Using deep gravel and bacteria to control nitrogen

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Re: Using deep gravel and bacteria to control nitrogen

Post by bekateen »

You guys get way more technical than I sometimes understand, but I do get lots out of this and it is fascinating to learn the crossover between aquariums, aquaculture and wastewater management.

The irony of this thread is that now I'm 100% in the opposite direction from where I started. Instead of using deep gravel, I now limit my tanks to a thin film of sand, just a few mm deep. I've had too many experiences in the meantime with sulfur gas deaths that I work hard to avoid its accumulation.

Cheers, Eric
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Re: Using deep gravel and bacteria to control nitrogen

Post by TwoTankAmin »

I ran a 50 gal. high tech planted tank for over a decade. Because of space considerations I opted to push my pressurized co2 into the intake of an Eheim Pro II 2026. I chose Eheim as it seemed the best choice to deal with gas passing though the impeller well. This filter has a max. flow rating of 950 l/h (247 gph). Moreover, I ran the spray bar vertically in the back right corner. The way the tank was scaped the actual volume of water was more like 35 gal. So my max turnover rate was about 7x/h. My best guess is with my media load it was more like 6x/h or even lower.

My plants did fine. The fish did so as well. It was the tank in which my Farlos spawned. The problem was the tank took up way to much of my weekly maint. time compared to its share of gals. and fish. I finally sold the co2 system and ultimately replaced the tank with a 75 which is now filled with L236s and no plants. But that Eheim is still the primary filter :)

I must be getting old, repeating myself in a thread. It must be that OBS and CRS again.
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Re: Using deep gravel and bacteria to control nitrogen

Post by dw1305 »

Hi all,
bekateen wrote: 15 May 2019, 14:03 ......and it is fascinating to learn the cross-over between aquariums, aquaculture and wastewater management.
It is really go with what you know.

I'm a pretty shoddy fish keeper, but I can mitigate for this by providing the fish with a high quality environment.
bekateen wrote: 15 May 2019, 14:03I now limit my tanks to a thin film of sand, just a few mm deep.
If I kept non-planted tanks that is all I would use. I've had a couple of high flow tanks (with lower planting levels) and I used a mix of sand and fine gravel in those, and let the flow sort it. The layer was pretty thin.

If you had an HMF (matten) filter that could perform a lot of the roles that would occur in the substrate (documented in Stephan Tanner's article).

It is the plant roots that allow you to have a deeper substrate, the rhizosphere is much more important factor than many people realize (which is documented in Myriophyllum aquaticum Constructed Wetland Effectively Removes Nitrogen in Swine Wastewater" (https://www.frontiersin.org/articles/10 ... 01932/full).)

Even with planting I don't have a huge depth of substrate.

cheers Darrel
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Re: Using deep gravel and bacteria to control nitrogen

Post by dw1305 »

Hi all,
A couple of papers have come my way that maybe of interest to some.

There is some further work on the oxygen dynamics within the rhizosphere of Lobelia dortmanna:

"The effect of Lobelia dortmanna L. on the structure and bacterial activity of the rhizosphere". K Lewicka-Rataj, A Świątecki, D Górniak - Aquatic botany, 2018. That one is behind a pay-wall, but I can email it if any-one really wants a copy?

There is also a really good, open source, review article on ammonia oxidation:

Laura E Lehtovirta-Morley "Ammonia oxidation: Ecology, physiology, biochemistry and why they must all come together:" <https://doi.org/10.1093/femsle/fny058> FEMS Microbiology Letters, Volume 365, Issue 9, May 2018. The author works for UEA in Norwich UK <https://people.uea.ac.uk/l_lehtovirta-m ... hinterests>.
AOA vastly outnumber AOB in most soil and aquatic environments, often by orders of magnitude (Leininger et al.2006). It is estimated that there are 1 × 10^28 AOA cells in the Earth's oceans and they are some of the most numerous living organisms on Earth, accounting for up to 40% of all prokaryotes in marine ecosystems and 1%–5% in terrestrial ecosystems (Karner, DeLong and Karl 2001; Leininger et al.2006). In contrast, AOB usually dominate numerically over AOA in wastewater treatment plants and occasionally in fertilised soils (Bates et al.2011; Mussmann et al.2011). Little is known about the abundance and diversity of comammox Nitrospira, although their abundance has been reported to be comparable to, or higher than the abundance of other ammonia oxidisers.......

