A survey in the Philippines in the early and mid-1990s (Lacierda et al.,2008) revealed that more than 100 chemicals and biological products are used in aquaculture production starting from pond preparation until harvest including chemicals for disease prevention and control. The survey comprises groups of chemicals and some are namely, soil and water conditioners, fertilizers, pesticides, probiotics and feed additives. A probable increase in the usage (volume) and number of chemicals throughout the years could be inferred since world aquaculture production and the number of species for culture also increased (Tacon & Metian, 2008). For health reasons of consumers and the safety regulations imposed by importing countries on aquaculture products, there is a pressing need to survey the chemicals used in aquaculture at present. In line with the promotion of food safety awareness in the region with regards to fish, the objective of this survey was to determine levels of commonly used antibiotics and pesticides in aquaculture that maybe present in aquaculture products such as fish and shrimps

Arnaiz, Margarita T.

Catacutan, Mae R.

Coloso, Relicardo M.

English

Southeast Asian Fisheries Development Center, Aquaculture Department Institutional Repository (SEAFDEC/AQD Institutional Repository )

IMPORTANT FINDINGS AND RECOMMENDATIONS ON CHEMICAL USE IN AQUACULTURE IN SOUTHEAST ASIA 5A survey in the Philippines in the early and mid-1990s (Lacierda et al.,2008) revealed that more than 100 chemicals and biological products are used in aquaculture production starting from pond preparation until harvest including chemicals for disease prevention and control.  The survey comprises groups of chemicals and some are namely, soil and water conditioners, fertilizers, pesticides, probiotics and feed additives.  A probable increase in the usage (volume) and number of chemicals throughout the years could be inferred since world aquaculture production and the number of species for culture also increased (Tacon & Metian, 2008).  For health reasons of consumers and the safety regulations imposed by importing countries on aquaculture products, there is a pressing need to survey the chemicals used in aquaculture at present. In line with the promotion of food safety awareness in the region with regards to fish, the objective of this survey was to determine levels of commonly used antibiotics and pesticides in aquaculture that maybe present in aquaculture products such as fish and shrimps.Chemistry of oxytetracyline (oTC) and oxolinic Acid (oXA)•    Oxytetracycline (OTC). Oxytetracycline or OTC is crystalline and yellow in color with a chemical formula of C22H25ClN2O9 and molecular weight of 496.89 (g/mol).  This antibiotic can cause reproductive toxicity (Category 2) and a possible risk to the unborn child. OTC is harmful when swallowed, can cause skin and eyes irritation and respiratory tract irritation if inhaled.  The LD50 oral in mouse is 6.696 mg/kg (Sigma-Aldrich.com). •   Oxolinic Acid (OXA). Oxolinic acid or OXA is solid, clear, and very faintly yellow in color with a chemical formula of C13H11NO5 and molecular weight of 261.23 (g/mol). This antibiotic is harmful if swallowed. In rats, oral ingestion causes acute toxicity at LD50 of 525 mg/kg while for dermal it is 2,000 mg/kg, and also carcinogenic when ingested and can cause damage to rat DNA (Sigma-Aldrich.com). •   Organochlorine Pesticides (OCPs). The standard of OCPs used in this study was composed of 20 specific pesticides with chemical formulae ranging from C6H6Cl6 to C12H8Cl60 and molecular weights of 290.83 g/mol to 422.92 g/mol.  These OCPs are manufactured chemicals and are not naturally occurring in the environment. These are harmful and toxic substances (restek.com).  