{"id":18986,"date":"2018-03-25T10:14:22","date_gmt":"2018-03-25T10:14:22","guid":{"rendered":"http:\/\/biomedpharmajournal.org\/?p=18986"},"modified":"2020-04-23T05:49:14","modified_gmt":"2020-04-23T05:49:14","slug":"isolation-identification-and-antibiotic-sensitivity-pattern-of-pyogens-from-pyogenic-pathogens","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol11no1\/isolation-identification-and-antibiotic-sensitivity-pattern-of-pyogens-from-pyogenic-pathogens\/","title":{"rendered":"Isolation Identification and Antibiotic Sensitivity Pattern of Pyogens from Pyogenic Pathogens"},"content":{"rendered":"

Introduction<\/strong><\/p>\n

Human skin acts as an excellent barrier to infection.\u00a0 Most bacteria live on our skin, in the nasopharynx, gastrointestinal tract and other parts of the body with little potential for causing disease because of the first line defense within the body.\u00a0 The surgical operation, trauma, burns, disease, nutrition and other factors affect the defenses.\u00a0 The skin barrier is disrupted by every skin incision and microbial contamination is inevitable, despite the best skin penetration.<\/p>\n

Skin and skin structure infections are common and range from minor pyodermas to severe necrotizing infections.\u00a0 Skin can be infected by a variety of microorganisms ranging from bacteria to fungus and parasites.\u00a0 Bacterial skin infections are the most common.\u00a0 The most common gram positive organisms are hemolytic Streptococcus<\/em> and Staphylococcus aureus.\u00a0 <\/em>The gram negative rods include Pseudomonas aeruginosa, Escherichia coli, Enterobacter species, Klebsiella species <\/em>and Proteus species <\/em>(Efstrtiou., 1989).1\u00a0The fungal organisms are Candida<\/em> species and moulds. There are many kinds of bacterial skin infections. The most common reported are impetigo, cellulitis, folliculitis, furunculosis, abscesses, scarlet fever, erysipelas, erythrasma, necrotizing fasciitis and some others.<\/p>\n

The aim of this study is to determine the prevalence of bacterial pathogens associated with a skin infection and their drug sensitivity pattern.<\/p>\n

Materials and Methods<\/strong><\/p>\n

Skin swabs were collected from a total of 100 patients with different kinds of skin infection.\u00a0 Samples were collected from patients in Medical College Hospital Trivandrum and KIMS Hospital Trivandrum, Kerala.<\/p>\n

Culture Media Used<\/strong><\/p>\n

Blood agar,Mac Conkey\u2019s agar,\u00a0 chocolate agar and Brain Heart Infusion Broth (BHI) for the bacterial isolation and identification.\u00a0 Muller – Hinton agar for Antimicrobial sensitivity testing.<\/p>\n

Isolation and Identification of Bacterial Isolates<\/strong><\/p>\n

The swabs are streaked directly to the labeled agar plates and incubate 370<\/sup>C for 24 hr. The primary identification of the bacterial isolates was made based on the colony appearance and hemolysis. Identification and characterization of isolates were performed on the basis of colony characteristic, hemolysis, Gram staining and biochemical tests using standard microbiological methods. Biochemical tests applied were standard catalase, Indole production, Citrate Utilization Urease and Triple sugar iron. Biochemical\u00a0 characterization and identification of the bacterial isolates were done (Cowan and Steel, 1985)2<\/sup><\/p>\n

Antibiotics Susceptibility Testing<\/strong><\/p>\n

Antibiotic susceptibilities of bacterial isolates were determined according to the method recommended by the Clinical and Laboratory Standards Institute and Kirby Bauer Disc Diffusion method.3,4<\/sup>\u00a0The inoculumwas prepared for each bacterial isolate by adjusting the turbidity to 0.5 McFarland standard and spread on Muller-Hinton agar plates. Antibiotic discs (Himedia, Mumbai, India) were placed on the agar plates and incubated overnight at 37\u00b0C for 24\u2009h. The zones of inhibition were measured in mm and the isolates were classified as sensitive, intermediate, and resistant according to CLSI tables and guidelines.5<\/sup><\/p>\n