....The notion that ammonia oxidation is problematic at low pH was overturned by the discovery of the first obligately acidophilic ammonia oxidiser, N. devanaterra (Lehtovirta-Morley et al.2011). Originally isolated from an acidic agricultural soil, this archaeon grows autotrophically in the pH range of 4–5.5 in laboratory culture with ammonium chloride as its sole energy source. Although Nitrosotalea is the only obligately acidophilic ammonia oxidiser described to date, it is unlikely to be the only microorganism performing nitrification......

.........The kinetic theory of the optimal pathway length suggested that the comammox process would lead to a higher yield but lower growth rate than incomplete ammonia oxidation, giving comammox organisms a competitive advantage when ammonia concentration is low (Costa, Pérez and Kreft 2006). It is interesting to note that the half-saturation constants of many AOA and comammox are roughly within the same range.
cheers Darrel
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Re: Using deep gravel and bacteria to control nitrogen

Post by dw1305 »

Hi all,
Honestly the last one for the moment, but there is also an update to Myriophyllum remediation of swine waste water paper (https://www.ncbi.nlm.nih.gov/pubmed/29056931).

It is "Enhancement of facultative anaerobic denitrifying communities by oxygen release from roots of the macrophyte in constructed wetlands (CW)" (https://www.ncbi.nlm.nih.gov/pubmed/29056931) It looks at Radial Oxygen Loss (ROL) in the rhizoplane (the section of the rhizosphere nearest the plant root).
......that the abundance of facultative anaerobic denitrifiers in the rhizoplane was the most of the three niches, that in the water (5–10 cm) was the less and that in the sediment was the least. .......Denitrifiers in the rhizoplane were mainly dominated by enriched Pseudomonas,Aeromonas, and Acinetobacter. The theoretical calculation of oxygen sources and consumption indicated that water re-aeration should support the oxygen demands for nitrification in the aerobic layer (0–5 cm), and the ROL could stimulate the growth of facultative anaerobic denitrifiers in the rhizoplane and water (5–10 cm) to achieve denitrification within CW systems.
cheers Darrel
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Re: Using deep gravel and bacteria to control nitrogen

Post by dw1305 »

Hi all,
Another interesting (to me anyway) paper has come my way, it isn't a plant based one, but looks in more detail at the microbial diversity in freshwater aquarium filters, and also provides some figures.

The paper is Sauder, L. et al. (2018) "“Candidatus Nitrosotenuis aquarius,” an Ammonia-Oxidizing Archaeon from a Freshwater Aquarium Biofilter" Appl. Environ. Microbiol. 84:e01430-18. (https://aem.asm.org/content/84/19/e01430-18).

I'm not sure what the access arrangement is for it, but if any-one wants a copy PM me.
Conclusions.

We report the cultivation and complete genome sequence of “Ca. Nitrosotenuis aquarius,” a mesophilic, autotrophic, ammonia-oxidizing archaeon that originates from a freshwater aquarium biofilter and belongs to the “Ca. Nitrosotenuis” lineage of the Thaumarchaeota.

“Ca. Nitrosotenuis aquarius” cells grow up to 3 µm, which is the longest cell length reported for AOA representatives. Cells are consistently slender (0.4 µm), maintaining a high surface area-to-volume ratio, which is characteristic of oligotrophic cells. “Ca. Nitrosotenuis aquarius” possesses several genes for protein glycosylation, which could modify flagella or S-layer proteins, and may promote enhanced surface adhesion.

Incubations of biomass from the freshwater aquarium biofilter where “Ca. Nitrosotenuis aquarius” originates demonstrate that AOA contribute to ammonia-oxidizing activity and support the previously reported numerical dominance of AOA in freshwater aquarium biofilters (1, 2).

This work suggests that, contrary to common belief in the aquarium industry, AOB are likely not primarily responsible for ammonia oxidation in aquarium biofilters. Laboratory cultures of AOA originating from freshwater aquaria, such as “Ca. Nitrosotenuis aquarius” cultures, are useful for future investigations of the ecology and physiology of freshwater AOA, with potential commercial applications in aquarium and aquaculture operations.
cheers Darrel
Last edited by dw1305 on 04 Sep 2019, 17:17, edited 1 time in total.
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Re: Using deep gravel and bacteria to control nitrogen

Post by bekateen »

Thanks, Darrel, for continuing to provide more info on this topic.

There are so many different (AMD similar) prokaryotes... I'm confident the discoveries will be coming for a long time.

Cheers, Eric
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Re: Using deep gravel and bacteria to control nitrogen

Post by Bas Pels »

As I am a fan of panted tanks - even with cichlids I try to keep plants - I use faily thick layers of sediment.

