a rule of thumb, tolerance values <0.1 or VIF statistics >10 indicates that potentially harmful multicollinearity is present.23 We simultaneously entered all independent variables into the models and obtained the Lindeman-Merenda-Gold metrics to quantify the amount of explained variance  attributable to each independent variable (R2) in the multiple regression model.24 The sum of R2 for all independent variables is equal to the overall R2 in the model.Second, we investigated the associations between the age- and sex-standardized direct estimates and a set of prefecture-level potential determinants using multiple regression models. We selected the following ecological factors as potential determinants based on a priori clinical knowledge, previous studies, and availability of data 13–15: .We also conducted three sets of additional analyses. First, we repeated the main analyses using data from the 2014 and 2015 FSMA to investigate the robustness of the study findings.25Second, we conducted the following subgroup analyses for the number of psychiatric admissions to assess the effect of the type of psychiatric admission: (1) patients experiencing a new long-stay (length of stay between ≥1 year and <5 years), (2) those experiencing an old long-stay (length of stay ≥5 years), and (3) those having a primary diagnosis of dementia (ICD-10 codes F00–F03 and G30–G31). Old long-stay patients were defined as those who had been admitted to a psychiatric bed before deinstitutionalization and remained hospitalized, while new long-stay patients were defined as those who began prolonged hospitalization despite efforts to prevent it.26,27 The 630 survey collects the number of “all” psychiatric admissions by prefecture, sex, age group, diagnostic group, and length of stay on June 30; however, this survey does not collect information on the status of public assistance. In this analysis, we calculated the age- and sex-standardized number of psychiatric admissions per 100,000 population using the direct method. The outcomes included the standardized number of (1) psychiatric admissions, (2) psychiatric admissions for a new long-stay, (3) psychiatric admissions for an old long-stay, and (4) dementia-related psychiatric admissions. Then, we computed Pearson correlation coefficients.Third, we compared the number of psychiatric admissions by prefecture between the FSMA (2016) and the 630 survey (2015) to examine the generalizability of the study results.28 Institutional review board approval was not required according to the guideline of the local ethics committee.The Statistics Act regulates primary data collection and the secondary use of data. Because all patient-level data were certified to be anonymous, our study design was not subject to the ethical guideline for medical research in Japan.A total of 46,559 psychiatric inpatients  were included in the main analyses Figure . The medThe number of psychiatric admissions per 100,000 population in 2016 was 36.6. There was a 7.1-fold difference between the prefectures with the highest (Nagasaki) and lowest (Nagano) numbers of admissions  and lowest (Fukui) costs  and lowest (Gifu) numbers of admissions for a new long-stay ; this was followed by the number of recipients of public assistance per 1,000 population (R2 = 17%) and the average length of stay in psychiatric beds ; this was followed by unemployment rate (R2 = 20%) and the number of psychiatric beds per 100,000 population , number of psychiatric admissions for a new long-stay (r = 0.74), number of psychiatric admissions for an old long-stay (r = 0.80), and number of dementia-related psychiatric admissions (r = 0.68).We observed strong positive correlations for all outcomes between the FSMA and the 630 survey Figure : the num29–32 For example, Watts et al found a 6-fold variation in the psychiatric admission rate by hospital service area in Northern New England.29 Fortney et al also found a substantial geographic variation in the county-level hospitalization rate for schizophrenia in 14 states of the United States.30 Our study is unique as it focused on recipients of public assistance in all of Japan.We found an over 7-fold geographic variation in the number and total medical cost of psychiatric admissions among recipients of public assistance between prefectures. Similar to our results, some previous studies in smaller areas found significant geographical variations in the numbers of psychiatric admission.3 Namely, the number of recipients of public assistance might be a proxy measure for the number of recipients with mental illnesses. Consequently, prefectures with a higher number of recipients of public assistance per population may have a greater demand for psychiatric admissions among recipients of public assistance. Another reason for this association is that case workers in prefectures with a higher caseload are less likely to be motivated to promote the patients’ discharge from hospitals to the community than those in prefectures with a lower caseload. This is because, in general, caseloads are lower for recipients receiving long-term inpatient care than those receiving ambulatory care.We observed that psychiatric admissions occurred more frequently in prefectures where the number of recipients of public assistance per population is higher. The simple reason for this association is that such prefectures are likely to have a greater demand for psychiatric inpatient care. Of note, in Japan, recipients of public assistance commonly suffer from mental illnesses .33 During the era of growing number of psychiatric beds, public assistance was a common expenditure programs for psychiatric inpatient care. The percentage of psychiatric admissions covered by public assistance relative to all psychiatric admissions ranged from 32% to 50% between 1960 and 1985.33,34 It can be assumed that most old long-stay patients in our study were hospitalized from this era, which was partially supported by the fact that over half of old-stay patients were aged ≥65 years.We found a significant association between the number of psychiatric admissions among recipients of public