SPE 61499
Acute Toxicity of Drilling Fluids Used in Mexican Offshore Facilities Tested with
Postlarvae White Shrimp (Litopenaeus Setiferus)
G. Muñoz-Mejía, L. E. Domínguez-Cuéllar, Mexican Petroleum Institute. V. M. Luna-Pabello and R. M. Nuñez-García,
University of Mexico
Copyright 2000, Society of Petroleum Engineers Inc.
This paper was prepared for presentation at the SPE International Conference on Health,
Safety, and the Environment in Oil and Gas Exploration and Production held in Stavanger,
Norway, 26–28 June 2000.
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Abstract
Environmental standards for the regulation of drilling fluids
used in petroleum facilities, land or offshore, do not exist in
Mexican legislation, specially their toxicity limits. Thus, type
and quality of these products and their toxicity are unknown,
mainly those of oil constituents, which are highly toxic to
aquatic biota and their early life cycles. In this study the LC50
(96 hours) of different drilling systems (generic fluids and
field muds) using white shrimp (Litopenaeus setiferus) in
postlarvae stage were evaluated. This organism was selected
because its regional distribution and economic importance.
Also Mysidopsis bahia was used, only for comparing results,
for assessing generic fluids because is applied as reference
specie by USEPA. In order to know the white shrimp
sensitivity, and its potential to be used as reference specie in
Mexico, additional tests using Sodium dodecyl sulfate were
run. A toxicity test procedure using L. setiferus is presented.
Results with both species shown similar responses to generic
fluids evaluations. Only one generic fluid was toxic for both
species while the others did not. In case of field muds only
two were toxic to L. setiferus because they were formulated
with oil. It is recommended to include Litopenaeus setiferus
postlarvae as a reference specie in Mexican Regulations for
drilling fluids due to its distribution in Mexican petroleum
offshore facilities and because of its replication and high
sensitivity (similar to Mysidopsis bahia) in toxicity tests.
Introduction
Nowadays the Sonda of Campeche has been the scenery of
very singular development for Mexico due to in this place
have been joined two productive activities of different
characteristics who represent an important foreign currency
attraction for the country. First, the petroleum activity has
permitted Mexico to be one of the most important petroleum
producers in the world (1); second, the fishery activity, which
for more than 20 years has represented a high fishing rate
because of high diversity of aquatic environments related to
the Sonda which are used for feeding and growing habitats of
a great diversity of economic importance species as fishes and
crustaceans.
In the case of crustaceans, the shrimp fishing in Mexico
represents the 60% of foreign currency obtained by marine
products exportation (2) and Litopenaues setiferus or Gulf
white shrimp is the more abundant specie in coastal Mexican
zone of the Gulf of Mexico and also has higher relationship
with estuarine environments (3); moreover, is an indicator
specie of the interface water-sediment events.
The aim of this study was to evaluate the potential of
Litopenaeus setiferus as test specie for toxicological analysis
of drilling fluids. To answer this, static acute toxicity tests
(LC50-96 hours) were carried on with seven generic drilling
fluids and six drilling muds used in offshore drilling facilities
in the Sonda of Campeche area and with Sodium dodecyl
sulfate as reference toxicant compound. For comparative
purposes, the generic drilling fluids also were evaluated with
Mysidopsis bahia, reference test specie used by United States
Environment Protection Agency (USEPA).
Materials and Methods
Seven generic drilling fluids were used in toxicity tests and
were prepared according with standard methods (4). After
collecting from the return flow-line localized under the shale
shaker, six field drilling muds also were tested: four were
water-based (bentonitic, salty bentonitic, lignosulfonate
inhibed and polimeric) and the other two were oil-based muds.
For the water-based drilling muds, the tests were run in the
suspended particle phase (SPP) following established
procedure (5). The oil drilling muds were tested by placing in
1000 ml flask, 100 ml/900 ml (mud/marine water) by shaking
(10 to 15 minutes) until mix was completed. The sample
obtained was used as initial solution for preparing test
concentrations.
2 G. MUÑOZ-MEJÍA, L.E. DOMÍNGUEZ CUÉLLAR, V. M. LUNA-PABELLO AND R. M. NUÑEZ-GARCÍA
Generic fluids were prepared just before of test starting at
ambient temperature (22 ± 2°C or 71.6 ± 3.6°F ). The field
muds were collected in high density polyethylene pails and
stored at 4 °C for less than a month since they were collected.
