lactobacillus acidophilus

Basic Characteristics

• Gram-Positive (1)

• Bacillus, rod-shaped (2)

• Commonly found in oral, vaginal and intestinal regions of many animals.

• Industrial Microbes that contribute to the production of cheese, yogurt and other products such as fermented milk, all stemming from production of lactic acid.

History

Lactobacillus Acidophilus was originally isolated by Ernst Moro from the human gastrointestinal tract and was designated Bacillus Acidophilus in 1900. Historically, L. acidophilus is the Lactobacillus species most often implicated as an intestinal probiotic capable of eliciting beneficial effects on the microbiota of the gastrointestinal tract (GIT) (3). This organism is used commercially in many different dairy products and as a dietary supplement because research has documented it as a normal resident of the human intestinal tract from its initial inoculation (4).

Domain: Bacteria

Phylum: Firmicutes

Class: Bacilli

Order: Lactobacillales

Family: Lactobacillaceae

Genus: Lactobacillus

Species: L. acidophilus

(https://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&id=891391&lvl=3&p=has_linkout&p=blast_url&p=genome_blast&lin=f&keep=1&srchmode=1&unlock)

What is the morphology of the cells?

The slide shows a picture of a pure culture and all of the microbes have a rod-shaped figure which is indicative of a bacilli morphology (5).

Are there any other visualizations of it?

If there are any published images, explain what microscope they use and if they used a stain

Yes, I found a publication on the use of Lactobacillus Acidophilus for the removal of Arsenic (III) from waste water (6). They included an image of the microbe they took using a Scanning Electron Microscope. The article did not mention the use of a stain and the image doesn’t show any color so I believe they did not use one.

Tests to Identify Microbe:

Test for Gram-Positive, Bacillus, growth on ordinary blood agar, glucose fermenter, Esculin Hydrolysis, Growth at 42 Degrees Celsius, Cellobiose,

Tests that could be done include: Blood Agar Plate, Mannitol Salt Agar, DNase Plate, Phenol Red Broth, Sulfur Indole Motility Test

(https://login.lp.hscl.ufl.edu/login?URL=http://web.gideononline.com/loginx.php?user=ufmicro)

According to Gideon Cutibacterium acnes, Bacillus Megaterium, Bacillus Subtilis all have similar test results.

Using primers in literature to run in In silico PCR amplification and BLAST to find 16S ribosomal RNA

Forward Primer: LbLMA1-rev (5′-CTC AAA ACT AAA CAA AGT TTC-3′)

Reverse Primer: A second primer R16–1 (5′-CTT GTA CAC ACC GCC CGT CA-3′)

Forward Primer Tm: 44.6 Degrees Celsius

Reverse Primer Tm: 55.9 degrees Celsius

% GC Forward Primer: 29%

% GC Reverse Primer: 60%

(https://pubmed.ncbi.nlm.nih.gov/12351242/)

full-length 16S rRNA sequence (in FASTA format) (11)

>NR_117062.1 Lactobacillus acidophilus strain VPI 6032 16S ribosomal RNA, partial sequence

GGGACGAACGCTGGCGGCGTGCCTAATACATGCAAGTCGAGCGAGCTGAACCAACAGATTCACTTCGGTG

ATGACGTTGGGAACGCGAGCGGCGGATGGGTGAGTAACACGTGGGGAACCTGCCCCATAGTCTGGGATAC

CACTTGGAAACAGGTGCTAATACCGGATAAGAAAGCAGATCGCATGATCAGCTTATAAAAGGCGGCGTAA

GCTGTCGCTATGGGATGGCCCCGCGGTGCATTAGCTAGTTGGTAGGGTAACGGCCTACCAAGGCAATGAT

GCATAGCCGAGTTGAGAGACTGATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCA

GCAGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGTTTTCG

GATCGTAAAGCTCTGTTGTTGGTGAAGAAGGATAGAGGTAGTAACTGGCCTTTATTTGACGGTAATCAAC

CAGAAAGTCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTA

TTGGGCGTAAAGCGAGCGCAGGCGGAAGAATAAGTCTGATGTGAAAGCCCTCGGCTTAACCGAGGAACTG

CATCGGAAACTGTTTTTCTTGAGTGCAGAAGAGGAGAGTGGAACTCCATGTGTAGCGGTGGAATGCGTAG

ATATATGGAAGAACACCAGTGGCGAAGGCGGCTCTCTGGTCTGCAACTGACGCTGAGGCTCGAAAGCATG

GGTAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGATGAGTGCTAAGTGTTGGGAGGTTT

CCGCCTCTCAGTGCTGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTC

AAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTT

ACCAGGTCTTGACATCTAGTGCAATCCGTAGAGATACGGAGTTCCCTTCGGGGACACTAAGACAGGTGGT

GCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTCATT

AGTTGCCAGCATTAAGTTGGGCACTCTAATGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACG

TCAAGTCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGACAGTACAACGAGGAGCAAGC

CTGCGAAGGCAAGCGAATCTCTTAAAGCTGTTCTCAGTTCGGACTGCAGTCTGCAACTCGACTGCACGAA

GCTGGAATCGCTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCC

CGTCACACCATGGGAGTCTGCAATGCCCAAAGCCGGTGGCCTAACCTTCGGGAAGGAGCCGTCTAAGGCA

GGGCAGATGACTGGGGTGAAGTCGTAACAAGGTAGCCGTAGGAGAACCTGCGGTTGGATCA

Lactobacillus Acidophilus is a non-pathogenic probiotic found in the intestines of many healthy people. They actually have great therapeutic potential for microbial equilibrium of the intestine, in food fermentation and used as a probiotic in food supplements. Lactobacillus Acidophilus has been studied to have inhibitory potential against pathogenic microorganisms and can be used to improve the safety of food products (12).

