In this study, we analyzed hydroxylated polychlorinated biphenyls (OH-PCBs) in urine of both PCB transport workers and PCB researchers. A method to monitor OH-PCB in urine was developed. Urine was solid-phase extracted with 0.1% ammonia/ methanol (v/v) and glucuronic acid/sulfate conjugates and then decomposed using β-glucuronidase/arylsulfatase. After alkaline digestion/derivatization, the concentration of OH-PCBs was determined by HRGC/HRMS-SIM. In the first sampling campaign, the worker's OH-PCB levels increased several fold after the PCB waste transportation work, indicating exposure to PCBs. The concentration of OH-PCBs in PCB transport workers' urine (0.55~11 μg/g creatinine (Cre)) was higher than in PCB researchers' urine (
Plastic resin pellets collected at Minh Chau island and Ba Lat estuary between 2007 and 2014 in Vietnam were analyzed for dichloro-diphenyl-trichloroethanes (DDTs), polychlorinated biphenyls (PCBs) and hexachlorocyclohexanes (HCHs). The study was carried out as part of the International Pellet Watch program for monitoring the global distribution of persistent organic pollutants (POPs). Higher levels of DDTs compared to PCBs indicated agricultural inputs rather than industrial discharges in the region. Most POP concentrations on both beaches decreased over the period, with the exception of HCH isomers. Though the concentration of DDTs showed a drastic decline on both beaches between 2007/2008 and 2014, DDTs accounted for 60-80% of total DDTs, suggesting that there is still a fresh input of these chemicals in the region. This study strongly recommends further investigations to track temporal and spatial patterns of POP levels in the marine environment using plastic resin pellets.
This study gives the first picture of whole RNA-Sequencing analysis of a PCB-degrading microbe, Rhodococcus jostii RHA1. Genes that were highly expressed in biphenyl-grown cells, compared with pyruvate-grown cells, were chosen based on the Reads Per Kilobase Million (RPKM) value and were summarized based on the criteria of RPKM ≥100 and fold change ≥2.0. Consequently, 266 total genes were identified as genes expressed particularly for the degradation of biphenyl. After comparison with previous microarray data that identified highly-expressed genes, based on a fold change ≥2.0 and p-value ≤0.05, 62 highly-expressed genes from biphenyl-grown cells were determined from both analytical platforms. As these 62 genes involve known PCB degradation genes, such as bph, etb, and ebd, the genes identified in this study can be considered as essential genes for PCB/biphenyl degradation. In the 62 genes, eleven genes encoding hypothetical proteins were highly expressed in the biphenyl-grown cells. Meanwhile, we identified several highly-expressed unannotated DNA regions on the opposite strand. In order to verify the encoded proteins, two regions were cloned into an expression vector. A protein was successfully obtained from one region at approximately 25 kDa from the unannotated strand. Thus, the genome sequence with transcriptomic analysis gives new insight, considering re-annotation of the genome of R. jostii RHA1, and provides a clearer picture of PCB/biphenyl degradation in this strain.
The coral reefs of the Persian Gulf are the most diverse systems of life in the marine environment of the Middle East. Unfortunately, they are highly threatened by local and global stressors, particularly oil pollutants. This is the first quantitative and qualitative study aimed at assessing the concentration and sources of n-alkanes and POPs (PAHs, PCBs and PCNs) in coral tissues, symbiotic algae (zooxanthellae), reef sediments and seawaters in coral reefs of Lark and Kharg in the Persian Gulf, Iran. This work was conducted on eight species of six genera and three families of hard corals and one family of soft coral. A significant variation in the concentration of ∑30n-alkanes and POPs (∑40PAHs, ∑22PCBs and 20PCNs) was found in the decreasing order: zooxanthellae > coral tissue > skeleton > reef sediment > seawater. The bioaccumulation of these compounds was 2-times higher in ahermatypic than in hermatypic corals, among which significant variations were observed in both sites. In Kharg, Porites lutea had the highest mean concentration of ∑30n-alkanes and ∑40PAHs in soft tissue, whereas the lowest values were in Platygyra daedalea. A contrasting trend was documented for ∑22PCBs and 20PCNs, with the highest level reported in soft tissue of P. daedalea and the lowest in P. lutea at Kharg. Compositional pattern of AHs and PAHs demonstrated the predominance of LMW-PAHs and n-alkanes. In skeleton and reef sediments, tetra, penta and tri-CBs were the most abundant PCBs congeners followed by di-CB > hexa-CB > hepta-CB > octa-CB,whiletri-CB > di-CB > tetra-CB > penta-CB > hexa-CB > hepta-CB > octa-CB was observed for soft tissue, zooxanthellae and seawater. The results of RAD test indicated significantly negative correlation between total concentration of these compounds with zooxanthellae density, the chlorophyll-a and C2 in corals at both reefs. This is the first report on levels, health assessment and source apportionments of POPs in zooxanthellae and a first step in the implementation of specific coral reef management measures.
