2.4.4: Results and Discussion

There were 5 time-points for VENUS
faecal samples collection – V1 (month 1), V3 (month 3), V5 (month 5), V7 (month
7) and RV (randomization visit). QPCR was conducted on all samples’ time-points,
however due to the differences in time-points for RV; RV’s were not included in
the results. As qPCR is a quantitative method, results could be analysed in the
form of prevalence. Prevalent bacteria species are species found in the NQ
(non-quantifiable) zone and quantifiable zone. The qPCR results collected were
segregated into two main groups – Bifidobacterium
species and other targets.

Graph
1: Prevalence of Bifidobacterium
species detected in VENUS’ qPCR samples

Graph
1 shows the different Bifidobacterium
species (Bifidobacterium breve,
Bifidobacterium longum and Bifidobacterium bifidum) detected in VENUS’
faecal samples over 4 different time-points, using qPCR. Although the number of
samples collected during each time-point varies, the number of subjects is considerably
well-distributed. Bifidobacterium are
expected to dominate healthy infants’ gut as it metabolizes HMOs (Human Milk
Oligosaccharides) to provide health benefits in hosts’
guts (O’Callaghan and Sinderen, 2016). Results have shown likewise, with
Bifidobacterium species being most
commonly detected in the qPCR assay. As seen from the graph, B.longum is the most prevalent (with a
prevalence of nearly 60% in V1), followed by B.breve and lastly B.bifidum.
As expected, these Bifidobacterium
species increases in prevalence over time as a general trend.

 

Graph
2: Prevalence of other targets detected in VENUS’ qPCR samples

Graph
2 shows the remaining targets (Bacteroides
genus, Staphylococcus aureus group, Clostridium difficile 16S, Clostridium perfringens 16S and Campylobacter species) detected in
VENUS’ samples over 4 time-points using qPCR. Absence of phylum Proteobacteria alongside high abundance
of Bacteroides implies a healthy gut
microbiota (Jandhyala
et al., 2015), hence Bacteroides are expected to be highly
prevalent in infants’ gut. S.aureus is
a commensal gut microbiota that may become pathogenic when the immune system
breaks down. C.difficile, C.perfringens and Campylobacter are expected to be low in counts due to their
pathogenic nature. According to graph 2, Bacteroides
and S.aureus has high prevalence in
infants’ gut with a general increase in Bacteroides
count over time. C.difficile is
shown to increase in prevalence over time. C.
perfringens has a generally high prevalence in infants’ gut, however they
are low in quantity (results not shown). Another observation is the low
prevalence and quantity (results not shown) of Campylobacter.

The results shown in graphs 1 and 2
indicates a healthy infant gut, as good gut microbiota like Bifidobacterium species is high in
prevalence and counts, while commensals are high in counts and pathogenic gut
microbiota are low in counts despite being highly prevalent.These results
collected imply that healthy infants’ guts are dominated by Bifidobacterium, Bacteroides and S. aureus.
They also imply that most of the infants could be BM-fed, due to the high
prevalence of Bifidobacterium in
their gut.

 

 

 

 

3: Reflection and Analysis of Internship Experience

3.1: Workplace Safety

Compulsory
trainings like WISE (Working in Safe Environment) Laboratory induction;
Laboratory induction; and Biological and Chemical Emergency Response conducted
before commencing laboratory work, equipped laboratory members with knowledge
of emergency protocols.

All laboratory
personnel and visitors must wear a (disposable) lab coat and shoe covers or covered
shoes before walking around the laboratory. Laboratory personnel must tie up
their long hair and don PPE (Personal Protective Equipment) such as gloves,
covered shoes and lab coats. These PPE would protect the wearers from injuries
such as chemical spillage, before leaving the laboratory, all personnel must
wash their hands with anti-bacterial soap to prevent bringing any hazardous
substances out of the laboratory.

To
reduce contamination between the laboratory, corridors and outside environment,
laboratory items were transported with cargo lifts while PPE were only worn in
the laboratory and when collecting laboratory items from the storage room. For
safety, thermal gloves were donned when retrieving items from the -80°C freezers
and autoclave machine.

The
laboratory has designated bins for biohazard waste, general waste and sharps. Non-sharp
items that are biological hazards, chemicals or items associated with potential
hazards were disposed in biohazard bins, while non-hazardous items like tip
boxes were disposed in general bins. Disposing these items separately prevents
any potential contamination and health effects on the cleaner as biohazards and
sharps require specific disposal methods.

