In Vitro Sensitivity Testing of Antimonium Tart on Culture of Klebsiella Pneumoniae

Sambhaji S. Phadatare, A. B. Jadhav, Rama Bhadekar

Keywords

Klebsiella pneumonia, Antimonium Tart, Potency, Turbidity, Micro-titer plate, Antibacterial Sensitivity

Abstract

“In Vitro Sensitivity Testing Of Homeopathic Remedy Antimonium Tart On Culture of Klebsiella Pneumoniae.”

1. pneumoniae can cause destructive changes to human lungs via inflammation and hemorrhage with cell death (necrosis) that sometimes produces a thick, bloody, mucoid sputum (currant jelly sputum). These bacteria gain access typically after a person aspirates colonizing oropharyngeal microbes into the lower respiratory tract. As a general rule, Klebsiella infections are seen mostly in people with a weakened immune system. Most often, illness affects middle-aged and older men with debilitating diseases. This patient population is believed to have impaired respiratory host defenses, including persons with diabetes, alcoholism, malignancy, liver disease, chronic obstructive pulmonary diseases, gluco-corticoid therapy, renal failure, and certain occupational exposures (such as paper mill workers). Many of these infections are obtained when a person is in the hospital for some other reason (a nosocomial infection). Feces are the most significant source of patient infection, followed by contact with contaminated instruments. The most common condition caused by Klebsiella bacteria outside the hospital is pneumonia, typically in the form of broncho-pneumonia and also bronchitis. These patients have an increased tendency to develop lung abscess, cavitations, empyema, and pleural adhesions. It has a death rate of about 50%, even with antimicrobial therapy. The mortality rate can be nearly 100% for people with alcoholism and bacteremia. In addition to pneumonia, Klebsiella can also cause infections in the urinary tract, lower biliary tract, and surgical wound sites. The range of clinical diseases includes pneumonia, thrombophlebitis, urinary tract infection, cholecystitis, diarrhea, upper respiratory tract infection, wound infection, osteomyelitis, meningitis, bacteremia and septicemia. For patients with an invasive device in their bodies, contamination of the device becomes a risk; for example, neonatal ward devices, respiratory support equipment, and urinary catheters put patients at increased risk. Also, the use of antibiotics can be a factor that increases the risk of nosocomial infection with Klebsiella bacteria. Sepsis and septic shock can follow entry of the bacteria in to the blood.

Most K. Pneumonia are resistant to antibiotics, so another therapy is required. I wondered if we could form similibiotics or energetic biotics, so I decided to do in- vitro sensitivity testing with Antim Tart. 30,200,1M, 50M commonly used potencies in practice. We perfomed 30 experiments from approximately Jan 2017 to October 2017. I got most positive results with Antim Tart. 30 and 200 potencies. This means no growth with Antim tart 30, 200 And Antim Tart. 1M  50M  growth.

Introduction

Homeopathy as a system of treatment has progressed remarkably during the last few decades. Since the beginning of the homeopathic era, high dilutions of homeopathic drugs have demonstrated their activity. Many theories are present to explain the anti-microbial activity of homeopathic drugs. In the present era  homeopathic medical practice proves its utility in all types of illnesses. Harmful stimuli like bacteria, fungal organisms, viruses and toxins cause reactions in the Vital Force. Their effects depend on the individual’s level of health as well as the strength of the harmful stimuli. Today all homeopaths may make use of Antimonium Tartaricum for complaints like upper respiratory tract infection, lower respiratory tract infection, pneumonia, bronchopneumonia, bronchitis, lung abscesses, pleural effusion etc.

It is a most useful drug listed in the highest grade for pneumonia and lower respiratory tract infection by Murphy’s and Boeiricke’s repertories. Many other repertories mention that this is a very useful drug for respiratory complaints. It is also used for pneumonia biliosa with hepatic congestion, pleura-pneumonia, when parts of lungs are hepatized and fine rales are heard over the hepatized part. Other symptoms include great oppression of breathing towards morning, must sit up to breathe, broncho-pneumonia, pneumonia catarrrhalis when second stage resolution has set in but fails , and oppression and prostration prevail,.