All the laboratory and field containers used were washed with
tap water, nitric acid (10% V:V), deionized water, acetone and
marine sinthetic water (6). Sodium dodecyl sulfate
(C12H25NaO4S) selected as a reference toxicant was Sigma
Chemical Co. with a 95% purity.
Mysidopsis bahia toxicity tests were run with juveniles
(age between 4-6 days old) cultured in the Drilling Fluids
laboratory of The Marine Region of Pemex Exploration and
Production. It was used a standard culture (6) and toxicity test
(4) methods.
Litopenaeus setiferus postlarvae from 7 to 34 days old,
produced in laboratory from wild parents collected around
Sonda of Campeche Bay, were used in toxicity tests. The
animals were sent by plane to our laboratory in Mexico, City
and they were maintained in a 1000 liters closed recirculating
system for a week before their use in toxicity tests. The water
for their maintenance and tests was prepared with artificial salt
with free content of phosphate and nitrate. Tap water was used
as water source and was vigorously reared and dechlorinated
with sodium thiosulfate before used. The chloride residual was
less than 0.01 mg/l. The acute toxicity test conditions for
drilling systems and reference toxicant used with Litopenaeus
setiferus, are shown in Table 1.
The LC50 values for the acute toxicity analysis of drilling
systems and reference toxicant, were calculated with the
Probit Method (7) using a LC50 software (8).
Results
A total of 29 acute toxicity tests with Litopenaeus setiferus
and Mysidopsis bahia were run; seven with each specie for
testing generic drilling fluids and fifteen only with L. setiferus
for testing field drilling muds (three water-based and two oil-
based) and reference toxicant (ten tests with Sodium dodecyl
sulfate).
According with the results obtained with M. bahia and L.
setiferus for generic drilling fluids (see Table 2), those species
have similar toxic response. Only the EPA-83-001 fluid was
toxic for those species because the LC50 values were below the
30,000 ppm limit (5, 9). The other six drilling fluids tested
were not toxic for the species selected.
Results obtained are almost in accordance with other paper
(4), in which the authors tested M. bahia with eight of the
seven generic drilling fluids that we have used. As in our
results, they also have found that the EPA-83-001 fluid, which
is KCl polymer, had a toxic response (LC50 of 27,000 ppm in
the SPP test) while the rest of generic drilling fluids were also
non toxic (LC50 above 30,000 ppm).
Our results could indicate that it is possible to use an
endemic specie of shrimp to do drilling fluid toxicity tests and
compare it with the USEPA reference specie (M. bahia). Also,
it is necessary for Mexico, where offshore exploration and
production activities are related to tropical waters, that drilling
SPE61499
fluids toxicity analysis can be made with endemic species
from the area such as Litopenaeus setiferus.
The LC50 results obtained with field drilling muds tested
with Litopenaeus setiferus postlarvae are shown in Table 3. As
can be observed, all the water-based drilling muds accomplish
with the indicated criteria, while the oil-based muds in spite of
could not been evaluated according with standard protocol (5),
because there is not phase separation after setting, the LC50
values were 0.02%. The results, show that only 0.0022 ml of
the total of the oil mud in 99.9978 ml of synthetic marine
water are enough to kill 50% of the exposed organisms, which
means highly toxic even there is no toxicity criteria for oil-
based muds. Fortunately it is prohibited to discharge them in
the sea according with national (10) and international (11)
regulations.
In order to know if certain specie is capable to be used as
toxicity reference organism, is needed to obtain the replication
of the results and the sensitivity of the tested specie (6). In this
study we used Sodium dodecyl sulfate as reference toxicant.
Ten toxicity tests were run since 1996 to 1998 and are shown
in Table 4. The LC50 results were in a range of 4.57 to 10.72
mg/l with a mean of 7.06 ± 1.102 mg/l. The precision of the
tests expressed as percent coefficient of variation (CV%) was
25.18%. This result was obtained nonetheless the animals used
became from wild parents from three different populations
collected on 1996, 1997 and 1998. Several researchers
(12,13,14,15) found in reference toxicants intra-laboratory
toxicity tests program a CV% ranged from 8% to 41%. In
accordance with that, our results accomplish with the
mentioned range and suggest that it is possible to use
postlarvae of Litopenaeus setiferus produced in laboratory
from parents captured in fiel, for toxicity tests and, in a
particular manner, for testing drilling fluids systems used in
Mexican offshore drilling facilities as well as water quality
indicator because of its high sensitivity almost similar to
Mysidopsis bahia.
