Effect of S-Girdling on Fruit Growth and Fruit Quality of Wax Apple

The study was performed to evaluate the effect of Sgirdling, fruit thinning plus bagging with 2,4-D application, fruit thinning plus bagging on growth and quality of wax apple fruit. Girdling was applied three week before flowering. The 2,4-D was sprayed at the small bud and petal fall stage. The effect of all treatments on fruit growth was measured weekly. The physical and biochemical quality characteristics of the fruits were recorded. The results showed that no significant effect on number of bud among treatments. S-girdling, 2,4-D application produced the lowest bud drop, fruit drop compared to untreated control. Moreover, S-girdling enhanced faster fruit growth producing the best final fruit length and diameter than the control treatment. It was also observed that Sgirdling greatly increased fruit set, fruit weight as well as total soluble solid, reduced fruit crack, and titratable acidity. In conclusion, S-girdling had a distinctive and significant effect on most of the fruit quality characteristics assessed. Application 2,4-D was also recommended as the industry norm to increase fruit set, and fruit quality in wax apple.

Interactions between Cells and Nanoscale Surfaces of Oxidized Silicon Substrates

The importance for manipulating an incorporated scaffold and directing cell behaviors is well appreciated for tissue engineering. Here, we developed newly nano-topographic oxidized silicon nanosponges capable of being various chemical modifications to provide much insight into the fundamental biology of how cells interact with their surrounding environment in vitro. A wet etching technique is exerted to allow us fabricated the silicon nanosponges in a high-throughput manner. Furthermore, various organo-silane chemicals enabled self-assembled on the surfaces by vapor deposition. We have found that Chinese hamster ovary (CHO) cells displayed certain distinguishable morphogenesis, adherent responses, and biochemical properties while cultured on these chemical modified nano-topographic structures in compared with the planar oxidized silicon counterparts, indicating that cell behaviors can be influenced by certain physical characteristic derived from nano-topography in addition to the hydrophobicity of contact surfaces crucial for cell adhesion and spreading. Of particular, there were predominant nano-actin punches and slender protrusions formed while cells were cultured on the nano-topographic structures. This study shed potential applications of these nano-topographic biomaterials for controlling cell development in tissue engineering or basic cell biology research.

A Study of Liver Checkup in Patients with Hepatitis C in the Region of Batna

Hepatitis C is an infectious disease transmitted by blood and due to hepatitis C virus (HCV), which attacks the liver. The infection is characterized by liver inflammation (hepatitis) that is often asymptomatic but can progress to chronic hepatitis and later cirrhosis and liver cancer. Our problem tends to highlight on the one hand the prevalence of infectious disease in the population of the region of Batna and on other hand the biological characteristics of this disease by a screening and a specific diagnosis based on serological tests, liver checkup (measurement of haematological and biochemical parameters). The results showed: The serology of hepatitis C establishes the diagnosis of infection with hepatitis C. In this study and with the serological test, 24 cases of the disease of hepatitis C were found in 1000 suspected cases (7 cases with normal transaminases and 17 cases with elevated transaminases). The prevalence of this disease in this study population was 2.4%. The presence of hepatitis C disrupts liver function including the onset of cytolysis, cholestasis, jaundice, thrombocytopenia, and coagulation disorders.

Response of Wax Apple Cultivars by Applied S-Girdling on Fruit Development and Fruit Quality

The study was carried out to evaluated effect of S-gridling on fruit growth and quality of wax apple. The study was laid in Random completed block design with four replicated. Four treatment were applied as follows: S-girdling, fruit thinning plus bagging with 2,4-D sprayed, fruit thinning plus bagging and the control treatment. 2,4D was sprayed at the small bud and petal fall stage. Girdling was applied three week before flowering. The effect of all treatments on fruit growth was measured weekly. Number of flower, fruit set, fruit drop, fruit crack, and fruit quality were recorded. The result indicated that S-girdling, 2,4D application produced the lowest bud drop, fruit drop compared to untreated control. S-girdling improved faster fruit growth producing the best final fruit length and diameter compared to untreated control. S-girdling also markedly enhanced fruit set, fruit weight, and total soluble solid, reduced fruit crack, titratable acidity. On the other hand, it was noticed that with 2,4-D application also increased the fruit growth rate, improved physiological and biochemical characters of fruit than control treatment. It was concluded that S-girdling was recommended as the industry norm to increase fruit set, fruit quality in wax apple. 2,4D application had a distinctive and significant effect on most of the fruit quality characteristics assessed.