Despite
the workplace safety standard being quite ideal, I have several suggestions.
Firstly, although gloved wrists or elbows were used to touch the laboratory
exit button when wearing gloves, I think that gloves should be removed before
touching the exit button – this would prevent contamination and potential
health hazards to the next personnel touching the button with their bare hands,
as they could potentially be bringing biohazards out of the lab, which would
render the intention of washing hands useless. Secondly, perhaps a separate lab
coat could be worn for biohazard waste disposal –because encountering
biohazards when changing biohazard bags is inevitable and if one’s lab coat
touches the biohazards and encounters laboratory samples, they could
potentially contaminate the samples. Lastly, although sick personnel entering
the labs don a face mask, I think that they should avoid entering the lab
unless absolutely necessary as these individuals could potentially contaminate
the samples or gain secondary infections from the samples due to their weakened
immune system.

 

 

 

3.2: Challenges and Solutions

Several
challenges were faced during my internship. Firstly, working in a Research and Innovation
(R&I) lab is unlike working in the NP labs which I was accustomed to. Being
a Research Assistant in Danone, experimental steps were carried out
independently after my mentor trained me. This was drastically different from
the dependency in NP, where we had technical support officers (TSOs) guiding
and watching over us in the laboratory; and partners or groupmates aiding in
the experimental process. In FYP (Final Year Project), experiments were
conducted with our partners. But, in internship I needed to be independent– learning
to work alone and handling the various tasks assigned to me. In FYP, there were
two pairs of hands and two brains; in internship although I could approach my
mentor for help if something unexpected happened with the experiments, there was
only 1 pair of hands. Therefore, I had to better manage my time to end work on
time. To complete the stipulated number of runs in a day, I arrived earlier and
learnt to better utilize my lunch break and the waiting time in-between runs.
Since the circumstances were unchangeable, I had to improve myself. Gradually,
with the improvement in time management, I also gained experience and independence.

Secondly,
although NP taught us the basic qPCR concepts, my internship made me realize
some misconceptions I had and how lacking my qPCR knowledge was when put to the
test. To ensure that I knew what I was doing and contribute better to the team,
I had to consistently read scientific articles on qPCR. Thankfully, my mentor gave
me a brief qPCR introduction and ISO-SOP trainings before the project
officially commenced.

Thirdly,
unlike NP’s practical sessions, there were occasions whereby runs were unable
to be conducted as the reagents ran out. As it was late December, the postal
order had a long processing time and reagents were low in stock at the company
we were ordering from, I discussed with my mentor how to counter this problem
and she told me that time could be maximized by running the Taqman qPCR runs for
another batch of samples first.

Another
challenge faced was the qPCR results being ‘unquantifiable’ or unsatisfactory. Although
the qPCR runs were somewhat repetitive, the results were often unpredictable.
Varying results could occur with the same steps and reagents. To solve this
challenge, frequent communication with my mentor and seeking her advice on the
runs were essential– as she had more experience. Thankfully, previous
experiences taught me not to give up at the sight of failure, but rather to seek
alternatives – such as re-diluting the 1010 standards from their
PCR-clean stock or simply repeating the runs. The PCR-clean standard for B.bifidum was also separated into four
tubes and stored at -80°C
to prevent degradation due to constant freezing and thawing. Through my
mentor’s guidance and my resilience, we obtained some useful results.

Occasionally,
there were minor issues such as the Hamilton machine being unable to connect to
the server or pipetting wrong volumes. So, I attempted to independently resolve
the problem – such as restarting the computer, reconnecting the USB cables,
etc. before seeking my mentor’s help in troubleshooting. Gradually, I also gained
independence.

Overcoming
these challenges helped me grow as a person and an intern, alongside teaching
me many valuable skills.

 

3.3: Applying Prior Knowledge

NP
has equipped me with several skills for my internship such as resilience,
teamwork, courage and responsibility. NP’s 6 core values – Responsibility,
Resilience, Respect, Integrity, Courage and Gratitude were taught via various
Educational Career Guidance modules and camps. Leadership camps taught me
resilience, teamwork and responsibility alongside the importance of setting
aside our differences and work as a team, encouraging each other to overcome
the challenges. Like the camp, every party in the SAT plays a different but crucial
role. My role is to conduct qPCR to analyze infants’ gut microbiota (repeating
the runs if necessary); then my mentor would analyze the results and generate
samples requiring repeats–  and I
will perform those repeats. The results may not always be ideal, but without
resilience and team work, there would not be results. If I had perceived qPCR as
tough and gave up at the sight of challenges, there would not have been a
conclusion to the runs. By being resilient and responsible enough to give my
all into the internship, I incorporated quality in my work and did not taint
Danone’s or NP’s reputation. By having realistic expectations of myself and
understanding what was required of me, I put in more effort to understand the
various technical concepts which aided in my learning. Open communication with
my mentor regarding my conceptual understanding also prevented
miscommunications from occurring alongside aiding in setting her teaching pace.