My largest tanks have a some 5 cm thick layer on average, even if both of them are unplanted, due to the herbivourous nature of the fishes involved (headstanders are quite similar to cows).

However, in spring ?I changed the soil in another tank, from 5 cm of sand into 10 cm of a clay- sand mixture. I wanted to have a field of Echionodorus argentinensis in full bloom. And this is a species needing a lot of food.

Plants are still a bit young, but are doing OK. However, after I entered some fishes (Tanichthys) I never saw them again. It could have been the soil, it could have been another cause, such as the temperature. The water was cold, early in spring.
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Re: Using deep gravel and bacteria to control nitrogen

Post by dw1305 »

Hi all,
bekateen wrote: 04 Sep 2019, 16:55.....There are so many different (AMD similar) prokaryotes... I'm confident the discoveries will be coming for a long time.
I'd definitely expect that they will find more. I'm particularly interested in whether they find a universal "core" assemblage of microbes, or whether that assemblage changes dramatically with differing ammonia loadings.

There is some interesting research in the UK by Laura E Lehtovirta-Morley (at UEA (https://people.uea.ac.uk/l_lehtovirta-m ... hinterests)):

It is summarised in "Ammonia oxidation: Ecology, physiology, biochemistry and why they must all come together" FEMS Microbiology Letters, Volume 365, Issue 9, May 2018 (https://academic.oup.com/femsle/article ... 58/4931719).
....AOA vastly outnumber AOB in most soil and aquatic environments, often by orders of magnitude (Leininger et al.2006). It is estimated that there are 1 × 10^28 AOA cells in the Earth's oceans and they are some of the most numerous living organisms on Earth, accounting for up to 40% of all prokaryotes in marine ecosystems and 1%–5% in terrestrial ecosystems (Karner, DeLong and Karl 2001; Leininger et al.2006). In contrast, AOB usually dominate numerically over AOA in wastewater treatment plants and occasionally in fertilised soils (Bates et al.2011; Mussmann et al.2011). Little is known about the abundance and diversity of comammox Nitrospira, although their abundance has been reported to be comparable to, or higher than the abundance of other ammonia oxidisers.......

....The notion that ammonia oxidation is problematic at low pH was overturned by the discovery of the first obligately acidophilic ammonia oxidiser, N. devanaterra (Lehtovirta-Morley et al.2011). Originally isolated from an acidic agricultural soil, this archaeon grows autotrophically in the pH range of 4–5.5 in laboratory culture with ammonium chloride as its sole energy source. Although Nitrosotalea is the only obligately acidophilic ammonia oxidiser described to date, it is unlikely to be the only microorganism performing nitrification in acidic soils......

.........The kinetic theory of the optimal pathway length suggested that the comammox process would lead to a higher yield but lower growth rate than incomplete ammonia oxidation, giving comammox organisms a competitive advantage when ammonia concentration is low (Costa, Pérez and Kreft 2006). It is interesting to note that the half-saturation constants of many AOA and comammox are roughly within the same range.
cheers Darrel
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Re: Using deep gravel and bacteria to control nitrogen

Post by dw1305 »

Hi all,
Bas Pels wrote: 04 Sep 2019, 20:04 As I am a fan of planted tanks - even with cichlids I try to keep plants ...........
I'd be very reluctant to have a system without plants, even if I had to have a spatial separation between fish and plants.

I don't have any proof, but I would guess that the planted over-tank trickle filters, that I used to use, would be more efficient than any of the other type of filter by an order of magnitude.
Bas Pels wrote: 04 Sep 2019, 20:04However, in spring ?I changed the soil in another tank, from 5 cm of sand into 10 cm of a clay- sand mixture. I wanted to have a field of Echionodorus argentinensis in full bloom. And this is a species needing a lot of food.

Plants are still a bit young, but are doing OK. However, after I entered some fishes (Tanichthys) I never saw them again. It could have been the soil, it could have been another cause, such as the temperature. The water was cold, early in spring.
I'm not sure, Tanichthys albonubes are pretty tolerant of cooler water. The clay shouldn't have added much BOD (it hasn't got any organic matter to decompose), but it could still potentially be an oxygen issue if the REDOX value of the substrate dropped far enough. There is also the possibility of metals being solubilised (by the reducing conditions in the substrate), but I think that is unlikely.

Once the plants have root development there shouldn't be any issues, as the roots would act as "oxygen conduits" into the substrate.

cheers Darrel
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Re: Using deep gravel and bacteria to control nitrogen

Post by MarcW »

Thanks for your input Darrel.

As someone that routinely kills java fern, it'd be great if you could let me know a few plants which could survive with low light and not planted in the substrate?