Enzymatic Saccharification of Dilute Alkaline Pre-treated Microalgal (Tetraselmis suecica) Biomass for Biobutanol Production

Enzymatic saccharification of biomass for reducing sugar production is one of the crucial processes in biofuel production through biochemical conversion. In this study, enzymatic saccharification of dilute potassium hydroxide (KOH) pre-treated Tetraselmis suecica biomass was carried out by using cellulase enzyme obtained from Trichoderma longibrachiatum. Initially, the pre-treatment conditions were optimised by changing alkali reagent concentration, retention time for reaction, and temperature. The T. suecica biomass after pre-treatment was also characterized using Fourier Transform Infrared Spectra and Scanning Electron Microscope. These analyses revealed that the functional group such as acetyl and hydroxyl groups, structure and surface of T. suecica biomass were changed through pre-treatment, which is favourable for enzymatic saccharification process. Comparison of enzymatic saccharification of untreated and pre-treated microalgal biomass indicated that higher level of reducing sugar can be obtained from pre-treated T. suecica. Enzymatic saccharification of pre-treated T. suecica biomass was optimised by changing temperature, pH, and enzyme concentration to solid ratio ([E]/[S]). Highest conversion of carbohydrate into reducing sugar of 95% amounted to reducing sugar yield of 20 (wt%) from pre-treated T. suecica was obtained from saccharification, at temperature: 40°C, pH: 4.5 and [E]/[S] of 0.1 after 72 h of incubation. Hydrolysate obtained from enzymatic saccharification of pretreated T. suecica biomass was further fermented into biobutanol using Clostridium saccharoperbutyliticum as biocatalyst. The results from this study demonstrate a positive prospect of application of dilute alkaline pre-treatment to enhance enzymatic saccharification and biobutanol production from microalgal biomass.

Energy Production Potential from Co-Digestion between Frozen Seafood Wastewater and Decanter Cake in Thailand

In this paper, a Biochemical Methane Potential (BMP) test provides a measure of the energy production potential from codigestion between the frozen seafood wastewater and the decanter cake. The experiments were conducted in laboratory-scale. The suitable ratio of the frozen seafood wastewater and the decanter cake was observed in the BMP test. The ratio of the co-digestion between the frozen seafood wastewater and the decanter cake has impacts on the biogas production and energy production potential. The best performance for energy production potential using BMP test observed from the 180 ml of the frozen seafood wastewater and 10 g of the decanter cake ratio. This ratio provided the maximum methane production at 0.351 l CH4/g TCODremoval. The removal efficiencies are 76.18%, 83.55%, 43.16% and 56.76% at TCOD, SCOD, TS and VS, respectively. The result can be concluded that the decanter cake can improve the energy production potential of the frozen seafood wastewater. The energy provides from co-digestion between frozen seafood wastewater and decanter cake approximately 19x109 MJ/year in Thailand.

In vivo Antidiabetic and Antioxidant Potential of Pseudovaria macrophylla Extract

This study has investigated the antidiabetic and antioxidant potential of Pseudovaria macrophylla bark extract on streptozotocin–nicotinamide induced type 2 diabetic rats. LCMSQTOF and NMR experiments were done to determine the chemical composition in the methanolic bark extract. For in vivo experiments, the STZ (60 mg/kg/b.w, 15 min after 120 mg/kg/1 nicotinamide, i.p.) induced diabetic rats were treated with methanolic extract of Pseuduvaria macrophylla (200 and 400 mg/kg·bw) and glibenclamide (2.5 mg/kg) as positive control respectively. Biochemical parameters were assayed in the blood samples of all groups of rats. The pro-inflammatory cytokines, antioxidant status and plasma transforming growth factor βeta-1 (TGF-β1) were evaluated. The histological study of the pancreas was examined and its expression level of insulin was observed by immunohistochemistry. In addition, the expression of glucose transporters (GLUT 1, 2 and 4) were assessed in pancreas tissue by western blot analysis. The outcomes of the study displayed that the bark methanol extract of Pseuduvaria macrophylla has potentially normalized the elevated blood glucose levels and improved serum insulin and C-peptide levels with significant increase in the antioxidant enzyme, reduced glutathione (GSH) and decrease in the level of lipid peroxidation (LPO). Additionally, the extract has markedly decreased the levels of serum pro-inflammatory cytokines and transforming growth factor beta-1 (TGF-β1). Histopathology analysis demonstrated that Pseuduvaria macrophylla has the potential to protect the pancreas of diabetic rats against peroxidation damage by downregulating oxidative stress and elevated hyperglycaemia. Furthermore, the expression of insulin protein, GLUT-1, GLUT-2 and GLUT-4 in pancreatic cells was enhanced. The findings of this study support the anti-diabetic claims of Pseudovaria macrophylla bark.