assistance and the number of psychiatric beds per population on the prefectural level, after controlling for several ecological factors. A similar association was observed for the number of psychiatric admissions not only for an old but also for a new long-stay; however, this association was weaker for dementia-related psychiatric admissions. The reason for the association among old long-stay patients is interpreted as a reflection of the “historical” phenomenon. Historically, the number of psychiatric beds in Japan steeply increased from 95,067 in 1960 to 247,265 in 1970, and then steadily increased to 333,570 by 1985.29,30 which is considered likely to occur in psychiatric admissions.29,30,35 Demand of inpatient psychiatric care might be induced until beds are full, irrespective of necessity. This is because physicians are in a strong position in which they can define the patients’ needs for the initiation and termination of inpatient psychiatric care. Given strong concerns by citizens and policy makers regarding the potential overuse of medical assistance, our findings should be of interest to local governments, particularly to those with high psychiatric admission rates for a new long-stay.On the contrary, the reason for the association among new long-stay patients may be interpreted as a reflection of the “supplier-induced demand” phenomenon,We identified a strong positive correlation in psychiatric admission rates between the FSMA and the 630 survey, suggesting that our results of the prefecture-level determinants of psychiatric admissions may be generalizable to all psychiatric inpatients. Namely, the extent and mechanism of the geographical variations in psychiatric admissions may not be unique to recipients of public assistance.Our findings have several limitations. First, our results were inherently vulnerable to confounding and ecological biases due to the nature of the study design. Second, we could not determine whether the admission was involuntary or not, because all claims data from admission to discharge are needed to identify the admission status. Involuntary admissions may be more associated with psychiatric bed availability than voluntary admissions. This is because the most common type of involuntary admission called the “hospitalization for medical care and protection” strongly considers the opinions of the patients’ family regarding admission and discharge, and they often prefer hospitalization for longer period. Third, we could not identify the exact length of hospitalization due to the nature of the data structure. Fourth, we could not incorporate ambulatory psychiatric services into our analyses because the medical costs for ambulatory services in recipients of public assistance who have chronic psychiatric illnesses are likely supported by another healthcare system  rather than medical assistance.We found a large geographical variation in the number and total medical cost of psychiatric admissions among recipients of public assistance. The number of recipients of public assistance per population and psychiatric bed availability may explain this variation. Our findings should encourage policy makers to assess the rationale for and consider strategies for reducing this variation.


In vitro anticancer activities showed compound 11 as the most promising inhibitor, and the computational docking further indicates its strong inhibitory activities towards some cancer-related receptors. Among the twenty-one modelled ligands, pyrazole-based compounds 7, 11, and 15 are the most promising inhibitors against the selected receptors followed by 18 and 21 which are derivatives of pyridine and phenanthroline, respectively. The presence of the carboxylic unit in the top five ligands that displayed stronger inhibitory activities against the selected receptors is an indication that the formation of noncovalent interactions such as hydrogen bonding and a strong electron-withdrawing group in these compounds are very important for their receptor interactions. The thermodynamic properties, the polarizabilities, and the LUMO energy of the compounds are in the same patterns as the observed inhibitory activities.The anticancer study of nitrogen-chelating ligands can be of tremendous help in choosing ligands for the anticancer metal complexes design especially with ruthenium(II). The inhibitory anticancer activities of some nitrogen-chelating ligands containing bis-pyrazole, bipyridine, and phenanthroline were studied using experimental screening against cancer cell and theoretical docking methods.  Othein rings –23. HoweThe selected receptors used for the docking studies are carbonic anhydrase II (CA-II), cathepsins B (Cat B) , two difThe synthesis and careful structural elucidation of these ligands have been reported in our previous works –45.in vitro anticancer activities of the ligands against the cancer cell line HT29 and normal cell line KMST were examined using the MTT colorimetric assay. All the ligands before their docking to the receptors were first optimized with DFT functional PBEPBE [The l PBEPBE  and the l PBEPBE . The docl PBEPBE  and Vinal PBEPBE  packagesx-, y-, and z-axes was set to 60 × 60 × 60 with each point separated by 0.375 Å. The Lamarckian genetic algorithm was chosen based on its efficiency and reliability in comparison with others like simulated annealing (SA) and generic genetic algorithm (GA) methods in AutoDock [The default parameters were used in the Vina docking package but with little modifications in Molegro and AutoDock dockings. The scoring function used in Molegro was MolDock because it takes care of the hydrogen bonding, intermolecular protein ligand, and intramolecular ligand interactions and has been successfully applying for molecular docking . The maxAutoDock , 52. TheAll the graphical representations of the docking results are prepared using the package Chimera .in vitro anticancer activities, and their results are presented in in vitro activities of these ligands were tested against the cancer cell line HT29 and the normal cell line KMST using the MTT colorimetric assay. The results clearly showed