Anubias do OK, but don't really seem to grow in my tanks, java fern is OK in some tanks but dies in others, maybe that's to do with temperature rather than lighting, it does OK in my lower tanks but not at the top of the rack where the water is 2-4c higher.
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Re: Using deep gravel and bacteria to control nitrogen

Post by dw1305 »

Hi all,
MarcW wrote: 05 Sep 2019, 15:02 .......As someone that routinely kills java fern, it'd be great if you could let me know a few plants which could survive with low light and not planted in the substrate?

Anubias do OK, but don't really seem to grow in my tanks, java fern is OK in some tanks but dies in others........
Have you still got the moss that came with the Asellus etc.? That should grow OK low light.

The best low light plant for me is Bolbitis heudelotii, it does much better in low nutrients and soft water than Java Fern (Microsorum pteropus).

I've also found that some of the selected cultivars of Java Fern (like "Needle Leaf" and "Trident") aren't anything like as good growers low tech as the ordinary variety.

Bolbitis isn't a quick grower, but it is a bit like Anubias, you don't really notice it growing but eventually it forms a reasonable clump (below).

Image

It isn't cheap to buy, and even more expensive as a "mother plant", (presumably because it isn't a quick grower even emersed), but your more than welcome to some of mine.

cheers Darrel
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Re: Using deep gravel and bacteria to control nitrogen

Post by MarcW »

Hi Darrel, yes I still have the moss it's in a tank with the Asellus, hopefully that'll grow to a decent sized clump.

Your comments about the java fern variants match with my experience, now I've thought about it, the narrow leaved version has all died, the few I have remaining are all the normal type and growing OK.

I've ordered some Bolbitis heudelotii to try out thanks for the tip.
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Re: Using deep gravel and bacteria to control nitrogen

Post by TwoTankAmin »

Here is what Tropica says about the heudelotti:
Bolbitis comes from West Africa, a fern with very beautiful transparent green leaves, 15-40 cm tall and wide. When planting do not cover the rhizome because it will rot, and it is best to plant Bolbitis heudelotii on a root or stone. Keep the plant in position with fishing line until it has gained a hold. Easy to propagate by splitting the horizontal rhizome. Growth can be increased considerably by supplying CO2, and is only optimal in soft, slightly acidic water.
from https://tropica.com/en/plants/plantdeta ... WS%29/4407

I seem to recall I tried to plant it where there was decent flow/circulation. I am not sure if that was an urban aquarium myth or good advice.

As for the AOA and AOB. I am not ready to accept the conclusions of Sauder. L. whose name appears on all of the studies I have read so far which conclude AOA are the dominant ammonia oxidizers in aquariums. Science has no idea how much ammonia a single cell of either organism is able to oxidize. The current science usually counts the number of of archaeal and bacterial amoA genes to determine numbers. I believe some AOB have more than one amoA gene. Further, the most recent research appears to indicate that there is an amoA gene in Nitrospira and this organism can oxidize ammonia to nitrate directly.
Although, AOA were numerically dominant over AOB, a presumed third ammonia-oxidizer was also present in the biofilter sand matrix. Identification of Nitrospira-like amoA (Figure 7B) in the biofilter and the strong correlation between the abundance of the Nitrospira nxrB uwm-2 gene and this Nitrospira amoA, suggests a complete ammonia-oxidizing Nitrospira spp. resides in the UWM biofilter. In fact, we found that the comammox amoA was the most abundant ammonia-oxidizing gene in the biofilter (on average 1.9X that of AOA amoA).
(Sorry I am not great at proper crediting, but the complete paper can be read here https://www.frontiersin.org/articles/10 ... 00101/full )

Original Research ARTICLE
Front. Microbiol., 30 January 2017 | https://doi.org/10.3389/fmicb.2017.00101

Freshwater Recirculating Aquaculture System Operations Drive Biofilter Bacterial Community Shifts around a Stable Nitrifying Consortium of Ammonia-Oxidizing Archaea and Comammox Nitrospira

Ryan P. Bartelme, Sandra L. McLellan and Ryan J. Newton*
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Re: Using deep gravel and bacteria to control nitrogen

Post by dw1305 »

Hi all,
TwoTankAmin wrote: 06 Sep 2019, 19:14I seem to recall I tried to plant it where there was decent flow/circulation. I am not sure if that was an urban aquarium myth or good advice.
I'm not sure. None of my plants are fixed to anything, so they tend to end up out of the direct flow from the filter.

