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Mesophilic acidification of gelatinaceous wastewater

Mesophilic acidification of gelatinaceous wastewater
Mesophilic acidification of gelatinaceous wastewater

Journal of Biotechnology 93(2002)99–108

Mesophilic acidi?cation of gelatinaceous wastewater

Herbert H.P.Fang a,*,Hanqing Yu b

a

Department of Ci 6il Engineering ,Centre for En 6ironmental Engineering Research ,The Uni 6ersity of Hong Kong ,Pokfulam Road ,

Hong Kong ,China

b

Department of Applied Chemistry ,Uni 6ersity of Science and Technology of China ,Hefei ,Anhui 230026,China

Received 26April 2001;received in revised form 19June 2001;accepted 31July 2001

Abstract

The in?uence of hydraulic retention time (HRT)and gelatin concentration on the acidi?cation of gelatinaceous wastewater in an up?ow anaerobic reactor was investigated at pH 5.5and 37°C.The degree of gelatin degradation increased with the HRT,from 84.1%at 4h to 89.6%at 24h,but decreased with the increase of the gelatin concentration in the in?uent from 65.2%at 2g-COD l ?1to 51.9%at 30g-COD l ?1.The degradation of gelatin followed the Monod kinetics with a maximum rate of 1.10g (g-VSS·d)?1and a half-rate constant of 0.23g l ?1.The overall production rate of VFA and alcohols decreased with HRT,from 0.33g (g-VSS·d)?1at 4h to 0.15g (g-VSS·d)?1at 24h,but increased with gelatin concentration in the in?uent,from 0.10g (g-VSS·d)?1at 4g-COD l ?1to 0.58g (g-VSS·d)?1at 30g-COD l ?1.The key acidi?cation products were acetate,propionate and butyrate,plus i -butyrate,valerate,i -valerate,caproate and ethanol in smaller quantities.Formate,methanol,propanol and butanol were found only in certain runs.Only 4.5–7.8%of COD in wastewater was converted to hydrogen and methane.The sludge yield was estimated as 0.32090.014g-VSS (g-COD)?1.?2002Elsevier Science B.V.All rights reserved.

Keywords :Acidi?cation;Gelatin;Hydraulic retention time;Protein;Volatile fatty acid

https://www.sodocs.net/doc/ed15126730.html, /locate /jbiotec

strength industrial wastewater.A number of high-rate processes have been successfully com-mercialized in the past decade (Lettinga,1995;Fang and Liu,2000).Most of the full-scale anaer-obic reactors were designed for treating wastewa-ters from the sugar,starch,and brewery industries,the main pollutants of which are car-bohydrates.However,many industrial and agricultural wastewaters also contain apprec-iable quantities of protein.Treating protein-rich

1.Introduction

Since the introduction of the anaerobic ?lter (Young and McCarty,1969),anaerobic processes have become viable for the treatment of high-

*Corresponding author.Tel.:+852-2859-2660;fax:+852-2559-5337.

E -mail address :hrechef@hkucc.hku.hk (H.H.P.Fang).

0168-1656/02/$-see front matter ?2002Elsevier Science B.V.All rights reserved.PII:S 0168-1656(01)00397-2

H.H.P.Fang,H.Yu/Journal of Biotechnology93(2002)99–108 100

wastewater often results in the formation of scum, which accumulates inside the reactor,and causes sludge washout(Lettinga and Hulshoff Pol, 1991).This problem has signi?cantly hindered the application of the anaerobic process to the treat-ment of wastewaters from dairy and slaughter industries.

In addition,proteins are degraded more slowly than carbohydrates.Anaerobic degradation of proteins is a complex process involving various groups of microorganisms.Proteins are?rst hy-drolyzed and degraded by proteolytic enzymes into peptides and individual amino acids(McIner-ney,1988).The peptides and amino acids are then acidi?ed into volatile fatty acids(VFA),hydro-gen,ammonium,and reduced sulfur.The VFA are further converted by acetogens into acetate and H2/CO2,both of which are lastly converted to methane by methanogens.The initial hydrolysis is the rate-limiting step in protein degradation(Gu-jer and Zehnder,1983),and the overall degrada-tion rate is slow(Harper and Pohland,1986; McInerney,1988).