Utilization of 3-N-trimethylamino-1-propanol by Rhodococcus sp. strain A4 isolated from Natural Soil

The aim of this study was to screen for microorganism that able to utilize 3-N-trimethylamino-1-propanol (homocholine) as a sole source of carbon and nitrogen. The aerobic degradation of homocholine has been found by a gram-positive Rhodococcus sp. bacterium isolated from soil. The isolate was identified as Rhodococcus sp. strain A4 based on the phenotypic features, physiologic and biochemical characteristics, and phylogenetic analysis. The cells of the isolated strain grown on both basal-TMAP and nutrient agar medium displayed elementary branching mycelia fragmented into irregular rod and coccoid elements. Comparative 16S rDNA sequencing studies indicated that the strain A4 falls into the Rhodococcus erythropolis subclade and forms a monophyletic group with the type-strains of R. opacus, and R. wratislaviensis. Metabolites analysis by capillary electrophoresis, fast atom bombardment-mass spectrometry, and gas chromatography- mass spectrometry, showed trimethylamine (TMA) as the major metabolite beside β-alanine betaine and trimethylaminopropionaldehyde. Therefore, the possible degradation pathway of trimethylamino propanol in the isolated strain is through consequence oxidation of alcohol group (-OH) to aldehyde (-CHO) and acid (-COOH), and thereafter the cleavage of β-alanine betaine C-N bonds yielded trimethylamine and alkyl chain.

Effect of Plasma Therapy on Epidermal Regeneration

The purpose of our study was to compare spontaneous re-epithelisation characteristics versus assisted re-epithelisation. In order to assess re-epithelisation of the injured skin, we have imagined and designed a burn wound model on Wistar rat skin. Our aim was to create standardised, easy reproducible and quantifiable skin lesions involving entire epidermis and superficial dermis. We then have applied the above mentioned therapeutic strategies to compare regeneration of epidermis and dermis, local and systemic parameter changes in different conditions. We have enhanced the reepithelisation process under a moist atmosphere of a polyurethane wound dress modified with helium non-thermal plasma, and with the aid of direct cold-plasma treatment respectively. We have followed systemic parameters change: hematologic and biochemical parameters, and local features: oxidative stress markers and histology of skin in the above mentioned conditions. Re-epithelisation is just a part of the skin regeneration process, which recruits cellular components, with the aid of epidermal and dermal interaction via signal molecules.

Effects of Microwave Heating on Biogas Production, Chemical Oxygen Demand and Volatile Solids Solubilization of Food Residues

This paper presents the results of the preliminary investigation of microwave (MW) irradiation pretreatments on the anaerobic digestion of food residues using biochemical methane potential (BMP) assays. Low solids systems with a total solids (TS) content ranging from 5.0-10.0% were analyzed. The inoculum to bulk mass of substrates to water ratio was 1:2:2 (mass basis). The experimental conditions for pretreatments were as follows: a control (no MW irradiation), two runs with MW irradiation for 15 and 30 minutes at 320 W, and another two runs with MW irradiation at 528 W for 30 and 60 minutes. The cumulative biogas production were 6.3 L and 8.7 L for 15min/320 W and 30min/320 W MW irradiation conditions, respectively, and 10.5 L and 11.4 L biogas for 30min/528 W and 60min/528 W, respectively, as compared to the control giving 5.8 L biogas. Both an increase in exposure time of irradiation and power of MW had increased the rate and yield of biogas. Singlefactor ANOVA tests (p

