International Research Journal of Biological Sciences ___________________________________ ISSN 2278-3202Vol. 4(5), 50-54, May (2015) Int. Res. J. Biological Sci.  International Science Congress Association        
50
 
A Spectrum of Bacterial Pathogens and its Antibiotic Susceptibility Pattern Isolated from Neonatal Sepsis in an NICU in a Government Pediatric Hospital Rao Pooja, K N Sowmya, Baliga Shrikala, M Radhakrishna and Bele Keerthiraj Kasturba Medical College, Mangalore, Manipal University, Karnataka, INDIA Kasturba Medical College, Light house, Hill Road, Mangalore, Manipal University, 575001, Karnataka, INDIA Available online at: www.isca.in, www.isca.me Received 12th March 2015, revised 14th April 2015, accepted 9th May 2015 Abstract  Neonatal sepsis being one of the major causes for mortality and morbidity in the NICU. This study will provide a data on the bacterial pathogens causing neonatal sepsis along with their antibiogram. To study the spectrum of significant bacterial isolates causing septicemia in neonates and to determine their antimicrobial sensitivity pattern. A total of 784 blood samples were screened for sepsis in newborns less than 28 days old in this retrospective study. The positive blood cultures were detected by BACTEC blood culture systems and antibiotic susceptibility testing was done by using Kirby-Bauer disk diffusion method. Out of 784 samples, 170 were positive blood cultures. Gram negative and gram positive organisms comprised of 79.94% and 18.17% respectively, out of which Burkholderiacepecia complex (30%), Klebsiellaspp.(15.5%) were the most common pathogens  followed by S.aureus (14.7%), Among the gram negative organism’s susceptibility for Imipenem, Meropenem and piperacillin/tazobactum was high. All Gram positive bacterial isolates showed 100% sensitivity to Netillin, vancomycin, teicoplanin, rifampacin. Gram negative species has been predominantly seen in neonatal sepsisand Carbapenems is considered one of the best options for empirical antibiotic therapy in our NICU.   Keywords: Neonatal sepsis, NICU.Introduction In neonates, sepsis incidence is increasing in the recent years. Neonatal septicemia refers to generalized infection with positive blood culture in the early28 days of neonates. Advances in early diagnosis and treatment have led to better prognosis of newborns in ICUs. It encompasses various diseases of new born including septicemia, meningitis, arthritis, pneumonia, osteomyelitis and urinary tract infection. 
According to National Neonatal Prenatal Database 2002-2003,  
neonatal septicemia in India comprised of 30/1000 live birth. 
 In rural India, clinical sepsis rates is ranging from 49 to 
170/1000 live births. Neonatal septicemia can be classified into 2 subtypes: Early onset neonatal sepsis (EONS) depending on the onset of symptoms before 72 hours and after 72 hours up to 28 days of life it is considered as late-onset neonatal sepsis (LONS). Risk factors for early onset sepsis includes prolonged ruptured membranes (� 18 hours), fetal distress, preterm delivery, history of Group BS treptococcus (GBS) infection in previous infant, GBS bacteriuria in this pregnancy. Late onset sepsis occurs due to prolonged hospitalization. e.g. preterm infant in a NICU, presence of foreign bodies e.g. intravenous catheters, end tracheal tubes, etc., cross infection by staff and parents, malformations such as urinary tract anomalies (e.g. vesico-ureteral reflux) or neural tube defects1,2.  