natgillin/translations-raw huggingface dataset:
The testes are dissected from each side of the fish.
Manj razgradljive snovi se preskusijo pri nižjih koncentracijah, tj. do 10 mg/l, če se ne pojavijo strupeni učinki.
The testes are generally heavily pigmented, rod shaped structures that are readily apparent at the lateral midline of the body.
Prednost lahko ima izvedba vzporednih preskusov z nizko (približno 2 mg/l) in visoko (približno 10 mg/l) koncentracijo preskusne snovi.
Use either of the following methods:
Vzporedno je treba določiti slepi kisik v steklenicah, ki ne vsebujejo niti preskusne niti referenčne snovi.
Using a pair of fine scissors, begin at the cloaca and make a 1-1,5 cm incision with a single snip angled at about 45 degrees.
Če je treba določiti zaviralne učinke, se v ločenih velikih steklenicah (odstavek 13) pripravijo naslednji nizi raztopin:
Use a scalpel to make a small incision in the side of the fish slightly posterior to the pelvis and just ventral of the lateral plates.
2 mg/l lahko razgradljive snovi, npr. katere koli navedene referenčne snovi;
The testes are removed using fine forceps and placed into a petri dish.
x mg/l preskusne snovi (x je običajno 2);
Each testis is covered with 100 μl freshly made Hank's final solution.
2 mg/l lahko razgradljive snovi plus x mg/l preskusne snovi.
The testes are finely diced by using a razor blade or scalpel.
Če se izkoristi možnost uporabe specifičnih analiz, se lahko izvede fizikalno-kemijski poskus za preverjanje, ali se je preskusna snov odstranila z abiotskimi mehanizmi, kot je hidroliza ali adsorpcija.
This will release sperm and give the Hank's solution a milky appearance.
Fizikalno-kemijski kontrolni preskus se lahko izvede z vnosom živosrebrovega (II) klorida (HgCl2) [11] (50– 100 mg/l) v dve bučki s preskusno snovjo, da se ustavi mikrobna dejavnost.
The fluid containing sperm is added into a tube, while trying not to include any pieces of testes tissue when pipetting.
Znatno zmanjšanje koncentracije specifične snovi med preskusom kaže abiotske mehanizme odstranjevanja.
800 μl of Hank's final solution are added into the tube and mixed well.
Število BPK-steklenic pri značilnem poteku preskusa
If required, the male can be preserved by fixing in 100 % ethanol or other desired fixative.
Pri značilnem poteku preskusa se uporabijo naslednje steklenice:
This is particularly important if the study is assigning parental origin of offsprings.
vsaj 8 steklenic s preskusno snovjo,
Although most of the stock solutions required can be made in advance, stock 5 and subsequently the final solution, should be made up fresh on the day of use.
vsaj 8 steklenic izključno z morsko vodo, ki je okrepljena s hranilnimi snovmi,
Stock 1
vsaj 8 steklenic z referenčno snovjo in po potrebi,
8,00 g
6 steklenic s preskusno in referenčno snovjo (kontrola strupenosti).
0,40 g
Takoj po pripravi je treba vsako raztopino izčrpati iz spodnje četrtine (ne z dna) ustrezno velike steklenice in z njo napolniti ustrezno skupino BPK-steklenic.
Distilled water (DW)
V kontrolah nič (čas nič) se takoj analizira raztopljeni kisik (odstavek 33) ali pa se shranijo za poznejšo kemijsko analizo z obarjanjem z MnCl2 (manganov (II) klorid) in NaOH (natrijev hidroksid).
100 ml
Preostale vzporedne BPK-steklenice se inkubirajo pri preskusni temperaturi (15–20 °C), shranijo v temi in umaknejo z inkubacijskega območja v ustreznih časovnih razmikih (npr. vsaj po 5, 15 in 28 dneh), pri čemer se v njih analizira raztopljeni kisik (odstavek 33).
Stock 2
Vzorci za specifično analizo se 15 minut membransko filtrirajo (0,2–0,45 μm) ali centrifugirajo (neobvezno).
HPO4 (anhydrous)
Če se ne analizirajo takoj, se hranijo pri 2–4 °C za največ 48 ur oz. pri – 18 °C za daljša obdobja (vzorec se pred shranjevanjem zakisa na pH 2, če je znano, da to ne bo vplivalo na snov).