Rapid Determination of Biochemical Oxygen Demand

Biochemical Oxygen Demand (BOD) is a measure of the oxygen used in bacteria mediated oxidation of organic substances in water and wastewater. Theoretically an infinite time is required for complete biochemical oxidation of organic matter, but the measurement is made over 5-days at 20 0C or 3-days at 27 0C test period with or without dilution. Researchers have worked to further reduce the time of measurement. The objective of this paper is to review advancement made in BOD measurement primarily to minimize the time and negate the measurement difficulties. Survey of literature review in four such techniques namely BOD-BARTTM, Biosensors, Ferricyanidemediated approach, luminous bacterial immobilized chip method. Basic principle, method of determination, data validation and their advantage and disadvantages have been incorporated of each of the methods. In the BOD-BARTTM method the time lag is calculated for the system to change from oxidative to reductive state. BIOSENSORS are the biological sensing element with a transducer which produces a signal proportional to the analyte concentration. Microbial species has its metabolic deficiencies. Co-immobilization of bacteria using sol-gel biosensor increases the range of substrate. In ferricyanidemediated approach, ferricyanide has been used as e-acceptor instead of oxygen. In Luminous bacterial cells-immobilized chip method, bacterial bioluminescence which is caused by lux genes was observed. Physiological responses is measured and correlated to BOD due to reduction or emission. There is a scope to further probe into the rapid estimation of BOD.

Biochemical Characteristics of Sorghum Flour Fermented and/or Supplemented with Chickpea Flour

Sorghum flour was supplemented with 15 and 30% chickpea flour. Sorghum flour and the supplement were fermented at 35 oC for 0, 8, 16, and 24 h. Changes in pH, titrable acidity, total soluble solids, protein content, in vitro protein digestibility and amino acid composition were investigated during fermentation and/or after supplementation of sorghum flour with chickpea. The pH of the fermenting material decreased sharply with a concomitant increase in the titrable acidity. The total soluble solids remained unchanged with progressive fermentation time. The protein content of sorghum cultivar was found to be 9.27 and that of chickpea was 22.47%. The protein content of sorghum cultivar after supplementation with15 and 30% chickpea was significantly (P ≤ 0.05) increased to 11.78 and 14.55%, respectively. The protein digestibility also increased after fermentation from 13.35 to 30.59 and 40.56% for the supplements, respectively. Further increment in protein content and digestibility was observed when supplemented and unsupplemented samples were fermented for different periods of time. Cooking of fermented samples was found to increase the protein content slightly and decreased digestibility for both supplements. Amino acid content of fermented and fermented and cooked supplements was determined. Supplementation was found to increase the lysine and therionine content. Cooking following fermentation decreased lysine, isoleucine, valine and sulfur containg amino acids.

Response of Wax Apple Cultivars by Applied GA3 and 2,4-D on Fruit Growth and Fruit Quality

The experiment was performed to evaluate the effect of GA3, 2,4-D on fruit growth and fruit quality of wax apple. The experiment consisted of Red A, Monulla, Atu, Red B cultivars. GA3 and 2,4-D were applied at the small bud and petal fall stage. Physiological, biochemical characters of fruit were recoded. The result showed application of GA3, 2,4-D greatly response in increasing fruit set for all treatment as compared to control. Fruit weight, fruit size were increased at 10 ppm 2,4-D in ‘Red A’, ‘Red B’, however it was also enhancing at 10 ppm GA3 in ‘Monulla’, ‘Atu’. For ‘Monulla’, ‘Atu’ fruit crack reduced by 10 ppm 2,4-D application, but ‘Red B’, ‘Red A’ gave least fruit crack at 10 and 30 ppm GA3, respectively. ‘Monulla’, ‘Atu’ and ‘Red B’ resulted in response well to 10 ppm GA3 on improving TSS, whereas application of 30 ppm GA3 greatly enhancing TSS in ‘Red A’. For ‘Atu’ titratable acidity markedly reduced by 10 ppm GA3 application, but spraying with 30 ppm GA3 greatly response in reducing titratable acidity in ‘Red A’, ‘Red B’ and ‘Monulla’. It was concluded that GA3, 2,4-D can be an effective tool to enhancing fruit set, fruit growth as well as improving fruit quality of wax apple.