At first we find sharp stiching pains, but pains cease and mucus rales are heard distinctly over the chest with extreme shortness of breath, rattling, hollow cough with hot moist hands, wweat on forehead, dysponea with desire to cough, chest is fall of mucus and inability to bring it up. Pneumonia of drunkards with bilious complication, even jaundice, meteorism, nausea and vomiting, typhoid complications, oedema pulmonum, impending paralysis of lungs or of heart. It is also suitable especially to infants and old or cachetic people.

Two unusual infections of note from Klebsiella are rhinoscleroma and ozena. Rhinoscleroma is a chronic inflammatory process involving the nasopharynx. Ozenais is a chronic atrophic rhinitis that produces necrosis of nasal mucosa and muco purulent nasal discharge. Klebsiella ranks second to E. coli for urinary tract infections in older people. It is also an opportunistic pathogen for patients with chronic pulmonary disease, enteric pathogenicity, nasal mucosa atrophy, and rhinoscleroma.

Klebsiella organisms are often resistant to multiple antibiotics. Current evidence implicates plasmids as the primary source of the resistance genes. Klebsiella with the ability to produce extended-spectrum beta-lactamases (ESBL) is resistant to many classes of antibiotics. The most frequent are resistance to amino glycosides, fluoroquinolones, tetracyclines, chloramphenicol, and trimethoprim/sulfamethoxazole. The WHO’s first global report on antibiotic resistance reveals antibiotics would not work for more than half the people treated.  Klebsiella pneumonia-News relase 30 April 2014 .

Klebsiella pneumoniae is member of K. genus of entrobacteriaceae   and belongs to normal flora of the human mouth and intestine. It is a Gram-negative, non motile, encapsulated, lactose-fermenting, facultative anaerobic, rod-shaped bacterium. Humans are the primary reservoir for K. pneumonia. Carrier rates of K. pneumonia range from 5 to 38 % in stool samples,  1 to 6 % in nasopharynx,  Klebsiella species are rarely on the skin. Higher rates of nasopharyngeal carriage have been noted in ambulatory patients. Carrier rates are markedly increased in hospitalized patients among whom reported rates are 77 % in the stool, 19% in pharynx and 42% on the hands. The higher rates of colonization are primarily related to the use of antibiotics which increases the prevalence. This is important clinically since in one report Klebsiella nosocomical infections was four times higher in stool carriers compared with non carriers. Similarly, in a study of 855 patients with Klebsiella pneumonia liver abscesses in Taiwan and 3420 age and sex-match controls ampicillin and amoxicillin use within the 30 days of diagnosis was associated with an increased risk of liver abscess. Klebsiella pneumonia infection is emerging as an important challenge in the health care system. The medical profession needs alternative medicines to control the Klebsiella pneumonia infections.

Aim-The aim of this study is to determine the sensitivity of  varied potencies of Ant.Tart against Klebsiella pneumonia.

Objectives-

1. To estimate the antimicrobial activity of varied potencies of Ant.Tart. against colonies of Klebsiella pneumoniae.
2. To find out the potency of Ant.Tart. having the most broad sensitivity against Klebsiella pneumonia infections.
3. After estimation of sensitivity of varied potencies of Ant. Tart. Can we can use this medicine as an antibiotic against Klebsiella pneumonia infections or its adjuvant medicine against Klebsiella pneumonia infections.

MATERIALS AND METHODS

Study settings–     This study will be done (in vitro) at the microbiology laboratory at Rajiv Gandhi institute of IT and Biotechnology, Bharati vidyapeeth Deemed University, Pune- 46.

Media and chemicals- All media will be procured from Hi Media Mumbai, India. All the reagents and chemicals used in this work were of AR grade and procured from MERK India.

Homoeopathic drug preparation– Homoeopathic drug Ant. Tart. In 30 CH, 200 CH, 1M, 50M with their liquid dilutions will be obtained from Dr.Willmar Schwabe India pvt. Ltd.

Organism –         Klebsiella Pneumoniae (Ncim ) will be used as standard culture to examine antimicroibial activity. other culture used in this study are clinical isolets procured from Bharati Hospital, Pune.

