Guide Questions in Writing the Critique

THE TITLE
Having entitled A Comparative Study of Some Functional Properties of
Lactobacillus and Enterococcus Isolated from Feces of Normo and HyperCholesterolemic Humans, it is long but informative in such a way that could be easily
understood when one is into the field of Microbiology. Having acquired from feces of
people with normal and high cholesterol levels, it aims to identify the functional
properties of both genera under variable conditions, including their importance in
maintaining the health of their host.
THE AUTHOR(S)
The author of the journal is Mara de Lourdes Reyes-Escogido, a Ph.D in
Microbiology, Molecular Biology and Biotechnology. She is a graduate of the University
of Guanajuato in Mexico and is under the Department of Medicine and Nutrition at San
Carlos Hospital. She has seven publications, including some studies pertaining to the
relationship between obesity and Lactobacillus probiotic intake.
KEYWORDS
The keywords associated with the journal are the following: Enterococcus;
Gastrointestinal microbiota; Hypocholesterolemic activity; and Lactobacillus. These are
helpful though one of them is too broad as bacteria other than Lactobacillus and
Enterococcus normally reside in the stomach.
THE ABSTRACT
The abstract implies that if there is imbalance of gastrointestinal microbiota,
several kinds of diseases (mainly metabolic) are most-likely to occur. Having explained
the purpose of the paper, it presents different conditions like acid pH, presence of bile
salts, antibiotic sensitivity, hydrolysis to bile salts and the resulting action such as
reduction of cholesterol. The researcher isolated, identified and characterized 26 strains
of Lactobacillus and 23 strains of Enterococcus from the feces of normo and
hypercholesterolemic humans and identified their reactions to certain conditions

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previously stated. The highest percentage of isolated and characterized Lactobacillus

strains was from the normocholesterolemic group, while in the hypercholesterolemic
group, most strains belonged to the Enterococcus. Also, Lactobacillus strains showed
greater resistance at pH 2.0, while strains of both genera showed similar survival rates
at pH 3.0 and in the presence of bile salts after 24 hours. The research informs one of
the benefits of these bacteria when they are in their normal flora, which is to reduce
cholesterol levels. To conclude, microbial imbalance involving the depletion of beneficial
bacteria could be detrimental to the health of the host.
THE INTRODUCTION
The introduction states that the gastro-intestinal tract contains native
microorganisms that can positively-influence health. This justifies the research as it also
connotes that a balanced and dynamic ecosystem exists and plays an important role in
various human body functions. Among the native flora, some species and strains could
lower cholesterol levels, although much work is done by the strains of Lactobacillus and
Bifidobacterium. Little information is obtained regarding the cholesterol-reducing
capacity of Enterococcus and the mechanism involved. The problem is researchable
considering that the benefits of such microorganisms are usually taken out in school and
in books, giving more emphasis on their pathogenicity instead. There is no research
question present, however, basing on the statements, it could be inferred that one of its
objective is to identify whether strains of Lactobacillus and Enterococcus can survive in
variable conditions at the gastro-intestinal tract after having acquired from feces of
normo and hypercholesterolemic individuals.
THE LITERATURE REVIEW
There is no section for the review of related literature, but citations are written in
the introduction. It mentions the positive views, and literally only has this two: that in the
beginning of the 20th century, Ely Metchnikoff proposed various theories about the
positive health effects of the consumption of Lactobacillus which are still accepted and
have contributed to the conviction that they perform a very important function. Marteau
et al. 2001 reported that Lactobacillus-Enterococcus comprise 6.6% of the bacterial
population in feces and that Enterococcus, which has been part of gut microbiomes
since at least the early Devonian period, comprises a little less than 1% of the human
gastrointestinal microbiota. It has no theoretical framework, leaving gaps in the knowing
which this research could fill.

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THE RESEARCH DESIGN

The journal does not present a rationale about the methods. Since it could be
inferred that it aims to identify whether strains of Lactobacillus and Enterococcus can
survive under variable conditions at the gastro-intestinal tract, the following are used:

Participants There are 65 adult, voluntary participants in the study with an age
range of 18-58 years. The general characteristics of the participants (such as
age, sex, body mass index, physical activity, nutrient consumption) and the blood
glucose and cholesterol levels are determined. This information is enough to
determine the participants condition and the possible risk factors associated with
their condition, if there is any.

