What better way to start Biomin ‘Aquaculture Days’ in mid-April than with a visit to a restaurant on top of an aquarium.

1. What better way to start Biomin
‘Aquaculture Days’ in mid-April
than with a visit to a restaurant on
top of an aquarium.
However, this was an aquarium
with a difference. This II-World War anti-aircraft gun emplacement and
battle tower, one of three built in Vienna and this one right in the heart of
the city, was converted in 1957 into the role it has today as an aquarium,
attracting over 350,000 visitors a year and becoming the second most
visited privately-operated sightseeing site in the country.
The dinner was proceeded with an introduction and welcome to the event
by marketing director of Biomin, Herbert Kneissl.
Aquaculture days
PHOTOSHOOT Biomin ‘Aquaculture Days’PHOTOSHOOT
28 | INTERNATIONAL AQUAFEED | May-June 2015

2. Over the next two days the group of
more than 40 aqua industry profes-
sionals from 14 countries heard 13
presentations and visited numerous
laboratories. The focus of the two-
days of visits and lectures was to
look at the detail behind research
and development being carried out
by Biomin into areas of fish nutrition
that provide farmers and processors
with financial benefits in both yield
and quality.
May-June 2015 | INTERNATIONAL AQUAFEED | 29

3. Day One
Biomin research and development facilities, the BIOMIN Research Center,
are integral in the research university campus at Tulln in Lower Austria,
but which is some 50km north-west of Vienna on the Danube River.
The first day saw the group visiting Biomin’s laboratories throughout the
university, that were dedicated to plant biotechnology, mycotoxin strate-
gies, disease challenges, phytogenics and pathogen control all centred on
aquaculture and aquatic species.
It was clear from the facilities dedicated to this research into products to
counter the impact of mycotoxin on fish species, in particular, that aqua-
culture nutrition and sustainability is at the forefront of this company’s
developments when it comes to new products.
The day was rounded off with discussions and a visit to the Göttweig
Abbey, a further 30km west along the Danube River, and near the town
of Krems.
Day Two
The venue for a series of lectures was held at the University of Veterinary
Medicine in Vienna the following day. The half-day seminar was intro-
duced by Professor Mansour El-Marbouti, who heads the Department
for Farm Animals and Veterinary Public Health and who has published
137 papers on topics relating to this discipline and has expertise in fish
diseases, fish bacteriology, fish virology and fish mycologoy.
Brief lectures covered topics from ‘Probiotics as modulators of the gastro
intestinal tract’, delivered by invited PhD student Benedict Standen of
Plymouth University in the UK: A presentation International Aquafeed will
be reporting on in an upcoming edition.
Biomin researchers explored the research behind several of its aquatic
products including the ‘Effective beneficial bacteria in aquaculture’ deliv-
ered through its product AquaStar and presented by Jutta Zwielehner.
The ‘Importance of phytogenic supplementation in aquafeeds’, by Otavio
Castro was another well-received presentation. And Pedro Encarnacao
Biomin’s aquaculture technical director based in Singapore, summarised
the morning session for delegates.
We should not overlook the presentation made by Jorge Dias on the
‘Trends in feed formulation and feed technology’ and the impact this will
have on meeting the growing demand of a world struggling to feed itself
adequately; another presentation International Aquafeed will be reporting
in more details upon.
The afternoon was taken up with delegates and staff from Biomin touring
the fish trials facilities at the University of Veterinary Medicine before
the group has one final meeting at the restaurant Viennese Heurigen
‘Weingut am Reisenberg’ overlooking the city of Vienna.
Overview
This was a most valuable visit with a balance of research explained, indus-
try issues addressed and a close look at the trials and tribulations that go
into product development. It also gave visitors time to network among
themselves and exchange knowledge and points-of-view, but overall it
brought home the message that aquaculture needs companies to com-
mit to research and development at this level if industry is to supply a
cost-effective food protein at affordable prices and in a consistent, safe
and efficient manner.
Many thanks to Biomin for its ongoing initiative to engage with the aqua-
culture industry at this level and in this depth.
The main task of our microbiology research team is to analyze
and develop new, innovative microbial feed additives for different
livestock species to improve animal performance and health.
Through years of research we have gained sound expertise in
the development of multispecies or multi-strain combinations of
beneficial bacteria, or probiotics.
These additives offer a good alternative to antibiotics or other
treatments without the apparent associated drawbacks.
Over time we have established a large strain collection with
plenty well-characterised probiotic strains, as well as a plethora of
livestock-specific and livestock-relevant pathogens. The ultimate
goal is to find strains that complement each other in their unique
actions.
In our research we combine techniques from classical microbiol-
ogy with modern state-of-the-art molecular biology techniques to
identify the best suitable probiotic strains for a distinct purpose.
To obtain a safe and acceptable biological feed supplement, the
development process covers all steps from in vitro (isolation,
identification, safety evaluation, characterization and assessment
for specific properties) to in vivo (alterations in host gene expres-
sion or microbial composition in the gut).
For aquaculture probiotics we focus on two different applications:
1) feed application to improve animal health via gut health and
2) pond application to improve water quality and indirectly aid
animal health.
For both applications, stability of probiotic strains under changing
environmental conditions is a prerequisite.
Thus, our in vitro work includes tests for pH, salt, bile, tempera-
ture tolerance and combinations thereof.
For feed application the main goal is to identify strains that
antagonize a broad spectrum of different pathogens.
Antagonism would ideally occur not only through classical mecha-
nisms, such as bacteriocin or acid production, but also through
alternative mechanisms, such as quorum quenching, that can
reduce a pathogen’s virulence.
For pond application the main goal is to identify strains able to
maintain water quality. To do so they need to efficiently degrade
organic matter and/or remove toxic waste compounds, such as
nitrate, nitrite, ammonia, or hydrogen sulfide.
In the laboratory we have several colorimetric test systems
available to define how these strains perform in vitro, especially
concerning removal of nitrogen compounds.
Microscopic life aquatic
- improving animal performance
by Barbara Weber, Microbiologist Responsible for
Aqua-related Activities, Biomin
Aquaculture days
PHOTOSHOOT +
30 | INTERNATIONAL AQUAFEED | May-June 2015