Result <\/strong><\/p>\n

A total of 100 patient\u2019s specimen was examined for different skin infections, 73 were found culture positive and 27 specimens were negative for growth.\u00a0 Out of which Gram Negative isolates were predominant (89%), followed by Gram Positive isolates. The most common isolates were Escherichia coli<\/em> (57.5%), the predominant isolate, second most was Proteus sp<\/em>. (31.5%) and the lowest percentage was recorded by Streptococcus pyogenes<\/em> (10.9%) (Table 1, Fig 1). Gram negative bacteria were the dominant isolates (89%) from skin samples compared to Gram Positive bacteria. Antibiogram results from the present study show that Escherichia coli <\/em>were more resistant to pencillin, cefotaxime while being least resistant to Cefaperazone\/Sulbactum and gentamicin. Proteus sp. was more susceptible to tested antibiotics compared to Escherichia coli<\/em><\/p>\n

Table 1: Bacterial Isolates from skin swab culture from different site collection<\/strong><\/p>\n\n\n\n\n\n\n
SL.No<\/strong><\/td>\nIsolates<\/strong><\/td>\nNumber<\/strong><\/td>\n%<\/strong><\/td>\n<\/tr>\n
1<\/td>\nEscherichia coli<\/em><\/td>\n42<\/td>\n57.5<\/td>\n<\/tr>\n
2<\/td>\nProteus species<\/em><\/td>\n23<\/td>\n31.5<\/td>\n<\/tr>\n
3<\/td>\nStreptococcus pyogenes<\/em><\/td>\n8<\/td>\n10.9<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n\n
\u00a0\"Figure<\/td>\nFigure 1: Percentage distribution of Bacterial isolates from Skin Infection.<\/strong><\/p>\n

 <\/p>\n

Click here to View figure<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

Escherichia Coli<\/em><\/strong><\/p>\n

Escherichia coli<\/em> is a\u00a0Gram-negative,\u00a0facultative,\u00a0rod-shaped\u00a0bacterium\u00a0of the\u00a0genus\u00a0Escherichia<\/em> that is commonly found in the lower\u00a0intestine\u00a0of\u00a0warm-blooded\u00a0organisms. Good growth occurs in ordinary media. Colonies are large, thick, greyish white, moist, smooth opaque or partially translucent discs. On Mac Conkey medium colonies are bright pink due to lactose fermentation. It ferments all the sugars and produes acid and gas. Four main types of clinical syndromes are caused by E. coli<\/em> – Pyogenic infection, Urinary tract infection, diarrhea and gastroenteritis.<\/p>\n

Proteus Species<\/em><\/strong><\/p>\n

Proteus<\/em>\u00a0bacilli are widely distributed in nature as saprophytes, being found in decomposing animal matter, sewage, manure soil, and human and animal feces. They are opportunistic pathogens, commonly responsible for wound infections, bronchopneumonia, cystitis and urolithiasis, septicemia. It\u00a0is Gram negative rod, motile, non-spore forming, non-encapsulated, facultative anaerobic. Cultures of Proteus bacilli have a characteristic putrefactive odour – Fishy odour.\u00a0 Swarming growth occur on solid culture media. Swarming does not occur on Mac Conkey medium, on which the smooth colourless colonies are formed. Proteus<\/em>\u00a0species do not usually ferment\u00a0lactose, but have shown to be capable lactose fermenters depending on the species in a triple sugar iron (TSI) test.\u00a0It is\u00a0oxidase negative but\u00a0catalase and\u00a0nitrate positive. It has the ability to degrade the urea to ammonia, by the production of the enzyme urease.<\/p>\n

Streptococcus pyogenes<\/em><\/strong> or Group A Streptococcus<\/em><\/strong>.<\/strong><\/p>\n

S. pyogenes<\/em>is the cause of many important human diseases, ranging from mild superficial skin infections to life-threatening systemic diseases. S.pyogenes <\/em>is the gram positive cocci arranged in chains or pairs. It is an aerobe and facultative anaerobe. It is exacting in nutritive requirement, growth occurs in media containing fermentable carbohydrates or enriched with blood or serum. On blood agar, the colonies are small, circular, semitransparent, low convex discs with an area of clear hemolysis around them.\u00a0 Streptococcus pyogenes<\/em> ferment lactose, sucrose, mannitol, glucose and produce acid.<\/p>\n

