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AAHM-321 MICROBIAL & PARASITIC DISEASES OF FISH AND SHELLFISH

Bioremediation, its role in disease management

Bioremediation
Bioremediation is a method of eliminating contamination from complex waste
by using micro-organisms, without using harmful chemicals. It is an innovative
technology that implements the use of beneficial micro-organisms called bio-
remediating agents for the optimization of water quality and soil by modifying
the harmful microbial load in ponds to boost the mineralization process of
organic loads and remove unwanted harmful substances. The intensification of
the aquaculture system gives rise to many problems associated with stress and
disease outbreaks, contamination of waterbodies due to the release of effluent
rich in organic load, and expensive disease treatment and control measures. By
using bioremediation, we can deal with a wide range of problems by lowering
the stress on cultured stock, successfully removing any contaminant from the
culture system, and using useful microorganisms to enhance the health of the
cultured fish stock, It also involves modifying environmental conditions so that
bacterial growth and degradation can proceed more rapidly and turn them into
harmless substances. It involves both in-situ and ex–situ approaches to treating
waste generated in the culture system from unconsumed feed and faecal matter.
Common bacteria employed are Bacillus spp. like Bacillus subtilis, Bacillus
licheniformis, Bacillus cereus, and Bacillus coagulans.
Applications of Bioremediation in aquaculture pond
There involve 3 major common methods on this bioremediation applications
which are biological methods, bacterial groups and algal biomass in the
bioremediation. Apart from this there occurs various technological
developments which are described in the following.
Common methods of bioremediation
Biological methods Bacterial groups Algal biomass
 IMTA- Integrated Anammox HRAP- High-rate algal
pond
 Multitrophic Aquaculture Bacteria to remove geosmin & MIB
 Oxidative bioremediation
 Microbial mats
 Zeolite
 Nitrifying bioreactor

Biological methods
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The technology named IMTA is not a new term and was followed by 3
productive decades and commonly called as Ecologically engineered
Aquaculture and ecological aquaculture and the name integrated aquaculture
was also came across by some researchers. In the advent of this invention, 2
types of the reactors have been described which are.
 In-Situ Stringed Bed Suspended Bioreactor (SBSBR)
 Ex-Situ Packed Bed Bioreactor (PBBR)
They form the key component for the establishment of Recirculating
Aquaculture Systems (RAS). Even though, great efforts have been made on the
examination of nitrifying biofilters for aquaculture practices, the research have
for the most part been concentrating on performance of an individual
component under specific operating conditions utilizing average ammonia
removal rate to depict the biofilter nitrification performance.
Algae a new green energy
Basically, there are two main types of algal pond systems: static algal ponds
and high-rate algal ponds (HRAP) which can be used for effluent treatment.
This HRAP which is a low-energy wastewater treatment made the second loop
for the water treatment of flow through or recirculating aquaculture systems.
They are characterized by a continuous water circulation and mixing in a culture
tank, either by a paddlewheel or by strong aeration, and by a short residence
time (days). The main advantage is that the outcome of this effluent treatment
which is wastewater solids can be used for recovering energy and the harvested
algal mass can be used for biofuel production. This perforated algal species can
assimilate nutrients from the wastewater and thereby providing both secondary
and partial tertiary-level treatment.
Bacterial groups
Anaerobic ammonium oxidation (Anammox)
An innovative technological advancement in the removal of ammonia nitrogen
is the Anammox. This is a bacterial consortium considering having a unique
property of removing ammonium ions instead of passing through a two-stage
process of aerobic nitrification and anaerobic denitrification, it combines nitrate
or nitrite with ammonia and form dinitrogen gas. The main advantage of this
innovation is that they don’t require any organic carbon source and moreover it
saves costs as less energy for aeration. The main problem in the freshwater
aquaculture ponds is the Geosmin and MIB which are terpene-type compounds
produced by algae. To elucidate this smell, many bacterial strains were tested
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and found that Rhodococcus wratslaviensis DLC-cam, Pseudomonas putida G1,

