Research proposal

Lactic acid bacteria and development of microbial agents

Introduction

Lactic acid bacteria have vital role in food industry as they have highest nutritional value
regarding food and food safety (Adeyemo & Onilude, 2014). Lactic acid bacteria improve
digestion of milk sugar (lactose), in this way they improve nutritional value of food. Lactic acid
bacteria (LAB) are Gram positive bacteria and do not produce spore ( non-spore forming), rod
shaped bacteria which grow an-aerobically difficult to grow on artificial media and grow
willingly on food substrates and lower the pH to a limit where the opposing organisms not able
to grow and it is main fermentation products of carbohydrate’s metabolism (Hayek & Ibrahim,
2013). LAB can also tolerate or survive in the presence of oxygen (aero tolerant anaerobes).
Fermentation of cassava into ‘pupuru’ manually by Rural women have some side effect like
increase level of microbes into fermented cassava product.

Therefore, it is essential to complete fermentation of product should be allow by LAB

(Sawadogo‐Lingani et al., 2007).LAB is non-respiring and metabolize carbohydrate through
fermentation rather than respiration. The functional food such as ‘pupuru’ that is derived from
the fermentation of food like cassava contains probiotic strains capable of preventing infectious
human diseases (Garneau, Martin, & Vederas, 2002), (Kaškonienė et al., 2017) . They also
control some factors including cancer, intestinal infection, diarrhea, level of serum cholesterol
and helping the body to sustain and fight infection. Live microorganism related to food
fermentations can supply useful functions in the gastro intestinal tract is reliable with the rising
view that middle health benefits of probiotic cultures can be assigned to a genus, instead of a
specific strains of a species (Hill et al., 2014).There is one of the most significant health benefit
of LAB species including (Lactobacillus species, Bifidobactarium sp and scharomyces), i.e. live
bacteria there utilization as probiotic food supplement. ‘Pupuru’ is fermented from cassava
product and is consumed by as a minimum of 4-6 million people in Nigeria and extra in other
African countries (Odetokun, AIYESAMNI, & Esuoso, 1998). LAB can be heritably engineered
in order to grow more rapidly than contaminants and also to reduce and raze growth and
development of microorganisms involving pathogens by producing antimicrobial agents
(Admassie, 2018).There are many bacteria which can destroy harmful microorganisms in a
process called biological control like ‘Rhizopus stolonifer’ which cause peach soft not disease
and frequently arise in post-harvest preservation of peach (Northover & Zhou, 2002) and is also
involved in significant losses of post-harvest food commodities. We originate Bacillus subtilis
fmbj strain, which was remote and identified in our laboratory (Liang, Lu, Zou, & Zhang, 2001),
has a strong inhibitory effect and can inhibit growth of pathogenic mycelium species ‘Rhizopus
stolonifer’. LAB have vital role in the fermentation industry of dairy, Fermented milk products
including Yoghurt, cheese, butter, sour milk and butter milk is formed during acid fermentation
of milk where lactose is converted to lactic acid to lower the pH of milk, this is accomplish
through utilization of lactic acid bacteria, and in arrangement with yeast they are frequently used
for fermentation of cereal products like dough (Muhialdin, Hassan, & Sadon, 2011).LAB also
produce lactic acid and it can destroy harmful microorganism and contaminants which cause
food spoilage.

LAB are useful as they are applied as obstacle against bacteria which are susceptible to acid, and
which are therefore economically eliminated from the acidic media as they are sensitive to acidic
environment (Ananou, Maqueda, Martínez-Bueno, & Valdivia, 2007). Lactic acid bacteria
(LAB) produce lactic acid which is used in many kind of food preservation, as curing agent, and
flavoring agent. LAB also produce lactic acid and it can destroy harmful microorganism and
contaminants which cause food spoilage, by lowering the pH to below 4. It is also used as
decontaminant in the preservation of cereals and yoghurt (Ananou et al., 2007) . LAB contains
antifungal activities (Schnürer & Magnusson, 2005).So in this way shelf life of fermented food is
prolonged. Raw materials caring high amount of mono and disaccharides and in some conditions
LAB are involved in fermentation of starch (Marco et al., 2017) . A starter culture is milk
containing lactic acid bacteria which convert milk sugar (lactose) into lactic acid so that milk
become sour or fermented. During Fermentation ATP are produced in the absence of oxygen
through the process of glycolysis that breaks the glucose into two pyruvate molecule producing
two ATP and two NADH molecules.

The applications may face various difficulties involving technological in less develop
countries as well as Consumer acceptability to new functional food may be affected by culture
and religion issues. There is need for more study to understand the basic probiotic properties
which will give more understanding regarding skills and acceptability in nutrition and health
sectors. Also I have no enough time and selective media to grow all kinds of bacteria which are
present and can grow in any kind of food products. I will use a plant polysaccharide called agar,
as many genera of bacteria can grow on this media and this media called plate count agar used
for isolation of bacteria. As a result of this study I would be able to understand all kind of genera
of bacterial species. I plan to study some bacteria which cause spoilage of many kind of food
products but unfortunately, I have no available media and lab for future study.

The basic purpose of this review is to give an overview of the main areas of resent
applications of lactic acid bacteria (LAB) in the improvement of human health, this research
technology still faces some of the Challenges and in order to provide some of the prospects as to
vital role they may play in the nearest future.

