natgillin/translations-raw huggingface dataset:
The stock cultures are grown in conical flasks containing the appropriate medium (volume about 100 ml).
Z običajno 2-parametrsko regresijsko enačbo je pri tej metodi potrebno prilagajanje 3 parametrov in zato zahteva več podatkovnih točk kot nelinearna regresija na podatkih, ki so normalizirani z uporabo vnaprej določenega odziva kontrole.
When the algae are incubated at 20 °C with continuous illumination, a weekly transfer is required.
Obratni intervali zaupanja
During transfer an amount of “old” culture is transferred with sterile pipettes into a flask of fresh medium, so that with the fast-growing species the initial concentration is about 100 times smaller than in the old culture.
Izračun intervalov zaupanja nelinearne regresije z obratno oceno je bolj zapleten in v navadnih statističnih računalniških programskih paketih ni razpoložljiv kot standardna možnost.
The growth rate of a species can be determined from the growth curve.
Približne meje zaupanja se lahko pridobijo s standardnimi programi nelinearne regresije s ponovno parameterizacijo (Bruce in Versteeg, 1992), ki vključuje predelavo matematične enačbe z želenimi ocenjenimi vrednostmi, npr. EC10 in EC50, kot parametri, ki se jih mora oceniti.
If this is known, it is possible to estimate the density at which the culture should be transferred to new medium.
(Naj bo funkcija I = f (α, β, koncentracija) in izkoristimo razmerja definicij f (α, β, EC10) = 0,1 in f (α, β, EC50) = 0,5, da nadomestimo f (α, β, koncentracija) z enakovredno funkcijo g (EC10, EC50, koncentracija).)
This must be done before the culture reaches the death phase.
Bolj neposredni izračun (Andersen idr., 1998) se izvede tako, da obdržimo izvirno enačbo in uporabimo Taylorjev raztezek pri srednjih vrednostih za ri in r0.
Pre-culture:
Nedavno so postale priljubljene metode vezanja.
The pre-culture is intended to give an amount of algae suitable for the inoculation of test cultures.
VIRI
The pre-culture is incubated under the conditions of the test and used when still exponentially growing, normally after an incubation period of 2 to 4 days.
Poglavje C.11 se nadomesti z naslednjim:
When the algal cultures contain deformed or abnormal cells, they must be discarded.
AKTIVNO BLATO, PRESKUS ZAVIRANJA DIHANJA (OKSIDACIJA OGLJIKA IN AMONIJA)
Appendix 5
Ta preskusna metoda je enakovredna Smernici za preskušanje OECD (TG) 209 (2010).
Data analysis by nonlinear regression
Opisana preskusna metoda ocenjuje učinek kemikalije na mikroorganizme iz aktivnega blata (večinoma bakterije) z merjenjem hitrosti dihanja (oksidacije ogljika in/ali amonija) v določenih pogojih pri različnih koncentracijah preskusne kemikalije.
General considerations
Ta metoda temelji na preskusu ETAD (Ecological and Toxicological Association of the Dyestuffs Manufacturing Industry – Združenja za ekologijo in toksikologijo industrije proizvodnje barvil) (1)(2), na prejšnji smernici OECD TG 209 (3) in na revidiranem standardu ISO 8192 (4).
The response in algal tests and other microbial growth tests — growth of biomass — is by nature a continuous or metric variable — a process rate if growth rate is used and its integral over time if biomass is selected.
Namen tega preskusa je omogočiti hitro presejalno metodo, s katero je mogoče oceniti vplive kemikalij na mikroorganizme aktivnega blata v biološki (aerobni) fazi čistilnih naprav za odpadne vode.
Both are referenced to the corresponding mean response of replicate non-exposed controls showing maximum response for the conditions imposed — with light and temperature as primary determining factors in the algal test.
Rezultati tega preskusa lahko služijo tudi kot kazalnik ustreznih koncentracij preskusnih kemikalij, ki niso zaviralne, za uporabo v preskusih biološke razgradljivosti (na primer poglavja C.4 A–F, C.9, C.10, C.12 in C.29 te priloge, OECD TG 302C).