To improve the process ef?ciency,a two-stage anaerobic process(Pohland and Ghosh,1971)has been developed for the treatment of protein-rich wastewaters.In such a process,hydrolysis and acidi?cation are carried out in the?rst reactor, the ef?uent of which is subsequently further treated in the second reactor for acetogenesis and methane production.Gelatin,a protein rich in animal connective tissue,is the main constituent in slaughterhouse and meat-processing wastewa-ters.Acidogenesis of gelatin in a continuously stirred tank reactor(CSTR)has been studied (Breure and van Andel,1984).Results show that pH is crucial to the acidogenesis ef?ciency and product distribution.Furthermore,gelatin hydrol-ysis was found to be suppressed by the presence of glucose,because the latter is the preferred substrate for the hydrolytic bacteria(Breure et al., 1986).However,little information is available on the acidogenesis of gelatin in wastewaters in con-tinuously fed up?ow reactors.This work was thus conducted to investigate the effects of hydraulic retention time(HRT)and gelatin concentration in wastewater on the acidi?cation of gelatin in a mesophilic up?ow anaerobic reactor.2.Materials and methods

2.1.Reactor and wastewater

The continuous experiment was conducted for 412days in a2.8l up?ow reactor(84mm inside diameter and500mm height)used previously for methanogenic anaerobic degradation(Fang et al., 1994).The reactor was water-jacketed and oper-ated at37°C.Synthetic wastewater was prepared by using gelatin as the sole carbon source,plus balanced nutrient and trace metals,following the formulation used in the previous study(Fang et al.,1994).Throughout the experiment,the pH of the mixed liquor was kept at5.590.1in order to suppress methanogenesis.The reactor was seeded with the sludge taken from a conventional methanogenic reactor treating dairy wastewater for a previous study(Fang and Chung,1999).The initial sludge concentration was10.8g l?1of volatile suspended solids(VSS).The sludge reten-tion was controlled at15days,by wasting one-?fteenth of the sludge in the reactor daily.

This study was conducted in two phases:in phase I,the in?uent COD(chemical oxygen de-mand)was kept at4g l?1,while the HRT was decreased stepwise from the initial24h to16,12, 8,6,and lastly4h;in phase II,the HRT was kept at12h,while the wastewater COD was increased stepwise from2g l?1to9,15,20,and lastly30 g l?1.The reactor was operated at each HRT or COD level for33–44days to ensure reaching steady state before changing the HRT or COD to the next level.

2.2.Analysis

The production of biogas was measured daily by the water displacement method.The contents of H2,CH4,CO2and N2in the biogas were analyzed by a gas chromatograph(Hewlett–Pack-ard,Model5890Series II)equipped with a ther-mal conductivity detector and a2m×2mm (inside diameter)stainless-steel column packed with Porapak N(80–100mesh).Injector and detector temperatures were kept at130and 200°C,respectively,while column temperature was increased from90to110°C.

H .H .P .Fang ,H .Yu /Journal of Biotechnology 93(2002)99–108101

The concentrations of individual acidogenic products in the ef ?uent were determined by a second gas chromatograph of same model equipped with a ?ame ionization detector and a 10m ×0.53mm HP-FFAP fused-silica capillary.The products were mostly VFA,including acetate,propionate,butyrate,i -butyrate,valerate,i -valer-ate,caproate and lactate,and alcohols,including methanol,ethanol,propanol and butanol.Ef ?uent samples were ?ltered through a 0.2m m membrane,acidi ?ed by formic acid,and mea-sured for free acids.The temperature of the column was initially 70°C for 4min,followed by 140°C for 3min,and lastly 170°C for 4min.The temperatures of injector and detector were both 200°C.Helium was used as the carrier gas at a ?ow rate of 25ml min ?1.The detectable levels were 1mg l ?1for individual VFA (from C2to C7)and 3mg l ?1for individual alcohols.The formate concentration was measured by the col-orimetric method (Lang and Lang,1972).Protein was measured by the Lowry –Folin method (Lowry et al.,1951).

Measurements of COD,pH,NH 3-N,and VSS were performed according to the Standard Meth-ods (APHA,AWWA and WEF,1992).