The photo above is in my kitchen tank, rain-water and low nutrients and that plant grew from a single rhizome with three or four leaves in ~five or six years.
TwoTankAmin wrote: 06 Sep 2019, 19:14As for the AOA and AOB. I am not ready to accept the conclusions of Sauder. L. whose name appears on all of the studies I have read so far which conclude AOA are the dominant ammonia oxidizers in aquariums. Science has no idea how much ammonia a single cell of either organism is able to oxidize. The current science usually counts the number of of archaeal and bacterial amoA genes to determine numbers. I believe some AOB have more than one amoA gene.
There is a fair bit of research from waste water treatment. "Unraveling the active microbial populations involved in nitrogen utilization in a vertical subsurface flow constructed wetland treating urban wastewater" (https://www.sciencedirect.com/science/a ... 9717301006). This is "The more important role of archaea than bacteria in nitrification of wastewater treatment plants in cold season despite their numerical relationships" (https://www.sciencedirect.com/science/a ... 5418306961)
TwoTankAmin wrote: 06 Sep 2019, 19:14......Freshwater Recirculating Aquaculture System Operations Drive Biofilter Bacterial Community Shifts around a Stable The more important role of archaea than bacteria in nitrification of wastewater treatment plants in cold season despite their numerical relationships Consortium of Ammonia-Oxidizing Archaea and Comammox Nitrospira

Ryan P. Bartelme, Sandra L. McLellan and Ryan J. Newton*
I'm pretty sure that people will carry on finding the complete nitrifier "COMAMMOX Nitrospira" wherever they look. My guess is also that people will keep on finding novel nitrifying micro-organisms, whether they are bacteria or archaea.

cheers Darrel
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Re: Using deep gravel and bacteria to control nitrogen

Post by TwoTankAmin »

I have one problem generalizing from waste water studies which find Nitrobacter is the dominant nitrite oxidizer. The one thing I think has been established beyond any doubt is that Nitrobacter is not found in any meaningful level, if found at all, in aquariums. It is suited to oxidizing higher levels of nitrite than is found in tanks. Nitrospira is what is regularly detected.

Combine this with the discovery of Nitrospira capable comammox and there is a whole host of microorganisms that are likely at work dealing with ammonia in our tanks. I have come to believe there is probably no single dominant bacteria or archaea across the universe of tanks. Rather each tank will have its own particular balance. I am also pretty sure the exact complement of nitrifying organisms in a tank can change over time as the ammonia loading and other parameters change.
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Re: Using deep gravel and bacteria to control nitrogen

Post by dw1305 »

Hi all,
TwoTankAmin wrote: 07 Sep 2019, 19:34 ......I have come to believe there is probably no single dominant bacteria or archaea across the universe of tanks. Rather each tank will have its own particular balance. I am also pretty sure the exact complement of nitrifying organisms in a tank can change over time as the ammonia loading and other parameters change......
I think that is where we are now.

I'll attempt to keep up with the scientific literature, but I probably won't have a proper trawl through it again before Christmas.

cheers Darrel
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Re: Using deep gravel and bacteria to control nitrogen

Post by dw1305 »

Hi all,
dw1305 wrote: 07 Sep 2019, 20:19 ..........I'll attempt to keep up with the scientific literature, but I probably won't have a proper trawl through it again before Christmas........
Another paper has come my way:

Zhao et al. (2015) "Microbial community and removal of nitrogen via the addition of a carrier in a pilot-scale duckweed-based wastewater treatment system" (https://www.sciencedirect.com/science/a ... 241401774X).

It is a plant/microbe bio-filtration paper, and it quantifies the net contributions, to nitrogen removal, of a floating plant (Duckweed - Lemna japonica) and microbial biofilm in a Duckweed based waste water treatment. It also investigates why the nitrogen balance might be slightly different when you use Water Hycanth (Eichornia crassipes) as a floating plant.

They attribute these differences in nitrification to the much larger root area of Eichornia, when compared to Lemna, with this expanded rhizosphere supporting a much larger volume of microbial biofilm.

What is really interesting is that when they added a plastic carrier to the effluent flow (to act as a surrogate "root surface" in the Lemna treatment), to give a comparable area for microbial biofilm adhesion.

They found that the addition of a carrier improves nitrogen (N) removal in a duckweed system and that abundant N-removal microbes on the carrier biofilm contribute to improved N removal.

cheers Darrel
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Re: Using deep gravel and bacteria to control nitrogen

Post by dw1305 »

Hi all,
TwoTankAmin wrote: 06 Sep 2019, 19:14.....As for the AOA and AOB. I am not ready to accept the conclusions of Sauder. L. whose name appears on all of the studies I have read so far which conclude AOA are the dominant ammonia oxidizers in aquariums.........
This is a paper where they didn't find any AOA, but they found a range of novel AOB.