The system is distributed or homogenous and the biomass can be viewed as a continuum without consideration of individual cells.
V tem primeru se preskus lahko izvede kot presejalni, podobno kot preskus za določanje območja delovanja ali mejni preskus (glej odstavek 39), pri čemer se upošteva samo celotno dihanje.
The variance distribution of the type of response for a such system relate solely to experimental factors (described typically by the log-normal or normal distributions of error).
Vendar je treba te informacije pri preskusih lahke biološke razgradljivosti (poglavji C.4 A–F in C.29 te priloge), za katere je koncentracija inokuluma znatno nižja od koncentracije, uporabljene pri tej preskusni metodi, vzeti z rezervo.
This is by contrast to typical bioassay responses with quantal data for which the tolerance (typically binomially distributed) of individual organisms are often assumed to be the dominant variance component.
Vsekakor odsotnost zaviranja pri tem preskusu dihanja ne pomeni samodejno, da bodo pogoji pri preskusu lahke biološke razgradljivosti iz poglavja C.4 A–F ali C.29 te priloge nezaviralni.
Control responses are here zero or background level.
V splošnem se zdi, da je bil preskus zaviranja dihanja, odkar je bil prvič objavljen, uspešno uporabljen, v nekaterih primerih pa je bilo poročano o lažnih rezultatih (2)(4)(5).
In the uncomplicated situation, the normalised or relative response, r, decreases monotonically from 1 (zero inhibition) to 0 (100 per cent inhibition).
Od koncentracije odvisne krivulje dihanja so včasih dvofazne, krivulje odmerek-odziv se popačijo in vrednosti EC50 so nepričakovano nizke (5).
Note, that all responses have an error associated and that apparent negative inhibitions can be calculated as a result of random error only.
Preiskave so pokazale, da do takih rezultatov pride, če aktivno blato v preskusih znatno nitrira in če ima preskusna kemikalija na oksidacijo amonija večji vpliv kot na splošno heterotrofno oksidacijo.
Regression analysis
Te lažne rezultate je zato mogoče preseči z dodatnim preskušanjem, pri katerem se uporabi ustrezen zaviralec nitrifikacije.
Models
Z merjenjem hitrosti privzema kisika v prisotnosti in odsotnosti takšnega zaviralca, npr. n-aliltiouree (ATU), je mogoče ločeno izračunati skupno, heterotrofno in nitrifikacijsko hitrost privzema kisika (4)(7)(8).
A regression analysis aims at quantitatively describing the concentration response curve in the form of a mathematical regression function Y = f (C) or more frequently F(Z) where Z = log C. Used inversely C = f– 1 (Y) allows the calculation of, ECx figures, including the EC50, EC10 and EC20, and their 95 % confidence limits.
Tako je mogoče določiti zaviralne učinke preskusne kemikalije na dveh procesih in na običajen način izračunati vrednosti EC50 za oksidacijo organskega ogljika (heterotrofno) in oksidacijo amonija (nitrifikacijo).
Several simple mathematical functional forms have proved to successfully describe concentration — response relationships obtained in algal growth inhibition tests.
Opozoriti je treba, da je v nekaterih redkih primerih zaviralni učinek n-aliltiouree delno ali popolnoma izničen zaradi kompleksacije s preskusnimi kemikalijami ali dodatki gojišču, npr. z ioni Cu++ (6).
Functions include for instance the logistic equation, the nonsymmetrical Weibul equation and the log normal distribution function, which are all sigmoid curves asymptotically approaching zero for C → 0 and one for C → infinity.
Ioni Cu++ so bistveni za bakterije vrste Nitrosomonas, vendar so v višjih koncentracijah strupeni.
The use of continuous threshold function models (e.g. the Kooijman model “for inhibition of population growth” Kooijman et al.
Potreba po nitrifikaciji pri aerobnem čiščenju odpadnih voda kot nujni korak pri odstranjevanju dušikovih spojin iz odpadnih voda z denitrifikacijo plinastih produktov je postala nuja, še posebej v evropskih državah;
1996) is a recently proposed or alternative to asymptotic models.
Evropska unija je zdaj za koncentracijo dušika v prečiščenih iztokih, odvajanih v sprejemne vode, določila nižje meje [2].