that the six ligands pose no threat to the normal cell line as their inhibitory activities (IC50) in KMST are found to be greater than 50 μm as shown in 11 and 14 are the most potent (<6.25 mm) against the cancer cell line HT29, while ligand 12 is the least active. Also, the same ligand 11 was found among the three best inhibitors of the selected receptors from the docking results , and three models of phenanthroline (phn) . Six of  results .11 and 15 have similar binding orientation in CA-II, Cat B, and HDAC7 based on the results obtained from the binding site interactions. Compound 11 is of interest because it shows promising anticancer activities in vitro, and ligand 15 is predicted as the best inhibitor of many of the receptors according to the results obtained from Molegro-QC and Molegro. The interaction of 11 with CA-II shows that all the docking methods gave similar orientation for the compounds except Molegro-QC (grey) that is slightly different in orientation (The results obtained from the different docking methods and packages: Molegro-QC (grey), Molegro (green), Vina (brown), AutoDock-QC (yellow), and AutoDock (cyan), are shown in entation .15 with CA-II, all of the methods locate the same binding site and give similar orientation of the ligand except Vina (brown) which locates a different binding site compared to others. The Molegro-QC and Molegro orientations of 15 are superimposed and likewise AutoDock and AutoDock-QC. All the five methods except Vina also locate the same binding site for the interaction of 11 with Cat B, and AutoDock prediction was found to be superimposed with that of Molegro-QC and Molegro. The results obtained from AutoDock predict different binding sites for the interaction of 15 with Cat B, while all other methods predict the same site for its binding. Another receptor of interest is HDAC7 in which strong inhibitory activities are displayed by 11 and 15. HDAC7 was predicted as one of the most targeted receptors for many of the ligands according to the results obtained from Molegro-QC and Molegro. Vina locates a completely different binding site for the interaction of 11 with HDAC7 when compared to the rest of the methods. The binding site orientation of 11 from all the methods besides Vina is very similar to that of Molegro-QC completely superimposed with that of AutoDock. All methods predicted a similar binding site orientation of the ligand 15, of which the Molegro-QC and Molegro orientations are found to be superimposed and in close orientation with those of Vina and AutoDock-QC, which are different from those of AutoDock. There is a significant difference between the results from AutoDock-QC and AutoDock methods in terms of the features of the ligand interactions with receptors just because of changes in the accuracy of the atomic charges. The correlation of the results of AutoDock-QC and AutoDock ranges from −0.32 to 0.46, while that of Molegro-QC and Molegro ranges from 0.96 to 1.00. It is obvious that the accuracy of atomic charges plays a very strong role in the determination of the ligand interaction with the receptors especially when using AutoDock. The results show that the reason for the inconsistency from Vina, AutoDock-DC, and AutoDock could be ascribed to different binding sites which were predicted for many of the ligand interactions with the receptors, while high consistency and similarity were obtained in Molegro-QC and Molegro because they both predicted the same binding site as cocrystallized inhibitors of the receptors. In addition, the differences in the ligand-receptor inhibitory energies obtained from AutoDock methods for many of the ligands are within the standard error margin of ∼2.177 kcal/mol [The orientation obtained from Molegro-QC (grey) gives the best interacting energy compared to others . Also inkcal/mol , 44 that7, 11, and 15 have the best inhibitory activities towards many of the receptors according to the results obtained from Molegro-QC and Molegro , Molegro-QC is then used for the analysis of the receptor interaction of ligands 7, 11, and 15. Although in many of the ligand interactions with the receptors, AutoDock and Vina gave similar features in their interaction, the ranking of the ligands according to their inhibitory interaction is vague. It has been pointed out that, to have a good ranking in AutoDock, the error values of ∼2.177 kcal/mol [The ligands  Molegro . Besideskcal/mol , 44 have7, 11, and 15 , 11 (green), and 15 (yellow) are found to be clustering around the same point and sharing common angles of interaction with the receptors CA-II, Cat B, DNA-2, Gyrase, HDAC7, HIS, Kinase, rHA, RNR, Top II, and TS  and polarizabilities W (range from 0.78 to 0.93). Other computed properties like LUMO (0.46 to 0.64), gap (0.32 to 0.60), Hyp (−0.20 to −0.40), and dipole (−0.22 to −0.37) have very low correlation (as shown in the parenthesis). The HOMO has the poorest correlation with the binding activities of the ligands which range from 0.00 to 0.18. The better correlation of the LUMO with the receptor interactions further supports the hypothesis that the lower the LUMO energy of inhibitors the easier the overlapping of it with the HOMO of the DNA . Howeverin vitro anticancer activities of the six ligands 5, 6, 9, 11, 12, and 14 against the cancer cell line HT29 are discussed. The result of the in vitro study shows that the ligands 11 and 14 are the most active ones compared to other ligands. Among the two, only the ligand 11 showed promising inhibitory activities towards the selected receptors. In many of the receptors, different docking methods locate similar binding sites and similar ligand orientations. The order of the ligand interactions with the receptors using Vina, AutoDock-DC, and AutoDock is inconsistent because they locate different binding sites in many cases and their energy difference is also within the range of the standard error of AutoDock , while the order from Molegro-QC and Molegro is highly consistent and similar.The inhibitory potentials of nitrogen donor ligands in their interaction with twelve cancer-related receptors were studied using