Wastewater treatment is an integral part of processing of sugar beets in the sugar industry. American Crystal Sugar Company (ACS) has five factories. Three of these factories, Moorhead (MHD), Hillsboro (HLB), and East Grand Forks (EGF) each have a 6.7 million gallon anaerobic contactor, aerobic basin, and ponds for processing of wastewater while the other two factories have lagoons and wetlands for the treatment of their wastewater.During the 2007/2008 campaign our efforts were focused on microbial issues in wastewater treatment at the MHD factory. Therefore, this paper deals with problems encountered with filamentous bacteria, poor settling in treatment of the high strength wastewater and studies to circumvent these problems. In addition some differences observed in the MHD and HLB anaerobic systems will also be discussed. Materials and Methods:A) Microbiology 1) Sample collection Weekly samples of wastewater were obtained aseptically in sterile screw cap containers from each of three locations: a) anaerobic influent from the new covered wastewater pond, b) anaerobic tank or anaerobic contactor, and c) aerobic basin or activated sludge system. These samples were observed microscopically at the ACS Technical Services Center Microbiology Lab. Samples from similar locations in the wastewater treatment systems at the Hillsboro and East Grand Forks factories were intermittently obtained for comparative purposes. 2) Microscopy and Photography i) Wet Mounts, Staining, Floc formation and Higher Life Forms -Separate wastewater (WW) wet mounts on slides were observed microscopically with or without a drop of lactophenol cotton blue and/or India ink stain. This was for determination of higher life forms (amoeba, flagellates, ciliates, rotifers) in the aerobic basin WW sample and presence of filaments and motility, tetrads, bacterial polysaccharides, and floc characteristics in other samples. The wet mounts allow living microorganisms to be observed as they appear in the environment. ii) Staining of Smears -Different WW samples were smeared on slides and air dried.Staining protocols for the Neisser and Gram stains were then carried out. This was for identification of filamentous bacterial types using a key provided by Environmental Leverage Inc. (www.EnvironmentalLeverage.com) (3). The Neisser stain provided a key for identification of bacterial filaments on the basis of branching, Neisser positive or negative, and other characteristics within this key. The gram stain provided a key for identification of bacterial filaments on the basis of whether it was Gram negative, Gram positive, or Gram variable and showed the presence or absence of sheath (with and without attached growth), sulfur granules, and other characteristics within that key. All above microscopy was carried out using an Olympus BHTU-001A microscope with the use of bright field, phase contrast, and dark field accessories. A Nikon Coolpix 4500 digital camera mounted on the microscope eyepiece tube was used for photographing of microscopic specimens. iii) Fecal c...
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