Nutritional and Physical Activity Assessment A previously-validatedm semiquantitative food frequency questionnaire (FFQ) is used to evaluate dietary
intake. It includes data regarding the consumption of 116 food items.
Isolation of Lactic Acid Bacteria Serial dilution techniques are used to isolate
the bacteria. Each sample suspension is prepared by adding 1 g of feces to 9 ml
of peptone water; the fecal matter is then suspended by vigorous stirring in a
vortex for at least 1 min.

Susceptibility to Antibiotics Disc diffusion method is used to evaluate the

behavior of the studied strains in the presence of the following antibiotics:
Ampicillin (10 g), Gentamicin (10 g), Kanamycin (30 g), Erythromycin (15
g), Clindamycin (2 g), Tetracycline (30 g), Chloramphenicol (30 g),
Streptomycin (10 g), and Neomycin (30 g).

Tolerance to Acid pH and Bile Salts (0.3% w/v) - Three different types of MRS
broth are prepared; two with 6M HCl, one adjusted at pH 2.0 and the other at pH
3.0; the third had MRS and bile salts (0.3% w/v) and was adjusted at pH 6.5.

Hydrolase Activity (Bile Salt Hydrolysis) Through using MRS agar

supplemented with sodium thioglycolate (0.2%) and taurocholic acid (0.2%).

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In-vitro Cholesterol Reduction Through an MRS broth supplemented with

cholesterol at a final concentration of 100 g/mL.

Molecular Identification Extracting genomic DNA from pure cultures after 24

hours of incubation, using a commercial kit.

Statistical Analysis Performing in triplicate; results are shown as mean SD

with p<0.05 considered statistically significant.

The most important ethical consideration is the use of human subject in this
study. Their participation is voluntary and informed consent. Such principle guarantees
that subjects are choosing to participate of their own free will and have been fullyinformed regarding the procedures of the research project and any potential risks. The
methods might not be 100% reliable as the researcher did not administer the tests again
over a period of time. The study is limited to assessing healthy individuals without recent
intake of alcohol and drugs.
DATA AND ANALYSIS or RESULTS AND FINDINGS
The results of people with normal blood glucose level (to which the number of
normal and high cholesterol levels are derived), other factors (weight and height in body
mass index), microscopy (Gram Staining) and biochemical tests (catalase & oxidase
tests) are emphasized. They are justified although some parts are lacking. Some data
are not taken into account, such as the consumption of nutrients and physical activity by
participants. It has found out that the intestinal microbiota is involved in several activities
such as energy production, synthesis of micronutrients, fermentation processes and
absorption of electrolytes (which regulates both nutrient acquisition and energy
extraction through the synthesis of enzymes involved in the metabolism of
carbohydrates, cholesterol and proteins). The table presents a number of constants (eg:
age, gender, weight, amount of glucose, lipids and proteins) with a corresponding,
varying statistical result.
THE DISCUSSION
The discussion involves interpretation through tables. Descriptively, it also
mentions certain range of normal and abnormal values. There is no direct research
question nor hypothesis and theoretical framework. Conclusions are appropriatelyAcebuche, Louisse T.

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drawn although it isnt separated from the discussions, and recommendation is

presented but lacking. All of the results are stated but not highlighted, thus it might be
difficult for readers to comprehend them textually and in long paragraphs. The result of
the distribution of strains in relation to normo and hypercholesterolemic hosts have been
associated to studies about Lactobacillus and Enterococcus strains presenting
resistance to one or more antibiotics, which may be intrinsic or acquired. Results are
vital as they help determine the possible conditions of participants prior to diagnosis of
certain conditions, if ever any develops.
CONCLUSION
Having written in the discussions, the conclusion denotes that bacteria can
remain in the gastrointestinal tract despite several changes in their environment. Both
the Lactobacillus and Enterococcus belong to the group of Lactic Acid bacteria, thus
they perform similar functions. When disease such as hypercholesterolemia occurs,
Enterococcus remains constant. However, Lactobacillus decreases, causing changes in
the gastrointestinal environment and eventually resulting to imbalance. Also, based on
the results, the researches concludes that Lactobacillus is more efficient than
Enterococcus in cholesterol reduction. Therefore, the restoration of the microbiota could
be an alternative for the control of total cholesterol levels and other related lipids such as
triglycerides, LDL, VLDL, and HDL, independently of the mechanism used. Going
through, these are the points drawn and one of the recommendations for future research
is making use of bile salt hydrolase (BSH), an enzyme produced by the bacteria.