3.Results and discussion

3.1.O 6erall performance

During anaerobic degradation,gelatin was con-verted into VFA and alcohols in the ef ?uent,plus H 2/CO 2in the biogas and biomass.Fig.1illus-trates:(a)HRT;(b)gelatin concentration in in ?u-ent;(c)gelatin concentration in ef ?uent;(d)total VFA and alcohol concentration in ef ?uent;and (e)biogas production rate throughout this study.Results in Fig.1c illustrate that over 84.1%of 4g-COD l ?1of gelatin in the wastewater was de-graded for HRT as low as 4h,and over 89.6%of gelatin up to 30g-COD l ?1in wastewater was degraded at 12h of HRT.Fig.1d illustrates that the ef ?uent VFA /alcohol concentration increased with both HRT and in ?uent gelatin concentra-tion.On treating wastewater containing 4g-COD l ?1of gelatin,the ef ?uent VFA /alcohol

concentration decreased from 1.67g l ?1at 24h of HRT to 0.96g l ?1at 4h of HRT;at 12h of HRT,the ef ?uent VFA /alcohol concentration in-creased from 0.76g l ?1at 2g l ?1of gelatin in wastewater to 7.76g l ?1at 30g-COD l ?1.

Fig.1e illustrates that the total biogas produc-tion rate increased with gelatin concentration in wastewater,but decreased when HRT https://www.sodocs.net/doc/ed15126730.html,pared to the conventional methanogenic pro-cess,the acidogenic process produces a much lower amount of gas due to the suppression of methane production.For example,in a previous

Fig.1.Operational conditions and performance of the acidi ?-cation reactor:(a)HRT;(b)gelatin concentration in in ?uent;(c)gelatin concentration in ef ?uent;(d)total VFA /alcohol concentration in ef ?uent;and (e)biogas production rate.

H .H .P .Fang ,H .Yu /Journal of Biotechnology 93(2002)99–108

102Fig.2.Effect of HRT on:(a)degradation ef ?ciency;and (b)speci ?c gelatin degradation rate.

this study were considerably lower than the value 3.653g (g-VSS ·d)?1found in another study treat-ing gelatin at pH 5.3,4h of HRT and 3.04g l ?1of gelatin in a complete-mix reactor (Breure and van Andel,1984).The lower rates of this study are likely due to the lack of mixing in the up ?ow reactor.However,the rates found in this study were still considerably higher than the protein degradation rate of 0.172g (g-VSS ·d)?1in the acidogenesis of a dairy wastewater using a similar up ?ow reactor (Yu and Fang,2000).This is due to the absence of carbohydrate in the wastewater of this study.Carbohydrates tend to suppress the synthesis of exopeptidases,a group of enzymes facilitating protein hydrolysis (McInerney,1988);their presence in dairy wastewater lowers the protein degradation rate.

The results of Figs.2a and 3a show that gelatin degradation ef ?ciency had a near-linear relation-ship with both HRT and gelatin concentration in wastewater.Based on linear regression analysis,it can be expressed as follows:

ef ?ciency =2.37×HRT ?0.63×S i +47.05r =0.947

(1)

where S i represents the gelatin concentration in wastewater.Eq.(1)shows that gelatin degrada-tion is more dependent on HRT (h)than S i (g l ?1).Therefore,longer HRT is required to increase the gelatin degradation ef ?ciency.

study on mesophilic methanogenesis of proteina-ceous wastewater using the same reactor (Fang et al.,1994),the gas production reached 16.70l (l ·d)?1at 10h of HRT and 10g l ?1of protein in wastewater,whereas the gas production of this acidogenic reactor was only 5.60l (l ·d)?1at 12h of HRT and 15g l ?1of protein in wastewater.

3.2.Gelatin degradation

In anaerobic degradation,gelatin is ?rst hy-drolyzed,and the products of this reaction are further fermented into acids and alcohols.It was found in this study that,gelatin degradation ef ?-ciency increased with HRT,from 84.1%at 4h to 94.3%at 24h (Fig.2a),but decreased with the increase of gelatin concentration in wastewater,from 98.9%at 2g-COD l ?1to 89.6%at 30g-COD l ?1(Fig.3a).The degradation ef ?ciency ranging from 84.1to 98.6%indicates that gelatin was easily hydrolyzed under acidogenic conditions.

