1. This cross-sectional study aims to identify common organisms causing infections in COPD and asthma patients by sputum culture and determine the antimicrobial susceptibility patterns of isolated microorganisms. 2. Sputum samples will be collected from 100 COPD and asthma patients experiencing acute exacerbations at a hospital in Jodhpur, India. Samples will undergo culture and identification of bacteria/fungi followed by antimicrobial susceptibility testing. 3. Preliminary results from previous studies suggest bacteria like H. influenzae, S. pneumoniae, and M. catarrhalis commonly cause COPD exacerbations, while studies of asthma patient microbiota show alterations compared to healthy individuals.

1. A CROSS SECTIONAL STUDY TO FIND COMMON ORAGANISM IN COPD
AND ASTHMA BY SPUTUM CULTURE &THEIR ANTIMICROBIAL SENSTIVITY
Principal Investigator
Dr INDRA SAINI
Resident Doctor
Department of Medicine
Guide
Dr. NARENDRA SINGH RAWAT
Senior Professor &Unit Head
Department of medicine

2. Introduction
• Asthma is a disorder, characterized by reversible airway narrowing and bronchial hyper- responsiveness (BHR) to
nonspecific agents as the main basic pathophysiology.
• An asthma exacerbation can happen as a consequence of a single cause, but more commonly will happen from a
mixture of causes leading to complex inflammatory pathways and initiation of bronchial narrowing.
• Bacterial infection contribute to airway wall remodeling through the activation of fibrosis by the release of growth
factors such as TGF beta, induced by bacterial LPS, Leading to fibroblast activation and release of extracellular matrix
protein.

3. • COPD is a heterogenous lung disease characterized by cough , dyspnea, sputum production and
exacerbation due to abnormality of airway or alveoli which lead to persistent and progressive
airflow obstruction.
• In the United States of America alone, approximately 24 million people suffer with COPD and have
emerged as the third leading cause of death.
Three classes of pathogens have been implicated as a cause of COPD exacerbation: respiratory viruses, atypical
bacteria, aerobic gram positive and gram negative bacteria.
Approximately 50% of COPD acute exacerbations are associated with the bacteria from lower respiratory tract.
The dominant bacteria isolated are H. influenzae, S. pneumoniae and M. catarrhalis. In advanced cases, P.
aeruginosa becomes prevalent.

4. AIM AND OBJECTIVES
Aim:
• To find the common organisms in COPD and Asthma patient by sputum culture and their antibiotic
sensitivity
Objectives:
1. To isolate and identify bacteria and fungal agents causing microbial infections in COPD and chronic
Severe Asthma patients.
2. To determine antimicrobial susceptibility pattern of the various microorganism in COPD and
Asthma patients.

5. MATERIAL AND METHODS
• This is a cross sectional study will be done at department of Medicine, Mahatma Gandhi Hospital
attached to Dr. S. N. Medical College, Jodhpur, Rajasthan, India.
• STUDY DESIGN: Cross sectional Study
• STUDY LOCATION: Department of Medicine, Mahatma Gandhi Hospital attached to Dr.
S.N. Medical College, Jodhpur
• STUDY PERIOD: 6 months after approval from ethical committee.

6. Sample size
• Sample size was calculated at 95% confidence interval and 10% relative allowable error using the formula for
sample size for estimation of a single sample proportion –
• 𝑁 =
(𝑍1−𝛼/2)2 𝑃 (1−𝑃)
𝐸2
• Where,
• 𝑍1−𝛼/2 = Standard normal deviate for 95% confidence interval (taken as 1.96)
• P = Expected proportion of sputum culture positivity was observed (taken as 48.7% as reported by P. Sharma23)
• E = Relative allowable error (taken as 10% of P)
• Sample size was calculated to be minimum of 100 subjects

7. INCLUSION CRITERIA
1. Age ≥18years
2. COPD patient defined according to GOLD guideline 2023.
3.Chronic severe Asthma patient with Acute exacerbation , increased dyspnea, increased sputum
production and purulence and fever.
EXCLUSION CRITERIA:
1. Patients having cavitary lesion like bronchiectasis, tuberculosis, malignancy, community
acquired pneumonia.
2. Previous antibiotic treatment in the last 15 days.
3. Patients managed in emergency or admission required in Intensive Care Unit.

8. METHODOLOGY:
After obtaining inform consent ,Patients will be educated about the difference between
sputum and oral secretion. Early morning samples will be obtained from cases that will be
clinically diagnosed as COPD And Asthma. Patients will be instructed to collect coughed
sputum into a sterile wide mouth container with a screw cap after rinsing the mouth twice
with plain water. Sample containers will be labelled after the collection and will be
transported to the laboratory immediately and processed within 30 minutes of collection.