Hüpeden, J. et al (2020) "Taxonomic and functional profiling of nitrifying biofilms in freshwater, brackish and marine RAS biofilters" Aquacultural Engineering 90.
In the freshwater biofilters >99 % of the total AOB reads were assigned to uncultured ammonia oxidizing bacteria with Nitrosomonas communis Nm2 as the closest relative (96 % sequence similarity to the 16S rRNA gene). .....................

In the freshwater biofilters >99 % of the total NOB reads were classified to the genus Nitrospira, which had Ns. defluvii as closest known relative (99 % similarity to the 16S rRNA gene)........

At each RAS, a site-specific nitrifying community was identified. The majority of reads was affiliated with unknown members of Nitrosomonas and Nitrospira, representing several potentially novel species within these genera, which were currently not possible to culture by traditional techniques. The comprehensive phylogenetic analysis to determine the closest taxonomically described species revealed a co-existence of different, Nitrosomonas-like AOB and Nitrospira-like NOB in all biofilters with one representative being the predominant ammonia or nitrite oxidizer at each RAS, respectively.......

The presence and in some cases dominance of AOA over AOB in aquaria and RAS biofilters was reported previously (Bartelme et al., 2017; Brown et al., 2013; Sakami et al., 2012; Sauder et al., 2011) and growth of AOA at low ammonia concentrations has been demonstrated for enriched and isolated representatives (Nicol et al., 2011; Sauder et al., 2012). In other studies, the quantity of AOA in RAS was reported negligible or zero (Foesel et al., 2008; Keuter et al., 2017). The load of ammonia was discussed as important factor for the coexistence of AOA and AOB in RAS, since AOA have a higher affinity towards ammonia than AOB and could be outcompeting AOB at lower concentrations (Brown et al., 2013; Martens-Habbena et al., 2009; Roalkvam et al., 2020). Mostly, AOA dominate in niches characterized by low nutrients, low pH and low DO (Erguder et al., 2009) as well as elevated temperature (Taylor et al., 2017). These parameters are not valid here and in our study, archaeal reads were not present in the 16S rRNA amplicon sequencing data set. In addition, no archaeal amoA sequences were found by specific PCR and no typical cells of AOA were detected via electron microscopic investigation of the biofilms
cheers Darrel
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Re: Using deep gravel and bacteria to control nitrogen

Post by bekateen »

Thanks Darrel, for keeping this up to date. I'm learning a lot.

Cheers, Eric
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Re: Using deep gravel and bacteria to control nitrogen

Post by dw1305 »

Hi all,
Last one for a bit.

This paper looks at the interaction between dissolved organic carbon (DOC), nitrification and the microbial assemblage. The paper is open source, and should be available to every-one:

Navada, S., Knutsen, M.F., Bakke, I. et al. (2020) Sci Rep 10.
Nitrifying biofilms deprived of organic carbon show higher functional resilience to increases in carbon supply
........ the organic carbon to ammonia nitrogen (C/N) supply ratio can influence resource competition between heterotrophic and nitrifying bacteria for oxygen and space. We investigated the impact of acute and chronic changes in carbon supply on inter-guild competition in two moving bed biofilm reactors (MBBR), operated with (R1) and without (R0) external organic carbon supply. The microbial and nitrifying community composition of the reactors differed significantly. Interestingly, acute increases in the dissolved organic carbon inhibited nitrification in R1 ten times more than in R0.

A sustained increase in the carbon supply decreased nitrification efficiency and increased denitrification activity to a greater extent in R1, and also increased the proportion of potential denitrifiers in both bioreactors...... Specifically, efficient removal of organic matter before the nitrification unit can improve the robustness of the bioreactor to varying influent quality.

Thus, maintaining a low C/N ratio is important in nitrifying biofilters when acute carbon stress is expected or when anoxic activity (e.g. denitrification or H2S production) is undesirable, such as in recirculating aquaculture systems (RAS).
Which I think can be digested as:

"a lot of of organic matter in your filter reduces nitrification, but increases denitrification".

Which partially explains why not using your filter as a syphon, not letting the filter media get too gungy and having a pre-filter, helps with nitrification.

