Indu Shukla

Antibacterial efficacy of the seed extracts of Melia azedarach against some hospital isolated human pathogenic bacterial strains.

OBJECTIVE To investigate the antibacterial potential of the polar and non-polar extracts of the seeds of Melia azedarach (M. azedarach) L. (Meliaceae) against eighteen hospital isolated human pathogenic bacterial strains. METHODS Petrol, benzene, ethyl acetate, methanol, and aqueous extracts at five different concentrations (1, 2, 5, 10 and 15 mg/mL) were evaluated. Disk diffusion method was followed to evaluate the antibacterial efficacy. RESULTS All extracts of the seeds demonstrated significant antibacterial activity against tested pathogens. Among all extracts, ethyl acetate extract revealed the highest inhibition comparatively. The present study also favored the traditional uses reported earlier. CONCLUSIONS Results of this study strongly confirm that the seed extracts of M. azedarach could be effective antibiotics, both in controlling gram-positive and gram-negative human pathogenic infections.

Chlamydia trachomatis infection in women with secondary infertility.

OBJECTIVE To assess the role of Chlamydia in secondary infertility in a prospective study. DESIGN Forty women with secondary infertility and 30 healthy term pregnant women of similar age composition were studied for past and present Chlamydia trachomatis infection. SETTING Women attending the outpatient Department of Obstetrics and Gynaecology with complaint of secondary infertility were enrolled as patients in the study. PATIENT(S) Forty women with secondary infertility formed the study group, and 30 healthy women served as the controls. INTERVENTION(S) Chlamydia IgG was detected by ELISA; titers of 1:320 or more were considered positive. Endocervical swabs were collected for culture on cycloheximide-treated McCoy cell lines, and ELISA was used to detect Chlamydia antigen. Hysterosalpingography was performed to assess tubal patency. MAIN OUTCOME MEASURE(S) A difference was expected between the prevalence of C. trachomatis infection in the infertile study subjects and fertile control group. RESULT(S) Immunoglobulin G antibodies were present in 22 (55%) women with secondary infertility, whereas positivity was seen among 2 (5.5%) controls. Tubal occlusion occurred in 16 (63.6%) cases positive for chlamydial antibody. Sensitivity of chlamydial IgG antibody as a diagnostic marker for infertility was 72.7%, and specificity was 44.4%. The majority of Chlamydia IgG antibody-positive cases, 17 (77.2%), were symptomatic. Unfavorable obstetric history was found in 16 (72.7%) cases. Active infection was found in 12 (30%) cases with one (3.3%) case of current infection occurring in the controls. CONCLUSION(S) Prevalence of past chlamydial infection is strongly statistically significant in women with secondary infertility. Current infection was also found statistically significantly in these women. Immunoglobulin G antibody detection is an effective and noninvasive tool for detection of Chlamydia and a more viable option than HSG in a developing country such as India. Screening of women with secondary infertility for C. trachomatis is strongly recommended to allow early therapeutic interventions.

Rising prevalence of antimicrobial resistance in urinary tract infections during pregnancy: Necessity for exploring newer treatment options

Background: Urinary tract infections (UTI) are one of the most common medical complications of pregnancy. The emergence of drug resistance and particularly the Extended-spectrum beta-lactamase production by Escherichia coli and methicillin resistance in Staphylococci, limits the choice of antimicrobials. Materials and Methods: Patients in different stages of pregnancy with or without symptoms of urinary tract infection attending the antenatal clinic of obstetrics and gynaecology were screened for significant bacteriuria, by standard loop method on 5% sheep blood agar and teepol lactose agar. Isolates were identified by using standard biochemical tests and antimicrobial susceptibility testing was done using Kirby Bauer disc diffusion method. Results: A total of 4290 (51.2%) urine samples from pregnant females showed growth on culture. Prevalence of asymptomatic bacteriuria 3210 (74.8%) was higher than symptomatic UTI 1080 (25.2%). Escherichia coli was the most common pathogen accounting for 1800 (41.9%) of the urinary isolates. Among the gram-positive cocci, coagulase negative species of Staphylococci 270 (6.4%) were the most common pathogen. Significantly high resistance was shown by the gram negative bacilli as well as gram positive cocci to the β-lactam group of antimicrobials, flouroquinolones and aminoglycosides. Most alarming was the presence of ESBL in 846 (47%) isolates of Escherichia coli and 344 (36.9%) isolates of Klebsiella pneumoniae, along with the presence of methicillin resistance in 41% of Staphylococcus species and high-level aminoglycoside resistance in 45(30%) isolates of Enterococcus species. Glycopeptides and carbepenems were the only group of drugs to which all the strains of gram positive cocci and gram negative bacilli were uniformly sensitive, respectively. Conclusions: Regular screening should be done for the presence of symptomatic or asymptomatic bacteriuria in pregnancy and specific guidelines should be issued for testing antimicrobial susceptibility with safe drugs in pregnant women so that these can be used for the treatment. For empirical treatment cefoperazone-sulbactum can be recommended, which is a safe drug, covering both gram positive and gram negative organisms and with a good sensitivity.