Isolation and Identification of an Acetobacter Strain from Iranian White-Red Cherry with High Acetic Acid Productivity as a Potential Strain for Cherry Vinegar Production in Foodand Agriculture Biotechnology

According to FDA (Food and Drug Administration of the United States), vinegar is definedas a sour liquid containing at least 4 grams acetic acid in 100 cubic centimeter (4% solution of acetic acid) of solution that is produced from sugary materials by alcoholic fermentation. In the base of microbial starters, vinegars could be contained of more than 50 types of volatile and aromatic substances that responsible for their sweet taste and smelling. Recently the vinegar industry has a great proportion in agriculture, food and microbial biotechnology. The acetic acid bacteria are from the family Acetobacteraceae. Regarding to the latest version of Bergy-s Mannual of Systematic Bacteriology that has categorized bacteria in the base of their 16s RNA differences, the most important acetic acid genera are included Acetobacter (genus I), Gluconacetobacter (genus VIII) and Gluconobacter (genus IX). The genus Acetobacter that is primarily used in vinegar manufacturing plants is a gram negative, obligate aerobe coccus or rod shaped bacterium with the size 0.6 - 0.8 X 1.0 - 4.0 μm, nonmotile or motile with peritrichous flagella and catalase positive – oxidase negative biochemically. Some strains are overoxidizer that could convert acetic acid to carbon dioxide and water.In this research one Acetobacter native strain with high acetic acid productivity was isolated from Iranian white – red cherry. We used two specific culture media include Carr medium [yeast extract, 3%; ethanol, 2% (v/v); bromocresol green, 0.002%; agar, 2% and distilled water, 1000 ml], Frateur medium [yeast extract, 10 g/l; CaCO3, 20 g/l; ethanol, 20 g/l; agar, 20 g/l and distilled water, 1000 ml] and an industrial culture medium. In addition to high acetic acid production and high growth rate, this strain had a good tolerance against ethanol concentration that was examined using modified Carr media with 5%, 7% and 9% ethanol concentrations. While the industrial strains of acetic acid bacteria grow in the thermal range of 28 – 30 °C, this strain was adapted for growth in 34 – 36 °C after 96 hours incubation period. These dramatic characteristics suggest a potential biotechnological strain in production of cherry vinegar with a sweet smell and different nutritional properties in comparison to recent vinegar types. The lack of growth after 24, 48 and 72 hours incubation at 34 – 36 °C and the growth after 96 hours indicates a good and fast thermal flexibility of this strain as a significant characteristic of biotechnological and industrial strains.

A Decision Support System for Predicting Hospitalization of Hemodialysis Patients

Hemodialysis patients might suffer from unhealthy care behaviors or long-term dialysis treatments. Ultimately they need to be hospitalized. If the hospitalization rate of a hemodialysis center is high, its quality of service would be low. Therefore, how to decrease hospitalization rate is a crucial problem for health care. In this study we combined temporal abstraction with data mining techniques for analyzing the dialysis patients' biochemical data to develop a decision support system. The mined temporal patterns are helpful for clinicians to predict hospitalization of hemodialysis patients and to suggest them some treatments immediately to avoid hospitalization.

The Effect of Silicon on Cadmium Stress in Echium amoenum

The beneficial effects of Si are mainly associated with its high deposition in plant tissue and enhancing their strength and rigidity. We investigated the role of Si against cadmium stress in (Echium C) in house green condition. When the seventh leaves was be appeared, plants were pretreated with five levels of Si: 0, 0.2, 0.5, 0.7and 1.5 mM Si (as sodium trisilicate, Na2(SiO2)3) and after that plants were treated with two levels of Cd (30 and 90 mM). The effects of Silicon and Cd were investigated on some physiological and biochemical parameters such as: lipid peroxidation (malondialdehyde (MDA) and other aldehydes, antocyanin and flavonoid content. Our results showed that Cd significantly increased MDA, other aldehydes, antocyanin and flavonoids content in Echium and silicon offset the negative effect and increased tolerance of Echium against Cd stress. From this results we concluded that Si increase membrane integrity and antioxidative ability in this plant against cd stress.

