NURS4301 Health Research Methods : Improve Human Health

Probiotics have over the years been attributed to tones of health benefits to the homo sapien species (Neethu et al, 2014). They first became a subject of interest in the 20^th century when Ellie Metchnikoff conducted a research to prove the theory that the long and healthy life of the Bulgarian farmers was due to their intake of “fermented milk and milk products by the fermenting bacillus (Lactobacillus) positive influence on the micro flora of the colon, decreasing toxic microbial activities” (Bibel, 1988). Since then many research studies have been conducted in an attempt to expound on the knowledge of these organisms. The research studies have been exemplary in deciphering the bio preservation advantages of the probiotics which seem to increase the shelf-life and safety of meat and milk products while conferring many health advantages to the human beings (Galvez et al, 2007). These findings provided the basis for the defining definition of probiotics as “the food ingredients which when administered in adequate amounts, confer a health benefit on the host” (Fuller, 1989). These microbes effectively preserve milk and other meat products by their production of lactic acid and other antimicrobial compounds (Neethu et al, 2014).

As food ingredients they help improve the taste of food by producing flavor compounds and also improve the nutritional quality of food (Neethu et al, 2014). Their health benefits in humans include the help in the control of serum cholesterol levels by production of the bile salt hydrolase (BSH) (Patel et al, 2014) and they are also beneficial in combating antibiotic resistance through microbial interference therapy (Parvez et al, 2006). Among the properties that render them effective in the gut include their salt, bile, and acid tolerance abilities with their capabilities to adhere to the gut mucosa and subsequently diffuse the antimicrobial compounds (Mourad and Mereim, 2008).

In understanding the properties that help probiotics in bio preservation and in increasing meat shelf-life, Neethu and his colleagues (2014) carried out a research that involved the culturing and isolation of the lactic acid bacteria of probiotics traits from different sources in different media and later examining them using various chemical test for salt, bile, and acid tolerance (Neethu et al, 2014). Their study design and sampling techniques was appropriate as it provided a representative study population of the microbes from various sources and measured the variables with easy to understand tests. The study identified the antimicrobial activity of the probiotics against some of the known human pathogens that cause food poisoning. This study, however, did little to identify the antimicrobial compounds produced by the probiotics and to correlate the role played by probiotics in the control of serum cholesterol levels.

Synopsis

Title: A research study to determine the antimicrobial products that are produced by the probiotics used in bio preservation, which help in the probiotic antimicrobial activity that suppresses the diseases by pathogens in humans and helps, improve human health.

Background: Many research studies have been conducted on the scope of the antimicrobial activity by the probiotics (Neethu et al, 2014) and on their mechanism of actions in microbial interference therapy (Sarita, 2016). The antimicrobial substances produced by these microorganisms have been determined to include “organic acids, hydrogen peroxide and bacteriocins” (Dunne et al, 1999) with their biological antimicrobial mechanism of action determined to involve the competition for nutrients with the pathogens that inhibit the growth of these organisms (Reid and Burton, 2002). The knowledge of the kind and nature of the bacteriocins (biologically active proteins) produced by these microorganisms is wanting and the aspect that I intent to decipher through this research.

Research Hypotheses: 

1. Probiotics are beneficial bio preservative.
2. Probiotics have antimicrobial activity.
3. Probiotics produce bacteriocins among many other antimicrobial substances.
4. Probiotics’ bacteriocins are of many different types.

Study Design: The research will make use of cultures of lactic acid bacteria that have probiotic traits which have been obtained from different probiotic microbial sources. These microbes will be grown in vitro in MRS media to isolate new strain of the probiotics (Lactobacilli) and the new strains obtained will be tested for the production of the various known bacteriocins. The study will take approximately two weeks to be completed and will be done in the nearby farm for collection of the milk samples and later in a fully equipped Molecular and Microbiology Research Laboratory in which the microbes’ culture will be obtained and the growth and isolation of new strains will be done with subsequent testing for the antimicrobial products.

Sampling Technique: The sample will be obtained in sterile bottles from different species, preferably from the milk of the cows and goats in the farm. These will be transported to the laboratory for the study. The samples will be from two cows and two goats in the farm.

Data Collection: Data will be collected from the tests carried out on the obtained probiotic strains via observation of the test results and subsequent documentation. These tests will be done to determine the presence and the kind of the bacteriocins produced by the probiotics by testing for the presence or absence of the bacteriocins produced by the other known gram positive microorganisms and by Archaea.

