Efficiency of the Biological System for Iron and Manganese Removals from Water

INTERNATIONAL JOURNAL OF SCIENTIFIC & TECHNOLOGY RESEARCH VOLUME 5, ISSUE 07, JULY 2016 ISSN 2277-8616 Efficiency Of The Biological System For Iron And Manganese Removals From Water

Fawazy G. Khedr, Khalid A. Shaaban, Said M. Ezzat, Mohamed Maher

Abstract: Fifty ground water samples were collected from five different localities in El-Sharkia Province, Egypt during 2014. Leptothrix discophora W19 was isolated from the collected ground water samples, purified and grown on nutrient agar media supplemented with 1mg/L of each of iron and manganese. Conventional identification of Leptothrix discophora W19 isolate following the key of Bergey's manual. Standard water containing different +2 +2 concentrations of Fe and Mn had been examined for both tested metals removal capacity using Leptothrix discophora W19 as a bioleaching. The biological system for iron and manganese removal had provided overall removal efficiency up to 95.25% and 95% for iron and manganese respectively.

Key words: Leptothrix discophora W19, Ground water, Iron, Manganese, Biological treatment, Bioleaching. ————————————————————

Introduction sampling cooled box to the laboratory to be examined. Water is the life secret of every living creature on earth. Ground water samples were collected from depth ranged There are two sources of water namely, surface water and between 35 to 65 m under earth’s surface. ground water. Fifty percent of the world's population depends daily on ground water for their drinking water [1]. Isolation and purification of Leptothrix discophora: In a global scale, ground water represents a potential Leptothrix discophora W19 was isolated from collected resource of drinking water. In Egypt, the ground water is ground water samples using nutrient agar medium widely used for drinking, agricultural and other domestic supplemented with 1 mg/L of each of iron and manganese purpose. The ground water reservoir in the Delta and Upper salts in combination. Inoculated plates were incubated at 37 0 Egypt is considered to be one of the biggest reservoirs in C for 3 d. Growing colonies were isolated and the most the world with stores capacity of about 400 km3 (120 km3 in active bacterial isolates were selected and subjected to Upper Egypt and 280 km3 in the Delta). However, most of morphological and biochemical examination according to underground aquifers are increasingly polluted by iron and methods of [7]. manganese, therefore, became useless resources of drinking water [2]. When iron and manganese are present Biological treatment of iron and manganese in in water as soluble form in drinking water supplies, then we water: will come across many objectionable problems related to The steps for bioleaching were previously described by [8]. their presence. The World Health Organization (WHO) has Leptothrix discophora W19 bacteria was grown in 250 ml approved the removal of iron and manganese when conical flask containing 100 ml of nutrient broth medium concentrations are higher than 0.3 mg/L and 0.4 mg/L supplemented with 3 mg/L of each of iron and manganese respectively. If concentrations are higher than these salts in combination. The incubation was done at 37 0C for standards, water must be treated before using it for drinking 3 d. Sand bed filter was prepared. The sand bed filter was purposes. If water is not treated then there can be different saturated with Leptothrix discophora W19, forming problems for water consumers and also for that municipality bioleaching system (Figure 1), to examine its efficiency for that delivers drinking water to consumers [3]. Fe+2 and Mn+2 removals from water. Standard water Iron/manganese-oxidizing bacteria have been recognized containing different concentrations of Fe+2 and Mn+2 (1, 2, 3, for their ability to deposit iron hydroxide or manganese 4 and 5 mg/L), separately had been examined for both oxide in structures outside its cells [4]. Leptothrix tested metals removal capacity. Water containing Fe+2 and discophora reacts with soluble iron or manganese, Fe+2 or Mn+2 was firstly aerated for 1 h and removal efficiency of Mn+2 , through an oxidation process that changes the iron Fe+2 and Mn+2 after aeration was determined then passed or manganese to an insoluble form, Fe+3 or Mn+4 [5]. The through the bioleaching system and efficiency of Fe+2 and processes available for iron and manganese removals are Mn+2 removal after bioleaching was also determined. either physico-chemically or biologically based. The Overall removal efficiency of Fe+2 and Mn+2 after abiotic and advantages of biological treatments compared with bioleaching step was determined. conventional physic-chemical treatments can be summarized as follows: No use of chemicals, higher Results filtration rates, the possibility of using direct filtration and lower operation and maintenance costs [6]. Location of ground water samples: Fifty ground water samples were collected from five Materials and Methods different localities in El-Sharkia Province, Egypt (Zagazig, Minia El-Qamh, Hehya, Faquos and Abo-Kabeer). Collection of ground water samples: Fifty ground water samples were collected from five Isolation and purification of Leptothrix discophora different localities in El-Sharkia Province. Localities of W19: collection were Zagazig, Hehya, Abo-Kabeer, Faqous and Nutrient agar medium supplemented with 1 mg/L of each of Minia El-Qamh. Samples were collected in cleaned sterile iron and manganese salts in combination was used for glass bottles and transported under aseptic conditions in isolation of Leptothrix discophora W19 from the examined