This retrospective study was undertaken to evaluate major bacterial isolates causing neonatal septicemia and their antibiogram pattern. Aims and Objectives: To study the spectrum of the significant bacterial isolates from positive blood culture causing septicemia in neonates. To determine the antimicrobial sensitivity pattern of the bacterial isolates obtained from positive blood culture causing septicemia in neonates. Material and Methods Study location: Kasturba Medical College Hospital. Study design: Retrospective study. Study Period: 10 months. February 2014 to November 2014. Sample size: All the Blood specimens of neonates of age group 1- 28 days were collected from February 2014 to November 2014. Blood for bacterial culture was aseptically collected and added to 2ml of blood BD Bactectm Pedsplustm/F culture bottle containing 40ml of processed water, soyabean casein digest broth 2.75% W/V and incubated at 37°C for 48 hours to 7 days in BD BACTEC system. Positive blood culture bottles were sub cultured onto 5% sheep blood agar, chocolate agar and Mac 
Research Journal of Biological Sciences ___________________________________________________________ ISSN 2278-3202  Vol. 4(5), 50-54, May (2015) Int. Res. J. Biological Sci. International Science Congress Association 
51
Conkey's agar and incubated at 37°C for 18 hours. Negative culture bottles were incubated for 7 days. The organism isolated was identified by standard biochemical test and the sensitivity to various antibiotics was assessed using the Modified Kirby - Bauer disk diffusion method on Mueller Hinton agar.Susceptibility testing for Streptococcus pneumoniaewas performed on Mueller Hinton agar supplemented with 5% sheep blood. Skin colonizers like coagulase negative staphylococcus & micrococcus were excluded from this study.  This study will provide adequate rational data on bacterial pathogens causing neonatal sepsis along with their antibiogram and hence in the prompt empirical management of neonatal septicemia.Results and Discussion During a period of 10 months, 784 blood samples were collected out of which170 positive bacterial blood culture samples were studied and Candida spp. was isolated from 21 samples. Among the 170 samples, EONS constituted about 44.4% while LONS formed about 53.53%, figure-1. The gram negative rods were the main cause of septicemia. Neonatal sepsis was predominant in males (64.7%) than females. The gram negative bacteria included Burkholderia cepacia complex, Klebsiella spp., Enterobacter spp., Acinetobacter spp., Pseudomonas spp. and among the gram positive bacteria Staphylococcus aureus, the major cause of gram positive bacterial sepsis. Gram negative and gram positive organisms comprised of 79.94% and 18.17% respectively, out of which Burkholderia cepacia complex (30%), Klebsiella spp (15.5%) were the most common pathogens followed by S.aureus (14.7%), Enterobacter spp. Acinetobacter spp., Pseudomonas spp, and less frequently isolated were other non-fermentative Gram Negative Bacilli and Enterococcus spp, figure-1. The antimicrobial susceptibility testing revealed that most of the Enterobacteriaceae family members were resistant to ampicillin (90.5%). Enterobacteriaceae family members showed high susceptibility to polymixin B and colistin. Among the gram negative organisms susceptibility for Imipenem, Meropenem, fluroquinolones and piperacillin/tazobactum were high, figure-2. All Gram positive bacterial isolates showed 100% sensitivity 
to Netillin, vancomycin, teicoplanin, rifampicin, figure-3. The 
prevalence of MRSA was found to be 52%. The overall 
prevalence of ESBL producers among E.coli and Klebsiella spp 
was found to be 27.7% and carbapenemase produces were about 
26%. Discussion: Neonatal sepsis is a life threatening emerging infection in the developing countries and it is estimated about 5 
million neonatal death occur every year world wide. The 
invasive procedures in the postnatal period and inadequate hand 
washing before and after handling babies also contributes to the 
neonatal sepsis in intensive care units. 