One of these is methoxychlor which can affect the organ systems such as the endocrine and reproductive systems.  Endosulfan on the other hand, can alter the gonad structure in juvenile and adult zebrafish and has been detected in significant levels in human milk (Wiley and Krone, 2001; Campoy et al., 2001). Sample Collection and Analytical Methods  Samples of aquaculture products such as fish and crustaceans from either of the following culture systems: ponds, cages or pens were obtained from the three major islands of the Philippines (Luzon, Mindanao & Visayas). Fresh samples were also obtained from wet markets in some localities visited. Live or fresh samples were immediately placed in styrofoam boxes with ice to avoid spoilage. Carefully labeled samples were packed and transported to SEAFDEC/AQD in Tigbauan and kept in the freezer until processed for the assays of residues of antibiotics and pesticides.  Survey of Antibiotic and Pesticide Residues in Aquaculture Products in the Philippines1BY MR CATACUTAN, RM COLOSO AND MT ARNAIZ6 COLOSO, CATACUTAN, ARNAIZFigure 1.1  The chromotogram (A) of the organochlorine pesticides (OCPs) standard showing the peaks and the retention time in minutes (B).BAFigure 1.2 Areas sampled in the Philippines showing (A) results of OTC (Oxytetratcycline) analysis, (B) results of OXA (Oxolinic acid) analysis, and (C) showing results of OCP analysis.B CAThe antibiotics, OTC and OXA were determined in samples by HPLC. For OTC assay, the extraction solvent system of metaphosphoric acid and dichloromethane (DCM) was used while for OXA, the extraction solvent system was acetonitrile and Na2SO4. The OCP in samples were determined by extraction using acetonitrile and purification with Florisil column chromatography and assayed by Gas Chromatography with electron capture detector (GC-ECD). Confirmatory tests for OTC, OXA and OCPs were done in some samples. The standard for the organochlorines showed peaks in the chromatograph with a range of retention time in minutes from 6.188 (alpha-BHC) to 12.179 (Endrin ketone)  (Figure 1.1A and 1.1B). Results and DiscussionA total of 69 aquaculture products were obtained and processed to determine/check for presence of residues of antibiotics (OTC and OXA), and pesticides (OCPs).  These comprised of the following: milkfish, tilapia, sea bass, snapper, grouper, rabbitfish, carp, catfish, silver perch, tiger shrimp, white shrimp and freshwater prawn. Some samples were positive for the antibiotics and OCPs residues and these comprised samples of low and high value fish species (Tables 1, 2 and 3).  The assessment of results were based on the values set for TWA (8-hr time weighted average) and PEL (Permissible Exposure Limits) values based on OSHA (Occupational Safety and Health Administration, US based) and MRLs (Maximum Residue Limits, Japan Food Chemical Research) (Table 4 and IMPORTANT FINDINGS AND RECOMMENDATIONS ON CHEMICAL USE IN AQUACULTURE IN SOUTHEAST ASIA 7Annex A).  The antibiotic OXA in one sample exceeded the PEL and MRL limits (P. vannamei) in Mindanao. In addition, the most common OCP in aquaculture products is Methoxychlor (Annex B) detected in samples from the three regions (Luzon, Visayas, and Mindanao).   In one sample (Macrobrachium sp.) from Luzon, the level of the Endosulfan I (0.01440 ppm) was found harmful based on PEL and MRL (Table 4).  The same sample contained a high level of Endrin ketone (0.02582 ppm) but there is no established PEL and MRL yet for this chemical, however, Endrin has PEL limit of 0.00642 and MRL limit of 0.005. The other OCPs detected that were below the MRL were, Heptachlor epoxide isomer B, Endrin, 4,4’-DDT and Trans-Chlordane.  