and Rhodococcus sp. T1 had the ability to transform geosmin or MIB. In
addition to these researchers are in the verge to improve the activity of these
organisms with a help of a key enzyme in the degradation of terpene-like
compounds. There is an option to raise a question on the bacterial toxicity of the
fishes. In context to that, they have found that, Rhodococcus wratslaviensis
DLC-cam was found to not be pathogenic against fish, even at very high
concentrations. These bacterial strains have the capacity to grow in the minimal
media with fish feed as the sole carbon source, a condition typical of
aquaculture environments. Finally, all three strains can be frozen without
significant loss in viability, which is desirable for storage and transport to a
treatment area.
How bioremediation in aquaculture helps in disease management
In aquaculture bioremediation is being used to regulate the following
parameters which helps in disease management.
 Water quality: by removing excess nutrients, organic matter, and
harmful substances such as ammonia, nitrites, sulphides, which can
induce stress to fish or weaken the immune system of cultured organisms
e.g., Phytoremediation, Biological filters.
 Nutrient cycling: by breakdown and recycling of organic waste such as
nitrogen and phosphorus through the activity of beneficial microbes and
converting it into input for growth of live feed for fish.
 Biological control: using beneficial microorganisms or organisms
competing with or prey upon harmful pathogens. For example, probiotics
containing beneficial bacteria can be added to the culture environment to
enhance the immune response, suppress the growth of disease-causing
bacteria, and use natural predators for specific pathogens.
In situ bioremediation involves treating the contaminated material at the site
while ex situ involves the removal of the contaminated material to be treated
elsewhere
Some examples of bioremediation technologies are bioventing, landfarming,
bioreactor, composting, bioaugmentation, rhizofiltration, and bio stimulation.
Each type of bioremediation has its specific applications and effectiveness
depending on the nature of the contaminants and the environment.
Biofilm/periphyton Based Bioremediation/Plant assisted Bioremediation.
Plant assisted bioremediation or periphyton based bioremediation involve use of
agricultural waste/byproducts such as sugar cane bagasse, paddy straw, rice
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husk, wheat corns on application it provides substrate for growth for Organisms
such as algae, zoogleal, filamentous bacteria which are source of food for fish
as well as play a key role in production of enzymes, degradation of organic
matter and environmental toxicants.
Applied periphyton Improve production as well as water quality. Sugar bagasse
is used in shrimp culture.
Green water technology-An innovative bio- augmentation technique
Involves the addition of beneficial microorganisms to the aquaculture water to
create a natural and balanced ecosystem that promotes the health and
productivity of the cultured organisms.
Involve use of photosynthetic algae to create natural environment for oxygen
production as well as nutrient removal from the system which male
environment favourable for growth. In these strains of beneficial microbes
(Probiotics) are also used to improve health of cultured system. Algae along
with associated bacteria in ecosystem and probiotic competes with disease
causing organisms for space, food and inhibit its growth thus provide an
optimum environment for growth of cultured species.
Bio-floc Based Bioremediation
Bio-floc technology is a technique of enhancing water quality in aquaculture
through balancing carbon and nitrogen in the system. It considered as a
resourceful alternative system since nutrients could be continuously recycled
and reused.
These microorganisms (bio-floc) have two major roles:
(i) maintenance of water quality, by the uptake of nitrogen compounds
generating “in situ” microbial protein.
(ii) nutrition, increasing culture feasibility by reducing feed conversion ratio and
a decrease of feed costs
In bio-floc complex species diversity provides an ecosystem where beneficial
organism reduces the chance of growth of disease-causing pathogen thus
providing optimum environment for growth of cultured species
Biotransformation of heavy metals
Commonly found heavy metals in the environment include lead, mercury,
arsenic, chromium, cadmium, nickel, zinc, and copper. Among these metals,
chromium (Cr) is one of the most toxic pollutants generated by anthropogenic
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activities. Product developed in which bagasse and coconut husk charred with
acid have been used to detoxify Cr(VI) which is more toxic into Cr(III) less
toxic form in acidic medium, due to their electron donating capacity.
Removal of heavy metals by using plants or other way improves the health of
culture system thus reduces the chance of occurrence of toxicity as well as its
bioaccumulation.
Enzymes in bioremediation
Different microbial enzymes have been reported to be helpful in the removal of
pollutants in the environment to make it health and fit for fish species.
pollutants like pesticides, dyes, heavy metals, xenobiotics can be removed.
Mechanisms such as elimination, oxidation, ring-opening, and reduction are
used by enzymes in bioremediation. Different factors which include
temperature, contact time, concentration and pH affect the potency of microbial
enzymes.
 Lipase produced by Bacillus pumilus is used to remove Oil content from
industrial wastewater.
 Oxidoreductase produced by Bacillus safenis for removal of Xenobiotics.
 Oxygenase produced by Pseudomonas species in pesticides removal.
 Of the several strains of bacteria, gram-positive Bacillus spp. is generally
more efficient in converting organic matter back to CO2.
 Other bacterial sp. of importance towards bioremediation are:
Nitrobacter, Pseudomonas, Enterobacter, Cellulomonas, and
Rhodopseudomonas spp. Bioremediators (biocontrol agents) apart from
improving water quality parameters, modify or manipulate the inherent
microbial communities and reduce the pathogenic microbes.
Ammonia build-up of in the culture systems are removed by nitrifiers by
oxidation of ammonia to nitrite and subsequently to nitrate.
Conclusion
Aquaculture production has expanded relentlessly in late years and is the
quickest developing sustenance creation part and has turned into an important
segment of national development. It was once viewed as an earth sound practice
on account of its polyculture and integrated aquaculture where they
incorporated the optimal utilization of land and water resources with restoring
the water quality. Due to intensification of the farming activities, we are in the
verge to remediate the present aquaculture systems. Bioremediation play an
important role in this process with the view to improve the water quality by
introducing macro and microorganisms and /or their products as additives,
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which are referred to as bio remediators or bioremediating agents. The

application of bio remediators in aquaculture shown promise, but further
investigations may be expected with propagation of molecular approaches to
analyse bacterial communities.