0,358 g
Določanje raztopljenega kisika
0,60 g
Koncentracija raztopljenega kisika se določi s kemijsko ali elektrokemijsko metodo, ki je nacionalno ali mednarodno priznana.
Stock 3
Obdelava rezultatov
0,72 g
Na priložene podatkovne liste se zabeležijo analitski rezultati (Dodatek 5).
50 ml
Biokemijska potreba po kisiku se izračuna kot razlika porabe kisika med slepim vzorcem in raztopino preskusne snovi v preskusnih pogojih.
Stock 4
Neto poraba kisika se deli s koncentracijo (m/v) snovi, da se biokemijska potreba po kisiku izrazi kot BPK/mg preskusne snovi.
MgSO4.7H2O
Razgradnja je opredeljena kot razmerje med biokemijsko potrebo po kisiku in po možnosti teoretično potrebo po kisiku (TPK) ali kemijsko potrebo po kisiku (KPK) ter izražena v odstotkih (glej odstavek 36).
1,23 g
Za vsak čas vzorčenja se za preskusno in referenčno snov izračunajo vrednosti biološke razgradnje z eno od naslednjih enačb:
Stock 5 (freshly prepared)
TPK teoretična potreba po kisiku (izračun, Dodatek 3),
0,35 g
KPK kemijska potreba po kisiku, določena s poskusom.
10 ml
občasno načina izračuna (odstotek teoretične potrebe po kisiku ali odstotek kemijske potrebe po kisiku) ne zagotovita enakih rezultatov;
If you already have some of the above salts but with different water content (i.e. 2H2O instead of anhydrous) you can still use it but first adjust weight based on molecular weight).
priporoča se uporaba teoretične potrebe po kisiku, ker v preskusu kemijske potrebe po kisiku nekatere snovi ne oksidirajo popolnoma.
For Hank's final solution combine in the following order:
Potek preskusa razgradnje se grafično prikaže na diagramu (glej primer v razdelku ‚Veljavnost in razlaga rezultatov‘).
stock 1
Če je na voljo dovolj podatkov, se iz krivulje biološke razgradnje izračunata lag faza (tL) in čas (t50) do 50-odstotne odstranitve od konca lag faze.
1,0 ml
Če se uporabi specifična analiza (neobvezno), se odstotek primarne razgradnje navede kot odstotek odstranitve specifične snovi v preskusnem obdobju (popravljen za analitske slepe vzorce).
stock 2
lokacija in opis mesta vzorčenja:
0,1 ml
značilnosti vzorca (datum vzorčenja, globina, videz, temperatura, slanost, raztopljeni organski ogljik (neobvezno), čas med odvzemom in uporabo v preskusu),
stock 3
uporabljena metoda za določanje kemijske potrebe po kisiku (če se je izvedlo),
8,6 ml
uporabljena metoda za meritve kisika,
stock 4
postopek disperzije za snovi, ki so v preskusnih pogojih slabo topne,
stock 5
uporabljena metoda za določitev števila heterotrofov v morski vodi (metoda štetja kolonij na ploščah ali alternativni postopek),
Mix well before use.
uporabljena metoda za določanje raztopljenega organskega ogljika v morski vodi (neobvezno),
Fertilisation
druge neobvezne metode, uporabljene za opis značilnosti morske vode (merjenje ATP itd.).
Large, gravid females are identified from the desired population;
analitski podatki, navedeni na podatkovnem listu (kot je priložen, Dodatek 5),
females are ready for squeezing only when you can see eggs protruding from the cloaca.
potek preskusa razgradnje, grafično prikazan na diagramu, ki kaže lag fazo (tL), naklon in čas (z začetkom ob koncu lag faze) do 50-odstotne odstranitve končne porabe kisika zaradi oksidacije preskusne snovi (t50).
Ready females have the characteristic “head up” posture.
Lag faza se lahko grafično oceni, kot je prikazano na priloženi sliki, ali se ustrezno šteje kot čas, potreben za 10-odstotno razgradnjo,
Gently run a finger or thumb down the side of the fish towards the tail to encourage the expulsion of an egg sack into a fresh petri dish.
odstotek razgradnje, izmerjen po 28 dneh.