This model assumes no effects at concentrations below a certain threshold EC0+ that is estimated by extrapolation of the response concentration relationship to intercept the concentration axis using a simple continuous function that is not differentiable in the starting point.
Za večino namenov zadošča že samo metoda ocenjevanja vpliva na procese oksidacije organskega ogljika.
Note that the analysis can be a simple minimisation of sums of residual squares (assuming constant variance) or weighted squares if variance heterogeneity is compensated.
Vendar pa je v nekaterih primerih za razlago rezultatov in razumevanje vplivov potrebna preiskava vpliva samo na nitrifikacijo ali pa na oboje, nitrifikacijo in oksidacijo organskega ogljika, pri čemer se to izvede ločeno.
Procedure
NAČELO PRESKUSNE METODE
The procedure can be outlined as follows:
Hitrost dihanja vzorcev aktivnega blata, hranjenega s hranili iz sintetičnih odplak, se meri po treh urah stika v zaprti celici, ki vsebuje kisikovo elektrodo.
Select an appropriate functional equation, Y = f(C), and fit it to the data by non-linear regression.
Ob upoštevanju realističnega scenarija izpostavljenosti so lahko primerni daljši časi stika.
Use preferably the measurements from each individual flask rather than means of replicates, in order to extract as much information from the data as possible.
Če se preskusna kemikalija hitro razkraja, npr. abiotsko s hidrolizo, ali če je hlapna in koncentracije ni mogoče zadovoljivo vzdrževati, je čas izpostavljenosti lahko krajši, npr. 30 minut.
If the variance is high, on the other hand, practical experience suggests that means of replicates may provide a more robust mathematical estimation less influenced by systematic errors in the data, than with each individual data point retained.
Občutljivost vsake šarže aktivnega blata je treba na dan izpostavljenosti preveriti s primerno referenčno kemikalijo.
Plot the fitted curve and the measured data and examine whether the curve fit is appropriate.
Preskus se običajno uporablja za določanje ECx (npr. EC50) preskusne kemikalije in/ali koncentracije brez opaznega učinka (NOEC).
Analysis of residuals may be a particular helpful tool for this purpose.
Zaviranje privzema kisika z mikroorganizmi, ki oksidirajo organski ogljik, je mogoče izraziti ločeno od tistega z mikroorganizmi, ki oksidirajo amonij, in sicer z merjenjem hitrosti privzema kisika v prisotnosti in odsotnosti n-aliltiouree, posebnega zaviralca oksidacije amonija v nitrit, ki na prvi stopnji poteka z nitrifikacijskimi bakterijami.
If the chosen functional relationship to fit the concentration response does not describe well the whole curve or some essential part of it, such as the response at low concentrations, choose another curve fit option — e.g., a non-symmetrical curve like the Weibul function instead of a symmetrical one.
V tem primeru se odstotek zaviranja hitrosti privzema kisika izračuna s primerjavo hitrosti privzema kisika v prisotnosti preskusne kemikalije s srednjo vrednostjo hitrosti privzema kisika ustreznih kontrol, ki ne vsebujejo preskusne kemikalije, v prisotnosti in odsotnosti specifičnega zaviralca, n-aliltiouree.
Negative inhibitions may be a problem with for instance the log — normal distribution function likewise demanding an alternative regression function.
Z določanjem hitrosti v zmeseh preskusne kemikalije, gojišča iz sintetičnih odplak in vode, pri čemer se zanemari aktivno blato, je mogoče zaznati kakršen koli privzem kisika, ki izvira iz abiotskih procesov.
It is not recommended to assign a zero or small positive value to such negative values because this distorts the error distribution.
INFORMACIJE O PRESKUSNI KEMIKALIJI
It may be appropriate to make separate curve fits on parts of the curve such as the low inhibition part to estimate EClowx figures.
Za pravilno razlago rezultatov morajo biti znani identifikacija (po možnosti številka CAS), poimenovanje (IUPAC), čistost, topnost v vodi, parni tlak, hlapnost in adsorpcijske lastnosti preskusne kemikalije.