The results, positive and negative values are indicated in sampled areas in the three regions of the country (Figure 1.2).  Among the antibiotics, OXA was detected in five samples compared to only 3 samples with OTC. The antibiotic OXA has no established PEL but has an MRL, such that the use of OXA has a basis for regulation based on MRL. For the OCPs, the most commonly found are Methoxychlor and groups of Endosulfan and Endrin. The antibiotic OXA is an effective antibacterial against infectious Aeromonas sp. and Flavobacterium columnare, while OTC is indicated for treatment of infectious diseases caused by Gram positive and Gram negative microorganisms such as Mycoplasma pneumonia, E. coli and Haemophilus influenza (Argent Chem Lab.). For fish, OXA is added in the feed at 30 mg/kg of fish per day; while OTC it is immersion in water at 200-700 mg/L of water for 2-6 hours depending on fish species (www.fda.gov). For waste disposal of both antibiotics, these substances can be mixed with a combustible solvent and burned in an incinerator equipped with afterburner and scrubber. For OXA, fish treated with this antibiotic may not be used for human consumption, and for OTC, to be used by qualified personnel for R & D and other authorized use only.The organochlorine endosulfan is cited as one of the most common insecticide used in Asian aquaculture (Rico et al., 2012).  In the Philippines it is used during pond preparation of brackish water ponds for milkfish and tiger shrimp culture at 0.1 ppm or at 8%. (Lacierda et al., 2008).  Also, endosulfan is used as pesticides in agricultural products such as beans, cabbage, eggplant and many others to control growth of insects.  The presence of this particular OCP in aquaculture products could be traced up from its use in pond preparation and in agricultural products and that endosulfan can stay in the soil for several years and is washed down into rivers during rainy season (Coloso, 2003).  In this present report, endosulfan was detected in aquaculture products from Luzon (harmful level) and Visayas.  The presence of other OCPs in aquaculture products could probably follow the same path since water from rivers are utilized in aquaculture ponds.Conclusion and RecommendationsThe antibiotics OXA and OTC were detected in aquaculture products while the most common OCP was Methoxychlor detected in samples from the three regions in the Philippines.  For the other OCPs, namely Endrin ketone/Endrin and Endosulfan I, only sample which was Macrobrachium from Luzon was detected to contain these chemicals in harmful levels based on PEL and MRL.  It is recommended that in future, heavy metals be also determined in aquaculture products and in feeds. AcknowledgementThe commendable technical assistance of Ms. Jessebel Valera is highly acknowledged. 8 COLOSO, CATACUTAN, ARNAIZTable 1.1 Levels of Oxytetracycline (OTC), Oxolinic Acid (OXA), and the Organochlorine Pesticides (OCPs) in fish samples from LuzonAquatic ProductNo. of Samples AnalyzedNo. of  (+) SamplesResidual Analysis (ppm)OTC OXA OCPAyungin or Silver perch 1 1 No spls No spls 0.00074 (Heptachlor epoxide isomerB)0.00093 (Endrin)Bangus or Milkfish 10 1 - - 0.00680 (Methoxychlor)Biya or Goby 1  No spls No spls -Carp (Big Head) 1  - - -Carp (Common) 1  - - -Dalag or Mudfish 1  No spls No spls -Kanduli or Catfish 1  1 - - 0.00353 (Endrin)0.00255 (4,4’-DDT)0.03255 (Methoxychlor)Shrimp (Macrobachium sp.) 1 1 No spls No spls 0.01440 (Endosulfan I)     0.02582 (Endrin ketone)Shrimp (P. monodon) 1  1 - - 0.00124 (trans-Chlordane)Siganid 2  - - -Tilapia (Nile) 9 1 - 0.00496 0.27146 (Methoxychlor)Tilapia (Red) 2 1 - - 0.27425 (Methoxychlor)Total Samples Analyzed 31  27 27 31Table 1.2 Levels of Oxytetracycline (OTC), Oxolinic Acid (OXA), and the Organochlorine Pesticides (OCPs) in fish samples from