Antibiotic Sensitivity Pattern of Bacterial Isolates<\/strong><\/p>\n

The commonest bacterial pathogen isolated from pyogenic infections followed by E.coli<\/em> (57.5%), Proteus<\/em> species (31.5%) and Streptococcus<\/em> pyogenes<\/em> (10.9%). The percentage of bacterial isolates towards Penicillin, Ampcillin, Cotrimoxazole, Cefaperazone, Cefotaxime, Netilmicin, Levofloxacin, Ofloxacin and Gentamicin were tabulated in Table 2 and Fig 2. The Gram negative pathogen, E.coli<\/em> shown maximum resistance towards Penicillin (95.25%), Ampcillin (85.71%), Cotrimoxazole (76.19%), Cefotaxime (80.95%), Netilmicin (61.9%) and Levofloxacin (57.14 %) where as Proteus<\/em> species have maximum resistance towards Cotrimoxazole (76.25%). In case of gram positive bacteria, Streptococcus pyogenes<\/em> were resistance towards Cotrimoxazole (62.5%), Cefotaxime (75%), Netilmicin (62.5%) and Gentamicin (87.5%).<\/p>\n

Table 2: Antibiotic Sensitivity Pattern of Bacterial Isolates.<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n
Antibiotics<\/strong><\/td>\nUnit<\/strong><\/td>\nEscherichia coli<\/em><\/strong><\/td>\nProteus species<\/em><\/strong><\/td>\nStreptococcus pyogenes<\/em><\/strong><\/td>\n<\/tr>\n
%S<\/strong><\/td>\n%R<\/strong><\/td>\n%S<\/strong><\/td>\n%R<\/strong><\/td>\n%S<\/strong><\/td>\n%R<\/strong><\/td>\n<\/tr>\n
Penicillin<\/td>\n1 Unit<\/td>\n4.76<\/td>\n95.2<\/td>\n56.52<\/td>\n4.3<\/td>\n100<\/td>\n0<\/td>\n<\/tr>\n
Ampcillin<\/td>\n10 mcg<\/td>\n9.52<\/td>\n85.71<\/td>\n69.56<\/td>\n30.4<\/td>\n100<\/td>\n0<\/td>\n<\/tr>\n
Cotrimoxazole<\/td>\n25 mcg<\/td>\n23.80<\/td>\n76.19<\/td>\n21.73<\/td>\n78.26<\/td>\n37.5<\/td>\n62.5<\/td>\n<\/tr>\n
Cefaperazone\/Sulbactum<\/td>\n30 mcg<\/td>\n90.47<\/td>\n9.5<\/td>\n95.6<\/td>\n4.3<\/td>\n–<\/td>\n–<\/td>\n<\/tr>\n
Cefotaxime<\/td>\n30 mcg<\/td>\n19.04<\/td>\n80.95<\/td>\n69.56<\/td>\n30.4<\/td>\n25<\/td>\n75<\/td>\n<\/tr>\n
Netilmicin<\/td>\n30 mcg<\/td>\n38.0<\/td>\n61.9<\/td>\n60.8<\/td>\n39.13<\/td>\n37.5<\/td>\n62.5<\/td>\n<\/tr>\n
Levofloxacin<\/td>\n5 mcg<\/td>\n42.85<\/td>\n57.14<\/td>\n82.60<\/td>\n17.39<\/td>\n50<\/td>\n50<\/td>\n<\/tr>\n
Ofloxacin<\/td>\n5 mcg<\/td>\n52.3<\/td>\n47.6<\/td>\n65.21<\/td>\n34.78<\/td>\n50<\/td>\n50<\/td>\n<\/tr>\n
Gentamicin<\/td>\n10 mcg<\/td>\n71.42<\/td>\n28.5<\/td>\n78.26<\/td>\n21.73<\/td>\n12.5<\/td>\n87.<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n\n
\u00a0\"Figure<\/td>\nFigure 2: Antibiotic Resistace Pattern of Bacterial isolates from Skin Infection.<\/strong><\/p>\n