Repeat on the other side and return the fish to its tank.
Respiracija v slepem vzorcu ne sme preseči 30 % kisika v preskusni steklenici.
The eggs can be spread out (forming a monolayer) using a fine paintbrush.
Če tega merila ni mogoče izpolniti s sveže odvzeto morsko vodo, je treba morsko vodo pred uporabo starati (stabilizirati).
It is important to try and expose as many eggs as possible to the sperm so maximising the surface area of the eggs is helpful.
Upoštevati je treba možnost, da lahko snovi, ki vsebujejo dušik, vplivajo na rezultate.
Keep the eggs humid by laying damp tissue around them (it is important the eggs do not touch water directly as this can prematurely harden the chorion preventing fertilisation).
Rezultati, pridobljeni z referenčnima snovema benzoatom in anilinom, morajo biti primerljivi z rezultati, pridobljenimi s krožnim preskusom (3) (odstavek 9).
There is a large variation in the number of eggs each female can produce but as an average, about 150 eggs should be easily obtained from a single gravid female.
Preskusna snov se lahko šteje za zaviralno za bakterije (pri uporabljeni koncentraciji), če je biokemijska potreba zmesi referenčne in preskusne snovi po kisiku manjša od vsote biokemijske potrebe ločenih raztopin obeh snovi po kisiku.
of sperm in Hank's mixture is spread evenly over the whole surface of the eggs using the paintbrush.
Zaradi razmeroma visokih preskusnih koncentracij v primerjavi z večino naravnih sistemov in zato neugodnim razmerjem med koncentracijami preskusne snovi in drugih virov ogljika se metoda šteje za predhodni preskus, ki se lahko uporabi za ugotovitev, ali je snov lahko biološko razgradljiva.
The eggs will quickly harden and change colour (within a minute) once fertilisation has begun.
V skladu s tem nizek rezultat ne pomeni nujno, da preskusna snov v morskem okolju ni biološko razgradljiva, ampak kaže, da bo za to ugotovitev potrebnega več dela.
If the estimated number of eggs is more than 150, repeat the procedure.
Na spodnji sliki je prikazan teoretičen poskus razgradnje, ki kaže izvedljiv način ocene vrednosti tL (dolžina lag faze) in t50, časovnega obdobja (ki se začne pri tL), potrebnega za 50-odstotno končno porabo kisika zaradi oksidacije preskusne snovi.
Similarly if the eggs don't harden within a minute add a bit more sperm.
Bonferronijev U-preskus
Adding more sperm does not necessarily improve fertilisation rate.
Poglavje C.4-B te priloge:
The eggs and the sperm solution should be left to “interact” for at least 15 minutes and the fertilised eggs should be placed into the exposure aquaria within 1,5 hours post fertilisation.
Določanje ‚dobre‘ biorazgradljivosti – Del III: Prilagojeni presejalni preskus OECD.
The procedure is repeated using another female until the desired number of eggs is collected.
Biorazgradnja – preskus zaviranja dihanja aktivnega blata
Spare few eggs from the last batch and fix them in 10 % acetic acid.
Poglavje C.4-E te priloge:
Counting and distributing eggs in test aquaria
Določanje ‚dobre‘ biorazgradljivosti – Del VI:
Eggs should be evenly distributed between each treatment level to avoid genetic bias.
Preskus z zaprto steklenico.
Each batch of fertilised eggs should be separated into equal size groups (as many as the treatment levels) by the use of a blunt instrument (i.e. wide-blade entomology forceps or use of an inoculation loop).
Določanje organskega ogljika v morski vodi
If you aim for 4 replicates per treatment, with 20 eggs each then you need to distribute 80 eggs per exposure aquaria.
Za določanje organskega ogljika v vzorcu vode organske spojine v vzorcu oksidirajo v ogljikov dioksid običajno z eno od naslednjih treh tehnik:
It is advisable to add an extra 20 % (i.e. 96 eggs per treatment level) until you are confident that you obtain 100 % fertilisation rates.
mokra oksidacija s persulfatom/ultravijoličnim obsevanjem,
Stickleback eggs are very prone to fungal infections outside the father-guarded nest.
mokra oksidacija s persulfatom/zvišano temperaturo (116–130 °C),
In this respect, treatment of all eggs with methylene blue during the first 5 days of the test is critically important.
sežiganje.