Antimicrobial assay- Antimicrobial assays will be performed using L-B (Luria Bertini) medium. For this the isolates and reference cultures were grow in LB medium to McForland standard 1-0.5 100 µl of culture was spread individually on LB plate containing base agar overplayed with soft agar. (ref) The wells were punctured in soft agar at equidistance using 3-5 mm bore. Drug potencies used for testing were 30CH, 200CH, 1M , 50M. The only solvent in one well on each plate is ethanol which serves as control. Plates were incubated at 37 degree C for 24 hours. At the end of incubation period diameter of zones of inhibition were measured. I have not got results from petri dish L-B (Lureia Bertini) medium so with the help of a microbiologist I changed method, that is dilution method.

Exclusion criteria- 1.Those isolated Klebsiella pneumonia organism cultures which are collected from HIV Tuberculosis patients.

2.Those Klebsiella pneumoniae organism which cannot be isolated from other microorganisms like any virus, bacteria, fungi.

 SLANT PREPARATION EXPERIMENT

PROCEDURE FOR COLLECTION OF SAMPLE OF K.PNEUMONIA

Firstly wash the test tube (CG-256 screw tubes round bottom) with help of distilled water and then take Luria Bertani broth solid powder from bottle. According to Hi-media preparation for 1000ml distilled water liquid slant test tube require 25 gram in 1000 ml distilled water.

Therefore,

25gram in 1000ml  then how many grams require 10 m slant test tube

X                         10ml

25    x    10    =     250

250 Upon 1000 = 0.25 gram for one test tube slant

Now we require agar-agar type 1. According to Hi-media Agar-agar type 1 for 100 ml distilled water 1.5 gram powder required.

Therefore,

1.5gram         in      100ml      then how many grams require 10 m slant test tube

X                         10ml

1.5    x    10    =     15

15 Upon 100 = 0.15 gram for one test tube slant

After that weighing with help of micro weight machine weight of Luria Bertani broth solid powder  Agar-agar type 1 powder taken. Poured both powder into the test tube slant then take 10 ml distilled water and shake well. Then with help of sterilizer.

PREPAIRED SLANT TEST TUBE FOR BACTERIAL COLLECTION

Next day after solidification of slant I went to the Bharati Hospital and inoculation of Klebsiella pneumonia was done.

Isolated K.Pnemonia Culture collected from Ett (Endo Trchial Tube) Secretion.

he slant test tube kept in INCUBATOR for 24 hours. After 24 hours it is ready for the culture experiment.

EXPERIMENT                  

( NUTRIENT BROTH DILUTION METHOD)

The experiment no 2 started with fresh culture which is prepaid above. Nutrient broth prepared in conical flask procedure mentioned as above by taking anti-contamination caution. . Then make 7 test tubes by putting 8.5ml nutrient broth and sterilize the 7 test tubes.

According to the above chart we put Nutrient Broth,Culture, Antim Tart 30CH, 200CH, 1M ,50M potencies and then distilled water, Alcohol 90% by taking anti-contamination caution with the help of micro pipette  droppers. I kept those test tubes in the incubator for 24 hours.

Result after  24 hours –

After performing  above  30 experiments : Observation and Results

My aim in this study was to determine the sensitivity of  varied potencies of Ant.Tart against Klebsiella pneumonia.

According to my judgement there was no growth of Klebsiella pneumonia bacteria for potencies 30 & 200.  Each experiment’s results are explained in the above chart.

Conclusion-

1. I cannot estimate the antimicrobial activity of varied potencies of Ant.Tart. against colonies of Klebsiella pneumonia. It required a micro titer plate that  can measure the anti microbial activity of medicine and alcohol separately .
2. Potency 30 & 200 of Ant.Tart. will have the most broad sensitivity against Klebsiella pneumonia infections but required micro titer plate experiment.
3. After performing the required micro titer plate experiment of the sensitivity of varied potencies of Ant. Tart., we can use this medicine as an antibiotic or similibiotic against Klebsiella pneumonia infections or its adjuvant medicine against Klebsiella pneumonia infections.