Calculate from the fitted equation (by “inverse estimation”, C = f– 1(Y)), characteristic point estimates ECx's, and report as a minimum the EC50 and one or two EClow x estimates.
Hlapnih kemikalij običajno ni mogoče zadovoljivo preskusiti, če niso upoštevani posebni varnostni ukrepi (glej odstavek 21).
Experience from practical testing has shown that the precision of the algal test normally allows a reasonably accurate estimation at the 10 % inhibition level if data points are sufficient — unless stimulation occurs at low concentrations as a confounding factor.
UPORABNOST PRESKUSNE METODE
The precision of an EC20 estimate is often considerably better than that of an EC10, because the EC20 is usually positioned on the approximately linear part of the central concentration response curve.
Preskusno metodo je mogoče uporabiti pri vodotopnih, slabo topnih in hlapnih kemikalijah.
So while the EC10 is normally obtainable with a sufficient accuracy it is recommended to report always also the EC20.
Če obstaja kakršna koli negotovost glede stabilnosti preskusne kemikalije ali njene hlapnosti, je treba za natančnejšo določitev koncentracije ECx predložiti dodatne analitske podporne podatke.
Weighting factors
REFERENČNE KEMIKALIJE
The experimental variance generally is not constant and typically includes a proportional component, and a weighted regression is therefore advantageously carried out routinely.
Za zagotovitev zanesljivosti preskusne metode in preskusnih pogojev ter za preverjanje občutljivosti vsake šarže aktivnega blata, ki se na dan izpostavljenosti uporablja kot mikrobni inokulum, je treba referenčne kemikalije redno preskušati.
Weighting factors for a such analysis are normally assumed inversely proportional to the variance:
Kot referenčna zaviralna kemikalija se priporoča 3,5-diklorofenol (3,5-DCP), saj je znan zaviralec dihanja in se uporablja pri mnogih vrstah preskusov zaviranja/strupenosti (4).
Many regression programs allow the option of weighted regression analysis with weighting factors listed in a table.
Kot referenčno kemikalijo pri zaviranju celotnega dihanja (9) je mogoče uporabiti tudi bakrov (II) sulfat pentahidrat.
Conveniently weighting factors should be normalised by multiplying them by n/Σ wi (n is the number of datapoints) so their sum be one.
Kot specifični referenčni zaviralec nitrifikacije se lahko uporabi N-metilanilin (4).
Normalising responses
MERILA ZA VELJAVNOST IN OBNOVLJIVOST
Normalising by the mean control response gives some principle problems and gives rise to a rather complicated variance structure.
Hitrost privzema kisika slepe kontrole (brez preskusne ali referenčne kemikalije) ne sme biti manjša od 20 mg kisika na gram aktivnega blata (suhe teže suspendiranih trdnih snovi) v eni uri.
Unless this error is negligibly small, weighting factors in the regression and confidence limits must be corrected for the covariance with the control (Draper and Smith, 1981).
Koeficient variacije hitrosti privzema kisika pri kontrolnih ponovitvah na koncu dokončnega preskusa ne sme biti večji od 30 %.
Note that high precision on the estimated mean control response is important in order to minimise the overall variance for the relative response.
V mednarodnem krožnem preskusu, ki ga je leta 2004 organizirala Mednarodna organizacija za standardizacijo ISO (4) in v kateri je bilo uporabljeno aktivno blato iz gospodinjskih odplak, je bilo ugotovljeno, da se vrednost EC50 kemikalije 3,5-DCP pri celotnem dihanju giblje med 2 mg/l in 25 mg/l, pri heterotrofnem dihanju med 5 mg/l in 40 mg/l in pri nitrifikacijskem dihanju med 0,1 mg/l in 10 mg/l.
This variance is as follows:
Če vrednost EC50 kemikalije 3,5-DCP ni v pričakovanem območju, je treba preskus ponoviti z aktivnim blatom iz drugega vira.
(Subscript i refers to concentration level i and subscript 0 to the controls)
Vrednost EC50 bakrovega (II) sulfata pentahidrata mora biti za celotno dihanje v območju med 53 mg/l in 155 mg/l (9).