REFERENCE LIST OR BIBLIOGRAPHY

The references follow an APA format but are not arranged chronologically.
Sources include books, other journals, foreign health-related departments (eg: ADA &
CSLI) and web sources. Majority of the sources are of the 21 st century, though there is
one that is published in 1994, a work by Huycke regarding the virulence of enterococci.

ABSTRACT

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There have been reports of an association between an imbalance of

gastrointestinal microbiota and several kinds of diseases, mainly metabolic diseases.
Lactobacillus and Enterococcus are two genera that form part of the native
gastrointestinal microbiota. In this work, we isolated, identified and characterized 26
strains of Lactobacillus and 23 strains of Enterococcus from the feces of normo and
hypercholesterolemic humans. We compared the behavior exhibited by all strains at
acid pH and in presence of bile salts, their sensitivity to antibiotics, and their ability to
hydrolyze bile salts and to reduce cholesterol in vitro. The highest percentage of all
isolated and characterized Lactobacillus strains was from the normocolesterolemic
group; in the hypercholesterolemic group, most strains belonged to the Enterococcus
genus. The Lactobacillus strains showed greater capacity to reduce cholesterol levels;
although this capacity has been related to bile salt hydrolase activity, four of the strains
we isolated showed no such activity, but still reduced cholesterol. All strains of both
genera isolated from normocholesterolemic participants showed a highest rate of
cholesterol reduction than those isolated from hypercholesterolemic participants.
Furthermore, the Lactobacillus strains showed greater resistance at pH 2.0, while
strains of both genera showed similar survival rates at pH 3.0 and in the presence of
bile salts after 24 h. These results support the evidence that a microbial imbalance
involving the depletion of beneficial bacteria could be detrimental to the health of the
host.

Reference:
Reyes-Escogido, M. L. (2016). A Comparative Study of Some Functional
Properties of Lactobacillus and Enterococcus Isolated from Feces
of
Normo and Hyper- Cholesterolemic Humans. J Bacteriol Parasitol
7: 274.
doi: 10.4172/2155- 9597.1000274

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ARTICLE CRITIQUE CHECKLIST

INTRODUCTION
Problem

Yes

No

N/A

Is there a statement of the problem or a qualitative topic of study?

____ ____

____

Does the problem of topic indicate a particular focus of study?

____ ____

____

Is the problem researchable, that is, can it be investigated

through the collection and analysis of data?
____

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Is the background information on the problem presented?

____

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Is the educational significance of the problem discussed?

____

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Does the quantitative problem statement indicate the

variables of interest and the specific relationship between
those variables that were investigated?
____ ____

____

Does the qualitative problem statement provide a general

indication of the research topic or issue?
____
When necessary, are variables directly or operationally defined?
____
Does the researcher have the knowledge and skill to carry out
the proposed research?
____

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____

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Review of the Related Literature

Is the review comprehensive?
____

____ ____

Are all references cited relevant to the problem under investigation?

____ ____

____

Are most of the sources primary; i.e., are there only a few

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or no secondary sources?

____

____

Have the references been analyzed and critiqued, and the

results of various studies compared and contrasted? That is, is
the review more than a series of abstracts or annotations?
____ ____

____

____

Is the relevancy of each reference explained?

____ ____

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Is the review well organized?

____
Does it logically flow in such a way that the references least
related to the problem are discussed first and the most related
references are discussed last?
____

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____

____

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Does it educate the reader about the problem or topic?

____ ____

____

Does the review conclude with a summary and interpretation

of the literature and its implications for the problem investigated?
____ ____
Do the implications discussed form an empirical or theoretical
rationale for the hypotheses that follow?
____

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____

Are references cited completely and accurately?