Conclusions
1. Toxicity of seven generic drilling fluids tested
simultaneously between Mysidopsis bahia and Litopenaeus
setiferus indicated a similar response for both species
according with the limit of 30,000 ppm in the SPP analysis
due to only the EPA-83-001 fluid was toxic while the others
did not.
2. Non acute toxic effects with Litopenaeus setiferus were
found in water-based drilling muds collected in field, while
the oil-based muds were highly toxic although there is not
toxicity criteria that can be applied to this type of muds.
3. Replication and sensitivity tests with Litopenaeus
setiferus using Sodium dodecyl sulfate, have shown a low
variability and, for instance, a constant response that highly
suggest this organism, in postlarvae stage, as a reference
specie for testing drilling systems and also, water quality
indicator in the Sonda of Campeche, Mexico. Moreover,
sensitivity of this organism is also like Mysidopsis bahia,
reference specie used by USEPA.
SPE 61499 ACUTE TOXICITY OF DRILLING FLUIDS USED IN MEXICAN OFFSHORE FACILITIES TESTED
WITH POSTLARVAE WHITE SHRIMP (LITOPENAEUS SETIFERUS)
Acknowledgments
We thank Ana Cosultchi-Marcú and Juan Antonio Morales-
Díaz de Vivar of Mexican Petroleum Institute for the valuable
support in the final revision of this manuscript and Alfredo
Flores-Izaguirre from Marine Region of Pemex Exploration
and Production for the toxicity tests analysis with Mysidopsis
bahia.
References
1. Petróleos Mexicanos.: “Anuario Estadístico de PEMEX”. Gerencia
Corporativa de Evaluación e Información de Petróleos
Mexicanos. México. D.F. (1997) 33.
2. Anónimo.: “Programa de Pesca y Acuacultura 1995-1996”.
SEMARNAP. Mexico, D. F. (1996) 96.
3. Navarrete del Proó, A. et al.: “La pesquería del Camarón en Alta
Mar, Golfo de México y Caribe Mexicano”. XXX Aniversario
del Instituto Nacional de la Pesca. Mexico, Series Pesquerías
relevantes. Tomo I, Crustáceos. (1992) 42.
4. Duke,T. W. et al.: “Acute toxicity of eight Laboratory-prepared
generic drilling fluids to Mysids (Mysidopsis bahia)”. EPA-
600/3-84-067. Gulf Breeze, FL. U.S.A. (1984) 27.
5. Environment Protection Agency. . “Oil and Gas Extraction Point
Source Category. Subpart A-Offshore Subcategory. Drilling
Fluids Toxicity Test”. 40 CFR 435 Ch. I. Appendix 2.
Washington, D.C. U.S.A. (1998). 301.
6. Weber, C. I.: “Methods for Measuring the Acute Toxicity of
Effluents and Receiving Waters to Freshwater and Marine
Organisms”. EPA/600/4-90/027F. Cincinnati, Ohio U.S.A.
(1993) 273.
Table- 1.- Acute toxicity test conditions used with Litopenaeus setiferus for
drilling systems and reference toxicant (Sodium dodecyl sulfate).
Variable
Test type
Duration (hours)
Light intensity (luxes)
pH (Units of pH)
Dissolved oxygen (mg/l)
Salinity (g/l)
Total ammonia (mg/l)
Temperature (°C or °F)
Photoperiod in hours (Light:darkness)
Test aquarium volume (ml)
Test solution volume (ml)
Postlarvae age (days)
Replicates number
Animals per replicate
Dilution water
Food
Response evaluated
Criteria for test accepting Surviving equal or higher than 90% in control aquariums
3
7. Finney, D. J.: “Probit Analysis”. Cambridge University Press.
Cambridge, England. (1971) 333.
8. Stephan, C. E.: “Methods for calculating an LC50. In: F. L. Mayer
and J. L. Hamelink (Eds.), Aquatic Toxicology and Hazard
Evaluation, ASTM STP 534”. Philadelphia, Pennsylvania, USA.
(1977) 65.
