Project 7: Antibiosis and effect of Cefalexin on lactobacilli and selected aerobic cells

Abstract: Antibiotics destroys pathogenic microorganisms and inhibits cell activity. In this study, the
effects of the antibiotic Cefalexin on lactic acid fermentation of lactobacilli and on aerobic cells were
determined. Three concentrations of Cefalexin were prepared (75%, 50% and 25%) and were added to a
mixture of fresh milk and yoghurt. The set-up with 50% concentration did not show curdling, implying
the inhibition of lactobacilli activity. The same three concentrations were also added to red blood cells,
hydrilla leaves and squamous cells. Red blood cells show crenation under 50% and 25% antibiotic
concentration. Hydrilla cells treated with 50% antibiotic concentration show plasmolysis. Squamous cells
show distortion under 50% and 25% antibiotic concentration. Using Cochran Q-test, the results show that
the milk and yoghurt mixture do not show the same response to varying concentrations of antibiotic. It
can be implied that 50% Cefalexin solution is effective in inhibiting bacterial activity. The aerobic cells
also show disturbance in cell morphology under 50% Cefalexin solution. This implies that high
concentration of antibiotics affect cell morphology and therefore affecting cell function as well. This
study contributes to the knowledge of the importance of dosages when taking antibiotics, since very high
concentrations not only kill pathogenic microorganisms but body cells as well.

I.

Introduction

glycolysis (pyruvate) in the TCA

II.

An antibiotic is a chemical or

cycle to produce much more energy

substance produced by one

currency in the form of ATP than

microorganism that is capable of

can be obtained from any anaerobic

destroying or inhibiting the growth

pathway. Aerobic respiration is

of another microorganism.

characteristic of eukaryotic cells

Antibiotics are used to destroy

when they have sufficient oxygen

pathogenic microorganisms causing

and most of it takes place in the

diseases in humans, important crop

mitochondria.

and animals useful to humans.

III.

IV.

Fermented food conveys health

Antibiotics blocks bacterial protein

benefits through lactic acid

synthesis thru interfering with the

fermentation. The fermentation

processes at the 30S subunit or 50S

process can transform the flavor of

subunit of the 70S bacterial

food from the plain and mundane to

ribosome.

a mouth-puckering sourness

Antibiotics have effects on aerobic

enlivened by colonies of beneficial

and aerobic cells. Aerobic

bacteria and enhanced

respiration, in the presence of

micronutrients. While fermented

oxygen, uses the end product of

food like yogurt, sauerkraut and

V.

kefir are well-known many other

bacterial infections such as

lesser-known foods also benefit

bronchitis, tonsillitis, ear infections,

from the lactic acid fermentation

skin infections, and urinary tract

process (McGruther, 2009).

infections (Lai & Wu, 2005).

Antibiotics are very important

Children and adults take this

chemicals which have been used to

antibiotic for mild acne and strep

cure many diseases (by killing or

infection. Cephalexin is also used

stopping the growth of bacteria)

for the treatment of heart diseases

such as bacterial meningitis,

due to its enhanced oral activity.

neurosyphilis, endocarditis, burn

VI.

VII.

This study is conducted to

wounds, skin infections, respiratory

determine the effects of antibiotics

and urinary tract infections,

on lactic acid fermentation by

pneumonia, anthrax, STDs, Lyme

Lactobacillus and on anaerobic and

disease, bronchitis, diarrhea

aerobic cells by observing changes

diseases, abdominal infections,

in its morphological structures. The

severe acne, and gastrointestinal

study will also determine the most

tract infections, hence this study is

effective concentration of antibiotic

very important to identify the

solution and will contribute to

mechanisms involved and on how

studies on appropriate dosages for

will it affect diseased persons and to

antibiotics.

know the right of amount dosage

VIII.

that a person should take.

IX.

Materials and Methods

Cephalexin, like the penicillins, is a

X.

Preparation of Antibiotic

beta-lactam antibiotic. By binding to

Solutions

specific penicillin-binding proteins

Three (3) different concentrations of

(PBPs) located inside the bacterial

Cephalexin solution were prepared

cell wall, it inhibits the third and last

as follows: 75%, 50% and 25%.

stage of bacterial cell wall synthesis.