Yi = Relative response = ri/r0 = 1 – I = f(Ci)
Preskusne posode in naprave
with a variance Var(Yi) = Var (ri/r0) ≅ (∂ Yi/∂ ri) · Var(ri) + ((∂ Yi/∂ r0)2 · Var(r0)
Uporabljena mora biti običajna laboratorijska oprema in naslednje:
and since (∂ Yi/∂ ri) = 1/r0 and (∂ Y I/∂ r0) = ri/r02
preskusne posode – na primer 1000-ml čaše za 500 ml reakcijske zmesi (glej št. 5 na sliki 1);
with normally distributed data and mi and m0 replicates:
druga možnost je uporaba BPK-steklenice s primerno prilagoditvijo v obliki obojke za zatesnitev kisikove elektrode na vrat steklenice (glej sliko 2 v Dodatku 3).
Var(ri) = σ2/mi
Priporočljivo je, da skozi obojko najprej potisnete pipeto ali stekleno cevko ali pa uporabite posode z navzven razširjenimi robovi; s tem pri vstavljanju kisikove elektrode preprečite izgubo izpodrinjene tekočine.
the total variance of the relative response Yi thus becomes
V obeh primerih je treba uporabiti metodo z magnetnim mešalom ali kakšno drugo metodo z mešalom, npr. metodo sonde za samodejno mešanje;
An alternative procedure is not to normalise the data and fit the absolute responses including the control response data but introducing the control response value as an additional parameter to be fitted by non linear regression.
naprava za prezračevanje:
With a usual 2 parameter regression equation, this method necessitates the fitting of 3 parameters, and therefore demands more data points than non-linear regression on data that are normalised using a pre-set control response.
po potrebi mora stisnjeni zrak preiti skozi ustrezen filter, ki odstrani prah in olje, ter skozi steklenice za pranje, ki vsebujejo vodo za vlaženje zraka.
Inverse confidence intervals
Vsebino posod je treba prezračiti s Pasteurjevimi pipetami ali drugimi napravami za prezračevanje, ki ne adsorbirajo kemikalij.
The calculation of non-linear regression confidence intervals by inverse estimation is rather complex and not an available standard option in ordinary statistical computer program packages.
Za zadovoljevanje potrebe po kisiku za blato in premagovanje težav s kemikalijami, ki se preveč penijo, so hlapne in se zato izgubijo ali pa jih je pri prezračevanju z razprševanjem zraka težko razpršiti, je mogoče uporabiti krožni stresalnik, ki deluje pri hitrostih kroženja med 150 in 250 vrtljaji na minuto in katerega stekleničke imajo prostornino na primer 2000 ml.
Approximate confidence limits may be obtained with standard non-linear regression programs with re-parameterisation (Bruce and Versteeg, 1992), which involves rewriting the mathematical equation with the desired point estimates, e.g. the EC10 and the EC50 as the parameters to be estimated.
Preskusni sistem običajno sestavlja več čaš, ki so nenehno prezračevane in nameščene zaporedno (npr. v 10- do 15-minutnih časovnih razmikih), nato pa so zaporedno analizirane.
(Let the function be I = f (α, β, Concentration) and utilise the definition relationships f (α, β, EC10) = 0,1 and f (α, β, EC50 ) = 0,5 to substitute f (α, β, concentration ) with an equivalent function g( EC10, EC50, concentration).
Lahko se uporabljajo tudi validirani instrumenti, ki omogočajo hkratno prezračevanje in merjenje hitrosti porabe kisika v zmeseh;
A more direct calculation (Andersen et al, 1998) is performed by retaining the original equation and using a Taylor expansion around the means of ri and r0.
merilnik vrednosti pH;
Recently “boot strap methods” have become popular.
centrifuga, splošna namizna centrifuga za blato, ki zmore hitrost 10000 m/s2.
Such methods use the measured data and a random number generator directed frequent re-sampling to estimate an empirical variance distribution.
Reagenti
REFERENCES
Vseskozi je treba uporabljati analitsko čiste reagente.