docking methods. Twenty-one derivatives of nitrogen-chelating ligands consisting of fifteen pyrazole, three bipyridine, and three phenanthroline derivatives were modelled. In addition, the experimental 7, 11, and 15 have better inhibitory activities towards many of the receptors and have similar angles of interaction with CA-II, Cat B, DNA-2, Gyrase, HDAC7, HIS, Kinase, rHA, RNR, Top II, and TS. This is an evidence that they have common residue interactions at the binding sites of each of the receptors. The receptor interactions of the three most promising ligands 7, 11, and 15 follow a different order based on their receptor preference. Apart from these three, 18 and 21 with bipyridine and phenanthroline moieties also show promising interaction with many of the receptors. The most common feature of the best five inhibitors is the carboxylic unit, which indicates that the carboxylic units enhance the binding site interaction of the ligands through formation of stronger HB interactions with the receptor residues. The thermodynamic properties have a high correlation with the docking results of the ligands' inhibitory activities. Other molecular properties, which have a high correlation with the ligands' inhibitory activities are polarizabilities. The LUMO of the ligands shows a good correlation with their receptor-binding interaction which further supports the hypothesis that the lower the LUMO energy of inhibitors the easier the overlapping of it with the HOMO of the DNA.The results from the Molegro-QC and Molegro showed that


The experiment was carried out in Viçosa, Minas Gerais, Brazil, and consisted of foliar applications of GA3 set in randomized blocks design, with four replicates and 6 treatments as following: NC (control with plants); 0 μM GA3; N1: 5 μM GA3; N2: 25 μM GA3; N3: 50 and N4: 100 μM GA3 per CWs, NC* (control with no plants): 0 μM GA3. The study was conducted over two crop cycles in the spring 2016. The parameters used to evaluate the performance of the Tifton 85 bermudagrass were its plant height, productivity, chlorophyll measurement, number of internodes, nutrients and Na removals. Chemical analyses of the effluents were conducted. In response to the application of GA3, the increase in height of Tifton 85 bermudagrass in the first crop cycle was higher than the increase in height in the second crop cycle. The decrease in plant growth in response to GA3 in the second crop cycle may be linked to the age of the plant tissue and climatic conditions. The greater growth of the plants cultivated in the CWs allows a more efficient removal of pollutants, using simple management and low cost. The results suggest that applying 50 μM of GA3 to the development of Tifton 85 bermudagrass provides higher dry matter yield and removal of nitrogen, phosphorus, and sodium for the first crop cycle in CWs. However, in the second crop cycle, the application of GA3 had no effect on dry matter production and nutrient removal by Tifton 85 bermudagrass in CWs.This study aimed to evaluate 1) the influence of gibberellic acid (GA The disposal of wastewater into rivers, lakes and other water bodies leads to a reduction in quality of water resources, which are of prime importance to human and aquatic life. The discharge of wastewater contains high levels of organic nitrogen and phosphorus compounds that enable growth of algae and aquatic plants, which are responsible for eutrophication of water bodies.Constructed wetland systems (CWs) are treatment systems planned and designed to use natural processes involving wetland vegetation, soils, and their associated microbial assemblages to improve water quality . In CWs—The efficiency of sewage treatment using CWs is obtained through the hydraulic interaction of the substrate, plants, microorganisms and solar radiation. Pollutant removal takes place in a support media, which is responsible for the filtration and the formation of the biofilm next to the roots of the plants. The microorganisms adhere to the biofilm, degrade carbonaceous and nitrogenous matter, and the plants absorb the nutrients and heavy metals, thus promoting phytoremediation .CWs have a high removal efficiency and appropriate effluent quality for final disposal, especially regarding organic pollutants , volatile suspended solids, and oils and greases). However, their efficiency of nitrogen (N) and phosphorus (P) removal is relatively low .The selection of wetland plants is crucial to the success of the wastewater treatment because the plants uptake macro and micronutrients from the CWs for their development. The macronutrients N and P are essential for plant growth and development. Nitrogen, besides constituting proteins, is required to produce the molecule chlorophyll. P is important for the energy reserve and structural integrity of tissues.Phragmites australis), cattail (Typha spp.), lakeshore bulrush (Schoenoplectus lacustris) and Canna indica are some of the most common wetland species studied in treatment wetlands [Pennisetum purpureum) and Tifton 85 bermudagrass (Cynodon spp.) [Cynodon dactylon) and a close tropical relative called stargrass (Cynodon nlemfuensis) with high zootechnical and economic potential due to its good palatability, digestibility, and expressive crude protein concentrations. In addition, it presents an attractive commercial value for the hay production, reaching higher values in regions with little forage availability.Reed  is one of the classes of growth phytohormones that influence plant growth, increase elongation and cell division, which is evidenced by increased length and number of cells [3 is influenced by several factors, such as environmental conditions, GA3 concentration, number of applications, and the season of application and species or cultivar [The growth rate of plants can be increased with the use of growth-regulating substances . Gibbereof cells . Howevercultivar .3 on plants grown in CWs. Therefore, it is necessary to investigate the effects GA3 has on productive characteristics of Tifton 85 bermudagrass, and pollutant removal efficiency in CWs.One of the roles played by plants in CWs is