Fig.2b illustrates that the speci ?c gelatin degra-dation rate treating 4g-COD l ?1of gelatin de-creased with the increase of HRT,from 0.85g (g-VSS ·d)?1at 4h to 0.25g (g-VSS ·d)?1at 24h.Fig.3b,on the other hand,illustrates that the speci ?c gelatin degradation rate at 12h of HRT increased with gelatin concentration in wastewa-ter,from 0.19g (g-VSS ·d)?1at 2g-COD l ?1to 1.49g (g-VSS ·d)?1at 30g-COD l ?1.The rates in

Fig.3.Effect of gelatin concentration in wastewater on:(a)degradation ef ?ciency;and (b)speci ?c gelatin degradation rate.

H .H .P .Fang ,H .Yu /Journal of Biotechnology 93(2002)99–108103

Fig.4.Gelatin degradation kinetics.

gelatin.In another study on acidi ?cation of gelatin using a complete-mix reactor (Breure and van Andel,1984),the R max and K values were estimated as 0.29g (g-VSS ·d)?1and 3.43g l ?1,respectively.The R max value is substantially lower than that in the present study.This could be due to the differences in reactor con ?guration and operational conditions.In the up ?ow reactor of the present study,the gelatin concentration de-creased as the wastewater ?owed upward,and the gelatin concentration in the ef ?uent was substan-tially lower than the average concentration in the reactor;but in a complete-mix reactor,the gelatin concentration in the ef ?uent equals that in the mixed liquor.

3.4.Production and distribution of VFA and alcohols

Acidi ?cation of gelatin produced not just VFA but also alcohols.Fig.5a illustrates that the over-all production rate of VFA and alcohols de-creased with HRT,from 0.33g (g-VSS ·d)?1at 4h to 0.15g (g-VSS ·d)?1at 24h.Fig.5b,on the other hand,illustrates that the production rate increased with gelatin concentration in wastewa-ter,from 0.10g (g-VSS ·d)?1at 4g-COD l ?1to 0.58g (g-VSS ·d)?1at 30g-COD l ?1.

Tables 1and 2summarize,respectively,the data of total VFA /alcohols concentration in the

3.3.Gelatin degradation kinetics

At steady state,the mass balance of gelatin in the acidogenic reactor may be expressed,assum-ing that the ef ?uent and waste gelatin concentra-tions are equal,as follows:RM =QS i ?(Q ?Q w )S e ?Q w S e (2)

or

R =Q (S i ?S e )/M

(3)

where Q and Q w are the ?ow rates (l d ?1)of ef ?uent and waste sludge,respectively;S i and S e are the in ?uent and ef ?uent gelatin concentrations (g l ?1),respectively;R is the speci ?c gelatin degradation rate (g (g-VSS ·d)?1);M is the total biomass (g-VSS).The gelatin degradation rate may follow the Monod model as:R =R max S e /(K s +S e )

(4)where R max is the maximum gelatin degradation rate (g (g-VSS ·d)?1);S e is the ef ?uent gelatin concentration (g l ?1);and K s is the half-rate con-centration (g l ?1).To best-?t the degradation data,R max and K s were determined as:R max =1.10g (g-VSS ·d)?1K s =0.23g l ?1

Fig.4illustrates that the degradation rates cal-culated from Eq.(4)using the aforementioned parameters ?t the measured data satisfactorily.In a recent study on acidi ?cation of lactose using a similar up ?ow reactor and operational conditions (Fang and Yu,2001),the R max and K values were found to be 4.39g (g-VSS ·d)?1and 1.97g l ?1,respectively.This suggests that acidi ?cation rate of lactose was signi ?cantly higher than that of

Fig.5.Speci ?c VFA /alcohol production rate at various:(a)HRTs;and (b)gelatin concentrations.

H .H .P .Fang ,H .Yu /Journal of Biotechnology 93(2002)99–108105

Fig.6.Ef ?uent propionate concentration at various gelatin concentrations.

production of propionate might only be achieved when less dilute substrate concentration is used.Tables 1and 2also show that production of alcohols was much lower than that of VFA.Etha-nol was the main alcohol produced,accounting for only 2–6%of total VFA /alcohols.Methanol was produced only in certain runs,and never exceeded 2.5%.Propanol and butanol were not detected when gelatin concentration was at 4–8g-COD l ?1.However,at 15g-COD l ?1and higher concentrations,propanol was produced.The proportions of propanol and butanol in-creased with gelatin concentration,but were inde-pendent of HRT.In the acidogenic reactors treating carbohydrate-rich wastewaters,alcohol concentrations could sometimes exceed those of VFA,especially when hydrogen partial pressure is higher than 30kPa (Dabrock et al.,1992;Jones and Woods,1986).However,results of this study show that acidi ?cation of gelatin produced mostly VFA and substantially lower amount of alcohols,as observed previously by others (Breure and van Andel,1984;Jain and Zeikus,1989).