82 IJSTR©2016 www.ijstr.org INTERNATIONAL JOURNAL OF SCIENTIFIC & TECHNOLOGY RESEARCH VOLUME 5, ISSUE 07, JULY 2016 ISSN 2277-8616 ground water samples. Growing colonies on the mentioned Discussion media were isolated and the most active bacterial isolates Heavy metal pollution in the aquatic environment has were selected and kept for further studies. Data in Table (1) become of great environmental concern since they are non- show that bacteria in 16% of ground water samples can biodegradable. Metals transferred in food chain as a result grew on medium supplemented with 1mg/L of each of iron of leaching from industrial wastewater, polluted soils and and manganese. water [11]. To avoid health hazards it is essential to remove these toxic heavy metals from wastewater before its Identification and confirmation of the most active disposal. Main sources of heavy metal contamination bacterial isolates: include urban industrial aerosols, solid wastes from Two bacterial isolates encoded W19 and W22 showed good animals, mining activates, industrial and agricultural growth on nutrient agar media supplemented with mixture chemicals. Heavy metals also enter the water supply from of iron and manganese were selected and identified. industrial and consumer water or even from acid rain which Results of bacterial characterization showed that the two breaks down soils and rocks, releasing heavy metals into selected bacterial isolates were rod-shaped, motile, Gram streams, lakes and ground water [12, 13]. Iron (Fe) and negative and sheath forming cells. They showed positive manganese (Mn) cause aesthetic, organoleptic and results with regard to oxidase test, catalase test, utilization operating problems when they are present in ground water. of glucose, mannitol, D-sorbitol, sucrose, glycerol but These metals consume chlorine in the disinfection process showed negative results regarding other tests listed in and promote bio-fouling and microbiological induced Table (2). The above data were confirmed by API 2O Gram corrosion in water networks [14]. When iron and negative kits (Biomereux, France) according to the manganese are present in water as soluble form in drinking manufacturer's instructions. By surveying literature [9] and water supplies, then we will come across many following the diagnostic key of [10], the two isolates were objectionable problems related to their presence. The World identified and classified as belonging to betaproteobacteria Health Organization (WHO) has approved the removal of Leptothrix discophora. iron and manganese when concentrations are higher than 0.3 mg/L and 0.4 mg/L respectively [3]. So if concentrations Biological treatment of iron and manganese in are higher than these standards, then water must be water: treated before using it for drinking purposes. If water is not Using sand bed filter saturated with Leptothrix discophora treated then there can be different problems for water W19, forming bioleaching system, an experiment was consumers and also for that municipality that delivers designed to examine efficiency of Leptothrix discophora drinking water to consumers. Leptothrix discophora bacteria W19 as bioleaching for biological treatment of iron and obtain carbon from the carbon dioxide (CO2) in the air and manganese in water (Figure 2). obtain energy from dissolved iron or manganese. These bacteria occur naturally in the soil and thrive when there is Iron removal from water using Leptothrix discophora adequate food (i.e., iron and/or manganese). The biological W19: activity of living of Leptothrix discophora bacteria is As shown in Table (3) and Figure (3), overall Fe+2 removal responsible for iron and manganese removal from water efficiency up to 95.25% was achieved. The major part of and many applications of biological iron and manganese removed Fe+2 (62% reduction of total Fe+2) occurred after system are being used in ground water engineering [15]. aeration step of first run with influent concentration 1 mg/L The most active bacterial isolates were subjected to due, probably to physic-chemical oxidation (abiotic morphological and biochemical examination according to oxidation). In the biofilm step, 31% removal developed on key of Bergey's manual and was identified as sheathed the bioleaching media whereas over all removal of 93% Leptothrix discophora. Sheath-forming bacteria are found in was achieved after the first run. Upon repeating the runs several disparate bacterial groups, including the non-fruiting with increase in influent concentration, increase of Fe+2 gliding bacteria, the cyanobacteria, and the methanogens. removal was observed where maximum values were In Bergey’s Manual of Systematic Bacteriology the following recorded after the fourth run (4 mg/L Fe+2 influent genera are described under the heading ―Sheathed concentration) in case of both