I'm a "nitrate factory" fan, nitrate (NO3-) can always be mopped up by plants and water changes.

cheers Darrel
Last edited by dw1305 on 09 Sep 2020, 16:42, edited 4 times in total.
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Re: Using deep gravel and bacteria to control nitrogen

Post by dw1305 »

Hi all,
bekateen wrote: 09 Sep 2020, 15:16Thanks Darrel, for keeping this up to date. I'm learning a lot.
Cheers, Eric
I am as well, but it's back to the old problem, trying to line up all the moving bits and then work out what they actually mean in practice.

I was thinking about this in terms of marine aquarists using Vodka, or similar, as a carbon source. I assume they do it (and it works by this mechanism) to increase denitrification in the deep sand bed.

Is it still in vogue? I assume some-one will know

cheers Darrel
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Re: Using deep gravel and bacteria to control nitrogen

Post by bekateen »

Here's a new video about the fish store in San Francisco where the owner uses the deep gravel and never does water changes. We've discussed earlier in this thread why that is not so simple, but it's still a cool pet store. Enjoy:

https://youtu.be/watch?v=rg1u-XVMU3Q
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Re: Using deep gravel and bacteria to control nitrogen

Post by dw1305 »

Hi all,
Three years on I'm going to resurrect this old thread, because it contains bits that are relevant. I've recently had some correspondence with Dr Newton at the University of Wisconsin (his is the lab. referenced by @TwoTankAmin earlier in the thread).

I contacted him after reading: Bartelme RP, Smith MC, Sepulveda-Villet OJ, Newton RJ. (2019). "Component microenvironments and system biogeography structure microorganism distributions in recirculating aquaculture and aquaponic systems". mSphere 4:e00143-19.
It is well worth a read and he very kindly both replied to my email and answered the questions I asked, which were:
1. If we planted an aquarium and waited for the plants to "grow in" over ~6 weeks, where does the initial inoculum of AOA and comammox Nitrospira come from? and

2. If we did need to add them? What would be the best source of initial inoculum?
and his reply was
1).... the water - most municipal water systems contain some number of nitrifiers, which then come out of your residence tap; 2) the plants - nitrifiers are also commonly associated with plants. Or, it could be they drift in from the air - seems less likely, but it is not impossible.

2. If you do need to add nitrifiers the best source is from an aquaponics or aquaculture system that is already running and removing ammonia. Some water or sediment/soil or part of the biobilter (if there is one) is an excellent starter. Without this source as an inoculum then you could add some roots from plants from any other tank that is running - these are likely to have nitrifiers associated with them. A small clipping put into the tank would be enough.

In some lab tests we found that adding previous material from a running biofilter could reduce ammonia oxidation start-up time from 2-3 weeks to 2-3 days. We also tested a commercial product of nitrifiers & it did decrease the time to ammonia oxidation start-up. It was slower than our biofilter material transfer, but much quicker than doing nothing. However, the microbes present in the system from the commercial product disappeared over a few weeks and were replaced by those more common to our system. So, it seems some products could help “jump-start” the process, but it will be a lot less predictable and ultimately may not determine what microbe succeed in the long run.
cheers Darrel
Last edited by dw1305 on 24 Jul 2023, 18:12, edited 1 time in total.
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Re: Using deep gravel and bacteria to control nitrogen

Post by TwoTankAmin »

@dw1305
TY for posting this Darrel.

Years ago I used to read the bacteria came into tanks from the air. I did not accept this as the normal route. I worked in Saudi Arabia int the late 1970s. I was there when it was 130F in the shade and you could not see your sweat because it evaporated as fast as you made it. There is no way floating bacteria were surviving in that dry air. We had a Brit who built a sw water tank and stocked it with fish and water from the Arabian Gulf. His tank had to cycle from the water not the air.

Here is a better explanation:

Bradley, Tyler C., Charles N. Haas, and Christopher M. Sales. 2020. "Nitrification in Premise Plumbing: A Review" Water 12, no. 3: 830. https://doi.org/10.3390/w12030830
Abstract
Nitrification is a major issue that utilities must address if they utilize chloramines as a secondary disinfectant. Nitrification is the oxidation of free ammonia to nitrite which is then further oxidized to nitrate. Free ammonia is found in drinking water systems as a result of overfeeding at the water treatment plant (WTP) or as a result of the decomposition of monochloramine. Premise plumbing systems (i.e., the plumbing systems within buildings and homes) are characterized by irregular usage patterns, high water age, high temperature, and high surface-to-volume ratios. These characteristics create ideal conditions for increased chloramine decay, bacterial growth, and nitrification. This review discusses factors within premise plumbing that are likely to influence nitrification, and vice versa. Factors influencing, or influenced by, nitrification include the rate at which chloramine residual decays, microbial regrowth, corrosion of pipe materials, and water conservation practices. From a regulatory standpoint, the greatest impact of nitrification within premise plumbing is likely to be a result of increased lead levels during Lead and Copper Rule (LCR) sampling. Other drinking water regulations related to nitrifying parameters are monitored in a manner to reduce premise plumbing impacts. One way to potentially control nitrification in premise plumbing systems is through the development of building management plans
full paper here https://www.mdpi.com/2073-4441/12/3/830