In the present study EONS comprised of 44.4% which was 
almost similar to the study done by Shaw et al (49.8%), while it was 95% in Shah et al, 55.3% in Vinod kumar et al. In our study Gram negative organisms were the major cause of neonatal sepsis about 79.9% and gram positive organisms about 18.17%. In a study conducted in tertiary care hospital Mangalore, gram positive bacteria (64.4%) were the main cause of sepsis compared to gram negative bacteria ( 34.56%). About 30% of the isolates were Burkholderia cepacia complex, the major isolate in our study. In comparison to other studies conducted in Pakistan and Lucknow, India, 
10152025
EONS(%)
LONS (%)
Figure-1 Distribution of different species of organisms depending on the onset of symptoms 
Research Journal of Biological Sciences ___________________________________________________________ ISSN 2278-3202  Vol. 4(5), 50-54, May (2015) Int. Res. J. Biological Sci. International Science Congress Association 
52
 
 
Figure-2 
Antibiotic susceptibility pattern of Gram negative Bacteria 
Figure-3 Antibiotic susceptibility pattern in Gram positive cocci E.coli and Klebsiella spp. was found to be the major cause of neonatal sepsis respectively10,11. Non fermentative gram negative bacilli like Pseudomonas spp, Acinetobacter spp and 
Stenotrophomonas maltophiliaare some of the opportunistic pathogens commonly isolated in NICUs with high multidrug resistance. B. cepacia can now be included in the list because of its high rate of transmissibility between in patients, health care staff and multiple drug resistance. In hospitals, B.cepacia has been found to contaminate various antiseptics, disinfectants, and nebulizer solution and dextrose solution12. In our NICU most of the neonates with B.cepacia sepsis were under mechanical ventilator support and with risk factors like prematurity and low 
Research Journal of Biological Sciences ___________________________________________________________ ISSN 2278-3202  Vol. 4(5), 50-54, May (2015) Int. Res. J. Biological Sci. International Science Congress Association 
53
birth weight. Group B Streptococcus which is one of the commonest pathogen to be isolated in western countries was not reported from our study13. A possible explanation might be due to the fact that GBS has a tendency to colonize the cervix and sexual partners of infected women can harbor GBS in their urethra and possibly act as reservoirs in certain societies. Therefore, differences in the ethnicity and socioeconomic status may contribute to the varying incidence of GBS infection among neonates in different populations14. The other commonly found organisms in our setup were Klebsiella spp. (15.5%), Enterobacter spp. (10.5%), Acinetobacter spp. (10.5%). S.aureus (14.5%) and amongless frequently isolated organisms were Enterococcus spp and other Non-fermentative gram negative bacilli. Male predominance (64.7%) as seen in our study were also found in other studies3,14,15. Antibiotic resistance is a global problem. The antibiogram pattern differs from country to country depending upon the variability of epidemiology of neonatal sepsis16. Our study shows high rate of resistance to first line of antibiotics. In our study gram negative bacteria showed high resistance to ampicillin (90.5%) and moderate sensitivity to gentamycin (64.4%) and third generation cephalosporins, ceftazidime (56.3%), cefotaxime (57.63%), figure-2. Our results showed contrast to the study conducted at Hyderabad, India having high resistance to Amikacin (13.6%) and moderate resistance to gentamycin (45%) and third generation cephalosporins like cefotaxime 73% 17. Increase in resistance was also seen in aminoglycosides and third generation cephalosporins which is being used in our setup as empirical therapy. Among the gram negative organisms susceptibility for Imipenem (86.13%), Meropenem (83.22%), piperacillin/tazobactum (76%) and fluroquinolones (74.5%) was high. But quinolones has shown increased side effects. Gram positive bacteria showed moderate sensitivity to Penicillin (57.14%) and high sensitivity to Amikacin (100%). All Gram positive bacterial isolates showed 100% sensitivity to netillin, vancomycin, teicoplanin, rifampicin which was similar to another study18. Hence, the changing antibiotic susceptibilities need continuous monitoring and re-evaluation for putting forth the guidelines for empirical treatment in NICUs. Conclusion Gram negative species are predominantly seen in neonates sepsis and Carbapenems and Beta lactam and Beta lactam inhibitor combinations is considered one of the best options for empirical antibiotic therapy in our NICU.  