VisayasAquatic ProductNo. of Samples AnalyzedNo. of  (+) SamplesResidual Analysis (ppm)OTC OXA OCPBangus or Milkish 12 1 - 0.00830 0.01745 (Heptachlor)0.03307 (Methoxychlor)Grouper 2 1 - 0.02004 - 1 0.01121Seabass 2  - - -Shrimp (P. monodon) 5  1 2.51844 - 0.00240 (Endosulfan II) - 0.00345 (Endosulfan sulfate) Siganid 1  - - -Snapper 1 - - -Tilapia 4 - - -Total Samples Analyzed 27  26 26 27Table 1.3 Levels of Oxytetracycline (OTC), Oxolinic Acid (OXA), and the Organochlorine Pesticides (OCPs) in fish samples from MindanaoAquatic ProductNo. of Samples AnalyzedNo. of  (+) SamplesResidual Analysis (ppm)   OTC OXA OCPBangus or Milkfish 5 1 0.04046 0.01006 0.03828 (Methoxychlor)  1   0.00187(Aldrin)Seabass 1  - - -Shrimp (P. monodon) 1  - - -Shrimp (P. vannamei) 1 1 0.78121 - -Siganid 1  - - No splsSnapper 1  - - -Tilapia 1  - - -      Total Samples Analyzed 11  11 11 10Notes for  Tables 1.1, 1.2 and 1.3:ppm = µg residue/g sample; No spls = Sample was not enough for the analysis; Red Values = samples exceed there MRlS for Agricultural chemicals (based on Japan); Black Values = samples Do NoT exceed there MRlS for Agricultural chemicals (based on Japan); “-“ = symbolizes absence of chemical contaminants in the samples (negative)Table 1.4 The list of Minimum Residue Limits (MRLs) for Agricultural Chemicals (Japan Food Chemical Research)*Chemical ContaminantsOSHA Exposure Limits set by US.(ppm TWA)MRLs for Agricultural Chemicals-The Japan Food Chemical Research(ppm) SalmoniformesAnguilliformesPerciformesOther fishShelled MolluscsCrustaceansOther Aquatic animalsAgricultural crops &  poultry supplyAntibiotics         Oxytetracycline (OTC) 0.02460 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.01-0.60Oxolinic Acid (OXA) No PEL established 0.1 0.1 0.06 0.05 - 0.03 - 0.02-20.0OC Pesticides          alpha-BHC No PEL established - - - - - - - 0.2gamma-BHC (Lindane) 0.04204 - - - - - - - 0.2beta-BHC No PEL established - - - - - - - 0.2delta-BHC No PEL established - - - - - - - 0.2Heptachlor 0.00327 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.006-10.0Aldrin 0.00335 0.1 0.1 0.1 0.1 0.1 0.1 0.1 ND, 0.006-0.20Heptachlor epoxide (isomer B) 0.00296 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.006-10.0trans-Chlordane 0.02983 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.002-0.50cis-Chlordane 0.02983 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.002-0.504,4'-DDE No PEL established 3.0 3.0 3.0 3.0 3.0 3.0 3.0 0.02-5.0Endosulfan I 0.00601 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004-30.0Dieldrin 0.01605 0.1 0.1 0.1 0.1 0.1 0.1 0.1 ND, 0.006-0.20Endrin 0.00642 0.005 0.005 0.005 0.005 0.005 0.005 0.005 ND, 0.005-0.104,4'- DDD No PEL established 3.0 3.0 3.0 3.0 3.0 3.0 3.0 0.02-5.0Endosulfan II 0.00601 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004-30.04,4'- DDT 0.06897 3.0 3.0 3.0 3.0 3.0 3.0 3.0 0.02-5.0Endrin aldehyde No PEL established No MRLs yetMethoxychlor 0.70734 - - - - - - - 0.01-7.0Endosulfan Sulfate No PEL established No MRLs yetEndrin ketone No PEL established No MRLs yet*Notes: Salmoniformes (such as salmon and trouts); Anguilliformes (such as eel); Perciformes (such as bonito, horse mackerel, mackerel, sea bass, sea bream and tuna); TWA= 8-hour time weighted average; PEL= Permissible Exposure Limits; TLV= Threshold Limit Values OSHA= Occupational Safety and Health Administration.10 COLOSO, CATACUTAN, ARNAIZ ANNEXMethoxychlor  • CAS ID #: 72-43-5  •  Affected Organ Systems: Endocrine (Glands and Hormones), Neurological (Nervous System), Reproductive (Producing Children) Cancer Effects: None Chemical Classification:  Pesticides (chemicals used for killing pests, such as rodents, insects, or plants) Summary:  Methoxychlor is a manufactured chemical that does