Albertsen, Mads

Karst, Søren Michael

McIlroy, Simon Jon

Nielsen, Per Halkjær

Stokholm-Bjerregaard, Mikkel

Enhanced biological phosphorus removal (EBPR) is widely used to remove phosphorus from wastewater. The process relies on polyphosphate accumulating organisms (PAOs) that are able to take up phosphorus in excess of what is needed for growth, whereby phosphorus can be removed from the wastewater by wasting the biomass. However, glycogen accumulating organisms (GAOs) may reduce the EBPR efficiency as they compete for substrates with PAOs, but do not store excessive amounts of polyphosphate. PAOs and GAOs are thought to be phylogenetically unrelated, with the model PAO being the betaproteobacterial “Candidatus Accumulibacter phosphatis” (Accumulibacter) and the model GAO being the gammaproteobacterial “Candidatus Competibacter phosphatis” (see Oehmen et al., 2007). Here, we report the discovery of a GAO from the genus Propionivibrio, which is closely related to Accumulibacter. Propionivibrio is often found in similar abundance to Accumulibacter in full-scale plants and has, until now, been mistakenly quantified as Accumulibacter using the canonical PAOmix FISH probes

    Aalborg UniversitetA GAO hiding among the PAOMcIlroy, Simon Jon; Albertsen, Mads; Stokholm-Bjerregaard, Mikkel; Karst, Søren Michael;Nielsen, Per HalkjærPublication date:2016Link to publication from Aalborg UniversityCitation for published version (APA):McIlroy, S. J., Albertsen, M., Stokholm-Bjerregaard, M., Karst, S. M., & Nielsen, P. H. (2016). A GAO hidingamong the PAO: The role of the Propionivibrio spp. in biological phosphorus removal. Poster session presentedat Microbial Ecology & Water Engineering and Biofilm IWA Specialist Conference, Copenhagen, Denmark.General rightsCopyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright ownersand it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights.            ? 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McIlroy1,3, Mads Albertsen1,3, Mikkel Stokholm-Bjerregaard1,2, Søren M. Karst1 and Per Halkjær Nielsen1 1Center for Microbial Communities, Department of Chemistry and Bioscience, Aalborg University, Aalborg, Denmark 2Krüger A/S, Aalborg, Denmark 3These authors contributed equally to this work  sjm@bio.aau.dk A	  6L	  lab	  scale	  sequencing	  batch	  reactor	  conﬁgured	  for	  EBPR	  was	  operated	  for	  3	  months	  using	  vola?le	  faPy	  acids	  (VFAs)	  as	  the	   carbon	   source.	   Biochemical	   transforma?ons	   indicated	   a	  mixed	  PAO-­‐GAO	  community.	  	  Metagenomics	   was	   applied	   to	   obtain	   representa?ve	  genomes	   from	   the	   abundant	   Accumulibacter	   and	  Propionivibrio	   species.	  Annota?on	   supported	  a	   typical	  GAO	  physiology	   for	   the	   Propionivibrio	   spp.	   where	   aerobically	  stored	   glycogen	   is	   predicted	   to	   provide	   the	   energy	   and	  reducing	   equivalents	   to	   support	   anaerobic	   uptake	   and	  storage	  of	  VFAs	  as	  PHAs	  (Fig.	  2).	  In	   situ	   staining	   revealed	   that	   both	  A c c umu l i b a c t e r	   s p p .	   a n d	  Propionivibrio	   spp.	   stored	   VFAs	   as	  polyhydroxyalkanoates	   (PHAs)	  under	   anaerobic	   condi?ons	   while	  only	   the	   former	   cycled	   polyP	   (Fig.	  3).	   This	   is	   consistent	   with	   the	  Accumulibacter	   and	   Propionivibrio	  behaving	   as	   PAO	   and	   GAO,	  respec?vely.	  Phylogene?c	   (Fig.	   1B)	   and	   FISH	   (Fig.	   1A)	   analyses	   revealed	  that	   the	   reactor	   was	   dominated	   by	   two	   popula?ons	  represen?ng	   related	   genera	   -­‐	   Accumulibacter	   and	  Propionivibrio.	   The	   PAOmix	   probes	   designed	   to	   target	   the	  Accumulibacter	   target	   both	   popula?ons	   Fig.	   1.	   A	   novel	  probe	  was	  designed	   to	   target	   the	  Propionivibrio	   (Prop207).	  The	   PAO651	   probe	   alone	   is	   recommended	   to	   target	   the	  Accumulibacter.	  	  Surveys	  of	   full	  scale	  EBPR	  systems	  reveal	  a	  high	  abundance	  of	  the	  Propionivibrio	  –	  some?mes	  in	  higher	  abundance	  than	  the	  Accumulibacter	  PAO.	  Fig.	  1.	  Phylogeny	  (16S	  rRNA	  gene)	  and	  FISH	  analysis	  of	  the	  reactor	  biomass	  community	  	  Fig.	  2.	  Metabolic	  model	  for	  the	  Propionivibrio	  GAO	  in	  EBPR	  systems	  Fig.	  3.	  In	  situ	  staining	  of	  the	  biomass	  STUDY	  PERSPECTIVES	  IDENTIFICATION	  OF	  A	  NOVEL	  GAO	  	  IN	  SITU	  ANALYSES	  GENOMIC	  ANALYSES	  20#μm#A.#Composite#FISH#B.#DAPI#stain##Accumulibacter#(PAO651)#=#Blue#Propionivibrio#(Prop207)#=#Red#PolyP#inclusions#=#yellow#Micropruina glycogenicaMicrolunatus phosphovorus PAO Micrococcus luteusAmaricoccus kaplicensisAcinetobacter lwoffiiPlasticicumulans acidivoransPropionivibrio dicarboxylicusRhodocyclus tenuisDechloromonas agitata0.10Defluviicoccus spp. GAO"Ca. Contendobacter odensis" GAO"Ca. Competibacter denitrificans" GAOAccumulibacter spp. PAOTetrasphaera spp. PAOUncultured Propionivibrio genome ActinobacteriaBetaproteobacteriaAlphaproteobacteriaGammaproteobacteriaProp207PAO468PAO846EUBmixPAO651PAO468PAO846EUBmixEUBmix targetting BacteriaProp207 targetting PropionivibrioPAOmix (PAO651, PAO468 & PAO846) targetting AccumulibacterB. Phylogeny of the PAO-GAOA. Composite FISH image of the SBR biomassB. Phylogeny of the PAO and GAO

A GAO hiding among the PAO:The role of the Propionivibrio spp. in biological phosphorus removal

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A GAO hiding among the PAO:The role of the Propionivibrio spp. in biological phosphorus removal

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