Lactobacillus Acidophilus can be found in the human oral, gastrointestinal, and vaginal microbial flora (13).

Lactobacillus is widely commercialized and has been recognized to have probiotic effects and is one of the most commonly suggested organisms for dietary use. It’s frequently added to yogurt and fermented milk products. Around 80% of yoghurts produced in the U.S. contain this bacteria (14).

Treatment

There has been evidence of resistance to trimethoprim by Lactobacillus Acidophilus. Acidophilus is actually used to treat or prevent several different kinds of infections and helps maintain normal consistency of bacteria in the body.

How do you grow it?

BSL Level

BSL 1 (15).

Type of Agar

Lactobacilli MRS Agar, which is composed of 55.0 g Lactobacilli MRS Broth and 1000 mL of water (15).

Aerobic/Anaerobic

Lactobacillus Acidophilus can be kept in either aerobic or anaerobic conditions, but are grown mainly in anaerobic conditions (16).

Procedure to Grow

1. Open vial according to enclosed instructions or visit www.atcc.org for instructions.

2. Rehydrate the entire pellet with approximately 0.5 mL of #416 broth. Aseptically transfer the entire contents to a 56 mL tube of #416 broth. Additional test tubes can be inoculated by transferring 0.5 mL of the primary broth tube to these secondary broth tubes.

3. Use several drops of the primary broth tube to inoculate a #416 plate and/or #416 agar slant.

4. Incubate at 37°C for 2448 (15).

5. Before Lactobacillus Acidophilus is grown. It must be stored in temperatures around -80 degrees Celsius or colder. It also shows enhanced growth in a media of Kulp (15).

First Documentation:

The first documentation was in the 1890’s by Dr. Ernst Moro (17).

(1) Mokoena MP. 2017. Lactic Acid Bacteria and Their Bacteriocins: Classification, Biosynthesis and Applications against Uropathogens: A Mini-Review. Molecules 22.

(2) Reid G. 1999. The scientific basis for probiotic strains of Lactobacillus. Appl Environ Microbiol 65:3763– 3766.

(3) K.M. Selle, ... W.M. Russell, in Encyclopedia of Food Microbiology (Second Edition), 2014

(4) W.R. Aimutis, in Encyclopedia of Food Microbiology (Second Edition), 2014

(5) Pyar H, Peh K-K. 2013. Isolation of probiotics Lactobacillus acidophilus from commercial yoghurt 13.

(6) Singh AL, Sarma PN. 2010. Removal of Arsenic(III) from Waste Water Using Lactobacillus acidophilus. Bioremediation Journal 14:92–97.

(7) Dubernet S, Desmasures N, Guéguen M. 2002. A PCR-based method for identification of lactobacilli at the genus level. FEMS Microbiology Letters 214:271–275.

(8) Rather IA, Choi K-H, Bajpai VK, Park Y-H. 2015. Antiviral mode of action of Lactobacillus plantarum YML009 on Influenza virus H1N1. Bangladesh J Pharmacol 10:475.

(9) Worku, Hable & Amenu, Kebde & Fesseha, Haben. (2020). The Role of Starter Culture in Producing Probiotic Yoghurt: Significance for Human Health -A Review. 8. 20-34. 10.18782/2322-0392.1286.

(10) Ahirwar SS, Gupta MK, Gupta G, Singh V. 2017. Screening, Isolation and Identification of Lactobacillus Species from Dental Caries of Children. IntJCurrMicrobiolAppSci 6:497–503.

(11) Dubernet S, Desmasures N, Guéguen M. 2002. A PCR-based method for identification of lactobacilli at the genus level. FEMS Microbiology Letters 214:271–275.

(12) Muhammad I, Ahmad AA, Shah T. 2018. Health Promoting and Disease Preventing Properties of Probiotics with Special Reference to Lactobacillus: A Review. J Prob Health 06.

(13) Pfeiler EA, Azcarate-Peril MA, Klaenhammer TR. 2007. Characterization of a Novel Bile-Inducible Operon Encoding a Two-Component Regulatory System in Lactobacillus acidophilus. JB 189:4624–4634.

(14) Pfeiler EA, Azcarate-Peril MA, Klaenhammer TR. 2007. Characterization of a Novel Bile-Inducible Operon Encoding a Two-Component Regulatory System in Lactobacillus acidophilus. JB 189:4624-4634.

(15) Lactobacillus acidophilus (Moro) Hansen and Mocquot (ATCC® 4356™). Lactobacillus acidophilus (Moro) Hansen and Mocquot ATCC ® 4356&tr.

(16) Jebur MSh. Therapeutic efficacy of Lactobacillus acidophilus against bacterial isolates from burn wounds. N Am J Med Sci. 2010;2(12):586-591. doi:10.4297/najms.2010.2586

(17) Weirich A, Hoffmann GF. Ernst Moro (1874-1951)--a great pediatric career started at the rise of university-based pediatric research but was curtailed in the shadows of Nazi laws. Eur J Pediatr. 2005 Oct;164(10):599-606. doi: 10.1007/s00431-005-1703-2. Epub 2005 Jun 2. PMID: 15931526.