The apparatus used are flame

Cell lysis is then mediated by

sterilized to prevent contamination

bacterial cell wall autolytic enzymes

of samples.

such as autolysins; it is possible that

cephalexin interferes with an

XI.

Antibiosis and effect on Lactic

autolysin inhibitor. Cephalexin is

acid fermentation

used to treat many different types of

Three (3) mL of fresh milk was

added to four test tubes. All tubes

antibiotic solution with different

were plugged with cotton to prevent

concentrations. The same procedure

contamination before and after

was repeated with Hydrilla leaves

addition of milk. Four drops of

and epithelial tissue obtained from

yoghurt were added next to each

the inner cheek lining of a student.

tube, careful not to touch the sides

XV.

of the tube. Lastly, eight drops of

XVI.

the antibiotic solution were added to

XVII. Results

each tube, one tube corresponding to

XVIII. Effect on Lactic Acid

a concentration as previously

fermentation. Among the three set-

mentioned, with one test tube

ups, the test tubes with 50%

serving as a control set-up. Three

antibiotic concentration had no curd,

replicates of each set-up were

implying the absence of lactic acid

performed. The experiment tubes

fermentation. The Q value for the

were arranged in a randomized

data obtained (see Appendices)

complete block design in test tube

computed is Q = 20.3125, using

racks and were stored in the

LOS 5% and df = 9, critical value =

laboratory for three days.

16.919. The null hypothesis that the

XII.
XIII.
XIV.

effect of antibiotics is the same for

Antibiosis and effects to selected

all samples is rejected, and the

cells

alternative hypothesis that the effect

Four (4) glass slides was filled with

of antibiotics is not the same for all

two drops of blood, smeared and

samples is accepted.

treated with 1-2 drops of the

XIX.

XX.
XXI.

Figure 1 Upper row: Red blood cells show their characteristic biconcave shape. Left is the control group and photo
on the right shows red blood cells treated with 75% antibiotic solution. Lower row: Red blood cells show
crenation. Left photo shows treatment with 50% antibiotic solution. Right photo shows treatment with 25%
antibiotic solution.

XXII. Effect on Aerobic cells.

XXIII. Red blood cells treated with 50%

and 25% Cephalexin solution show
crenation. The two solutions are
hypertonic solutions for red blood

cells; water inside the cells moved

out into the external environment.
XXVI.
XXVII.
XXVIII.
XXIX.
XXX.
XXXI.

XXIV.
XXV.

XXXII.
XXXIII.
XXXIV.
XXXV.
XXXVI.
XXXVII.
XXXVIII.
XXXIX.
XL.
XLI.
XLII.

XLIII.
XLIV.
XLV.

Figure 2 Hydrilla cells show no observable changes except for cells treated with 50% antibiotic solution showing
plasmolysis (lower row, left). 50% Cefalexin solution is a hypertonic solution and caused water inside the plant
cells to diffuse out of the cytoplasm, causing plasmolysis. Upper left photo shows the control group, upper right
shows treatment with 75% solution and lower right shows treatment with 25% solution.

XLVI. The cells of the hydrilla plant

solution is a hypertonic solution for

treated with 50% antibiotic solution

the cells, causing water to diffuse

show plasmolysis. Treatment with

out of the cytoplasm and the

75% and 25% antibiotic solution did

membrane to shrink away from the

not result in any significant

cell wall.

observable changes. 50% antibiotic

XLVII.

XLVIII.
XLIX.
L.
LI.
LII.
LIII.
LIV.
LV.
LVI.
LVII.
LVIII.
LIX.
LX.
LXI.
LXII.
LXIII.
LXIV.
LXV.
LXVI.
LXVII.
LXVIII.
LXIX.
LXX.
LXXI.
LXXII.

Figure 3 Squamous cells show distorted shapes under the microscope (lower row). These cells were subjected to
50% and 25% antibiotic solution, respectively. The upper row, with the control group on the left and cells treated
with 75% antibiotic solution on the right, show no significant observable changes; the cells are flat and scale-like.

LXXIII.