Taking
Basic Conversational Japanese (BCJ) as an elective taught me that courage
coupled with resilience went a long way. Learning a foreign language was not
easy but, my interest in the subject led to perseverance– that coupled with my
teacher’s patience, allowed me to successfully grasp BCJ. Though we may have
conducted PCR in our practical sessions and learnt the qPCR in modules such as
Medical Microbiology and Forensic Science, qPCR was novel to me. The star of my
internship program may be foreign to me however; the process of learning
something foreign was not. Hence, I gathered my courage, resilience and learnt
how to conduct qPCR with aid from my patient mentor.

 

FYP,
like internship, was mandatory for all 3rd year students. Before beginning
our FYP, there were many things to research and read up on. Following that,
there were several challenges, which taught us about teamwork, resilience, time
management and gratitude. Like FYP, internship was full of new experiences, requires
research prior to embarking on the tasks, and requires good time management to
plan and finish the tasks by the stipulated deadline.  Most importantly, resilience and independence
was required. FYP had prepared me for independence but internship was the real
test of it. Thankfully, FYP taught me the importance of planning before jumping
straight into an experiment and I incorporated planning into my internship. By
planning the structure of the runs with my mentor, I could conduct the experimental
runs more efficiently and effectively. FYP has also exposed me to the knowledge
of gut microbiota, which aided me in being able to grasp certain concepts and
terms which would have otherwise seemed foreign in my internship.

 

3.4: Influence on Career Choice

This
past 4 months at Danone was life-changing to say the least. Not only had I
gained newfound knowledge and experienced working in the local life science
industry, I also understood myself better. Although I enjoyed the various
aspects of my internship, one of the highlights was conducting the various
experiments in the lab, as I realized that I may have an aptitude in laboratory
work due to my organizational skills and I feel in my element when working in
the lab. If I know what to do, and given sufficient time to plan my schedule, I
am able to complete the stipulated tasks on time which makes me feel
accomplished.

This
internship gave me exposure to what would it be like working in a science team
supporting the clinical studies (CS) team, as most of the lab experiments I conducted
aims to find results that proves or disproves the hypothesis made by the CS team
in their VENUS study – and to be honest, I am glad to be in this team as
knowing that the qPCR results would be used in research to improve many
infants’ health is motivational.

Discussion
with my mentor opened my eyes to the differences in working at academia
laboratories and industrial laboratories. The former would contain a wider job
scope and more in-depth knowledge while the latter is more repetitive and is
only specific in a certain area. As I prefer routine work, I would prefer the
latter.

The
internship has further reinforced my passion for Biomedical Science. Through
various platforms such as laboratory work, Marketplace events and inductions, I
realized that I genuinely enjoy learning about things associated with human
health.

Although I am still uncertain if I
want a career in biomedical research, this internship experience evoked my
interest in gaining more laboratory experiences if an opportunity arises before
concluding if I am suitable for research. Regardless of whether I end up in the
R industry, this internship has made me recognize that I would ultimately
pick a career path with the purpose of improving the lives of many, be it in
terms of behind-the-scene works such as research or physically helping others.

 

4: Acknowledgment Letter to the
Company

The 4-month
internship with Danone Nutricia Research was an unforgettable journey of
learning and development. I am utterly grateful to Danone for giving me the opportunity
to interact with so many lovely colleagues and patient researchers who guided
me throughout this journey.

Firstly, I
would like to thank my company supervisor, Dr. Juandy Jo and my mentor, Claudia
Lee Seok Koon, for their limitless support and guidance during my internship.
Thank you for the patience when teaching me the qPCR concepts and giving
ISO-SOP trainings, answering all my queries, mentoring me, teaching me skills
such as critical thinking and for giving me this opportunity to be involved in
a research project with such a great purpose. Thank you for making this
internship so insightful and memorable. I would also like to thank my mentor
for helping me integrate into the novel environment with ease, transition into
working life at Danone, and taking care of my well-being. Thank you for being a
friend and a mentor to me. Secondly, I would like to thank the two scientists
in the laboratory – June Low Su Yin and Goh Chee Yong for answering any queries
I had when my mentor and supervisor were not around. Lastly, I would like to
thank the lovely colleagues in Danone for being so friendly and helpful
throughout my entire internship period. Their kindness has left a huge imprint
on me and I am glad I had the opportunity to interact with them.