A stock solution of methylene blue is prepared at 1 mg/ml and added to the exposure aquaria to give a maximum final concentration of 2,125 mg/l.
Sproščeni CO2 se kvantificira z uporabo infrardeče spektrometrije ali titrimetrije.
Sticklebacks should not be exposed to methylene blue once hatched so the system should be free of methylene blue by day 6.
Druga možnost je redukcija CO2 na metan, ki se kvantificira s plamensko ionizacijskim detektorjem (FID).
The eggs are inspected daily and any dead or unfertilised eggs are recorded as such.
Metoda s persulfatom/ultravijoličnim obsevanjem se običajno uporablja za analizo ‚čiste‘ vode z nizko vsebnostjo delcev.
The eggs should never be outside water until they hatch even for very brief periods.
Zadnji metodi se lahko uporabljata za večino vzorcev vode, pri čemer je metoda s persulfatom/zvišano temperaturo najprimernejša za vzorce nizke stopnje, sežigalna tehnika pa za vzorce z vsebnostjo nehlapnega organskega ogljika (NVOC) veliko nad 1 mg C/l.
Hank's Buffered Salt Solution (HBSS):HBSS is needed to preserve the sperm whilst preparing for fertilisation.
Motnje
C.42 BIODEGRADABILITY IN SEAWATER
Vse tri metode so odvisne od izločevanja ali izravnave anorganskega ogljika, ki je prisoten v vzorcu.
GENERAL INTRODUCTION
Najpogosteje uporabljena metoda za izločevanje anorganskega ogljika je spiranje CO2 iz zakisanega vzorca, čeprav se s tem izgubijo tudi hlapne organske spojine (1).
This test method is equivalent to OECD Test Guideline (TG) 306 (1992).
Za vsako matriko vzorčenja je treba zagotoviti popolno izločitev ali izravnavo anorganskega ogljika, glede na vrsto vzorca pa je treba poleg nehlapnega organskega ogljika določiti hlapni organski ogljik (VOC).
When the original test methods were developed, it was not known to what extent results from the screening tests for ready biodegradability using freshwater, and sewage effluent or activated sludge as inoculum, could be applied to the marine environment.
Visoke koncentracije klorida so razlog za manjšo učinkovitost oksidacije z uporabo metode s persulfatom/ultravijoličnim obsevanjem (2).
Variable results on this point have been reported (e.g. (1)).
Vendar se lahko ta motnja prepreči z uporabo oksidacijskega reagenta, spremenjenega z dodajanjem živosrebrovega (II) nitrata.
Many industrial waste waters, containing a variety of chemicals, reach the sea either by direct discharge or via estuaries and rivers in which the residence times are low compared with the period necessary for complete biodegradation of many of the chemicals present.
Priporoča se, da se za oceno posamezne vrste vzorca, ki vsebuje klorid, uporabi največja dovoljena prostornina vzorca.
Because of the growing awareness of the need to protect the marine environment against increasing loads of chemicals and the need to estimate the probable concentration of chemicals in the sea, test methods for biodegradability in seawater have been developed.
Visoke koncentracije soli v vzorcu, analiziranem z metodo sežiganja, lahko povzročijo prekritje katalizatorja s soljo in čezmerno korozijo sežigalne cevi.
These investigations were unsuccessful.
Primer ustrezne metode
The test medium therefore is natural seawater pre-treated to remove coarse particles.
Nehlapen organski ogljik se določi z oksidacijo s persulfatom/ultravijoličnim obsevanjem in nato kvantifikacijo sproščenega CO2, pri čemer se uporabi nedisperzivna infrardeča spektrometrija.
In order to assess ultimate biodegradability with the Shake Flask Method, relatively high concentrations of the test substance have to be used because of the poor sensitivity of the dissolved organic carbon (DOC) analytical method.
Oksidacijski reagent se prilagodi v skladu s predlogi iz (2), kot je opisano v navodilih proizvajalca:
This in turn necessitates the addition to the seawater of mineral nutrients (N and P), the low concentrations of which would otherwise limit the removal of DOC.