nutrient removal. Nutrient uptake affects plant growth rate. However, there are no references in the scientific literature regarding the effect of GA3 in the development of Tifton 85 bermudagrass grown in CWs; and 2) the plant's capacity to remove nutrients and sodium (Na) from synthetic municipal wastewater (SMW).This study aimed to evaluate 1) the influence of GAThe experiment was carried out in a greenhouse located at the experimental area of the water resources reference center belonging to the Department of Agricultural Engineering at the Federal University of Viçosa,  Viçosa, Minas Gerais state, Brazil. The climate, according to the Köppen climate classification, is Cwa .2, cultivated with Tifton 85 bermudagrass , which were then submitted to SMW applications  with 26.50 cm height, 23.58 cm diameter, with a surface area of 0.0437 mications .The CWs were filled up to 21.2 cm height with gneissic gravel 0  and a free edge of 5.3 cm. Prior to filling the CWs, the rocks were immersed in a bleach solution and rinsed with running water.3 are listed in The different treatments and their respective doses of GA3, and one treatment control with no plants to assess the effluent, totaling 24 experimental plots.A randomized complete block design was used, with four replications and five treatments with Tifton 85 bermudagrass exposed to different doses of GAThe SMW was used in the experiment to guarantee greater control of chemical and organic compounds, and for sanitary purposes, since the operation of CWs and handling of SMW was done manually.The SMW used in this study was modified from Nopens ; the con-1 [Municipal wastewater is characterized by a high concentration of salts, especially K and Na. The concentrations of K and Na in the influent of the present study are similar to those found at the municipal wastewater treatment plant of Janaúba, Minas Gerais . The con-1 .The CWs worked as a batch system, being fed from the top. Influent SMW was applied until the reactors were filled up to the point where the upper layer of gravel was reached. Then the volume of input of each CWs was recorded. SMW remained in reactors during a cycle time, then the SMW was collected from the bottom to characterize a vertical flow.SMW was changed every 7 days (cycle time) during the acclimatization period of the experiment, and every 3.5 days after that. The cycle time in batch systems is analogous to the hydraulic residence time (HRT) in continuous systems.The salts and the other ingredients were diluted into drinking water to prepare SMW; weekly evaluations of the parameters of the influent were conducted, totaling 11 analyses. The averages and standard deviations of the actual SMW composition in the present study are shown in After the acclimatization period, the differentiation of treatments with HRT of 3.5 d began and the average rates of SMW application were calculated .Tifton 85 bermudagrass sprigs, formed by a segment of stolon with two buds with leaves and roots removed, were collected and standardized at 0.10 m. Then four sprigs were planted in each reactor.3 (2 drops of spreader in 25 ml GA3 solution) to facilitate the contact and absorption of the phytohormone by plants. The solutions were applied on the day of the standardization cut and at the end of the first cycle.The stabilization period lasted 48 days. At the end of this period, the plants were cut at a standard height of 0.10 m above the level of the support media. 0.1% (v/v) A spreader was added to the different doses of GA3; the second application was seven days after the plants being cut. The experiment was carried out for 89 days.The first and second cycles lasted 18 and 23 days respectively and were carried out in the spring 2016 (southern hemisphere). The end of each cycle was pinpointed by when the Tifton 85 bermudagrass started lodging. Both cycles received two applications of the different doses of GAThe weather conditions  were meaThe weather conditions inside the greenhouse were not controlled, so in the first and second crop cycles, the average temperatures were 27.9 and 30.2° C, and the relative humidity was 58% and 56%, respectively.The parameters analyzed in Tifton 85 bermudagrass and their methodologies are shown in An estimate of chlorophyll content in leaves was determined at the end of each cycle using a chlorophyll meter. First, the chlorophyll meter was calibrated according to the specifications of the equipment, then SPAD readings were measured on one leaf per stem, on two distinct stems per reactor, on the middle third of the stem and the leaf blade.The analyses of TN in the aerial part of the plants were quantified by the Kjeldahl semi-micro method with the addition of salicylic acid, adapted from Kiehl . AbsortiTN, K, TP or Na—nutrient or Na accumulation (g), CNutri—content of nutrients or Na in dry matter of the aerial part of Tifton 85 bermudagrass (g g-1); DMY—dry matter of the aerial part of Tifton 85 bermudagrass.Based on the analyses results, the nutrients/Na accumulation by Tifton 85 bermudagrass was calculated using TN, K, TP or Na—nutrient or Na uptake (g m-3), ATN, K, TP or Na−nutrient or Na accumulation (g), Vol—CWs volume (m3).The nutrients/Na uptake by Tifton 85 bermudagrass was calculated using TN, K, TP or Na–nutrient or Na uptake rate (g m-3 d-1), TN,K,TP or NaU – nutrient or Na uptake (g m-3), Ct—cycle time (d).The nutrients/Na uptake rate by Tifton 85 bermudagrass was calculated using oxidation-reduction potential, pH, electrical conductivity, evapotranspiration, COD, total nitrogen, total phosphorus, K, Na and total dissolved solids were performed to understand the current reactions and the nutrients and Na removal efficiency in CWs. The parameters analyzed are presented in In the experiments, influents and effluents were evaluated weekly, i.e., every two cycle times (HRT of 3.5 d). Analyses of c) was determined in each plot and after each cut (0.10 m). ETc was calculated as the difference between the volumetric inflow and outflow divided by the surface area of the plot for data collected during the exchange of SMW, c (mm d−1), Qin is the plot volumetric