9. Jones, F. V.; et al.: “Drilling fluids firms respond to EPA toxicity
concerns”. Technology. J. Oil Gas. (1986) 71.
10. Leyes y Códigos de Mexico.: “Ley General del Equilibrio
Ecológico y la Protección al Ambiente y disposiciones
complementarias” Colección Porrúa. México, D. F. (1996) 656.
11. Environment Protection Agency. “Oil and Gas Extraction Point
Source Category. Subpart A-Offshore Subcategory”. 40 CFR
435 Ch. I. Washington, D.C. U.S.A. (1998) 294.
12. Jop, K. M., et al.: “Use of hexavalent chromium as a reference
toxicant in aquatic toxicity tests”. In: T. M. Poston and R. Purdy
(Eds.). Aquatic Toxicology and Environmental Fate, ASTM STP
921, American Society of Testing and Materials, Philadelphia,
Pennsylvania, U.S.A. (1986) 390.
13. Dorn, P. B. And Rogers, J. H.: “Variability associated with
identification of toxics in National Pollutant Discharge
Elimination System (NPDES) effluent toxicity tests”. Environ.
Toxicol. Chem. (1989) 8:893.
14. Hall, W. S. et al.: “Acute toxicity of industrial surfactants to
Mysidopsis bahia”. Arch. Environ. Contam. Toxicol. (1989)
18:765.
15. Cowgill, U. M. et al.: “The reproducibility of the three brood
Ceriodaphnia test using the reference toxicant sodium lauril
sulfate”. Arch. Environ. Contam. Toxicol. (1990) 19:513.
Conditions
Static non-renewal test
96
600-1000
8-8.5
>3.0
30-32
<0.1
27 ± 1.0 °C or 80.6 ± 1.8 °F
16:8
2000
1500
16-51
3
10
Synthetic
300 Artemia nauplii three times at day
Mortality after 96 hours
4 G. MUÑOZ-MEJÍA, L.E. DOMÍNGUEZ CUÉLLAR, V. M. LUNA-PABELLO AND R. M. NUÑEZ-GARCÍA SPE61499

Table 2.- Acute toxicity results (LC50-96 hours) of generic drilling fluids (SPP)
with Mysidopsis bahia and Litopenaeus setiferus.
Mysidopsis bahia Litopenaeus setiferus
Generic Drilling
LC50-96 hours (5) LC50-96 hours (5)
Mud EPA Criteria EPA Criteria
(ppm) (ppm)
EPA-83-001 23,952 Toxic 12,800.4 Toxic
EPA-83-002 434,109. No toxic >1,000,000 Non toxic
EPA-83-003 131,598 No toxic 131,598.6 Non toxic
EPA-83-004 >1,000,000 No toxic >1,000,000 Non toxic
EPA-83-005 >1,000,000 No toxic 260,936.3 Non toxic
EPA-83-006 >1,000,000 No toxic >1,000,000 Non toxic
EPA-83-007 942,611 No toxic 144,752.90 Non toxic

Table 3- Acute toxicity results (LC50-96 hours) of field

water and oil-based muds using Litopenaeus setiferus.
LC50 -96 hours (5)
Drilling Mud EPA Criteria
(ppm)
Bentonitic >1,000,000 Non toxic
Salty bentonitic >1,000,000 Non toxic
Inhibid Lignosulfonate 33,700 Non toxic
Polimeric >500,000 Non toxic
Oil mud 1 0.02%* There is no criteria
Oil mud 2 0.02%* There is no criteria
* LC50 values expressed as percentage because it was used a volumetric relation of 1:9 of oil
mud and synthetic marine water.

Table 4- LC50-96 hours of Sodium dodecyl sulfate (mg/l) with Litopenaeus

setiferus and correlation coefficient obtained in each test
LC50-96 hours
Test Number Date of the test Correlation coefficient (r)
(mg/l)
001 08-sep-1996 6.58 0.8352
002 27-sep-1996 5.83 0.7548
003 2-oct-1996 10.72 0.9214
004 17-sep-1997 4.57 0.9537
005 02-oct-1997 6.84 0.9819
006 26-may-1998 7.10 0.9555
007 28-may-1998 7.39 0.8778
008 2-jun-1998 9.52 0.9026
009 9-jun-1998 6.18 0.9330
010 15-jun-1998 7.78 0.8956