The part about which starter bacteria has been put forward by Dr. Hovanec for years. In one of his patent applications and I think also in one of his papers they did a test with a bacterial starter and they found that bacteria was gone once the tank was cycled.
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Re: Using deep gravel and bacteria to control nitrogen

Post by bekateen »

Thanks Darrel and TTA. All good info. From Darrel's post, this affirms my experience that a "good squeeze" from a dirty sponge filter into a new clean tank with a new sponge filter can cycle the tank in days. For the most part, that's all I do now.

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Re: Using deep gravel and bacteria to control nitrogen

Post by cwindram »

I plan to try a deep substrate as an experiment in a tank that I'll be setting up soon. I'd seen that video by Aquarium Coop on Ocean Aquarium in San Francisco, and found it fascinating. Maybe a bit off-topic, but I have always used plants to control nitrogen... all of my tanks have live plants, and there is never any measurable nitrate in them, in fact I sometimes add a general fertilizer to help grow the plants. I've got a recent setup (50 gals) going that's almost exclusively emergent plants, and this leaves lots of swimming space in the tank. No cats are visible in the photo, but this tank has Platydoras, Mochokiella, Microglanis, Tatia, and a fish that is probably
a Pseudopimelodus.
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Re: Using deep gravel and bacteria to control nitrogen

Post by bekateen »

cwindram wrote: 07 Dec 2022, 22:18I'd seen that video by Aquarium Coop on Ocean Aquarium in San Francisco, and found it fascinating.
Hi cwindram,

This thread was started after I visited Ocean Aquarium (a couple of years before the Aquarium Coop video) and I talked to the owner about his tanks. He was encouraging me to give it a try. I think he has a really unique system, and it definitely works well for him, but I think there are a few aspects which are difficult to recreate in many home tanks.
1) Most of his tanks are packed with plants, esp. java moss and other floating stem plants, to the point that it's often difficult to see what's in the tank. That's fine if that's the effect you want, but if you want a tank with open area or water current, that becomes less practical.
2) While he doesn't do outright water changes, he does do a lot of water replacement. First, his tanks are mostly uncovered, so he's often topping the tanks with clean water to compensate for evaporative water loss. Second, he does sell fish, and when he does, he removes water for bagging and has to replace it.

Obviously, none of these steps amount to the water changes we do in most home tanks, but my experience (and if you read this whole thread, you learned this) is that I'm a relatively bad plant keeper because my tanks mostly have high current and low low light. So for me, the model doesn't work as well.

I love your emergent plants in your tank. How are you supporting them in the tank? I presume there are poles (it looks like bamboo) holding the baskets in place, but am I seeing that right?

Thanks,
Eric
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Re: Using deep gravel and bacteria to control nitrogen

Post by aquaholic »

I have not read through all the posted pages on this topic but my city council water often has high levels of ammonia, partly because of chloramine dose fluctuations and partly because of cleaning activities. There is no prior warning, no indication how long this lasts and with several fish rooms, having to delay a scheduled water change day creates problems.

I solved this issue very easily with a slow constant drip (automatic) water change that displaces old water out. No timers, valves, mechanical devices. There are multiple other benefits to constant drip but this solved the poor water quality issue instantly and removes nitrate build up issue at the same time.

I run a few hundred tanks on central filtration systems, all bare bottom but do keep several 55 gallon drums full of gravel to keep biofilm - aquatic stability. They have not been disturbed for almost 15 years.
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Re: Using deep gravel and bacteria to control nitrogen

Post by dw1305 »

Hi all,
bekateen wrote: 07 Dec 2022, 21:21 .........From Darrel's post, this affirms my experience that a "good squeeze" from a dirty sponge filter into a new clean tank with a new sponge filter can cycle the tank in days. For the most part, that's all I do now......
I think you are right Eric, I'm guessing that nearly all people use the "good squeeze" method if they can.

We've also had some correspondence with Dr Tim Hovanec (on UKAPS) fairly recently, which again may be of interest to some members.

cheers Darrel
Last edited by dw1305 on 31 Jan 2023, 15:28, edited 1 time in total.
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