3.5.Ammonium production

Ammonium was produced during the acidi ?ca-tion of protein.As shown in Table 3,the ef ?uent ammonium concentration increased with gelatin concentration,but decreased with the increase of HRT,as expected from the gelatin degradation patterns.The ammonium concentration in the ef ?uent ranged from 0.23to 2.50g l ?1,which is substantially below the threshold level of 5g l ?1above which it becomes toxic to acidogens (Breure and van Andel,1984;Koster and Let-tinga,1988).Thus,the production of ammonium

ef ?uent plus percentages of individual VFA and alcohols at various HRT and gelatin concentra-tions in wastewater.It shows that acetate was the main acidi ?cation product,accounting for 17.8–35.1%of total VFA /alcohols,with an average of 27.4%.The next important VFAs were propi-onate,butyrate and i -butyrate ranging from 10.6to 20.3%,9.0to 15.0%and 10.6to 14.7%,respec-tively.Valerate,i -valerate and caproate and etha-nol were found at lower percentages.Formate,methanol,propanol and butanol were found only in certain runs,at less than 5%,whereas lactate was not detected in all runs.

The product distribution was dependent upon the operational parameters.For instance,the ace-tate concentration increased with both HRT and gelatin concentration in wastewater;but by con-trast,the concentration of propionate in the ef ?uent was independent of HRT,but increased with the gelatin concentration,as illustrated in Fig.6.Since the methanogenesis of propionate is slower compared with acetate and butyrate,pro-pionate was an undesirable intermediate product in the two-stage anaerobic process (Cohen et al.,1984;Harper and Pohland,1986).The engineer-ing implication of this observation is that low

Table 3

Ef ?uent ammonia concentrations at various HRTs and gelatin concentrations in wastewater HRT (h)NH 3-N (g l ?1)Gelatin in wastewater (g-COD l ?1)NH 3-N (g l ?1)40.29590.01520.23390.0170.34990.0260.42490.031460.38690.028890.96490.0570.42490.03112 1.44390.078150.40990.0301620 1.79190.0950.44890.037

24

30

2.50190.106

H .H .P .Fang ,H .Yu /Journal of Biotechnology 93(2002)99–108

106Fig.7.Partial pressures of hydrogen,methane and carbon dioxide at various:(a)HRTs;and (b)gelatin concentrations.

detectable at 16h.Fig.7b illustrates that on treating gelatinaceous wastewater at 12h of HRT,the hydrogen partial pressure increased with gelatin concentration,from undetectable at 2g-COD l ?1to 10kPa at 30g-COD l ?1.The meth-ane partial pressure followed an opposite trend to that of hydrogen,while carbon dioxide partial pressure was steady,ranging from 50to 69kPa.The total COD of hydrogen and methane ac-counted for only 4.5–7.8%of COD in wastewa-ter.It increased with HRT,but decreased with the increase of gelatin concentration.The COD con-version to hydrogen and methane in acidi ?cation was considerably lower than the 86–90%found in a methanogenic reactor treating proteinaceous wastewater (Fang and Chung,1999).3.7.Sludge yield

Sludge yield in an anaerobic treatment system can be estimated based on COD balance (Fang et al.,1994).COD is a wastewater parameter indi-rectly measuring the amount of electrons that are available for oxidation.In a strict anaerobic pro-cess,no electron acceptor is added to the system.In such a case,although the COD in the in ?uent is transformed into VFA,alcohols,hydrogen,methane and biomass,the overall COD should remain unchanged.As a result,the COD differ-ence between in ?uent and ef ?uent should be equal to the COD in biogas,i.e.hydrogen and methane,and COD in the biomass.The biomass COD can,thus,be estimated from the other three terms of COD,all of which can be accurately measured.Assuming the chemical formula of sludge to be C 5H 7NO 2,each gram of biomass is equivalent to 1.42g of COD,and the sludge yield can be estimated accordingly.

appears to have no in ?uence on the acidi ?cation of gelatin in this study.This is also due to the acidic condition in the acidogenic reactor,under which little ammonium is converted to the more toxic form of ammonia.