8,2 g HgCl2 in 9 6 g Hg(NO3)2 · H2O se raztopi v več sto mililitrih vode z reagentom z nizko koncentracijo ogljika;
It is also necessary to add the nutrients in the Closed Bottle Method because of the concentration of the added test substance.
20 g K2S2O8 se raztopi v raztopini živosrebrove soli;
Hence, the methods are not tests for ready biodegradability since no inoculum is added in addition to the micro-organisms already present in the seawater.
zmesi se doda 5 ml HNO3 (koncentracija);
Neither do the tests simulate the marine environment since nutrients are added and the concentration of test substance is very much higher than would be present in the sea.
reagent se razredči na 1000 ml.
For these reasons the methods are proposed under a new subsection “Biodegradability in Seawater”.
Motnja, ki jo povzroči klorid, se prepreči s 40 μl vzorca za 10-odstotni klorid in 200 μl vzorca za 1,9-odstotni klorid.
APPLICATION
Tudi aktualno (vsebuje opis sistema avtoanalize):
The results of the tests, which would be applied because the pattern of use and disposal of the substance in question indicated a route to the sea, give a first impression of biodegradability in seawater.
Biološka razgradnja v morski vodi
If the result is positive (> 70 % DOC removal; > 60 % ThOD — theoretical oxygen demand), it may be concluded that there is a potential for biodegradation in the marine environment.
PODATKOVNI LIST
However, a negative result does not preclude such a potential but indicates that further study is necessary, for example, using as low a concentration of the test substance as possible.
LABORATORIJ:
In either case, if a more definitive value for the rate or degree of biodegradation in seawater at a particular site is required, other more complex and sophisticated, and hence more costly, methods would have to be applied.
DATUM ZAČETKA PRESKUSA:
For example, a simulation test could be applied using a concentration of test substance nearer to the likely environmental concentration.
PRESKUSNA SNOV:
Also, non-fortified, non-pre-treated seawater taken from the location of interest could be used and primary biodegradation could be followed by specific chemical analysis.
Naziv:
For ultimate biodegradability, 14C-labelled substances would be necessary in order that the rates of the disappearance of soluble organic 14C and the production of 14CO2 at environmentally realistic concentrations could be measured.
Koncentracija založne raztopine:
CHOICE OF METHODS
mg/l kot snov
The selection of which method to use depends on a number of factors;
Začetna koncentracija v mediju, to:
the following Table is given to help the selection.
MORSKA VODA:
While substances of water solubility below the equivalent of about 5 mg C/l cannot be tested in the Shake Flask Method, at least, in principle, poorly soluble substances may be tested in the Closed Bottle Method.
Vir:
Table
Datum odvzema:
Advantages and disadvantages of the shake flask and closed bottle test
Globina odvzema:
METHOD
Videz ob odvzemu (npr. moten itd.):
ADVANTAGES
Slanost ob odvzemu:
DISADVANTAGES
Temperatura ob odvzemu:
SHAKE FLASK
DOC ‚x‘ ur po odvzemu:
simple apparatus except C analyser
Tretiranje pred preskusom (npr. filtriranje, sedimentacija, staranje itd.):
60 d duration is not a problem
Število mikrobnih kolonij
no interference from nitrification
izvirni vzorec:
can be adapted for volatile substances
kolonije/ml
needs C analyser
na začetku preskusa:
uses 5-40 mg DOC/1, could be inhibitory
Druge značilnosti:
DOC determination is difficult at low concentrations in seawater (chloride effect)
DOLOČITVE OGLJIKA:
DOC sometimes high in seawater
Analizator ogljika:
CLOSED BOTTLE
Št. bučke
simple apparatus
DOC po n dneh (mg/l)
simple end determination
Preskus:
uses low concentration of test substance (2 mg/l) thus less chance of inhibition
morska voda s preskusno snovjo, okrepljena s hranilnimi snovmi
easily adapted for volatile substances
sredina, Ca(t)
could be difficult to maintain air-tightness of bottles
sredina, Cb(t)
wall growth of bacteria can lead to false values
Slepi vzorec:
blank O2 uptake values can be high especially after 28 days;
morska voda brez preskusne snovi, okrepljena s hranilnimi snovmi
could be overcome by ageing the seawater
sredina, Cc(t)
possible interference from O2 uptake by nitrification
sredina, Cd(t)
SHAKE FLASK METHOD
sredina,Cblt
This method is a seawater variant of the Modified OECD Screening Test described in Chapter C.4B of this Annex (2).