Methodology
 Review of literature

(Jannah, Saraswati, Handayani, & Pujiyanto, 2018) conducts an experiment in order to

conclude the antimicrobial action of lactic acid bacteria remote from hen gastrointestinal tract
with the importance of their probiotic properties. Bacterial colonies producing obvious zone on
MRSA and 0 .5% CaCO3, Gram positive and catalase negative were remote as LAB. Some
species (10 isolates) were checked for their capacity to inhibit growth of E-coli and
staphylococcus aureus and for acid pH and bile acid forbearing. Results showed that all the
selected isolates that producing antimicrobial compound inhibits the growth of E-coli and
staphylococcus in case of both supernatant and supernatant plus 2M NAOH, and still growing
in the medium of pH 2% and 0.1 % bile salt. It reflects the potential use of LAB from hen
gastrointestinal tract for probiotics in food.

(Adeyemo, 2018) Made his research aims at determining the antimicrobial substances
formed from LAB remote from samples of ‘pupuru’ in opposition to food borne pathogens.
Samples of ‘pupuru’ were composed from ondo state. Nigeria and other species of LAB were
remote from them.LAB were tested by using different morphological and biochemical tests.
They were experienced in opposition to pathogenic organisms by using Agar well diffusion
method. As a result, antimicrobial substances formed also monitored about five days. A total of
18 LAB were isolated and recognized as L. casei, L. delbrueckii, L. brevis, L. plantarum,
L.pentosus, L. fermentum and L. acidophilus. As a result, antimicrobial substances formed as;
Lactic acid, Diacetyl and Hydrogen peroxide ranges from (0.0110-0.0196g per litter). Maximum
production was obtained from L. plantarum and L. acidophilus; and the slightest being L.
pentosus. During fifth day decline in pH was observed in fermenting cassava and ranges from
5.8-3.5. LAB species isolated from ‘pupuru’ had significant inhibitory effect on food borne
pathogen. Four species were checked for antagonistic action and L.plantarum showed maximum
zone of inhibition on staphylococcus aureus (24mm) and pseudomonas aeruginosa (20mm)
correspondingly whereas antimicrobial action lessen with time. Antimicrobial action of LAB was
shown to noticeably (p<0.05) compare with ordinary antibiotics used as control. LAB has
highest antimicrobial activity against food borne pathogen and this potential can be applied as
starter in food industry in order to hinder food spoilage microorganisms and another
contaminant.

(Admassie, 2018) Lactic acid bacteria (LAB) is a large group of bacteria having
properties of lactic acid production, that is end product of fermentation. Biotechnology is the
process of development of useful products by using living organisms. some of the applications
involved in this research like biotechnology in food fermentation in order to improve taste, color,
aroma, shelf life, texture, quality and nutritional value of food. Daily use of LAB, fermented
food improve the immune system and reinforce the body in opposition to pathogenic bacterial
infection, bacteriocins produced by LAB may become a prospective drug candidates in order to
replace antibiotic to treat numerous drug resistance pathogen in the future. Modern fermentation
technology designed to obtain fastidious compound using microbial metabolism as chemical
machinery. Hence LAB fermentation is not only a monetary significance, but it also promotes
human health and increase food or feed production throughout the world.

(Vasiee, Mortazavi, Tabatabaei-yazdi, & Dovom, 2018) conduct an experiment in order

to recognize the predominant LAB isolates. Overall 75 gram positive and catalase-negative
isolates were subjected to alliance and identifying by using 16s rDNA sequencing, and
carbohydrate fermentation correspondingly. As a result, species found were bulk of strains
belonged to the genus Lectobacillus, and isolated into five species L. plantarum (25.33%), L.
fermentum (22.66%), L. pentosus (12%), L. brevis (10.66%), and L. diolivorans (1.33%).
Inconsequential inhabitants of other genera involve: Enterococcus (18.66%), Leuconostoc
citreum (5.33%) and Pediococcus (3.99%). The entire strains of L. plantarum, and L. brevis were
subjected to rep-PCR typing. Elevated intra-species varieties of L.plantarum and L.brevis
isolates were found. As a result, the poly-phasic strategies, depending on molecular system, are
essential for steadfast and precise studying of microbial composition of fermented food.

(Qian, Long, Pan, Li, & Zhao, 2018) In this research a lactic acid bacterium was
isolated from yak yoghurt in order to determine its probiotic properties after culturing on MRS
medium, a bacterium was identified as LAB by discoloration and microscopic study. And the
LAB by extra study identified as Lectobacillus plantarum by physiological and biochemical
experimentation.LAB also identified as L. plantarum by 16s DNA determination, and LAB was
named as L. plantarum YS2 (LP-YS2). Its acid opposition activity (68.05%) is higher than
L.bugaricus(LB,30.35%). LP-YS2 also showed elevated bile salt confrontation activities
(19.85%, 15.01% and 8.35%) as compare to LB ((3.08%, 1.87% and 1.44%) in 0.3%, 0.5% and
1% bile salt solution. The hydrophobicity results also showed that LP-YS2 (72.35%) had higher
hydrophobicity than LB (45.67%). LP-YS2 contains excellent significance in food and
pharmaceutical development.

(Vinusha, Deepika, Johnson, Agrawal, & Rakwal, 2018) made his research in order to
understand the proteomic which plays an important role in biological cell functioning. In this
study proteomic of probiotics, with reference to LAB are reviewed with the view of
understanding i) proteomic map, ii) mechanism of adaptation to insensitive gut surroundings
like low pH and bile acid, iii) role of cell outside protein in adhering to intestinal epithelial cells,
and iv)as a tool to answer fundamental cell functions. We also reviewed several analytical
methods used to carry out proteome analysis, in which 2D-MS and LC-MS/MS approaches were
found to be adaptable technique to carry out high throughput samples analyses even for a
complex gut samples. Further, we present prospect road map of under-standing gut microbes
combining meta-proteomics, meta-genomics, meta-transcriptomics and -matabolomics.

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