Comparison of ANFIS and ANN for Estimation of Biochemical Oxygen Demand Parameter in Surface Water

Nowadays, several techniques such as; Fuzzy Inference System (FIS) and Neural Network (NN) are employed for developing of the predictive models to estimate parameters of water quality. The main objective of this study is to compare between the predictive ability of the Adaptive Neuro-Fuzzy Inference System (ANFIS) model and Artificial Neural Network (ANN) model to estimate the Biochemical Oxygen Demand (BOD) on data from 11 sampling sites of Saen Saep canal in Bangkok, Thailand. The data is obtained from the Department of Drainage and Sewerage, Bangkok Metropolitan Administration, during 2004-2011. The five parameters of water quality namely Dissolved Oxygen (DO), Chemical Oxygen Demand (COD), Ammonia Nitrogen (NH3N), Nitrate Nitrogen (NO3N), and Total Coliform bacteria (T-coliform) are used as the input of the models. These water quality indices affect the biochemical oxygen demand. The experimental results indicate that the ANN model provides a higher correlation coefficient (R=0.73) and a lower root mean square error (RMSE=4.53) than the corresponding ANFIS model.

Antioxidant Responses to Different Exposure Regimes of Kazakhstan Light Crude Oil in Livers of Male Albino Rats

Biochemical investigations were carried out to assess the effect of different exposure regimes of Kazakhstan crude oil (KCO) on hepatic antioxidant defense system in albino rats. Contaminants were delivered under two different dosing regimes, with all treatments receiving the same total contaminant load by the end of the exposure period. Rats in regime A injected with KCO once at a dose of 6 ml/kg bw while in regime B injected multiply at a dose of 1.5 ml/kg bw on day 1, 3, 5 and 8. Antioxidant biomarkers were measured in hepatic tissue after 1, 3, 5 and 8 days. Significant induction was observed in serum aminotransferases (ALT, AST) (p

Biosynthesis and In vitro Studies of Silver Bionanoparticles Synthesized from Aspergillusspecies and its Antimicrobial Activity against Multi Drug Resistant Clinical Isolates

Antimicrobial resistant is becoming a major factor in virtually all hospital acquired infection may soon untreatable is a serious public health problem. These concerns have led to major research effort to discover alternative strategies for the treatment of bacterial infection. Nanobiotehnology is an upcoming and fast developing field with potential application for human welfare. An important area of nanotechnology for development of reliable and environmental friendly process for synthesis of nanoscale particles through biological systems In the present studies are reported on the use of fungal strain Aspergillus species for the extracellular synthesis of bionanoparticles from 1 mM silver nitrate (AgNO3) solution. The report would be focused on the synthesis of metallic bionanoparticles of silver using a reduction of aqueous Ag+ ion with the culture supernatants of Microorganisms. The bio-reduction of the Ag+ ions in the solution would be monitored in the aqueous component and the spectrum of the solution would measure through UV-visible spectrophotometer The bionanoscale particles were further characterized by Atomic Force Microscopy (AFM), Fourier Transform Infrared Spectroscopy (FTIR) and Thin layer chromatography. The synthesized bionanoscale particle showed a maximum absorption at 385 nm in the visible region. Atomic Force Microscopy investigation of silver bionanoparticles identified that they ranged in the size of 250 nm - 680 nm; the work analyzed the antimicrobial efficacy of the silver bionanoparticles against various multi drug resistant clinical isolates. The present Study would be emphasizing on the applicability to synthesize the metallic nanostructures and to understand the biochemical and molecular mechanism of nanoparticles formation by the cell filtrate in order to achieve better control over size and polydispersity of the nanoparticles. This would help to develop nanomedicine against various multi drug resistant human pathogens.

Evaluation Biofilm Sewage Treatment Plant

The research study is carried out to determine the efficiency of the Biofilm sewage treatment plant which is located at the Engineering Complex-s. Wastewater analyses have been carried out at the Environmental Engineering laboratory to study the six parameters: Biochemical Oxygen Demand BOD, Chemical Oxygen Demand COD l, and Total Suspended Solids TSS, Ammoniac Nitrogen NH3-N and Phosphorous P which have been selected to determine the wastewater quality. The plant was designed to treat 750 Pe (population equivalent) at hydraulic retention time of 5 hours in the aerobic zone. The results show that Biofilm wastewater treatment plant was able to treat sewage successfully at different flow condition. The discharge has fulfilled the Malaysia Environmental of Standard A water quality. The achieved BOD removal is more than 85%, COD is more than 80%, TSS is more than 80%, NH3-N is more than 70%, and P was more than 70%. The Biofilm system provides a very efficient process for sewage treatment and it is compact in structure thus minimizes the required land area.