However,methanogens are more vulnerable to the ammonia toxicity and the increase of pH in the methanogenic reactor would convert ammo-nium into toxic ammonia.Thus,controlling the ammonium concentration could be critical in feeding the acidi ?ed ef ?uent to the methanogenic reactor in the two-stage process.

3.6.Gas production

Fig.7a illustrates that on treating gelatin at 4g-COD l ?1,the hydrogen partial pressure de-creased with the increase of HRT,becoming un-

Table 4

Sludge yields at various HRTs and gelatin concentrations in wastewater HRT (h)Yield (g-VSS (g-COD)?1)Gelatin in wastewater (g-COD l ?1)Yield (g-VSS (g-COD)?1)0.32690.015420.31190.01840.32390.02060.31690.0200.32390.008890.32090.007120.32290.018150.31690.0120.31090.00716200.33090.0210.30190.011

24

30

0.33890.013

H.H.P.Fang,H.Yu/Journal of Biotechnology93(2002)99–108107

Table4summarizes the estimated yield of gelatin-acidifying sludge at various HRTs and gelatin concentrations.The average yield was 0.32090.014g-VSS(g-COD)?1,which is com-parable to that of acidogenic sludge reported in literature ranging from0.230to0.324g-VSS(g-COD)?1(Cohen et al.,1980;Zoetemeyer et al., 1982;Kissalita et al.,1989;Fang and Yu,2001; Yu and Fang,2000).

4.Conclusions

At pH 5.5and37°C,the degree of gelatin degradation increased with the HRT,from84.1% at4h to89.6%at24h,but decreased with the increase of the gelatin concentration in the in?u-ent from65.2%at2g-COD l?1to51.9%at30 g-COD l?1.The degradation of gelatin followed the Monod kinetics with a maximum rate of1.10 g(g-VSS·d)?1and a half-rate constant of0.23 g l?1.The overall production rate of VFA and alcohols decreased with HRT,from0.33g(g-VSS·d)?1at4h to0.15g(g-VSS·d)?1at24h, but increased with gelatin concentration in in?u-ent,from0.10g(g-VSS·d)?1at4g-COD l?1to 0.58g(g-VSS·d)?1at30g-COD l?1.The key acidi?cation products were acetate,propionate and butyrate.Only4.5–7.8%of COD in wastewa-ter was converted to hydrogen and methane.The sludge yield was estimated as0.32090.014g-VSS (g-COD)?1.

Acknowledgements

The authors wish to thank the Hong Kong Research Grants Council for the support of this study(HKU7004/98E),and H.H.P.F.wishes to thank the Croucher Foundation for the Senior Research Fellowship.

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Zoetemeyer,R.J.,Arnoldy,P.,Cohen,A.,Boelhouwer,C., 1982.In?uence of temperature on the anaerobic acidi?ca-tion of glucose in a mixed culture forming part of a two-stage digestion process.Water Res.16,313–321.

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明胶提纯工艺 北京希涛技术开发有限公司 、

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Dear teacher and colleagues: my topic is on “spare time”. It is a huge blessing that we can work 996. Jack Ma said at an Ali's internal communication activity, That means we should work at 9am to 9pm, 6 days a week .I question the entire premise of this piece. but I'm always interested in hearing what successful and especially rich people come up with time .So I finally found out Jack Ma also had said :”i f you don’t put out more time and energy than others ,how can you achieve the success you want? If you do not do 996 when you are young ,when will you ?”I quite agree with the idea that young people should fight for success .But there are a lot of survival activities to do in a day ,I want to focus on how much time they take from us and what can we do with the rest of the time. As all we known ,There are 168 hours in a week .We sleep roughly seven-and-a-half and eight hours a day .so around 56 hours a week . maybe it is slightly different for someone . We do our personal things like eating and bathing and maybe looking after kids -about three hours a day .so around 21 hours a week .And if you are working a full time job ,so 40 hours a week , Oh! Maybe it is impossible for us at

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