Squamous cells treated with

50% and 25% antibiotic solution

from their characteristic flat and

scale-like shape.

show distorted shapes, different

LXXIV.
LXXV. Discussion
LXXVI.

In as much as the history of

medicine is concerned, the

discovery of penicillin is considered

a miracle for saving millions of lives

and curing almost every sort of

yielded after inoculation of milk

diseases there was at the time.

samples with bacteria and addition

Bacteria posed a great risk, causing

of antibiotic solutions varied for

infection and chronic illnesses that

each concentration (Figure ). The

even led to death. Consequently,

absence of curd after a few days of

people became desperate that it was

incubation was observed at 50%

once thought it can protect them

concentration of antibiotic, whereas

from bacteria-caused diseases and

the rest including the control group

that taking indiscrete amounts

have undergone curdling. Curd

would help them recover faster.

formation was a sign that the

Never then would one thought about

bacteria werent killed by the

the possible side-effects of

antibiotic, regardless of the

excessive intake of antibiotics.

concentration.

LXXVII.

In this activity, we aim to

LXXVIII. As for the red blood cells, only

determine the effects of antibiotics

25% and 50% antibiotic solution

at different concentrations to lactic

induced crenation. It can be deduced

acid bacteria. Generally, these are a

therefore that the prepared antibiotic

group of bacteria that are naturally

solution was hypertonic. This result

found in the gastrointestinal tract

in contrast to erythrocytes function

and are often the primary

as drug carriers along with oxygen

ingredients of dairy products,

distribution could possibly imply

having been generally recognized as

that antibiotics may have different

safe (GRAS). To determine which

effects in vitro and in vivo.

concentration is most efficient in

eliminating bacteria, three different
concentrations of antibiotic
solutions were used respectively,
25%, 50%, 75%, 100%, and a
control group. Lactobacillus, a
beneficial bacterium usually added
to yoghurt was chosen for this
experiment as it can be easily
obtained, and is less harmful
compared to other genus. Results

LXXIX.

Similarly, squamous cells

obtained from the cheeks exhibited

distortion when added with 25% and
50% antibiotic solution (Figure 3).
The effect of varied antibiotic
concentrations may imply which
concentrations are considered safe
and harmful for sensitive cells such
as those that lines the mouth.

LXXX. Antibiotics are applied in plant

responses, and culminate in cell

culturation to eliminate invading

lysis (Kohanski, Dwyer & Collins,

bacteria that may infect plants.

2010). The specific mechanism by

Generally, Cephalexin belongs to a

which antibiotics can be lethal to

group of antibiotics called beta-

bacteria was not determined in this

lactans which are considered non-

activity, however, in a study

toxic to plants, however in some

conducted by Kohanski, Dwyer,

cases their break down products in

Hayete, & et. al. in 2007, three of

culture medium can influence plant

the major classes of antibiotics

cell growth (Grzebelus & Skop,

stimulated the production of harmful

2014). As seen in this activity,

hydroxyl radicals in both Gram

hydrilla treated with 50% antibiotic

negative and positive bacteria,

solution caused plasmolysis and

thereby inducing cell death.

protoplasmic streaming (Figure 2).

Nonetheless, it has to be noted that

It can be concluded therefore that

these antibiotics do not produce

phytotoxicity of antibiotics depends

hydroxyl radicals. Formation of

upon concentration and the type of

reactive oxygen species (ROS) such

plant. In comparison, Cephalexin is

as hydroxyl peroxide, hydroxyl

used most effective to animal cells

radicals, and peroxyl radicals which

and may have different effects to

can interact to biological molecules

plant cells.

contribute to cell damage (Belenky

LXXXI.

Antibiotics are composed of

different families and every each

et. al., 2015).

LXXXII.

Other than the production of

one act at a very specific target,

ROS, antibiotics were also

inducing different effects on the

discovered to induce metabolic

bacteria (Michel, 1978). Cephalexin,

perturbations, mainly affecting the

an antibiotic that belongs to the

TCA cycle activity and oxidative

family of cephalosporin, acts

phosphorylation (Belenky et.al.,

specifically by targeting the

2015). Increased levels of coenzyme

bacterias cell wall (Cephalexin,

A involved in oxidative metabolism

n.d.). Successful treatment with a

indicated that fermentation was

cell wall synthesis inhibitor can

inhibited, whereas bacterial

result in changes to cell shape and

respiration accelerated, and this

size, induce cellular stress

induction is critical to antibiotics

lethality (Belenky et.al., 2015).

present. However, intake in the

Fermentation is an essential event in

absence of precautionary measures

the production of ATP and

can lead to many side-effects; the

regeneration of NAD for anaerobic

most common mild type of which is

organisms. Only two are produced

diarrhea. When excess amount of

and both of which are utilized for

antibiotics is taken in, it can kill the

other processes including

good bacteria found in the

conversion of milk into yoghurt.

gastrointestinal tract thus causing

The addition of antibiotics that

imbalance in the systems

brings about oxidative stress to the

environment. Taking up food with

bacteria forces them to undergo

probiotics is usually advised to

respiration; however, since they are

counter-balance the situation.

obligate anaerobes, they do not

adapt to such conditions and rather
killed.
LXXXIII. Possible explanation why the

LXXXV.