inflow (mm3 d−1), Qout, is the measured plot volumetric outflow (mm3 d−1), and SA is the measured plot surface (mm2).The Tifton 85 bermudagrass evapotranspiration  and were calculated using e—corrected effluent concentration (g m-3), CE—effluent concentration (g m-3), Vf−final volume collected (L), Vi−initial volume applied (L).To obtain the actual value of the mass reduction in each parameter evaluated, the effluent concentrations of each CWs were corrected by ET0—influent concentration (g m-3); Ce—corrected effluent concentration (g m-3).The removal efficiency of SMW pollutants was calculated using 2) and that expressed the behavior of the phenomenon. The regression coefficients were subjected to t-test at the 5% level of significance. When the regression model presented the level above 5%, the probability found for the model was used. When the regression model did not explain the phenomenon, the Dunnett’s test was used to compare the GA3 treatments with the control.All numerical data were subjected to variance analysis . When th3  and Tifton 85 bermudagrass SPAD index were significantly influenced by various doses of GA3 . In the 3 doses. A square root model regression was set for each cycle. For the 1st cycle, the model presented p = 0.1192 by t-test.3 is to promote elongation of the stem [3) with the control, it is observed an increase of 35.10% and 20.90% in plant height for the first and second cycles, respectively.One of the roles of GAthe stem . This ef3 showed lower growth than the other treatments  2682.60 kg ha-1 yield, i.e., 33.80% lower than the yield obtained with the treatment of 50 μM of GA3.The DMY of aerial parts of Tifton 85 bermudagrass in the first cycle presented a square root behavior for the various doses of GA3 . The hig3 indicated that the phytohormone plays an important role in the absorption of water and nutrients and processing the accumulation of carbohydrates within plant tissues. The same was observed by Zang during foliar application of GA3 and evaluation of dry matter in blueberry leaves [The increase of the DMY in aerial parts of Tifton 85 bermudagrass with the exogenous application of increasing doses of GAy leaves .3 stimulates cell division and cell elongation and the exogenous addition of this phytohormone leads to an increase in plant growth, improving the availability of endogenous GA3. This view is supported by Kaur [rabidopsis thaliana plants treated with GA3 showed a higher shoot development compared to the control treatment [The GA by Kaur  and Tuna by Kaur , workingreatment .3 is a viable practice to improve the development of plants, as well as to create conditions to increase the removal of nutrients (pollutants).Plants have an important role in the removal of nutrients in CWs, however, the removal capacity can be improved if crop conditions that allow rapid growth and dry matter production are created . In view3 was observed, indicating a lower chlorophyll content in the treatment without exogenous application of GA3.As for the SPAD index evaluated in the first cycle , an incr3 improved chlorophyll levels in plants grown in CWs.Since the plants were grown in adverse conditions , this lower amount of chlorophyll in the control could be attributed to the formation of proteolytic enzymes such as chlorophyllase, an enzyme responsible for the degradation of chlorophyll , which m3 for DMY and SPAD index in the second cycle may be related to the period of application of the phytohormone. Some studies reported that GA3 applied during the spring season promotes a high forage growth and that response to the hormone is reduced in summer due to the high temperatures [3 in the second cycle are the age of the plant, space restriction for root growth, and a lower GA3 response in older tissues.The absence of effect of GAeratures . As can 3 compared to the treatment without the application of GA3  differed from the control level without GA3 application.The accumulation of TP in the aerial part of Tifton 85 bermudagrass is presented in 3 in a maize crop under salinity conditions increased foliar TP [3 compared to the control promoted a greater accumulation of TP for the N1 treatment. From the environmental point of view, it is an advantage since it showed that the removal of TP in CWs can be improved with an application of GA3.The treatment with exogenous application of GAoliar TP . The higoxidation-reduction potential (ORP) of 127.2 mV. Only in three peaks ; between 400–200 mV, the presence of O2 and nitrate (NO3-); and below 200 mV indicates low oxygen presence tending to anoxic/facultative conditions [In function of the ORP evaluated in the present study, it was possible to indicate that the conditions of the CWs were anoxic/facultative. A classification for the ORP of the medium, ORP above 400 mV has the predominance of Onditions .-1. Except for the NC*, the treatments presented a similar behavior for the parameter pH and EC. In both cycles the pH and EC were higher for the NC*, this allows to understand the importance of the removal of the pollutants (nutrients and Na) and other salts by the plants.After collecting the effluents, the pH and EC were determined, and their values are presented in The increase in pH in the different treatments may be related to the use of carbon dioxide by plants and algae .The pH values ranged from 6.5 to 8.2 for treatments and control plots, what indicated that SMW (influent and effluent) presented ideal conditions for the survival of microorganisms and for the degradation of organic matter . The optc was calculated through the determination of the volume of the input (influent) and the output (effluent) (-1 and the system with Tifton 85 bermudagrass grass showed an average ETc of the treatments of 5.2 mm d-1.The daily average evaporation and ETffluent) . SubsequIt is possible to observe the importance of plant cultivation in CWs since they transform the SMW into water into the atmosphere through transpiration. Thus, in addition to removing the pollutants (nutrients), CWs with plants reduce the volume of sewage for the final disposal.3- and ammonium (NH4+); the ammonium