____
____

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____

Yes

No

N/A

____

____

Hypothesis
Are specific questions to be answered listed or specific
hypotheses to be tested stated?
____
Does each hypothesis state an expected relationship or difference?
____ ____

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If necessary, are variables directly or operationally defined?

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Is each hypothesis testable?

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METHOD
Participants Are the size and major characteristics of the
population studied described?
____
Are the accessible and target populations described?
____ ____

____

If a sample was selected, is the method of selecting

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the sample clearly described?

____
Does the method of sample selection suggest any
limitations or biases in the sample?
____

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For example, is stratified sampling used to obtain sample subgroups?

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Are the size and major characteristics of the sample described?

____ ____

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Does the sample size meet the suggested guideline for minimum
sample size appropriate for quantitative analyses?
____

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Instruments
Do instruments and their administration meet guidelines
for protecting human subjects?
____

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Have needed permissions been obtained?

____
Is the rationale given for the selection of the instruments
(or measurements) used?

____

Is each instrument described in terms of purpose, content,

validity, and reliability?
____

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Are the instruments appropriate for measuring the intended variables?

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Does the researcher have the needed skills or experience to
construct or administer an instrument?
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Is evidence presented to indicate that the instruments are
appropriate for the intended sample? (For example, is the
reading level of an instrument suitable for sample participants?)
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If appropriate, are subtests reliabilities given?

____
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If an instrument was developed specifically for the study, are the

procedures involved in its development and validation described?
____ ____
Yes
If an instrument was developed specifically for the study, are
administration, scoring or tabulating, and interpretation procedures
fully described?
____

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No

N/A

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Is the correct type of instrument used for data collection?

____ (For example, using a norm-referenced instrument when a
criterion-referenced one is more suitable.)

____

____

Design and Procedures

Are the design and procedures appropriate for examining
the research question or testing the hypotheses of the study?
____ ____

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Are the procedures described in sufficient detail to permit them

to be replicated by another researcher?
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Do procedures logically relate to each other?

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Are instruments and procedures applied correctly?

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If a pilot study was conducted, are its execution and results

described as well as its impact on the subsequent study?
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Are control procedures described?
____

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Did the researcher discuss or account for any potentially

confounding variables that he or she was unable to control?
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Is the application of the qualitative method chosen described in detail?

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Is the context of the qualitative study described in detail?
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RESULTS
Are appropriate descriptive statistics presented?
____
Was the probability level at which the results of the tests of
significance were evaluated specified in advance of the data analysis?
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Was every hypothesis tested?
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If parametric tests were used, is there evidence that the researcher

avoided violating the required assumptions for parametric tests?
____ ____
Are the tests of significance described appropriate, given the
hypotheses and design of the study?
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Was the inductive logic used to produce results in a qualitative

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study made explicit?

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Are the tests of significance interpreted using the appropriate

degrees of freedom?
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Are the results clearly described?
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Are the tables and figures (if any) well organized and easy to understand?
____ ____

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Are the data in each table and figure described in the text?
____ ____

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DISCUSSION, CONCLUSIONS, OR RECOMMENDATIONS

Yes

No

Is each result discussed in terms of the original hypothesis or topic

to which it relates?
____ ____
Is each result discussed in terms of its agreement or disagreement
with previous results obtained by other researchers in other studies?
____ ____

N/A
____
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Are generalizations consistent with the results?

____ ____

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Are the possible effects of uncontrolled variables on the results discussed?

____ ____

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Are theoretical and practical implications of the findings discussed?

____ ____

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Are recommendations for future action made?

____ ____

____

Are the suggestions for future action based on practical significance

or on statistical significance only; i.e., has the author avoided confusing
practical and statistical significance?
____
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ABSTRACT OR SUMMARY
Is the problem restated?

____ ____

Are the number and type of subjects and instruments described?

____ ____

____
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Is the design used identified?

____

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Are procedures described?

____

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Are the major results and conclusions restated?

____ ____

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--------------------------------www.faculty.unlv.edu/

Acebuche, Louisse T.

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September 16, 2016