Chapter C.11 is replaced by the following:
Voda
ACTIVATED SLUDGE, RESPIRATION INHIBITION TEST (CARBON AND AMMONIUM OXIDATION)
Uporabljati je treba destilirano ali deionizirano vodo, ki vsebuje manj kot 1 mg/l raztopljenega organskega ogljika (DOC), razen če je navedeno, naj se uporablja neklorirana vodovodna voda.
This test method is equivalent to OECD test guideline (TG) 209 (2010).
Hranila iz sintetičnih odplak
This test method describes a method to determine the effects of a chemical on micro-organisms from activated sludge (largely bacteria) by measuring their respiration rate (carbon and/or ammonium oxidation) under defined conditions in the presence of different concentrations of the test chemical.
Gojišče mora biti pripravljeno tako, da vsebuje naslednje sestavine v navedenih količinah:
The test method is based on the ETAD (Ecological and Toxicological Association of the Dyestuffs Manufacturing industry) test (1) ( 2), on the previous OECD TG 209 (3) and on the revised ISO Standard 8192 (4).
pepton
The purpose of the test is to provide a rapid screening method to assess the effects of chemicals on the microorganisms of the activated sludge of the biological (aerobic) stage of waste-water treatment plants.
16 g,
The results of the test may also serve as an indicator of suitable non-inhibitory concentrations of test chemicals to be used in biodegradability tests (for example Chapters C.4 A-F, C.9, C.10, C12 and C.29 of this Annex, OECD TG302C).
mesni ekstrakt (ali primerljiv rastlinski ekstrakt)
In this case, the test can be performed as a screening test, similar to a range-finding or limit test (see paragraph 39), considering the overall respiration only.
11 g,
However, this information should be taken with care for ready biodegradability tests (Chapter C.4 A-F and C.29 of this Annex) for which the inoculum concentration is significantly lower than the one used in this test method.
sečnino
Indeed, an absence of inhibition in this respiration test does not automatically result in non-inhibitory conditions in the ready biodegradability test of Chapters C.4 A-F or C.29 of this Annex.
3 g,
Overall, the respiration inhibition test seems to have been applied successfully since it was first published, but on some occasions spurious results were reported, e.g. (2) (4) (5).
natrijev klorid (NaCl)
Concentration related respiration curves are sometimes bi-phasic, dose-response plots have been distorted and EC50 values have been unexpectedly low (5).
0,7 g,
Investigations showed that such results are obtained when the activated sludge used in the test nitrifies significantly and the test chemical has a greater effect on the oxidation of ammonium than on general heterotrophic oxidation.
kalcijev klorid dihidrat (CaC12, 2H2O)
Therefore, these spurious results may be overcome by performing additional testing using a specific inhibitor of nitrification.
0,4 g,
By measuring the oxygen uptake rates in the presence and absence of such an inhibitor, e.g. N-allylthiourea (ATU), the separate total, heterotrophic and nitrification oxygen uptake rates can be calculated (4) (7) (8).
magnezijev sulfat heptahidrat (MgSO4, 7H2O)
Thus, the inhibitory effects of a test chemical on the two processes may be determined and the EC50 values for both organic carbon oxidation (heterotrophic) and ammonium oxidation (nitrification) may be calculated in the usual way.
0,2 g
It should be noted that in some rare cases, the inhibitory effect of N-allylthiourea may be partially or completely nullified as a result of complexation with test chemicals or medium supplements, e.g. Cu++ ions (6).
brezvodni kalijev dihidrogen fosfat (K2HPO4)
Cu++ ions are essential for Nitrosomonas, but are toxic in higher concentration.
destilirano ali deionizirano vodo do 1 litra.
The need for nitrification in the aerobic treatment of wastewaters, as a necessary step in the process of removing nitrogen compounds from wastewaters by denitrification to gaseous products, has become urgent particularly in European countries;
Vrednost pH te raztopine mora biti 7,5 ± 0,5.
the EU has now set lower limits for the concentration of nitrogen in treated effluents discharged to receiving waters [2].
Če se pripravljeno gojišče ne uporabi takoj, ga je treba najdlje en teden hraniti v temi pri temperaturi od 0 °C do 4 °C ali v pogojih, ki ne povzročajo nikakršnih sprememb v njegovi sestavi.