ion can also be adsorbed by plants roots. The sum of these N forms is described as TN [The behavior of TN and TP concentrations in the effluent is shown in ed as TN . The preed as TN , 43.In the first cycle, the COD removal was higher for the CWs without plants (NC*), and the CWs with the treatment levels N3 and N4 were the ones with the lowest COD removals compared to the two controls .3 can stimulate root growth [The treatments N3 and N4 were those that obtained higher plant height and DMY in comparison with NC (1st cycle) . As a ret growth . Togethet growth .3) did not differentiate from the NC control. This lower COD efficiency removal in treatments with the presence of Tifton 85 bermudagrass may be related to the release of root exudates and plant senescence.In the second cycle, the NC* treatment presented a lower COD concentration when compared to N1 and N4. The treatments that received gibberellin  worked with a high rate of TN application, around 25.70 g m-2 d-1 TN, via swine wastewater [The TN removal in the present study was higher than the values observed by Matos (2008), Fia (2011) and Matos (2010) , 34, 47.-2 d-1 when treating municipal wastewater with TN application rates equal to 9.20 g m-2 d-1 in CWs with Tifton 85 bermudagrass. These values of application and removal rates were higher than the ones in the present study [Jesus (2016) observed removals of 0.79 g mnt study .-2 d-1 by the Tifton 85 bermudagrass treated with 50 μM of GA3 was higher than that observed by Matos (2008), Fia (2011), Matos (2010) and Jesus (2016). These authors verified removal rates of 0.05, 0.08, 0.04 and 0.04 g m-2 d-1 of TP of aerial parts of Tifton 85 bermudagrass, respectively [For different application rates and different influents , the TP ectively , 47, 48.3 showed potential for nutrient removal via biomass of Tifton 85 bermudagrass cultivated in CWs in the treatment of SMW, it would be interesting to conduct further field testes with higher load applications.Although the use of GA3 in spring and another in autumn resulted in the increased dry mass of azevem and white clover [3 strongly depend on the time of year in which the plants were treated. And for perennial crops such as Tifton 85 bermudagrass, GA3 may be interacting with a fixed strategy for seasonal plant growth [An application of GAe clover . Thus, rt growth .The gibberellic acid promoted higher growth in height of Tifton 85 bermudagrass in constructed wetland systems.3 provided higher dry matter yield and nitrogen removal by Tifton 85 bermudagrass in constructed wetland systems for the first crop cycle.The application of 50 μM of GA3 was the best dose for the global removal of P and Na from synthetic municipal wastewater in the first crop cycle of Tifton 85 bermudagrass in constructed wetland systems.The application of 50 μM GAThe gibberellic acid applied in the second crop cycle had no effect on dry matter yield and nutrient removal by Tifton 85 bermudagrass grass in constructed wetland systems.S1 TableResults of giberelic acid addition to batch constructed wetlands systems.(XLSX)Click here for additional data file.


The aim of the study was to evaluate the efficiency of the Hankins and Howe (HH46), Valadares Filho (V06), and Marcondes (M12) equations for predicting the physical and chemical composition of dairy crossbred bulls carcasses, as well as the chemical composition of their empty bodies.This study was conducted using 30 dairy crossbred bulls. One group of five animals was slaughtered at the beginning of the experiment, and the remaining were slaughtered 112 days later. Animals were distributed in a completely randomized design into treatments consisting different levels of concentrate . The physical and chemical compositions of the cattle were obtained from the right half of the carcass and using samples taken between the 9th and 11th ribs of the left half of the carcass. The estimated and experimentally determined values were compared using the correlation and concordance coefficient, as well as the mean square error of prediction (MSEP) and its components.The HH46 equations were better at estimating the amount of muscle plus fat in the carcass. The amount of bone in the carcasses could not be well estimated by the HH46 and M12 models. The M12, HH46, and V06 equations were worst at estimating the amounts of protein, ether extract, and water in the carcass, respectively. In the empty body, the amounts of protein and water were well estimated by the HH46 equations. Protein, ether extract, and water were accurately estimated by the V06 equations, and ether extract by the M12 equations.The physical and chemical composition of dairy crossbred bull carcasses, as well as the chemical composition of their empty bodies, can be predicted using the equations tested here. The amount of bone in these carcasses could not be accurately predicted. Bos taurus taurus, which has a different body composition than zebu or crossbred [To reduce the time and cost spent on the complete dissection of cattle carcasses, Hankins and Howe  proposedrossbred , and therossbred .Work by Marcondes et al , PaulinoThe use of crossbred Holstein×Gir for milk production is common in tropical regions. This cross produces animals that are more tolerant to tropical conditions while maintaining good productivity. Thus, male calves from Holstein×Gir crosses are an important alternative meat source in Brazil, as well as in other parts of the world. Despite the importance of this genetic group, there is little available data on Holstein×Gir crossbred cattle –14. HereThis work was conducted with the approval of the Animal Use Ethics Committee of Federal Rural University of Pernambuco (UFRPE), Recife, Pernambuco, Brazil (Permit Number: 23082.015634/2012–41).Thirty crossbred dairy bulls, with an average initial body weight (BW) of 339.1 (standard deviation = 35.4 kg) were used. First, animals were weighed, identified, treated against internal and external parasites, and supplemented with injectable vitamin A, D, and E.After a 30 day adaptation period, in which animals were fed the same diet , five animals were