9. Processing of sputum sample
1. After obtaining sputum sample, do direct smear formation for gram staining and culture of
sputum sampling in TG media for 24 hour and if there is turbidity in growth media then we go
for incubation in incubator at 370centigrade temprature for 10-12 hour then the bacterial growth
put on blood agar and MacConkey agar and observe type of bacterial colony
2. In blood agar we look for whether the bacterial colony hemolytic or Non-Haemolytic by zone of
hemolysis . hemolytic colony we look for whether alpha or beta or gamma hemolysis

10. • In MacConkey agar we look whether colony lactose fermenting or non-lactose fermenting
3. 10% KOH mount for fungal growth
• Then after again do gram smear from colony to look whether gram positive or negative cocci or
bacilli.
• Then after obtaining bacterial colony antimicrobial susceptibility done using Kirby-Bauer disk
diffusion test.

11. REVIEW OF LITERATURE
• Miravitlles M et al (1999) to assess microorganisms causing the acute exacerbations of COPD are distributed unevenly among
patients with different degrees of severity, with patients more severely affected showing a greater incidence of Pseudomonas and
H influenzae. Besides FEV1, other easy-to-obtain clinical data, such as active smoking and the time since the last exacerbation,
may help in choosing the appropriate empiric treatment for exacerbations.
• Zhang Q. Illing et al (2012) to concluded bacterial colonization of the lower airways is a common occurrence in patients with
chronic stable severe asthma, but this was not related to the degree of airway wall thickness measured radiologically. Bacterial
colonization of asthmatic airways may not be the primary driver of airway wall remodeling, but it could be involved in other
asthmatic processes.
Marri PR et al (2013) studied all sputum samples contained 5 major bacterial phyla: Firmicutes, Proteobacteria, Actinobacteria, Fusobacterium,
and Bacteroidetes, with the first 3 phyla accounting for more than 90% of the total sequences. Proteobacteria were present in higher proportions in
asthmatic patients (37% vs 15%, P < .001). In contrast, Firmicutes (47% vs 63%, P = .17) and Actinobacteria (10% vs 14%, P = .36) were found
more frequently in samples from non asthmatic subjects, although this was not statistically significant. Hierarchical clustering produced 2
significant clusters: one contained primarily asthmatic samples and the second contained primarily non asthmatic samples. In addition, samples
from asthmatic patients had greater bacterial diversity compared with samples from non asthmatic subjects. They concluded patients with mild
asthma have an altered microbial composition in the respiratory tract that is similar to that observed in patients with more severe asthma.

12. Ahmed Aya H. et al (2020) to evaluate the bacterial organisms are responsible for about one- third of bronchial asthma exacerbation, with
predominance of Gram-negative bacteria.Amikacin, Quinupristin/ Dalfopristin, Linezolid, Vancomycin, and Tigecycline, Tobramycin,
Ciprofloxacin, Levofloxacin, and Gentamicin were the most common sensitive antibiotics.
Sharma P et al (2017) to assess acute exacerbation of COPD is associated with bacterial infections, profile of which varies in various
geographical areas. To improve the morbidity and mortality with such exacerbations it seems logical to assess the bacteriological profile of
AECOPD in an area from time to time along with the antibiotic resistance pattern of the organism. Moreover, judicious use of antibiotics
based on sputum culture and antibiogram seems to be the best strategy to prevent resistance pattern. While, production of mucopurulent and
purulent sputum, are already known to be associated with bacterial infection, SPO2 at presentation might be evaluated further in future
studies as a guiding tool to suspect bacterial cause of AECOPD.

13. Clinical Performa
• Name
• Age
• Sex
• Address
• Locality (Rural/Urban)
• Occupation

14. 1. Total Leucocyte Count………Platelets………SGOT…….SGPT
2. Blood Urea…………S Creatinine…….
3. Lipid Profile…..
4. CRP………………
5. T. Bili. / Cong………..
6. T. Protein Albumin……..
7. Sodium……….Potassium…….
8. HIV
9. HBsAg
10.HCV
11.ABG
12.Chest X Ray
13.ECG

15. CHIEF COMPLAINT DURATION
• Shortness of birth
• Cough
• Fever
• Loss of appetite
• Specific if progress
MEDICAL HISTORY DURATION
• COPD
• ASTHMA
DRUG HISTORY

16. Vitals and General examination
 Pulse –
 Blood Pressure
 Respiratory Rate –
 SpO2 (pulse oximetry)
 Consciousness
 Pallor
 Icterus
 Cyanosis
 Lymphadenopathy
 Clubbing
 Oedema