4. The year 1960 is commonly regarded as the starting point that led to the dis-
covery and study of mycotoxins. While many in the mycotoxin research com-
munity are familiar with the turkey “X” disease that year when turkeys in
England died from aflatoxin contamination, few know about the epizootic of
hepatoma in trout occurred in California that same year, later found to be
caused by aflatoxins.
At the BIOMIN Research Center, we take advantage of the fact that mycotox-
ins are natural molecules.
Mycotoxins are degradable in the environment, and a goal of our research is to
get natural mycotoxin degradation and detoxification reactions to work in the
gastrointestinal tract of animals.
Our molecular biology team has isolated strains of fumonisin-degrading bacteria,
and studied the molecular biology and biochemistry of fumonisin degradation
in detail. We found that a cluster of genes has been shaped and optimised by
evolution over countless years specifically to break down fumonisins and use
them for bacterial cellular metabolism.
Transcription of the genes is turned on in the presence, and turned off in the
absence of fumonisins, and availability of fumonisins accelerates bacterial growth.
One particular enzyme of the degradation pathway, fumonisin esterase, is now
available and registered in the EU as the world’s first mycotoxin degrading feed
enzyme, FUMzyme®.
Fumonisin esterase removes two side chains from fumonisins by hydrolytic
cleavage.
This cleavage drastically reduces the affinity of the mycotoxin to the eukaryotic
target protein, ceramide synthase. An important part of developing FUMzyme®
for use as feed enzyme for gastrointestinal fumonisin detoxification was to
develop biomarkers of fumonisin exposure and fumonisin effects.
We established sophisticated methods, based on liquid chromatography and
mass spectrometry, to measure concentrations of fumonisin-derived metabo-
lites, and metabolites involved in ceramide synthase activity in samples from
animals.
These methods allow to study dose and effect of fumonisins and of FUMzyme®,
and were already used to tune FUMzyme® formulation and dose for pigs and
poultry. Aquatic species should be next, and we already know that fumonisin
esterase activity at low temperature is still very high.
We have also made microbes for detoxification of zearalenone and deoxyni-
valenol available as feed additives. Most recently, we isolated a bacterial strain
which breaks down ergopeptines, the predominant class of ergot alkaloids.
Aquatic species are sensitive to mycotoxins, and aquaculture is young and
open to science and innovation. For these reasons, application of mycotoxin
detoxification biotechnology in aquaculture is a particularly interesting challenge
for us at Biomin.
The use of plant-derived feed for aquaculture continues
to grow, exposing those species to the risk of mycotoxin
contamination.
While previously conducted fish trials have demonstrated
numerous negative impacts of mycotoxins on aquaculture
species, this information is scarce compared to what is
known regarding terrestrial animals, particularly livestock.
One objective of the Biomin Research Center’s cell
biology team is to better understand the risks that
mycotoxins pose to aquaculture and the effectiveness of
detoxification products. The research effort comprises
several assays and techniques based on cell cultures.
Working with cell lines is faster and less cost-intensive
than working with whole animals, especially when it
comes to analyses of different concentrations and com-
binations of mycotoxins. Ethical considerations further
support use of these techniques when possible.
Furthermore, cell culture systems offer a more sensitive
and reproducible method for preliminary screening of
toxicity of mycotoxins and elucidating their modes of
actions.
For example, the adherent cell line RTgill-W1, derived
from a primary culture of rainbow trout gill fragments,
can be used as suitable cell culture model to test effects
of mycotoxins in fish cells.
Cytotoxicity studies with the most abundant and most
carcinogenic mycotoxins, DON and AFB1 respectively,
showed impaired cell viability with regard to total protein
content and lysosomal integrity of the cells.
To investigate the effect of mycotoxins on the intestinal
epithelial barrier integrity, an in vitro model has been
developed measuring the transepithelial electrical resist-
ance (TEER), currently using a porcine intestinal cell line.
Decreased TEER values indicate a weakened cell barrier
function against pathogens or toxins. This sophisticated
cell culture model currently representing the porcine
gut will be transferred to a newly-developed fish cell line
called RTgutGC, isolated from the intestine of a rainbow
trout.
In the near future the team hopes to establish a cell
culture model representative of fish gut to gain insight
into effects of mycotoxins on the gut permeability and
the effects thereof.
Research gathers pace to combat mycotoxins
in aquaculture - Turkeys, trout and 1960
by Dieter Moll, Research Team Leader Molecular Biology, BIOMIN
Plant-derived feed for aquaculture
- Investigations at the cellular level
by Barbara Novak, Scientist, BIOMIN
May-June 2015 | INTERNATIONAL AQUAFEED | 31