One might wonder why

pharmaceutical stores prohibit

selling of antibiotics without
prescription. This is mainly because

bacteria introduced with 25%, 75%,

antibiotics should only be taken

and 100% is that bacteria can

upon the doctors advisory and most

develop resistance to antibiotics.

importantly to prevent bacterial

According to Korkohen (2010),

resistance otherwise all the

there is a close relation between the

endeavours poured upon the

amount of antimicrobials used and

discovery of such drugs will end up

the rate of resistance to these

in vain.

substances and thus the misuse of

antibiotics was believed to be the
principal cause of antibiotic
resistance problem. Furthermore,
antibiotic resistance genes can be
passed from one bacterium to
another.
LXXXIV. Antibiotics contribution in the
field of medicine is highly
recognized both in the past and at

LXXXVI.
LXXXVII. V.

Conclusion

LXXXVIII.
LXXXIX. Antibiotics inhibit cell activity.
The milk and yoghurt mixture did
not show curdling under 50%
antibiotic solution treatment. The
milk and yoghurt mixture do not

show the same response to varying

concentrations of antibiotic. Red
blood cells show crenation under
under 50% and 25% antibiotic
concentration. Hydrilla cells treated
with 50% antibiotic concentration
show plasmolysis. Squamous cells
show distortion under 50% and 25%
antibiotic concentration. It can be
implied that 50% Cefalexin solution

XC.
XCI.
XCII.
XCIII.
XCIV.
XCV.
XCVI.

is effective in inhibiting bacterial

activity. The aerobic cells also show
disturbance in cell morphology

XCVII.
XCVIII.

under 50% Cefalexin solution. High

concentration of antibiotics affects

XCIX.

cell morphology and therefore

C.

affecting cell function as well.

CI.
CII.
CIII.
CIV.

Conclusion

CV.
CVI.

Antibiotics inhibit cell activity. The milk and yoghurt mixture did not show curdling under
50% antibiotic solution treatment. The milk and yoghurt mixture do not show the same
response to varying concentrations of antibiotic. Red blood cells show crenation under under
50% and 25% antibiotic concentration. Hydrilla cells treated with 50% antibiotic
concentration show plasmolysis. Squamous cells show distortion under 50% and 25%
antibiotic concentration. It can be implied that 50% Cefalexin solution is effective in
inhibiting bacterial activity. The aerobic cells also show disturbance in cell morphology under
50% Cefalexin solution. High concentration of antibiotics affects cell morphology and
therefore affecting cell function as well.

CVII.
CVIII.
CIX.

CX.
CXI.
CXII.
CXIII.
CXIV.
CXV.
CXVI.
CXVII.
CXVIII.
CXIX.
CXX.
CXXI.
CXXII.
CXXIII.
CXXIV.
CXXV.
CXXVI.
CXXVII.
CXXVIII.
CXXIX.
CXXX.
CXXXI.
CXXXII.
CXXXIII.
CXXXIV.
CXXXV.
CXXXVI.
CXXXVII.
CXXXVIII.
CXXXIX.
CXL.
CXLI.
CXLII.
CXLIII.
CXLIV.
CXLV.
CXLVI.
CXLVII.
CXLVIII.
CXLIX.
CL.
CLI.
CLII.
CLIII.

CLIV.
CLV.
CLVI.
CLVII.
CLVIII.
CLIX.
CLX.
CLXI.
CLXII.
CLXIII.
CLXIV.
CLXV.
CLXVI.
CLXVII.
CLXVIII.
CLXIX.
CLXX.
CLXXI.
CLXXII.
CLXXIII.
CLXXIV.
CLXXV.
CLXXVI.
CLXXVII.
CLXXVIII.
CLXXIX.
CLXXX.
CLXXXI.
CLXXXII.