Early identification of organisms causing neonatal sepsis and appropriate use of antibiotics to minimize the morbidity, mortality andemerging resistance in bacteria is essential to control neonatal sepsis in neonatal intensive care units. References 1.Huda H.A., Goma Edet E. and Udoc Usha Rajaram,  Neonatal Septicemia in Al-Jahra Hospital, Kuwait: Etiologic Agents and Antibiotic Sensitivity Patterns Med Principles Pract, 10, 145–150 (2001) 2.Poonam S, Parminder K and Aruna A, Staphylococcus aureus- the predominant pathogen in the neonatal ICU of a tertiary care hospital in Amritsar, India, JCDR, 7(1),66-69 (2003) 3.Vergnano S, Sharland M, Kazembe P, Mwansambo C and Heath PT, Neonatal sepsis: An international perspective, Arch Dis Child Fetal Neonatal Ed90, F220–F224 (2005) 
4.Thaver D and Zaidi AK, Burden of neonatal infections in 
developing countries: A review of evidence from 
community-based studies, Pediatr Infect Dis J, 28, S3-9 
(2009) 
5.Paolucci M, Landini MP and Sambri V, How can the 
microbiologist help in diagnosing neonatal sepsis?,Int J 
Pediatr, 120139 (2012) 6.Nalini A., Neelam K. and Varsha G, Antimicrobial susceptibility of isolates from neonatal septicemia, Jpm. J. Infect. Dis., 57, 273-275 (2004) 7.Rahman S, Hameed A, Roghani MT and Ullah Z, Multidrug resistant neonatal sepsis in Peshawar, Pakistan, Arch Dis Child Fetal Neonatal Ed, 87, F52-F54 (2002) 8.Roy I, Jain A, Kumar M and Agarwal SK, Bacteriology of neonatal septicaemia in a tertiary care hospital of northern, India. Indian J Med Microbiol, 20, 156-9 (2002)9.Afroza S., Neonatal sepsis, A global problem an overview, Mymensingh med J.Jan, 15(1), 108-14 (2006) 10.Mustafa M and Ahmed SL, Bacteriological profile and antibiotic susceptibility patterns in neonatal septicemia in view of emerging drug resistance, J Med Allied Sci; 4(1), (2014) 11.Karthikeyan G and Premkumar K., Neonatal sepsis: Staphylococcus aureus as the predominant pathogen. Indian J Pediatr, 68, 715–17. (2001) 12.CDC, Trends in Perinatal Group B Streptococcal Disease - United States, MMWR. 2009, 58, 109–12 (2000-2006) 13.Nidal S. Younis, Neonatal sepsis in Jordan: Bacterial isolates and susceptibility pattern, Rawal Med J, 36(3), 169–72 (2011) 14.Jones AM, Dodd ME and Webb AK, Burkholderiacepacia: current clinical issues, environmental controversies and ethical dilemmas, EurRespir J., 17, 295-301 (2001)
Research Journal of Biological Sciences ___________________________________________________________ ISSN 2278-3202  Vol. 4(5), 50-54, May (2015) Int. Res. J. Biological Sci. International Science Congress Association 
54
15.Ahmed NU, Chowdhury MA, Hoque M and Darmstadt GL, Clinical and bacteriological profile of neonatal septicemia in a tertiary level pediatric hospital in Bangladesh, Indian Pediatr, 39, 1034-1039 (2002)16.Shaw CK, Shaw P and Thapaliala A., Neonatal sepsis bacterial isolates and antibiotic susceptibility patterns at a NICU in a tertiary care hospital in western Nepal: A retrospective analysis, Kathmandu Univ Med J, 5, 153-160 (2007)17.Vinodkumar CS, Neelagund YF, Kalsurmath S, Banapurmath S, Kalappannavar NK and Basavarajappa KG, Perinatal risk factors and microbial profile of neonatal septicemia: A multicentred study, J Obstet Gynecol India; 58, 32-4 (2008) 18.Arpita Jigar Shah, Summaiya A. Mulla and Sangita B. Revdiwala, Neonatal Sepsis: High Antibiotic Resistance of the Bacterial Pathogens in a Neonatal Intensive Care Unit of a Tertiary Care Hospital, Journal of Clinical Neonatology, 1(2), (2012)
<end>