17. SYSTEMIC EXAMINATION
• Cardiovascular
• Respiratory
• CNS
• Per abdomen

18. Result of sputum culture
Organism Positive culture Number (%)
Negative culture
Total

19. Variable N (%among total
patient)
Culture positive cases
N (%)
p value
Age group
Smoking
Ex –smoker
smoker
Residence
Rural
urban
COPD
ASTHMA
ABG
With acidosis
without
Leukocytosis
Yes
No
CRP
Positivity
Negative

20. INFORMED CONSENT FORM
STUDY TITLE:“ A CROSS SECTIONAL STUDY TO FIND OUT COMMON ORGANISM IN COPD AND ASTHMA PATIENT BY
SPUTUM CULTURE AND THEIR ANTIMICROBIAL SENSITIVITY
• Patient ID number.
• Patient's name:
• Date of Birth/Age:
• I confirm that dated__________ have read and understood the information sheet for the above study and have had the opportunity to ask questions.
• I understand that my participation in the study is voluntary and I am free to withdraw at any time, without assigning any reason, without affecting my
medical care or legal rights.
• I understand that, the ethics committee and regulatory authorities will not need my permission to view my health records in connection with the current
study and any other research conducted in relation to it, even if I withdraw from the trial. I agree with this approach. However, I understand that my
identity will not be revealed in any information released or published to third parties.
• I agree not to restrict the use of any data or results that may be generated for this study, provided that such use is for scientific purposes only.
• I agree to participate in the above study.
Signature (or thumb impression) of the patient

21. • Representative___________________________
• Date ___________________________
• Signatory's name ___________________________
• Signature of the investigator ___________________________
• Study investigator’
s name/phone no. ___________________________
• Signature of witness ___________________________
• Date ___________________________
• Name and address of the witness ___________________________
• I, _____________________S/D/W/o___________________ consent to be a subject of the proposed study.
• I have been informed well enough about the nature of study : Title: "A CROSS SECTIONAL STUDY TO FIND OUT COMMON ORAGANISM IN ASTHMA AND COPD BY SPUTUM
CULTURE AND THEIR ANTIMICROBIAL SENSTIVITY”
• and I am consenting to the study by my own will without any pressure or force whatsoever after understanding everything.
• Therefore, I will be myself responsible for any harm, untoward effects or benefits whatsoever due to the study and I will not make any claims afterward.
Signature (or thumb impression) of the patient
Date

22. सूचित सहमचत प्रपत्र
अध्ययन शीर्षक: "थूक कल्िर और संवेदनशीलता द्वारा सीओपीडी और अस्थमा रोगी में सामान्य जीव का पता लगाने क
े चलए एक क्रॉस सेक्शनल
अवलोकन अध्ययन।
• रोगी आईडी नंबर।
• रोगी का नाम:
• जन्म चतचथ/आयु:
• मैं पुचि करता हं चक dated__________ उपरोक्त अध्ययन क
े चलए सूिना पत्र पढा और समझा है और प्रश्न पूछने का अवसर चमला है।
• मैं समझता हं चक अध्ययन में मेरी भागीदारी स्वैच्छिक है और मैं चकसी भी समय, चबना कोई कारण बताए, अपनी चिचकत्सा देखभाल या कानूनी अचिकारों को प्रभाचवत
चकए चबना वापस लेने क
े चलए स्वतंत्र हं।
• मैं समझता हं चक, नैचतकता सचमचत और चनयामक अचिकाररयों को वतषमान अध्ययन और इसक
े संबंि में चकए गए चकसी भी अन्य शोि क
े संबंि में मेरे स्वास्थ्य ररकॉडष
को देखने क
े चलए मेरी अनुमचत की आवश्यकता नहीं होगी, भले ही मैं परीक्षण से हट जाऊ
ं । मैं इस दृचिकोण से सहमत हं। हालांचक, मैं समझता हं चक तीसरे पक्ष को
जारी या प्रकाचशत चकसी भी जानकारी में मेरी पहिान प्रकट नहींकी जाएगी।
• मैं इस अध्ययन क
े चलए उत्पन्न होने वाले चकसी भी डेटा या पररणामों क
े उपयोग को प्रचतबंचित नहीं करने क
े चलए सहमत हं, बशते चक ऐसा उपयोग क
े वल वैज्ञाचनक
उद्देश्यों क
े चलए हो।
• मैं उपरोक्त अध्ययन में भाग लेने क
े चलए सहमत हं।
• रोगी क
े हस्ताक्षर (या अंगूठे का चनशान)
•

24. THANK YOU