randomly chosen as the reference group. The remaining animals were randomly distributed in a completely randomized design consisting five treatments , confined for 112 days, and slaughtered.Diets were formulated according to National Research Council  for a weAt the end of the experiment, the animals were slaughtered after 16 h of fasting. Procedures followed Instruction No. 3 (17 Jan 2000) of the Ministry of Agriculture, Livestock and Food Supply, which regulates humanitarian slaughter [9–11), according to the methodology described by Hankins and Howe [To determine the empty body weight (EBW), the gastrointestinal tract was emptied, washed, and its weight added to the remaining parts of the empty body . To obtain the cold carcass weight, the carcass of each animal was divided into two half carcasses, which were weighed and then cooled in a cold chamber at 4°C for 24 h. From the left half of the carcass, the section between the 9th and 11th ribs , and water content, organs and viscera, muscle and fat, leather, bone, tail, head, and limbs were dried in stove at 105°C (method 24.002 ).Later, a pre-degreasing was performed on the samples through successive washings with petroleum ether, to obtain the pre-defatted dry matter (PDDM). Then, samples were ground in a ball mill for further analysis and for quantification of the ether extract  and nit9–11, as well as other parameters, were used to design these equations (The equations proposed by Hankins and Howe  (HH46), quations .The data used to test the equations are shown in The body composition estimated by the equations of HH46, V06, and M12 were compared with the values observed using the regression model:0 and β1 = intercept and inclination coefficient of the linear equation, under the following statistical hypothesis:Where X = predicted value; Y = observed value; βIf the null hypotheses were not rejected, the prediction model accurately estimated the evaluated body composition.The efficiency of the prediction models was also evaluated using the mean square error of the prediction (MSEP) and its components :2, MaF = (sx–sy)2, MoF = 2sxsy(1–r), where x are the predicted values; y are the observed values; MSEP is the mean square error of the prediction; SB is the squared bias; MaF is the magnitude of random fluctuation component; MoF is the model of random fluctuation component; sx and sy are the standard deviations of the predicted and observed values, respectively; and r is the Pearson linear correlation coefficient between the predicted and observed values. Accuracy and precision of the model were evaluated by estimating the correlation and concordance coefficient (CCC) or the reproducibility index, described by Tedeschi [http://nutritionmodels.com/mes.html).SB = (x–y)Tedeschi . Models The equation proposed by HH46 was best at estimating the composition of the muscle tissue plus adipose tissue in the carcass when compared with M12 when the CCC was observed . The int0 and β1 parameters (p<0.05). Both equations overestimated the amount of bone tissue in the carcass for the majority of the evaluated samples . However, the M12 model have problems with intercept and slope was unable to adequately estimate the equation parameters (p<0.05).When estimating the EE, the M12 and V06 returned about equal CCC and MSEP values. Using the HH46 equation returned high CCC and MSEP values, and the coefficient of inclination was not properly estimated (p<0.05). Most of the EE concentration was overestimated by the HH46 equation .0 and β1 parameters of the linear equation.When estimating the amount of water in the carcass, the V06 equation returned a higher CCC value and lower MSEP value than the other equations. However, the intercept and the slope were not well estimated using V06 (p<0.05). On the other hand, this problem was not observed for the HH46 and M12 equations, which both returned high CCC values . The M12When estimating the chemical components in the empty body, all of the tested equations returned high CCC values .The MSEP was similar between M12 and V06 equations for EE and P, with most of the MSEP relative to MoF, indicating that most of this error was unrelated to the equation.0 of the equation, this was supported by the SB value for this equation. Thus, V06 equation were better, because most of the MSEP was not relative to the model (MoF = 39.39).When estimating water content, the MSEP of the V06 equation were smaller than the M12 equation. The highest proportion MSEP of the M12 model was associated with the deviation in the βThe performance of the HH46 equation when estimating the amount of muscle plus fat in the carcass might be connected to the antagonism of these two components, leading to a positive result when these two variables are added together . HH46 deThe introduction of internal fat in the M12 equation may have interfered in its efficiency at estimating muscle plus fat in crossbred dairy bulls, since this animal category tends to store dietary energy excess in the form of internal fat .The low efficiency of the HH46 and M12 equations at estimating the amount of bone tissue may be related to the genetic group used in the study. Costa e Silva et al  found thNone of the evaluated models could adequately estimate all of the chemical components in the carcass . HoweverWe found that the V06 equations are suitable for estimating EE and water in the empty bodies of crossbred dairy cattle , which iIn crossbred dairy bulls, the amount of fat plus muscle in the carcass can be estimated using the HH46 equations. However, the amount of bone tissue in the carcass cannot be estimated from the HH46 and M12 models. The M12, HH46, and V06 models were worst at estimating the amounts of protein, EE, and water in the carcass, respectively. In the empty body, the amount water are best estimated using the equation proposed by V06; protein, and EE, are best estimated by the V06 and M12 equations. Overall, we propose that for dairy crossbred bulls, the carcass physical and chemical composition (except bone content) and empty body chemical composition can be predicted using the equations tested in this work.