Statistical Analyses:

1. Descriptive Statics:This will make use of tables to compare the results from the two species of animals used to obtain the cultures of the microbes and to compare the results of the strains derived from these cultures and how they tested for the various bacteriocins tests.
2. Inferential Statistics:This will involve the drawing of conclusions based on the data obtained on each strain of the probiotics examined.

Ethics Application: This proposal needs to be submitted to the Australian National University Human Research Ethics Committee.

Limitations of the Study: The biases that might occur during this study include:

1. Possible contamination of the samples during the collection and transportation process
2. Possible inappropriate isolation of the strains in the laboratory
3. Possible occurrence of inconclusive test results

Significance of the Study: This study will help in the understanding of the antimicrobial activity and potential of the probiotics which might act as a guide in the expansion of the production and use of probiotics with potent and many bacteriocins production potential to help improve human health by minimizing the effects of the human pathogens.

Justification of proposed synopsis

I have chosen a study design that will involve the collection of fresh samples from the farm. Culture of the lactic acid bacteria will be obtained from the milk samples in vitro in the laboratory and strains of the lactobacillus bacteria depending on their culture morphological characteristics will be isolated and grown in MRS medium to obtain new strains. This study design is appropriate as it will oversee the generation of new and uncontaminated strains of the probiotics that will be a step forward in attaining quality results and hence good and correct inferences.

My sampling technique will make use of sterile bottles to collect the milk samples. The milk will be from two different species of the farm animals: cows and goats. I will use two cows and two goats as a representative of the study population and to act as a control in case of any contamination in the sample collection process. This sample technique will oversee the appropriate representation of the study population as it widens the range of the probiotic strains likely to be obtained in the cultures and hence a comprehensive study of the lactobacillus strains and types of bacteriocins that they produce.

The data collection method involves the observation of the test results from the test tubes containing the lactobacillus strains. This observation will be recorded as positive for strains that test positive for a specific type of bacteriocins and as negative for the strains that do not demonstrate the expression of the bacteriocins in question. This method of data collection is easy and will make the creation of comparison tables and the drawing of conclusions from the obtained data equally easy.

My chosen descriptive statistical analysis is the use of comparison tables for the cultures obtained from the two different animal species and for the different strains of the microbes obtained from the cultures. The inference statistical analysis will involve simple conclusions of the data describing a single strain of the lactobacilli. I picked on these statistical analyses because they are easy to follow and understand.

My Choice of Human Research Ethics Committee(s) is the Australian National University Human Research Ethics Committee as the research will be conducted with affiliation to the University and hence will involve student participation.

References:

Bibel, D. J. (1988). Ellie Metchnikoff’s Bacillus of Long Life. ASM News. 54: 661-665

Dunne C, O’Murphy L, Flynn S, O’Mahony L, O’Halloran S, Feeney M, Morrissey D, Thornton G, Fitzgerald G, Daly C, Kiely B, Quigley EMM, O’Sullivan GC, Shanahan F and Kevin J, (1999). Probiotics: from Myth to Reality: A Demonstration of Functionality in Animal Models of Disease and in Human Clinical Trials. Antonie van Leeuwenhoek, 76:279-292.

Fuller, R. (1989). Probiotics in Man and Animals: A Review. J Appl Bacteriol. 66: 365-378

Galvez, A., Abriouel, H., Lopez, R. L., and Omar, N. B. (2007). International Journal of Food Microbiology. 120: 51-70

Mourad, K. and Mereim, K. H. (2008). Gracsas y aceites. 59, 3, 218-24

Neethu, K., Christy, G., Charles, L. P., and Bhaskara, R. K. V. (2014).Bio preservation of Meat by Probiotics Bacteria from Dairy Products. Der Pharmacia Lettre, 2014, 6(6): 226-271

Patel, A., Prajapati, J. B., Holst, B., and Ljungh, A. (2014).Food Biosciences. 27-33

Parvez. S., Malik, K. A., Kang, S. A. H., and Kim, Y. H. (2006). Journal of Applied Microbiology. 100: 1171-1185

Reid, G. and Burton, J. (2002). Use of Lactobacillus to Prevent Infection by Pathogenic Bacteria. Microbes Infect. 4: 319-324

Sarita, A. B. (2016). Studies on Health Promoting and Disease Preventing Properties of Lactobacilli Isolated from Various Sources and their Microencapsulation. UGC-Faculty Recharge Programme of the University Grant Commission