not occur naturally in the environment. Pure methoxychlor is a pale-yellow powder with a slight fruity or musty odor. Methoxychlor is used as an insecticide against flies, mosquitoes, cockroaches, chiggers, and a wide variety of other insects. It is used on agricultural crops and livestock, and in animal feed, barns, grain storage bins, home gardens, and on pets. Methoxychlor is also known as DMDT, Marlate®, or Metox®.ReferencesArthu r, J. R., Lavilla-Pitogo, C. R. and Subasinghe, R. P. 2000.Use of Chemicals in Aquaculture in Asia. Proceedings of the meeting on the Use of Chemicals in Aquaculture in Asia, 20-22 May 1996, Tigbauan, Iloilo, Philippines, aquaculture Department, SEAFDEC/AQD. 235 pp.FAO.  1995. Code of Conduct on Responsible Fisheries, FAO, Rome. 41 pp.Camp oy, C., Jimenez, M., Olea-Serrano, M.F., Moreno-Frias, M. Canabate, F., Olea, N. Bayes, R. and Molina-Font, J.A. 2001. Analysis of organochlorine pesticides in human milk: preliminary results. Early Hum. Dev. 65:S 183-190.Carig nan, G., Carrier, K and Sved, S. 1993. Assay of Oxytetracycline residues in salmon muscle by Liquid Chromatography with Ultaviolet detection. J. of AOAC International. 76 (2) 325-328.Colos o, R.M. 2003.Endosulfan: a hidden menace. SEAFDEC Asian Aquaculture.V-25.April-June issue, 1-6pp.Colos o. R. and Borlongan, I. 1999. Significant organotin contamination of sediment and tissues of milkfish in brackish water ponds. Bull. Environ. Contam.Toxicol. 63:297-304.Lond ershausen, M. 2009. Presentation Title “Aquaculture in the light of regulatory requirements in import countries” presented during a conference on “Innovations to Sustain Sea-food Supply” on March 12, 2009, BITEC, Bangkok.Lacie rda, E., de La Pena L. & Lumanlan-Mayo, S. 1996. Use of Chemicals in Aquaculture in the Philippines.  In: Proceedings of the Meeting on the Use of Chemicals in Aquaculture in Asia (JR    Arthur, CP Lavilla-Pitogo, CR Subasinghe, Eds.), 20-22 May 1996, Tigbauan, Iloilo, Philippines, Aquaculture Department, Southeast Asian Fisheries Development Center, Philippines. 155-184 p.Lacie rda, E., Corrre V., Yamamoto, A., Koyama, J. and Matsuoka, T. 2008. Current status on the Use of Chemcials and Biological Products and Health Management Practices in Aquaculture Farms in the Philippines.Mem. Fac. Fish. Kagoshima Univ. 57: 37-45.MFR D Report. 2008. Japanese Trust Fund II. Project on Research and Analysis of Chemical Residues and Contamination in Fish and Fish Products (2004-2008), Technical Compilation of Heavy Metals, Pesticide Residues, Histamine and Drug Residues in Fish and Fish Products in Southeast Asia. p 212.Rico, A., Satapornvanit, K., Haque, M., Min, J, Nguyen P., Telfer, T. and van den Brink, P. 2012. Use of chemicals and biological products in Asian aquaculture and their potential environmental risks: a critical review. Reviews in Aquaculture 4: 75-93.Taco n, A. and Metian, M. 2008. Global overview on the use of fish meal and fish oil in industrially  compounded feeds: Trends and future prospects. Aquaculture 285:146-158.Traini ng Manual on Pesticide Residue Analysis, 6-15 June 2005, Singapore. MFRD. Singapore (Modified).Willey , J.B. and Krone, P.H. 2001. Effects of endosulfan and nonylphenol on the primordial germ cell population in pre-larval zebrafish embryos Aquat. Toxicol. 54: 113-123.

Survey of antibiotic and pesticide residues in aquaculture products in the Philippines

https://repository.seafdec.org.ph/bitstream/10862/5875/3/5875-CatacutanMR2015.pdf

Survey of antibiotic and pesticide residues in aquaculture products in the Philippines

Abstract

Similar works

Full text

Available Versions

Southeast Asian Fisheries Development Center, Aquaculture Department Institutional Repository (SEAFDEC/AQD Institutional Repository )