abiotic oxidation (74%) and Bacteria‖ [16]: Sphaerotilus, Leptothrix, Crenothrix, bioleaching step (81.73%) with 95.25% over all Fe+2 Phragmidiothrix, Haliscomenobacter, and Lieskeella. These removals. taxa are distinguished primarily on the basis of morphological criteria. Leptothrix is a sheathed filamentous Manganese removal from water using Leptothrix bacterium that can generally be found in different types of discophora W19: aquatic environments with sufficient organic As shown in Table (4) and Figure (4), removal efficiencies matter. Leptothrix bacteria are known to be capable of up to 95.0 % of Mn+2 was achieved with the use of oxidizing both iron and manganese, unlike other sheathed bacteria [17]. Previous studies reported presence of Leptothrix discophora W19 as bioleaching in combination with abiotic oxidation. Maximum Mn+2 removal efficiency (33 Leptothrix spp. in ground water resources and their isolation %) was recorded after third run with influent concentration from hand pumps placed on the ground water wells [18]. (3 mg/L) without bioleaching. Meanwhile, application of The most popular form of biological treatment for drinking bioleaching step increased over all removal to 93.66%. This water is the fixed-bed biofilm system. This system includes indicates that the removal of Mn+2, unlike the Fe+2 removals, a media such as sand, anthracite and/or GAC, which will takes place mainly through the bioleaching by biotic support biological growth. The most common placement of oxidation, while the majority of Fe+2 removal capacity a fixed-bed biofilm system is just before final disinfection achieved abiotically after aeration. where it oxidizes organic and inorganic contaminants, as 83 IJSTR©2016 www.ijstr.org INTERNATIONAL JOURNAL OF SCIENTIFIC & TECHNOLOGY RESEARCH VOLUME 5, ISSUE 07, JULY 2016 ISSN 2277-8616 well as removing particles [19]. In an application Tables: experiment, Standard water containing different concentrations of Fe+2 and Mn+2 (1, 2, 3, 4 and 5 mg/L) had Table (1): Bacterial growth on nutrient medium been examined for both tested metals removal capacity supplemented with a combination of 1mg/L of each of iron using Leptothrix discophora W19 as a bioleaching. Overall and manganese: removal efficiency up to 95.25% was achieved in case of +2 +2 Fe after abiotic and bioleaching steps. Maximum Fe Nut. Med. +2 removal was observed after the fourth run (4 mg/L Fe No. of Nut. Med. No. of Supplemented influent concentration) in case of both abiotic oxidation Water Supplemented with Water with (74%) and bioleaching step (81.73%) with 95.25% over all Sample iron and manganese Sample iron and Fe+2 removals. The major part of removed Fe+2 could be manganese due to the abiotic oxidation [20, 21]. Accordingly, [22] 1 -ve 26 -ve mentioned that biological Fe+2 removals were likely to be 2 -ve 72 -ve supplementary to conventional physic-chemical removal in 3 -ve 72 -ve the presence of oxygen. [23] reported that the biological 4 -ve 72 -ve removal of iron is very efficient over a long operational 5 -ve 03 -ve period and residual iron concentrations below 10μg/L can 6 -ve 03 +ve be constantly achieved. Removal efficiencies up to 95 % of 7 -ve 07 -ve +2 Mn were also achieved with the use of Leptothrix 8 -ve 00 -ve discophora W19 as bioleaching in combination with abiotic 9 -ve 03 -ve +2 oxidation. Significantly high biological Mn removal 10 -ve 03 +ve ranging between 88.64% (fourth run) and 92.85% (first run) 11 +ve 03 -ve had been observed compared to the abiotic physic- 12 -ve 02 -ve chemical oxidation taking place ranging between 23.00% 13 -ve 02 -ve (second run) and 33% (third run). Mn+2 oxidation in 14 +ve 02 -ve freshwater is thought to be from a combination of bacterial 15 -ve 33 +ve oxidation and chemical oxidation with oxygen [24]. Aeration +2 16 -ve 33 -ve does not oxidize organically bound Mn , but in situ +2 17 -ve -ve treatment is considered a good primary treatment for Mn 37 removals [25]. Mn+2 removals from drinking water sources, 18 -ve 30 -ve minimizing chemical oxidants that could form unwanted by- 19 +ve 33 -ve products [26]. Biological oxidation of Mn+2 is a bioleaching 20 -ve 33 -ve process that has not been fully explored, although it is 21 +ve 33 -ve believed that Mn+2 oxidation causes Mn+4 oxide 77 +ve 32 -ve accumulations on bacterial surface, attached to the 70 -ve 32 -ve leaching media. This accumulated manganese was to be 73 -ve 32 -ve removed together with excess bacteria and biofilm [27]. The 73 -ve 33 -ve removal of Mn+2, unlike Fe+2 removal, takes place mainly -ve: No bacterial growth. +ve: Bacterial growth occurs. through the bioleaching by biotic oxidation, while the majority of Fe+2 removal capacity was achieved abiotically Table (2): Cultural, morphological and biological after aeration. characteristics of the most active bacterial isolates:

Bacterial characteristics Isolate Gram Catalase Coagulase Oxidase Urease staining W - + - + - 19 W22 - + - + - Voges Methyl Indole Citrate H S Proskauer 2 red test utilization production

W19 - - - - - W22 - - - - - Gelatin D- Motility Glucose Mannose liquifaction sorbitol

W19 + - + + - W22 + - + + - Maltose Mannitol Sucrose Glycerol W - + + + 19 W22 - + + + +: Positive results. -: Negative results.

Table (3): Removal efficiency of Fe+2 at different influent Figures: concentrations:

After % % Influent After Ru aeratio Remov Removal Total concentrat bioleachi n n al through % ion ng No (avera After the remov (average, (average, . ge, aeration bioleachi al mg/L) mg/L) mg/L) ng 1 1 0.38 62.00 0.07 81.57 93.00 2 2 0.74 63.00 0.12 83.78 94.00 3 3 0.94 68.66 0.17 81.91 94.33 4 4 1.04 74.00 0.19 81.73 95.25 5 5 1.40 72.00 0.25 82.14 95.00

% Removal after aeration = 100 – (Concentration after aeration X 100 / Influent concentration).

% Removal through the bioleaching = 100 – (Concentration after bioleaching X100/Concentrat ion after aeration).

Total % removal

= 100 – (Concentration after bioleaching X 100 / Influent Figure (1): Sand bed filter saturated with Leptothrix concentration). discophora W (bioleaching system). 19 +2 Table (4): Removal efficiency of Mn at different influent concentrations:

% After % influent remov After aeratio removal Total Ru concentrat al bioleachi n through % n ion After ng (avera the remov No. (average, aeratio (average, ge , bioleachi al mg/L) n mg/L) mg/L) ng

1 1 0.70 30.00 0.05 92.85 95.00 2 2 1.54 23.00 0.12 92.20 94.00 3 3 2.01 33.00 0.19 90.54 93.66 4 4 2.73 31.75 0.31 88.64 92.25 5 5 3.77 24.60 0.36 90.45 92.80 % Removal after aeration = 100 – (Concentration after aeration X 100 / Influent concentration).

% Removal through the bioleaching = 100 – (Concentration after bioleaching X 100 / Concentration after aeration).

Total % removal = 100 – (Concentration after bioleaching X 100 / Influent concentration).

Figure (2): Sand bed filter saturated with Leptothrix discophora W19 (bioleaching) for biological treatment of iron and manganese. 85 IJSTR©2016 www.ijstr.org INTERNATIONAL JOURNAL OF SCIENTIFIC & TECHNOLOGY RESEARCH VOLUME 5, ISSUE 07, JULY 2016 ISSN 2277-8616

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