Malaysia is a developing country in Southeast Asia, with rapid industrial and economic growth. Speedy population growth and aggressive consumerism in the past five decades have resulted in environmental pollution issues, including products containing polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs). PCBs and PBDEs are classified as persistent organic pollutants (POPs) by the Stockholm Convention due to their persistence, bioaccumulation in the environment and toxicity to humans and wildlife. These compounds are known to cause liver dysfunction, thyroid toxicity, developmental neuro-toxicity and possibly cancer. PCBs in air, mussels, pellets, seawater, fresh water, and human breast milk samples were analyzed in Malaysia, while studies on the pollution level of PBDEs in Malaysia were conducted on mussels, soils, leachate and sediment samples. PCBs in breast milk collected from Malaysia was the highest among Asian developing countries, with mean concentration of 80 ng/g lipid weight. On the other hand, the mean concentration of PCBs in mussels collected from Malaysia recorded the second lowest, with 56 ng/g and 89 ng/g lipid weight in two studies respectively. The concentrations of PBDEs in mussels taken from Malaysia fall in the range of 0.84-16 ng/g lipid weight, which is considerably low compared to 104.5 ng/g lipid weight in Philippines and 90.59 ng/g in Korea. Nevertheless, there are limited studies on these compounds in Malaysia, particularly there is no research on PBDEs in breast milk and sediment samples. This review will summarize the contamination levels of PCBs and PBDEs in different samples collected from Asian countries since 1988 until 2010 with a focus on Malaysia and will provide needed information for further research in this field.
Contamination of persistent organochlorines (OCs) such as PCBs (polychlorinated biphenyls), DDT and its metabolites (DDTs), HCH (hexachlorocyclohexane) isomers (HCHs), chlordane compounds (CHLs), and HCB (hexachlorobenzene) were examined in mussels collected from coastal waters of Asian countries such as Cambodia, China, Hong Kong, India, Indonesia, Japan, Korea, Malaysia, Philippines, Far East Russia, Singapore, and Vietnam in 1994, 1997, 1998, 1999, and 2001 to elucidate the contamination status, distribution and possible pollution sources and to assess the risks on aquatic organisms and human. OCs were detected in all mussels collected from all the sampling sites investigated. Considerable residue levels of p,p(')-DDT and alpha-HCH were found in mussels and the concentrations of DDTs and HCHs found in mussels from Asian developing countries were higher than those in developed nations suggesting present usage of DDTs and HCHs along the coastal waters of Asian developing countries. On the other hand, lower concentrations of PCBs detected in mussels from Asian developing countries than those in developed countries indicate that PCBs contamination in mussels is strongly related to industrial and activities. To our knowledge, this is a first comprehensive report on monitoring OCs pollution in the Asia-Pacific region.
This study reports the concentrations and congener partners of polychlorinated biphenyls (PCBs) in commercially available paints. Polycyclic-type pigments containing dioxazine violet (pigment violet (PV) 23, PV37) and diketopyrrolopyrrole (PR254, PR255) were found to contain PCB-56, PCB-77, PCB-40, PCB-5, and PCB-12, and PCB-6, PCB-13, and PCB-15, respectively, as major congeners. Dioxazine violet is contaminated with by-products during synthesis from o-dichlorobenzene, which is used as a solvent during synthesis, and diketopyrrolopyrrole is contaminated with by-products during synthesis from p-chlorobenzonitrile. The concentration of PCBs in paint containing PV23 or PV37 was 0.050-29 mg/kg, and toxic equivalency (TEQ) values ranged 1.1-160 pg-TEQ/g. The concentration of PCBs in paint containing PR254 or PR255 was 0.0019-2.4 mg/kg. Naphthol AS is an azo-type pigment, and PCB-52 was detected in paint containing pigment red (PR) 9 with 2,5-dichloroaniline as its source. PCB-146, PCB-149, and PCB-153 were identified from paint containing PR112 produced from 2,4,5-trichloroaniline, as major congeners. These congeners have chlorine positions similar to aniline, indicating that these congeners are by-products obtained during the synthesis of pigments. The concentrations of PCBs in paints containing PR9 and PR112 were 0.0042-0.43 and 0.0044-3.8 mg/kg, respectively. The corresponding TEQ for PR112 was 0.0039-8.6 pg-TEQ/g.
A new species of semiterrestrial gecarcinucid freshwater crab, Terrathelphusa secula, is described from Danum Valley in Sabah, East Malaysia, eastern Borneo. In the form of its carapace, third maxilliped, and male second gonopod, it most closely resembles T. ovis Ng, 1997, and T. telur Ng, 1997, from eastern Sarawak and Brunei, respectively. It differs markedly from these and other congeners in its proportionately much wider carapace, and a male first gonopod that is strongly curved and sickle-shaped.
The search for a high temperature lead-free solder replacement for high temperature leaded solder eutectic alloy has been an evolving process as the threat of a regional lead ban became a reality in July 2006. The advantages and disadvantages of lead-free solder in terms of manufacturing, performance and reliability have been increasingly revealed through companies’ Research and Development (R&D), industrial consortia and university researchers. Materials and component design are the primary criteria to focus on the development for the current generation of high temperature lead-free solder alloys. According to the current status of high temperature lead free soldering, there are many unsolved technical problems such as explanation on the lift-off phenomenon, establishment of high temperature lead-free plating technology, construction of a database of physical properties (solder, parts, PCBs), standardization of high temperature solder materials evaluation technology, and most importantly, the best candidate material for high temperature solder. Clearly, high temperature soldering is one of the unsolved problems of the century in lead-free soldering. Moreover, most of the questions still remain unanswered by researchers. This paper reviews research conducted on the Bi-Ag solder alloy, which is one of the candidate alloys that has been proposed as an alternative for high temperature lead-free solder.
Polychlorinated biphenyls (PCBs) were monitored in surface water collected in the Selangor River basin, Malaysia, to identify the occurrence, distribution, and dechlorination process as well as to assess the potential adverse effects to the Malaysian population. Ten PCB homologs (i.e., mono-CBs to deca-CBs) were quantitated by using gas chromatography-mass spectrometry (GC/MS). The total concentration of PCBs in the 10 sampling sites ranged from limit of detection to 7.67 ng L(-1). The higher chlorinated biphenyls (tetra-CBs to deca-CBs) were almost not detected in most of the sampling sites, whereas lower chlorinated biphenyls (mono-CBs, di-CBs, and tri-CBs) dominated more than 90 % of the 10 homologs in all the sampling sites. Therefore, the PCB load was estimated to be negligible during the sampling period because PCBs have an extremely long half-life. The PCBs, particularly higher chlorinated biphenyls, could be thoroughly dechlorinated to mono-CBs to tri-CBs by microbial decomposition in sediment or could still be accumulated in the sediment. The lower chlorinated biphenyls, however, could be resuspended or desorbed from the sediment because they have faster desorption rates and higher solubility, compared to the higher chlorinated biphenyls. The health risk for the Malaysia population by PCB intake that was estimated from the local fish consumption (7.2 ng kg(-1) bw day(-1)) and tap water consumption (1.5 × 10(-3)-3.1 × 10(-3) ng kg(-1) bw day(-1)) based on the detected PCB levels in the surface water was considered to be minimal. The hazard quotient based on the tolerable daily intake (20 ng kg(-1) bw day(-1)) was estimated at 0.36.
Nineteen pairs of air and seawater samples collected from the equatorial Indian Ocean onboard the Shiyan I from 4/2011 to 5/2011 were analyzed for PCBs and HCB. Gaseous concentrations of ∑(ICES)PCBs (ICES: International Council for the Exploration of the Seas) and HCB were lower than previous data over the study area. Air samples collected near the coast had higher levels of PCBs relative to those collected in the open ocean, which may be influenced by proximity to source regions and air mass origins. Dissolved concentrations of ∑(ICES)PCBs and HCB were 1.4-14 pg L⁻¹ and 0.94-13 pg L⁻¹, with the highest concentrations in the sample collected from Strait of Malacca. Fugacity fractions suggest volatilization of PCBs and HCB from the seawater to air during the cruise, with fluxes of 0.45-34 ng m⁻² d⁻¹ and 0.36-18 ng m⁻² d⁻¹, respectively.
Historical trends of the accumulation of polybrominated diphenyl ethers (PBDEs) and polychlorinated biphenyls (PCBs) in a typical tropical Asian environment were investigated using radio-dated sediment cores from Manila Bay, the Philippines and from the upper Gulf of Thailand. Vertical profiles indicated earlier usage of PCBs than of PBDEs which coincided with their industrial production. The increasing concentrations of total PBDEs and PCBs toward the surface suggested an increased consumption of PBDEs; and possible leakage of PCBs from old machineries into the aquatic environment in recent years. Current input of PCBs to the catchment of Manila Bay was supported by the analyses of air samples and plastic resin pellets. The vertical profiles of total PBDEs in the cores (i.e., rapidly increasing concentrations corresponding to the mid-1980s until mid-1990s, followed by a decrease until the early 2000s, and increasing again toward the surface) likely corresponded to the rapid economic growth in Asia in the 1990s, the Asian financial crisis in 1997, and the economic recovery since early 2000s. BDE-209 was predominant especially on the surface layers. BDEs 47 and 99 generally decreased toward the surface, reflecting the phase-out of the technical penta-PBDE products and the regulation by the Stockholm Convention in recent years. Increasing ratios of BDE-202/209, 206/209, 207/209 and decreasing % of BDE-209 down the core layers may provide evidence for the anaerobic debromination of BDE-209 in the sediment cores. Inventories in ng/cm(2) of total PCBs were higher than total PBDEs (92 vs. 34 and 47 vs. 11 in the Philippines; 47 vs. 33 in Thailand). However, the doubling times indicated faster accumulation of total PBDEs (6-7 years) and BDE-209 (6-7.5 years) than of PCBs (8-11 years). Furthermore, the temporal increase in BDE-209 was comparable to or faster than those reported in other water bodies around the world.
Polychlorinated biphenyl (PCB) was detected as isomer groups (congener numbers 28, 52, 101, 118, 138, 153 and 180) in the coastal water and sediment of four stations around Shadegan wetland protected area in the northwestern part of the Persian Gulf. Total PCB concentration range was 8-375 ng/L in water and 3.4-50.2 μg/g in sediment. Concentration of different congeners and chromatogram indicates that the source of PCB in this area can be Clophen A60; it used for long time in Iranian electronic industries. Other chlorinated hydrocarbons such as lindane, DDT and their metabolites were also present in the samples.
Public concern about the adverse environmental and human health impacts of organochlorine contaminants led to strict regulations on their use in developed nations two decades ago. Nevertheless, DDT and several other organochlorine insecticides are still being used for agriculture and public health programs in developing countries in Asia and the South Pacific. As a consequence, humans in this region are exposed to greater dietary levels of organochlorines. In this review, published information on organochlorine concentrations in foodstuffs from South and Southeast Asia and Oceanic countries has been compiled. Foodstuffs that contribute to human exposures and dietary intakes of organochlorines were examined, and the data compared with those reported from more developed nations. Among various developing countries in Asia, considerable information on organochlorines in foodstuffs has been available from India since the late 1960s. DDT and HCH were the major insecticides in Indian foodstuffs. Concentrations of these insecticides have declined more than two orders of magnitude in farm products, such as food grains and vegetables, in two decades. Milk and milk products are the major sources of dietary exposure to DDT and HCH in India. The residues of these insecticides in dairy products were close to or above the MRLs of the FAO/WHO. Dietary intake of DDT and HCH by Indians was > 100 fold that in more developed nations. Sporadic incidences of greater concentrations (> 1 microgram/g) of aldrin, dieldrin, and heptachlor have been measured in Indian vegetables. Untreated surface waters could be a potential source of DDT and HCH exposure. In most Southeast Asian countries DDT was the common contaminant in animal origin foodstuffs. The higher percentage of p,p'-DDT in meat and fish from Southeast Asian countries, except Japan and Korea, indicated the recent use of DDt in vector control operations. Dietary intakes of DDt and HCH in Southeast Asia were an order of magnitude less than those of Indians but 5- to 10 fold greater than in more developed nations. In addition to DDT, aldrin and dieldrin were prominent in meat collected from Thailand and Malaysia. Aquatic food products from more industrialized countries, such as Japan, South Korea, Hong Kong, and Taiwan, contained significant levels of PCBs. In South Pacific countries, particularly in Australia and New Zealand, chlordanes and PCBs were the most prevalent organochlorines in foodstuffs. Food contamination by DDT, HCH, aldrin, and dieldrin was less than in developing countries in Asia but greater than in the U.S. and Japan. Intake of PCBs in Australia was greater than in the U.S. Meat and fish were the major sources of organochlorine exposure by Australians. Human dietary intake of organochlorines has been declining more slowly in developing countries in Asia. Current intakes were at least 5- to 100 fold greater than those in more developed nations, suggesting a greater risk from organochlorine exposure. Factors such as malnutrition, common among rural poor in developing nations, can increase these risks. Of greatest concern is the magnitude of exposure to organochlorines to which infants and children are subjected through human and dairy milk. The estimated intake of DDT by infants was at least 100 fold greater than the ADI of the FAO/WHO. In addition to DDT, excessive exposures to HCH and dieldrin may cause potential health effects in infants because they are more vulnerable to toxic effects. The design and implementation of appropriate epidemiological studies and their integration with monitoring of human, food, and environmental samples would be a major step in assessing the risks of organochlorine residues in foods and controlling or eliminating them. With the continued globalization of trade in food products, and the concomitant risk that food contaminated through point-source pollution may be widely distributed, identification of sources and their control should be matters of
Field and laboratory studies were conducted to estimate concentration of potential contaminants from landfill in the underlying groundwater, leachate, and surface water. Samples collected in the vicinity of the landfill were analyzed for physiochemical parameters, organic contaminants, and toxic heavy metals. Water quality results obtained were compared from published data and reports. The results indicate serious groundwater and surface water contamination in and around the waste disposal site. Analysis of the organic samples revealed that the site contains polychlorinated biphenyls and other organo-chlorine chemicals, principally chloro-benzenes. Although the amount of PCB concentration discovered was not extreme, their presence indicates a potentially serious environmental threat. Elevated concentrations of lead, copper, nickel, manganese, cadmium, and cobalt at the downgradient indicate that the contamination plume migrated further from the site, and the distribution of metals and metals containing wastes in the site is nonhomogeneous. These results clearly indicate that materials are poorly contained and are at risk of entering the environment. Therefore, full characterization of the dump contents and the integrity of the site are necessary to evaluate the scope of the problem and to identify suitable remediation options.
This paper presents the findings of a study on adsorption of dichlorodiphenythreechloroethen (DDT) and polychlorinated biphenyls (PCBs) on three nanomaterials including Multi walled Carbon Nanotube (MWNT), nano-clay and nano-alumina. DDT and PCBs are of significant concern due their high toxicity and long environmental half-lives. Experiments were conducted using batch adsorption procedures at different DDT and PCBs concentrations, from 10 to 60 mg/L. The amounts of MWNT, nano-clay and Nano-alumina used were 0.25%, 0.50%, 0.75%, 1%, 2% and 10%. The adsorption of PCBs solution onto the MWNT, nano-clay and nano-alumina was characterized by an initial rapid adsorption which eventually became constant within 22, 20, and 17 hours, respectively. The adsorption of DDT solution onto the MWNT, nano-clay and nano-alumina was also characterized by an initial rapid adsorption which gradually became constant within 22, 22 and 16 hours, respectively. Results of this study indicated that MWNT was a better adsorbent material compared to nano-clay and nano-alumina for both contaminants in this study. While at 10% of MWNT 88.9% and 77% of DDT and PCB were removed by MWNT, respectively. The effect of pH and temperature were also investigated.
A total of 127 and 177 seafood samples from Malaysia were analyzed for polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDD/Fs) and dioxin-like polychlorinated biphenyls (dl-PCBs), respectively. The World Health Organization-toxic-equivalency quotients (WHO-TEQ) of PCDD/Fs varied from 0.13 to 1.03 pg TEQ g(-1), whereas dl-PCBs ranged from 0.33 to 1.32 pg TEQ g(-1). Based on food-consumption data from the global environment monitoring system-food contamination monitoring and assessment programme, calculated dietary exposures to PCDD/Fs and dl-PCBs from seafood for the general population in Malaysia were 0.042 and 0.098 pg TEQ kg(-1) body weight day(-1), respectively. These estimations were quite different from the values calculated using the Malaysian food-consumption statistics (average of 0.313 and 0.676 pg TEQ kg(-1) body weight day(-1) for PCDD/Fs and PCBs, respectively). However, both of the dietary exposure estimations were lower than the tolerable daily intake recommended by WHO. Thus, it is suggested that seafood from Malaysia does not pose a notable risk to the health of the average consumer.
An average 50 ml breast milk samples were collected from 21 lactating primiparous mothers (range 25 to 45 years, mean 33 years), 4-8 weeks after delivery in Penang Island, Malaysia. The geometric mean concentration of the most toxic congeners, 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) was 0.14 pg WHO2005-TEQ g-1 zlipid. The most abundant congeners of PCDD/Fs were octachlorodibenzo-p-dioxin (OCDD) (5.9-75.4%), followed by 1,2,3,4,6,7,8-heptachlorodibenzo-p-dioxin (1,2,3,4,6,7,8-HpCDD) (1.1-30.7%). The geometric mean level of total dioxins and dl-PCBs was 2.2 pg WHO2005-TEQ g-1 lipid, significantly lower than those in developed countries or highly contaminated areas. The total dioxins and dl-PCBs in pg WHO2005-TEQ levels in breast milk were significantly correlated with years of residence at potential contaminated site. The average daily intake of 11.8 pg WHO2005-TEQ kg-1 body weight was estimated for a breastfed infant at 6 months of age. This demonstrates the exposure risk to infants, especially from Penang region, to these pollutants from human milk intake are potentially high during the lactation period.
Despite the ubiquity of microplastics (MPs) in aquatic environments and their proven ability to carry a wide variety of chemicals, very little is known about the impacts of virgin or contaminant-loaded MPs on organisms. The primary aim of this study was to investigate the impacts of virgin or phenanthrene (Phe)-loaded low-density polyethylene (LDPE) fragments on a suite of biomarker responses in juvenile African catfish (Clarias gariepinus). Virgin LDPE (50 or 500µg/L) were preloaded with one of two nominal Phe concentrations (10 or 100µg/L) and were exposed to the fish for 96h. Our findings showed one or both Phe treatments significantly increased the degree of tissue change (DTC) in the liver while decreased the transcription levels of forkhead box L2 (foxl2) and tryptophan hydroxylase2 (tph2) in the brain of C. gariepinus. Exposure to either levels of virgin MPs increased the DTC in the liver and plasma albumin: globulin ratio while decreased the transcription levels of tph2. Moreover, MPs modulated (interacted with) the impact of Phe on the DTC in the gill, plasma concentrations of cholesterol, high-density lipoprotein (HDL), total protein (TP), albumin, and globulin, and the transcription levels of fushi tarazu-factor 1 (ftz-f1), gonadotropin-releasing hormone (GnRH), 11 β-hydroxysteroid dehydrogenase type 2 (11β-hsd2), and liver glycogen stores. Results of this study highlight the ability of virgin LDPE fragments to cause toxicity and to modulate the adverse impacts of Phe in C. gariepinus. Due to the wide distribution of MPs and other classes of contaminants in aquatic environments, further studies are urgently needed to elucidate the toxicity of virgin or contaminant-loaded MPs on organisms.
Rhodococcus jostii RHA1 (RHA1) degrades polychlorinated biphenyl (PCB) via co-metabolism with biphenyl. To identify the novel open reading frames (ORFs) that contribute to PCB/biphenyl metabolism in RHA1, we compared chromatin immunoprecipitation chip and transcriptomic data. Six novel ORFs involved in PCB/biphenyl metabolism were identified. Gene deletion mutants of these 6 ORFs were made and were tested for their ability to grow on biphenyl. Interestingly, only the ro10225 deletion mutant showed deficient growth on biphenyl. Analysis of Ro10225 protein function showed that growth of the ro10225 deletion mutant on biphenyl was recovered when exogenous recombinant Ro10225 protein was added to the culture medium. Although Ro10225 protein has no putative secretion signal sequence, partially degraded Ro10225 protein was detected in conditioned medium from wild-type RHA1 grown on biphenyl. This Ro10225 fragment appeared to form a complex with another PCB/biphenyl oxidation enzyme. These results indicated that Ro10225 protein is essential for the formation of the PCB/biphenyl dioxygenase complex in RHA1.