 <\/p>\n

Click here to View figure<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

Discussion<\/strong><\/p>\n

Pyogenic infection is referred to bacterial infection that leads to severe local inflammation with pus. The invasion and multiplication pathogens in tissue will cause cell damage and leads to loss of integrity of tissue and skin. This will leads to subcutaneous infection to life threatening complications. The present study is aimed to isolate the bacterial pathogens which cause pyogenic infection and study their antibiotic resistance pattern. In this study, both gram positive and gram negative pathogens were isolated from a total of 100 samples. The predominant pathogens were gram negative bacteria. It was agreed with a previous studies Ghosh et al<\/em>6\u00a0<\/sup>and Zubair et al<\/em>,7<\/sup>\u00a0in their studies the aerobics growth of pus culture the dominance pathogens were Gram negative bacterias. E<\/em>.coli<\/em> (57.7%), one of the most commonest and predominant pathogen and Proteus species (31.5%) and followed by gram positive pathogens Streptococcus pyogenes (10.9%). A previous report states that out of 59.3% of gram negative bacteria, the predominant pathogen was E<\/em>.coli<\/em> (21.7%), Klebsiella<\/em> (16.8%), Pseudomonas<\/em> aeruginosa (7.5%), Proteus<\/em> species (7.1%) and Acinetobacter<\/em> species (6.7%) where as gram positive bacteria (40.7%) like Staphylococcus<\/em> aureus<\/em> (37.2%), Coagulase negative Staphylococcus<\/em> aureus<\/em> (1.3%) and Streptococcus<\/em> pyogenes<\/em> (2.2%) (Mantravadi et al<\/em>.,2015).8<\/sup>\u00a0From another report it was found that E.coli<\/em> (Basu et al<\/em>., 2009)9<\/sup>\u00a0and Pseudomonas<\/em> (Raza et al<\/em>., 2013)10\u00a0<\/sup>were the most predominant gram negative pathogen occur in wound infections. The antibiotic resistance pathogens were rapidly increased due to the frequent use of antibiotics. Now a day it became great difficulty to manage or control the pyogenic pathogen and one of the major problems faced by the physicians (Singh et al<\/em>., 2013).11\u00a0<\/sup>In this study the gram positive pathogen, Streptococcus pyogenes<\/em> shows resistance towards Gentamicin (87.5%), Netilmicin (62.5%), Cotrimoxazole (62.5%) and sensitive to Penicillin (100%) and Ampicillin (100%) and intermediate towards Levofloxacin (50%) and oflaxacin (50%). These findings were similar to those of Manthravadi et al,<\/em>8\u00a0<\/sup>and Rao et al<\/em>.12<\/sup>\u00a0 In other hand, most of the gram negative pathogens were highly resistance towards Sulfamethoxazole, Cephalosporin, Fluroquinolones and sensitive to aminoglycosides. These findings were agreed with the previous studies (Mantravadi et al<\/em>., 2015).8<\/sup>\u00a0The combination of antibiotics Cefaperazone + Sulbactum shows maximum sensitivity of about 90-95%. It was correlated with the previous studies done by Javeed et al<\/em>.13<\/sup>\u00a0Rao et al<\/em>,12<\/sup>and Anguzu and Olila.14<\/sup><\/p>\n

Conclusion<\/strong><\/p>\n

This study revealed the presence of skin infection caused by bacteria, those were capable of causing various human illness. The bacterial isolates screened in various skin infections were Escherichia coli<\/em> (57.5%), Proteus <\/em>species (31.5%), Streptococcus <\/em>pyogenes <\/em>(10.9%).\u00a0 The bacterial isolates from the skin infection in this study predominately were Escherichia coli<\/em>, compared with to others. Bacterial isolates exhibited high to moderate levels of resistance against different classes of antibiotics.<\/p>\n

References<\/strong><\/p>\n

    \n
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  2. Cowan S.T, Steel K.J.\u00a0 Manual for the identification of medical bacterial. 2nd<\/sup> Cambridge University Press London.\u00a01993.<\/li>\n
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  12. DVMVSVR R, Basu R, Biswas D.B. Aerobic bacterial pro\u00adfile and antimicrobial susceptibility pattern of pus isolates in a south Indian tertiary care hospital. IOSR J Dent Med Sc<\/em>i. 2014;13(3):59\u201362.
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  14. Anguzu J.R, Olila D. Drug sensitivity patterns of bacterial isolates from septic post-operative wounds in a regional referral hospital in Uganda.2007;7(3):148\u201354.<\/li>\n<\/ol>\n","protected":false},"excerpt":{"rendered":"

    Introduction Human skin acts as an excellent barrier to infection.\u00a0 […]<\/p>\n","protected":false},"author":9,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[55],"tags":[],"class_list":["post-18986","post","type-post","status-publish","format-standard","hentry","category-vol11no1"],"_links":{"self":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/18986","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/users\/9"}],"replies":[{"embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/comments?post=18986"}],"version-history":[{"count":6,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/18986\/revisions"}],"predecessor-version":[{"id":32185,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/18986\/revisions\/32185"}],"wp:attachment":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/media?parent=18986"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/categories?post=18986"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/tags?post=18986"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}