VREDNOTENJE NEOBDELANIH PODATKOV:
It was finalised as a result of a ring test organized for the European Commission (EC) by the Danish Water Quality Institute (3).
Št. bučke
In common with the accompanying marine Closed Bottle Method, the results from this test are not to be taken as indicators of ready biodegradability, but are to be used specifically for obtaining information about the biodegadability of substances in marine environments.
Izračun rezultatov
A pre-determined amount of the test substance is dissolved in the test medium to yield a concentration of 5-40 mg dissolved organic carbon (DOC)/l.
% razgradnje po n dneh
If the limits of sensitivity of organic carbon analyses are improved, the use of lower concentrations of test substance may be advantageous, particularly for inhibitory substances.
Podobne oblike se lahko uporabijo, če razgradnji sledi specifična analiza ter za referenčne snovi in kontrole strupenosti.
The solution of the test substance in the test medium is incubated under agitation in the dark or in diffuse light under aerobic conditions at a fixed temperature (controlled to ± 2 °C) which will normally be within the range 15-20 °C.
ABIOTSKA RAZGRADNJA (neobvezno)
In cases where the objective of the study is to simulate environmental situations, tests may be carried out beyond this normal temperature range.
Koncentracija DOC (mg/l) v sterilnih kontrolah
The recommended maximum test duration is about 60 days.
Če je med D1 in D2 velika razlika, se srednja vrednost ne izračuna.
Degradation is followed by DOC measurements (ultimate degradation) and, in some cases, by specific analysis (primary degradation).
Izračun teoretične biokemijske potrebe po kisiku
The organic carbon content of the substance must be established, its volatility must be such that significant losses do not occur during the course of the test and its solubility in water should be greater than the equivalent of 25-40 mg C/l.
Teoretična biokemijska potreba po kisiku snovi CcHhClclNnNanaOoPpSs molekulske mase MW se izračuna glede na:
Also, the test substance should not significantly adsorb onto glass surfaces.
Ta izračun pomeni, da se C mineralizira v CO2, H v H2O, P v P2O5 in Na v Na2O.
Information on the purity or the relative proportions of major components of the test substance is required in order that the results obtained can be interpreted, especially when the result lies close to the “pass” level.
Halogen se izloči kot vodikov halid in dušik kot amoniak.
Information on the toxicity of the test substance to bacteria, for example as measured in short-term respiration rate tests (4), may be useful when selecting appropriate test concentrations and may be essential for the correct interpretation of low biodegradation values.
Primer:
However, such information is not always sufficient for interpreting results obtained in the biodegradation test and the procedure described in paragraph 18 is more suitable.
glukoza C6H12O6, MW = 180
Suitable reference substances must be used to check the microbial activity of the seawater sample.
Molekulske mase soli, razen soli alkalnih kovin, se izračunajo ob predpostavki, da so bile soli hidrolizirane.
Sodium benzoate, sodium acetate and aniline are examples of substances which may be used for this purpose.
Predpostavlja se, da je žveplo oksidiralo na stopnjo + 6.
The reference substances must be degraded within a reasonably short time span, otherwise it is recommended that the test be repeated using another seawater sample.
natrijev n-dodecilbenzensulfonatC18H29SO3Na, MW = 348
In the EC ring test where seawater samples were taken at different locations and at different times of the year (3), the lag phase (tL) and time to achieve 50 per cent degradation (t50), excluding the lag phase, were 1 to 4 days and 1 to 7 days respectively for sodium benzoate.
V primeru snovi, ki vsebujejo dušik, se lahko dušik izloči kot amoniak, nitrit ali nitrat, ki ustreza različnim teoretičnim potrebam po kisiku.
For aniline the tL ranged from 0 to 10 days, whilst the t50 ranged from 1 to 10 days.
Če je bila v primeru sekundarnega amina z analizo opažena popolna tvorba nitrata:
REPRODUCIBILITY AND SENSITIVITY OF THE METHOD
Začetna koncentracija v mediju morske vode:
The reproducibility of the method was established in the ring test (3).
TPK ali KPK: