BIOFOULING IN A SEAWATER REVERSE OSMOSIS PLANT ON THE RED SEA COAST, SAUDI ARABIA1

Mohamed O. Saeed, A.T. Jamaluddin and I. A. Tisan

Saline Water Conversion Corporation P.O.Box 8328, Al-Jubail -31951, Saudi Arabia Tel: + 966-3-343 0012, Fax: + 966-3-343 1615 Email: rdc@swcc.gov.sa

D. A. Lawrence and M. M. Al-Amri

Satellite Plants Department, SWCC, PO Box 6538, Jeddah 21452

Kamran Chida
DuPont Permasep Co., PO Box 1049, Jeddah 21431

ABSTRACT This paper reports on a study that investigated the environmental and pretreatment impact on biofouling in a seawater reverse osmosis (SWRO) plant. The effect of a pretreatment chemical (chlorine), and certain alterations of chemical dosing on membrane biofouling was investigated. The paper also reports on the biofouling potential of the source water, and the effect of chlorination on this biofouling potential. Experiments were carried out to study biofouling in a SWRO Plant, on the Red Sea coast, under a set of four pretreatment modes. These included the normal operation mode, where coagulant is dosed immediately before the media filter and where sodium metabisulfite (SBS) is dosed after the media filter. Secondly, the operation with the coagulant dosing point shifted back to the pressure side of the seawater intake pump. Thirdly, with the SBS dosing point shifted to after the micron cartridge filter, and fourthly while the plant was operating without chlorination/dechlorination. Bacterial generation time and biofilm attachment slides were used to evaluate biofouling. Generation (doubling) times were lowest (higher multiplication capacity) nearest the intake, and they increased gradually along the pretreatment line, becoming the highest, closest to the membranes and in the brine reject. When the SBS was shifted, chlorine removal became closer to the membranes. Following this, generation time in the water samples, taken after the dual media filter (ADMF), after the micron cartridge filter (AMCF) and immediately before the membranes, decreased significantly, reflecting more biofouling potential in the membranes. This correlates well with operational data
1

Published in Desalination, 128 (2000) 177-190.

where the SBS shift has resulted in doubling the frequency of membrane cleaning. The generation times were higher when no chlorine was used, indicating less membrane biofouling potential. Water samples from the plant's intake in the sea had 24 hgeneration time values less, but close to those of chlorinated seawater. This indicated high nutrient load and questionable water quality of the intake. The bacterial attachment to the biofilm slides showed the general trend exhibited by the generation time of planktonic bacteria. The biofilm formation in the brine was generally to a lesser extent than the preceding sampling stations ADMF and AMCF, indicating removal of nutrients along the pretreatment line. However, when the SBS dosing point was shifted closer to the membranes, the number of attached bacteria in the brine increased significantly. Bacteria attached to the biofilm slides randomly and in microcolonies with vesicles in between. Although chlorination enhances biofouling, the current experiments indicate that the questionable quality of source water is one cause of the operational problems in this plant. Key words : Biofouling; Chlorination; Bacteria; Generation time; Water quality. 1. INTRODUCTION

A seawater reverse osmosis (SWRO) plant at Al-Birk, in the southern region of the Red Sea coast of Saudi Arabia, is facing major operational problems which were thought to stem from biofouling. These problems include: increased P build up across

membranes at alarming rates that necessitate frequent membrane cleaning (at times cleaning is needed at 10 days intervals or less), increased membrane flux decline rates, elevated permeate conductivity, and clogging of media and micron cartridge filters. The main cause for the decline in SWRO plant performance in the Middle East has been identified as topical biofilm accumulation on membranes, aided particularly by the large surface areas of the membranes [1]. The bacteriological activity, and hence biofouling problems in SWRO plants, vary with each site. There are site dependent factors which govern the degree of biological activity, such as: temperature, nutrient load, pollution, and the depth and location of the intake. The temperature of the southern region of the Red Sea around the intake of the SWRO plant in Al-Birk is high. For example, it varied between 27.1 to 34.5oC during

the period of March -November, 1997, the period in which this study was undertaken. As indicated by the high bacterial multiplication rate, the nutrient load of the intake seemed to be high, while water currents are minimal and the water column over the intake seems stagnant [current study]. All these factors are conducive to higher bacterial activity. Biofouling of the hollow fine fibers (HFF) membranes which are in use at AlBirk plant has been reported [2,3&4]. Enhanced aftergrowth rates of bacteria were observed by Winters [5]. In conclusion, the biofouling of SWRO plants with surface intake is too well known to need further documentation. To control the biofouling in these plants, chemical

disinfection is used. Conventionally, chlorine has been used as a chemical disinfectant. It was reported [6] that chlorine degrades humic acids and similar high molecular weight compounds present in coastal seawater to smaller molecules that can be assimilated by bacteria. The chlorine suppressed bacterial activity, but when the

sodium metabisulfite (SBS) is added to remove chlorine, the surviving bacteria quickly take advantage of the nutrients furnished by the degradation of larger molecules and enter into a cycle of tremendous growth (termed by some as "aftergrowth"). The significant increase in the biomass of bacteria after dechlorination continued with slime development on the surfaces of the piping and RO membranes [7]. To overcome the aftergrowth problem encountered upon dechlorination, alternate disinfectants have been proposed such as chloramine and copper sulfate [8]. The Umm Lujj SWRO plant, northern Red Sea, Saudi Arabia, has been operating since 1986 with copper sulfate as disinfectant without unexpected operational troubles or membrane fouling. Another approach is to stop chlorination/dechlorination altogether. The no chlorine RO operation has been tried in a commercial facility, operating in warm water with high total dissolved solids. The performance of the plant has been improved and membrane cleaning, which had reached an alarming rate, was cut down significantly [9]. An SWRO plant in the Caribbean was operated successfully without ever using chlorination/dechlorination [10]. Several other plants receiving surface seawater feed of differing characteristics with regard to salinity and temperature have operated

successfully without chlorine. Intermittent chlorination is used only to protect the intake structures from marine fouling [9]. The reported success of the no chlorine RO operation has led to speculation that a similar operation may be an avenue to overcome the biofouling problem at Al-Birk SWRO Plant. This approach looks attractive, particularly when considering the

involvement of M/s DuPont in the reported successful operation without chlorine [9]. DuPont is the manufacturer and supplier of the Permasep B-10 single bundle permeators used at Al-Birk. A joint project between the Saline Water Conversion Corporation (SWCC), represented by its Research and Development Center, and DuPont, was drafted to study the operational problems of the SWCC Al-Birk SWRO Plant. This part of the experiments deals with biofouling which is a major task in this research.

2. (a).

OBJECTIVES Investigate status and the potential of biofouling in seawater, and pretreated feed water at different operational modes with chlorination/dechlorination, and without chlorine.

(b). Suggest possible measures to alleviate the biofouling problem.

3.

EXPERIMENTAL

Experiments were designed to study the biofouling in the SWCC Al-Birk SWRO Plant. 3.1 Project Phases

The study was divided into 4 operational phases with respect to chemical pretreatment as follows: 3.1.1 Plant Operating as is

In this mode of regular operation, the coagulant "magnifloc C-573" is injected just ahead of the coarse sand filters, chlorine (Calcium hypochlorite) is dosed in the intake

pit, while sodium metabisulfite (SBS) is dosed to remove chlorine after dual media filters ahead of micron cartridge filters.
3.1.2 Coagulant Dosing Point Shift

The dosing of coagulant was shifted from before coarse sand filters up stream to the pressure side of the seawater pump (a distance of about 85 m from the regular dosing point mentioned above).
3.1.3

SBS Dosing Point Shifts

The SBS dosing point was shifted from before to after the micron cartridge filter (a distance of about 13 m ahead of the high pressure pump). A second point was selected by moving the SBS dosing to a location just ahead (1 m) of the high pressure pump. This shift was meant to ascertain the effect of maintaining chlorinated flow over the low pressure section between the micron cartridge filter and the high pressure pump. This piping section was suspected to be an additional biofouling source.
3.1.4

Plant Operating Without Chlorination/Dechlorination

The coagulant dosing point remained as in 3.1.2 above. For all phases; analyses were carried out using water samples for enumeration of bacteria, and biofilm samplers with studs (coupons) were fixed along the pretreatment line to study bacterial attachment. Water samples were taken from 6 sampling stations: (1) The pressure side of the seawater intake pump, (2) Before the media filter, (3) After the media filter, (4) After the micron cartridge filter, (5) Ahead of the membranes (not sampled during coagulant shift), (6) The brine reject of B-10 permeators. The biofilm samplers were fixed in 4 locations as follows: (1) The pressure side of the seawater intake pump, (2) After the media filter, (3) After the micron cartridge filter, (4) Brine reject.

3.2

Bacteria in Water Samples

In the chlorinated sampling stations, water samples were taken in sterile plastic sampling bags containing sodium thiosulfate as dechlorinator, while plain sterile bags without the dechlorinator were used to collect water samples from dechlorinated sampling stations. Bacteria were counted immediately after sampling and this count was designated 0-h count. Further counting was carried out after 24h (24-h count) and 72 h (72-h count) following the incubation of samples at a temperature of 30 oC in a thermostatically controlled incubator. The samples were first mixed well on a vortex mixer, and a pour plate count in marine agar was employed to reveal the colony forming units (CFU) [11,12]. Briefly; the samples were first serially diluted in ten-fold steps in filtered (5 m) and heat sterilized seawater. Three dilutions from each sample were plated to enable a suitable number of bacteria to be counted. For each dilution, three replicates of 1 ml were seeded in separate sterile petri dishes. Thirteen milliliters of Molten marine agar at 46 oC were added to each dish. In each dish, the agar was mixed well with the 1 ml sample. The dishes were then allowed to cool for 30 min at room temperature and then incubated (inverted) in the constant temperature incubator. The colony forming units were counted after 96-h of incubation.

Water samples from the vicinity of the plant intake in the sea at 5 m depth, which is the depth of the intake chamber's inlet, were also analyzed for comparison with chlorinated intake seawater.

Zero-hour counts were used as a base to calculate the generation time for 24h and 72 h of incubation as per the following formula [13]: Generation time (h) = t K / (ln Nt - ln Nt0), where: t = 24 for 24 h - generation time and 72 for 72 h - generation time, Nt = count at 24 or 72 h and Nt0 count at 0h. The generation time reflects the speed of bacterial multiplication and was used as an index of nutrient load of water samples. K = 0.693

3.2

Biofilm Bacteria

Biofilm samplers were installed in four locations as mentioned above. Each sampling unit contained six holders, each with a stud (coupon) of glass slide measuring 2.5 x 2.2 cm. In each sampling station, water was diverted to flow through the sampler at a rate of 10 l/min. After 20 days the slides were retrieved and the biofilm attached was scraped off for enumeration of attached bacteria. The slide was aseptically removed from the holder, placed into a sterile petri dish, and then flooded with a measured quantity of sterile seawater. The biofilm was scraped first by a sterile dissecting knife and then by a sterile cotton swab. The slide was removed and the biofilm suspension was transferred to a test tube and mixed vigorously on a vortex mixer. The suspension was then serially diluted and grown in marine agar, employing the pour plate technique as described. Following 96 h of incubation at 30 oC the number of bacteria (CFU) was obtained and expressed as CFU/cm2. A scanning electron microscope was used to reveal bacteria attached to biofilm slides. Slides with intact biofilms were air dried and placed on a brass plug using double sided carbon (conductive) tape. They were then coated with 30 nm of gold, using JEOL-1100E ion sputtering device. The specimens (slides) were placed in the JEOL model JSM-5300LV scanning electron microscope and were observed at 2000, 3500, and 5000 magnification using 20 KV accelerating voltage at 10 mm working distance.

3.3

Analysis of Data

Data was subjected to rigorous statistical comparisons using an analysis of variance to reveal differences between means and LSD test to distinguish differing means.

4. 4.1

RESULTS Bacteriological Analysis of Water Samples

Bacterial generation times (doubling times) were computed for all phases of the project and compared statistically (Table 1). The general trend is that the time needed for bacterial cells to divide (generation time), increases along the pretreatment line, is lowest in the chlorinated seawater feed and is highest in the brine reject. The table also shows that generation times from the no chlorine phase are consistently higher than

from the other three phases when the plant was running with chlorination/ dechlorination. Table 2 shows the bacterial density and the generation times of water samples from Al-Birk SWRO Plant intake site at a depth of 5 m. The water currents around the intake were minimal and the water column seemed stagnant. A comparison of the generation time of the intake samples and those of chlorinated seawater shows longer generation times for the intake samples than the chlorinated water samples (Table 2). Table 3 gives a comparison of bacterial counts and generation times during the two SBS dosing shifts. In chlorinated seawater the 0-h count was low when compared to sampling stations with no chlorine. However, upon dechlorination there was a rapid bacterial growth which resulted in lower generation times as compared to samples with no chlorine (whether chlorine was removed prior to these sampling stations or no chlorine was used). 4.2 Biofilm Studies

Table 4 presents bacterial attachment to biofilm sampling slides at four stations along the pretreatment line during the four phases of the project. The table shows there was a 2-log reduction of bacteria attached to biofilm slides after the micron cartridge filter (MCF) when this location became chlorinated following the SBS dosing shift. The number of attached bacteria was consistently and significantly lower in the brine than either after the dual media filter (DMF) or the MCF, when the SBS dosing point was before the MCF. However, when the SBS dosing point was shifted to after MCF, significantly more bacteria were attached in the brine than after DMF or after MCF. In this phase (SBS shift), the number of bacteria attached to the slides in the brine reject was higher than those attached in the same sampling station during the first two phases of the project, i.e., when the SBS dosing point was before the MCF. During the no chlorine phase, the bacterial attachment was steady at 5 logs in all sampling stations except in the after DMF where it registered an increase of 1 log over the other sampling stations.

Figure 1 shows scanning electron micrographs of the bacteria attached to microscope slides at different stations during various phases of the project. The biofilm is not a homogeneous continuous lawn, rather, it is an aggregation of cells in twisting patches creating a network of vesicles or channels between them. Cells are rod shaped, more or

less uniform in size, and many take a curved (vibrioid) shape. Many cells show biopolar bodies and possess capsules.

5.

DISCUSSION

Biofouling is not a problem unique to a particular SWRO plant but is a major concern in many such plants operating on surface seawater feed. The Al-Birk SWRO Plant is more vulnerable to biofouling because the geographic location is in a hot climate which is more conducive to biofouling. One way to study this biological activity is by quantitative estimation of bacterial biomass. Whilst the estimation of numbers of free bacteria in water (planktonic) may initially appear a relatively simple task, it is in practice fraught with problems. There is a wide natural variation in the number of bacteria in seawater because many factors make their numbers change constantly, such as light, temperature, tide, currents, turbidity and nutrients. An added variation source along the pretreatment line is the untimed detachment of bacteria from biofilm in the system into water lines and possibly in the water samples. In this study the variation in the same location was very noticeable, giving a standard deviation of close to the average count. Other studies have shown a difference in the count of more than one log in seawater. This was encountered even in consecutive samples [12].

The constant variation in number makes comparison by bacterial density in water column unreliable. Since generation time deals with an initial and a later count of the same sample it becomes a very suitable tool for comparison. However, a general trend in bacterial count is significant. The 72-h counts were in the vast majority of samples more than the 24-h counts in all phases except the no chlorine phase where it was less. This is clear evidence that with no chlorine in water, the nutrients were exhausted following the first 24-h of growth. This has lead to a significant decrease in the biofouling potential during the no chlorine phase. Plant performance data also support the above-mentioned evidence of decreased biofouling potential when no chlorine was used. The average rise in P across the membranes was 0.1 bars/day during the no chlorine phase compared to an average rise of 0.3 bars/day during the chlorinated phases. The membrane flux decline rate was on an average <0.40%/day compared to 0.86% in earlier phases when the plant was operated with chlorine.

When generation times of the four phases of the project are compared (Table 1), a general trend could be noticed. There is an increase in generation times from the intake water towards the membranes. This means bacteria attached to pipings and filters or floating in the water column are removing nutrients from the flowing water prolonging the multiplication time of bacteria. Therefore, control of nutrients is one way of ultimately controlling membrane biofouling. Bott [14] investigated biofilm

accumulation using a flow-through system, where bacteria from a fermenter are supplied on heat exchanger surfaces. After the biofilm had reached a plateau, he cut down the supply of new bacteria but kept the nutrients at the same level as before. The level of the plateau remained the same. However, when he supplied bacteria in a suspension of decreased nutrients, the plateau was significantly lower.

The chlorinated source water (intake seawater) has very high bacterial multiplication capacity (low generation time) when compared to its generation time during the no chlorine phase (Table 1). This is a general trend which has been reported. Surface chemical conditioning precedes the biofilm formation [15]. This is accomplished by adsorption of organic molecules from seawater. humic acids and glycoproteins. These organic molecules include

During an RO chemical pretreatment, these large

organic molecules are degraded into simple molecules which could be assimilated by bacteria [6]. Such degradation could be more pronounced at Al-Birk because of the prevailing high temperatures during the study (27.1-34.5 oC). Applegate et al., [6], showed degradation of humic acid to be much greater in a temperature of 35 than 18oC. Thus, temperature at Al-Birk is conducive to humic acid degradation. It must be remembered that the RO feed water and brine reject temperatures are always higher than that of seawater feed. These temperatures reached 36.7 and 37.5 oC for the RO water and the brine, respectively. Increasingly, a higher rate of fouling is obtained with the increase of operation temperature up to 40 oC. This was indicated by the increase of fouling deposit and the decline in parallel flux across the RO membranes [16]. During the no chlorine phase, the bulk of nutrient was removed in the dual media filter. This is indicated by the sharp increase in the generation time of the DMF when compared to that of the intake seawater and to those of subsequent sampling stations (Table 1). This effective property of the dual media filter could be encouraged by enhancing the

metabolic activity of the residence bacteria. The intake water is acidified to a pH of about 6.2 before the coarse sand filter. This pH is inhibitory to growth of bacteria from the Red Sea [17]. Scaling deposit is not likely to occur at a higher pH and, if possible, the pH could be raised to about 7.0. Also, chlorination (continuous or in back-washing) would disrupt the balance of the population of bacteria in the dual media filter. A balanced population of bacteria would act as an effective biofilter for removing nutrients from the process stream.

Table 1 also shows a sharp significant decrease in generation time during the SBS shifting to after the MCF in three sampling stations (i.e., after the dual media filter, after the micron cartridge filter, and just ahead of the first stage membranes) when compared to corresponding values in the base line phase. With the SBS dosing shift, chlorine is brought closer to the membranes, and based on humic acid degradation reports, nutrients are also brought closer to the membranes. Another source of nutrients is thought to be dead bacterial cells from the biofilm in the piping between the SBS injection point and the membranes. It was noticed during the study that this piping section was not clean. Sloughing of dead cells and the degradation of larger molecules attached to piping (the conditioning surface) and free in water will furnish a good source of nutrients for bacteria in the membranes. The effect of this piping section was not found to be significant because, in general, the generation time during the second SBS dosing shift decreased instead of increasing. For this reason during the SBS shifting phases, membrane cleaning frequency is doubled following the sharp decrease in generation time. This cleaning was necessitated by a phenomenal build-up of P across the permeaters at a rate of approximately 0.3bars/day. It is therefore advisable to move the SBS dosing point as far as possible from the membranes.

A comparison of generation times between chlorinated raw seawater and nonchlorinated seawater from the intake (Table 2) showed that the difference between the 24 h-generation time though significant, is not outstanding (2.47 to 2.88 h). At 72h, the two generation times differed noticeably (6.78 to 9.58 h). This indicates two things: first, yes, chlorination is adding nutrients to water, and second, the nutrient load of the intake water is high. Then not only chlorination, but the source water for the plant is a source of nutrients. It is interesting to note that near shore surface water samples from

Al-Birk showed less nutrient content than the water samples from the intake, as reflected by generation times. This is evident from generation times of five samples taken from the near shore. The 24 h-generation time for the intake was 2.88 as compared to 3.14 for near shore, while the 72 h-generation time for the intake and near shore was 9.58 and 10.65 h, respectively. The intake is situated in a ditch deeper than the shallow surrounding water. It is used as a water way by fishing boats to get to the open sea and back to their docking place on shore. The fishing fleet consists of around 300 boats and they make a profound disturbance of the water column over the intake. Also, the intake ditch acts as a dumping ground where materials from the surrounding shallower sea settle down into the ditch. In this setting it is easy for the intake chamber and the intake pipe, which extends about 300m offshore, to get fouled and ultimately clogged should the chlorination is abandoned. Therefore, shifting of the SBS dosing point closer to the intake, but leaving the intake pipe chlorinated, is probably an alternative to be weighed against no chlorine in the intake section of the plant. Another alternative that would protect the intake section is intermittent or shock dosing chlorination. Plants which operated successfully without chlorine had adopted different strategies for controlling biological growth in the intake pipe. Some chlorinated for 6 to 8 h per week with a residual chlorine level of 1 mg/l, some twice a week, and others had found no need for intermittent chlorination at the intake [18].

During the no chlorine phase, the unfavorable location of the intake was complicated by extremely bad weather. The sea was very rough and the source water was loaded with silt. The finer part of the silt was able to pass the coarse sand and the dual media filtration causing choking of the micron cartridge filters. During a micron cartridge filter replacement, it was noticed that the filter elements did not have fouling smell. Instead, the smell was that of ordinary sea mud. Bacterial fouling smell was a

characteristic odor of replaced micron cartridge filter during the chlorinated phases of the project. Fine suspended particles reaching the micron cartridge filter are thus a problem to be reckoned with in this plant. In a word, the no chlorine operation at Al-Birk SWRO plant will result in less biofouling. It remains to be seen whether the nutrient and silt load of the intake will still support significant biofouling and microfiltration problems for the plant. This could

further be verified by running the plant without chlorination during a period of favorable weather, e.g., November- April. The biofilm slide study (Table 4) gives a direct picture of the dynamics of the biofilm formation. The bacterial growth studies showed that the shortest

generation/multiplication time is about 2.5 h. Since the residence time of water passing through the plant is too short (about 13 min), a significant bacterial growth will not occur to clog filters or membranes by mere density of unattached cells. Biofouling is thus a biofilm problem. Results of the bacterial attachment showed that attachment in chlorinated intake water was in the order of 103 to 104CFU/cm2 which is generally one to three orders of magnitude lower than that of dechlorinated water. Chlorination thus, as any other biocide, does not sanitize the water and the surviving bacteria, whether few or more, will attach to surfaces and form biofilm. Once attached, some bacteria will continue to multiply and support the biofilm, irrespective of the biocide. Biofilm organisms are much more difficult to eradicate than suspended cells because they are protected by the biofilm matrix [19] and they also become biochemically different and resistant [20]. During the first two project phases (termed normal operation and coagulant dosing shift), all biofilm sampling stations were dechlorinated except the chlorinated intake stations. The number of attached bacteria was fewest in the chlorinated intake water. As chlorine was removed, rapid bacterial attachment occurred in the sampling stations thereafter (Table 4). The numbers were higher in the after dual media filter station (first station after dechlorination) and they gradually decreased becoming lowest in the brine. This trend was similar to what had been observed in water samples (planktonic bacteria) for generation times. As nutrients decreased along the pretreatment line, less bacteria attached. When the SBS dosing point was shifted to after the micron cartridge filter, all sampling stations became chlorinated except the brine reject. Consequently the number of

attached bacteria decreased but it increased in the brine. This also conforms with findings from analyses of generation times for water samples. It indicates that as the SBS dosing point was moved forward, closer to the membranes, more nutrients became

available to the immediate sampling station which was the brine. Also, during the SBS dosing shift phase and when the three chlorinated sampling stations (intake, media filter and micron cartridge filter) were compared (Table 4), it could be seen that there was more bacterial attachment following the dual media filter. The same was true for the no chlorine phase where biofilm slides from after the DMF possessed more bacteria than all other sampling stations. There appears to be some nutrient entrapment in the media filter. Possibly the media filter could be manipulated to act as a biofilter to remove the bulk of nutrients from the RO feed.

The arrangement of bacteria on the biofilm slide (Fig.1) is typical of what has been reported [21]. It shows random arrangement of cells into patches or microcolonies. The empty spaces between cell aggregates are channels which flush the cells and transport wastes and nutrients. Whether the homogeneous appearance of cells is an indication that the biofilm is formed of a limited number of species remains to be seen.

Analyses of experimental results indicate chlorination is a factor enhancing biofouling at this SWRO plant. They also indicate that the source water is not ideal for smooth operation. Other pretreatment chemicals should not be excluded from contributing to biofouling. There are examples where antiscalants and organic coagulants have enhanced the fouling rate [22,23]. In this regard, it should be noticed that the RO plants in the Caribbean reverted successfully to seawells in place of surface intake. No chemical pretreatment is ever used even though the bioactivity in the seawells was identical to that in the open seawater [22]. Since sanitization by means of biocide is impossible, and since RO membranes are characterized by enormous surface area a single bacterium reaching a membrane could colonize and form a biofilm. Therefore, biofilms are unavoidable and the best alternative is to manage or learn to live with them. It is sufficient for a RO Plant Manager to prolong the time of pressure build-up across filters and membranes so that he brings down washing and cleaning frequencies to a level that would make the plant consistently available.

Beside judicious use of chemical pretreatment, biofouling control strategies should include careful monitoring and manipulation of other physical parameters. RO success

has been achieved by lowering the conversion rate or by increasing the brine velocities through the RO membranes [22].

6. 1.

CONCLUSIONS The numbers of bacteria in the intake feed water and along the RO pretreatment line show a great deal of variability. This makes the comparison of different pretreatment stages using the bacterial load in the water, unreliable. This

difficulty is easily overcome by using aftergrowth rates, in terms of generation times, to evaluate biofouling potential. Biofilm slides were found useful for direct measurement and for monitoring the effect of chlorination-dechlorination on biofilm formation. 2. Along the pretreatment line, bacteria remove nutrients effectively resulting in significant reduction in multiplication capacity and biofouling potential. 3. 4. Chlorination enhances bacterial growth, and, consequently, biofouling potential. The closer the dechlorination point to the membranes, the higher is the fouling of membranes, i.e., chlorination/dechlorination or chemical pretreatment is adding nutrients to RO water. 5. Bacteria are capable of biofilm formation in chlorinated water but to a lesser extent than dechlorinated water. Thus when the SBS dosing point is shifted past the media and micron cartridge filters, biofouling in these filters becomes less extensive, but upon dechlorination a huge aftergrowth occurs creating serious biofouling problems. 6. The level of nutrients and silt in the sea at the intake site is high. This

questionable water quality makes successful operation without chlorine difficult. No-chlorine operation of plants with surface intake may not be suitable in this type of location. 7. During rough seas a tremendous load of silt is brought into the intake water. This silt load is an added problem to the filtration system.

7.

ACKNOWLEDGMENT

Mr. John O'Hara carried out the electron microscopy photography, Dr. Ata M. Hassan and Mr. M. AK. Al-Sofi reviewed the text and Mr. Rafique M. Imambakhsh typed the manuscript.

Table 1.

Comparison of generation times of bacteria* at 24 and 72 hours of growth (incubated at 30 oC) for various phases of the project (n=15) PROJECT PHASE

Sampling Station Intake seawater Before coarse sand filter After Dual media filter After micron cartridge filter Ahead of 1ststage membranes Brine reject of 1ststage

Base line (as is) 24 h 72 h 2.47 0.194,aa 2.66 0.39aa 5.42 0.89a 9.62 3.54ab 7.70 2.41ab 10.86 3.60a 6.78 0.95dd 6.18 0.87dd 14.77 1.92d 21.02 6.34dd 18.35 4.56dd 23.19 2.93d

Coagulant dosing shift2 24 h 72 h 2.29 0.21aa 2.40 0.34aa 5.06 0.69a 7.93 2.69a Not sampled 9.12 2.67a 5.97 0.66ee 5.78 0.69dee 13.36 1.56d 18.62 4.69dd Not sampled 22.43 5.91dd

SBS dosing shift3 24 h 72 h 2.29 0.49aa 3.90 0.90ee 2.68 0.89aa 5.11 0.55ee 2.86 0.67bb 5.69 1.09ee 2.92 0.91bb 5.32 0.99ee 4.71 1.03b 12.61 2.25e 11.50 3.07a 28.48 5.90e

Without chlorination 24 h 72 h 6.49 1.67b 11.08 2.36cc 18.00 4.15f 36.45 6.53ff Not sampled 10.89 1.83bcc 43.34 7.92ff 11.85 3.00acc 36.64 8.90ff 10.05 2.45acc 57.34 11.58ff

* Generation time, (g) = t k / (ln Nt- ln Nt0 ) ; where t =24 for 24h-generation time and 72 for 72h-generation time, K = 0.693, Nt count at 24 or 72 h and Nt0 count at 0h.
1

Baseline ( as is ) plant operating with the coagulant dosing before the coarse sand filter and the SBS dosing point after the dual media filter. Coagulant dosing point shifted back to the pressure side of the intake pump . Sodium metabisulfite ( SBS ) dosing point shifted to after the micron cartridge filter Standard deviation.

a,b,c,d,e,f

For the same sampling station within different phases (horizontal rows) and for the same generation time (24 or 72 h ) , means sharing any common single letter superscript are not different; others are different . For the same project phase and within different sampling stations (vertical columns), means (of either 24 or 72h generation times) sharing any 2 common letter superscripts are not different ; others are different (P 0.05) ; Analysis of Variance and LSD test.

Table 2.

Comparison of bacterial count (colony-forming units/ml) and generation timesb(hours) of the intake area and chlorinated intake feed seawater (n=5) 0-h count 6.46x102 2.69x102 24-h count 1.93x105 2.36x105 24 h - generation time 2.88 0.31b 2.47 0.19c 72-h count 1.47x105 4.41x105 72 h-generation time 9.58 0.25 6.78 0.95c

Sampling station Intake raw water Chlorinated intake water

a

Sample taken from 7 m depth close to the intake chamber in the open sea. Each generation time is statistically different from its corresponding one , (P 0.05); LSD test. c Standard deviation.
b

Table 3. Comparison of bacterial count (colony-forming units/ml) and generation times (hours) when the sodium metabisulfite (SBS) dosing point was shifted to two locations after the micron cartridge filter (n=15) Sampling Station 0-h Count 24-h Count 24-h Generation Time1 72-h Count 72-h Generation Times

After dual media filter 1st shift2 1.06x105 2.86 0.67*a 3.87x105 5.69 1.09*a 1.08x102 nd 3 2 4 a 5 2 shift 0.46x10 7.88x10 2.80 0.63 3.02x10 5.78 0.76a After micron cartridge filter 1st shift 2.34 0.43b 3.47x105 5.32 0.99b 0.51x102 3.99x104 nd 2 4 c 5 2 shift 0.16x10 3.93x10 2.38 0.49 2.80x10 5.21 0.76b st Ahead of 1 stage membranes 1st shift 4.71 1.03d 7.29x105 12.61 2.25c 1.54x104 4.91x105 nd 4 5 e 5 2 shift 1.76x10 3.70x10 5.75 0.75 5.01x10 13.81 1.25c st Brine reject of 1 stage 1st shift 11.50 2.67f 8.86x105 28.48 5.90d 1.43x105 6.55x105 nd 4 5 g 6 2 shift 5.89x10 4.22x10 7.83 0.59 1.07x10 17.09 1.07e 1 For each sampling station, means with same letter superscript are not different while those with different ones are different. (P 0.05) ; Analysis of variance and LSD test. 2 Shifted to a point 13 m ahead of the high pressure pump. 3 Shifted to a point just ahead (1 m) of the high pressure pump. Standard deviation Table 4. Bacterial attachment to biofilm sampling slidesa at four stations along the pretreatment line Sampling Stationb Bacteria ( CFU) / cm2 After After micron dual media filter cartridge filter (8.15 0.18)x106 (4.70 0.50)x105 (1.17 0.12)x105 (2.20 0.30)x106 (1.57 0.03)x106 (7.55 1.05)x105 (5.77 0.23)x103 (1.70 0.33)x105

Project phase1 Normal operation Coagulant dosing shift SBS dosing shift No Chlorine
a

Intake Seawater (7.88 0.22)x103 (2.22 0.09)x104 (1.24 0.11)x104 (1.23 0.34)x105

Brine reject of 1st stage (1.50 0.13)x105 (3.43 0.12)x105 (1.30 0.07)x106 (3.45 0.99)x105

Glass slides (coupons) measuring 2.5 x 2.2 cm fixed in biofilm samplers at each of four sampling station as indicated in table. b Within each project phase (horizontal rows), number of bacteria attached at each sampling station is different from others. In the same sampling station during different phases of the project (vertical columns), the number of attached bacteria is different from others (P 0.05); Analysis of variance and LSD test. Standard deviation

Fig. 1a.

An Electron micrograph of a biofilm slide place in SBS-dechlorinated feed after the micron cartridge filter showing cells attached in irregular patches. Some cells with bipolar bodies appearing as white spots at the poles of the cells. Sheathes around the cells could also be noticed. Cells are rod-shaped and many are curved (bar=5 m).

Fig.1b. A micrograph from a slide placed in SBS dechlorinated feed after the micron cartridge filter showing more cells attached than in above case, Fig.1a. Also, cells take more elongated shape than in the above figure (bar = 5 m).

Fig.1c. A micrograph from a slide in the chlorinated intake water during the first phase of the project. Cells are few and smaller in size when compared to cells from dechlorinated stations in Fig. 1a & b above. Note the formation of a base layer on which cells are attached (bar = 5 m).

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1. 2. 3.

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