Antibacterial activity of leaves extracts of Trifolium alexandrinum Linn. against pathogenic bacteria causing tropical diseases

OBJECTIVE To investigate antibacterial potential of Trifolium alexandrinum (T. alexandrinum) Linn. against seven gram positive and eleven gram negative hospital isolated human pathogenic bacterial strains responsible for many tropical diseases. METHODS Non-polar and polar extracts of the leaves of T. alexandrinum i.e., hexane, dichloromethane (DCM), ethyl acetate (EtOAc), methanol (MeOH) and aqueous (AQ) extracts at five different concentrations (1, 2, 5, 10 and 15 mg/mL) were prepared to evaluate their antibacterial value. NCCL standards were strictly followed to perform antimicrobial disc susceptibility test using disc diffusion method. RESULTS Polar extracts demonstrated significant antibacterial activity against tested pathogens. EtOAc and MeOH extracts showed maximum antibacterial activity with higher inhibition zone and were found effective against seventeen of the tested pathogens. While AQ plant extract inhibited the growth of sixteen of the test strains. EtOAc and MeOH plant extracts inhibited the growth of all seven gram positive and ten of the gram negative bacterial strains. CONCLUSIONS The present study strongly confirms the effectiveness of crude leaves extracts against tested human pathogenic bacterial strains causing several tropical diseases. Since Egyptian clover is used as a fodder plant, it could be helpful in controlling various infectious diseases associated with cattle as well.

blaCTX-M, blaTEM, and blaSHV in Enterobacteriaceae from North-Indian tertiary hospital: high occurrence of combination genes

OBJECTIVE To delineate the frequency of occurrence of bla(CTX-M), bla(TEM), and bla(SHV) in Enterobacteriaceae from North-Indian tertiary hospital. METHODS A random collection of a subset of 45 Escherichia coli (E. coli) and 28 Klebsiella pneumoniae (K. pneumoniae) that was resistant to a third generation cephalosporin and obtained during 2007-2008 was selected for detailed screening for bla(CTX-M), bla(TEM), and bla(SHV) by monoplex PCRs. The isolates demonstrating the presence of bla(CTX-M) alleles were characterized for the specific CTX-M-genogroup by using a multiplex PCR. RESULTS Resistance to cefoperazone, ceftazidime, ceftriaxone, cefotaxime, cefoxitin and piperacillin was 100% each in K. pneumoniae isolates, whereas these resistance-rates for E. coli isolates were 93.1%, 83.8%, 91.9%, 93.6%, 97.3% and 97.1%, respectively. Concomitant resistance to aminoglycosides, quinolones and aztreonam was also noticed. Presence of any of the bla genes (bla(CTX-M), bla(TEM), and bla(SHV)) was noticed in a total of 28 (38.4%) isolates of the 73 isolates studied. Many isolates demonstrated occurrence of these genes in various combinations. bla(CTX-M), bla(TEM), and bla(SHV) were noticed in 28.8%, 10.9% and 13.7% isolates, respectively. Multiplex PCR in bla(CTX-M) harboring isolates demonstrated the presence of CTX-M-Genogroup-1 and sequencing for the specific CTX-M-type revealed presence of CTX-M-15 type. RAPD typing showed wide diversity in isolates. CONCLUSIONS This is amongst the premier report describing the simultaneous occurrence of bla(TEM), bla(SHV), and bla(ampC) in Indian Enterobacteriaceae and that wider dissemination of these genes, as demonstrated by diversity of isolates, raises concern and emphasizes a need for extensive search for the presence of these gene pools in Indian subcontinent.

Increasing antimicrobial resistance among uropathogens: Is fosfomycin the answer?

Introduction: Urinary tract infection (UTI) is one of the most common infectious diseases in clinical practice. The choice of antibiotics for the treatment of UTI is limited by the rising rates of antibiotic resistance. There is an urgent need to discover new effective treatment solutions. Fosfomycin may be an interesting alternative to the currently used treatments of UTIs. Materials and Methods: The study was conducted over 6 months period (January to June 2013) in Department of Microbiology, JNMCH, AMU, Aligarh. A total of 1840 urine samples were submitted. Culture and sensitivity was done as per standard microbiological procedures. Methicillin-resistant Staphylococcus aureus (MRSA), high-level aminoglycoside resistance (HLAR), extended spectrum beta-lactamases (ESBL), AmpC and metallo-beta-lactamases (MBL) production was detected. Results: Culture was positive in 504 (27.4%) cases. Gram-negative etiology was identified in 390 (73%) cases. ESBL production was detected in 154 (37.1%) while 82 (21.6%) were Amp C. No, MBL was detected. Among Gram-positive bacteria, 68 (51.5%) were MRSA, while 4 (13.3%) were vancomycin resistant enterococci (VRE). HLAR was seen in 53.3% of enterococci. Fosfomycin was effective in 100% of MRSA, VRE, ESBL, HLAR, and overall, susceptibility to fosfomycin in AmpC producers was extremely high (99%). Norfloxacin and cotrimoxazole were not proved effective as only three isolates were sensitive to norfloxacin, while all Gram-negative isolates were resistant to cotrimoxazole. Pseudomonas species showed 65% and 75% susceptibility to colistin and polymixin B, respectively. Conclusion: Fosfomycin has emerged as a promising option, especially in cases involving multi-drug-resistant pathogens in which previous antibiotics have failed to cure the infection.

Emergence of coryneform bacteria as pathogens in nosocomial surgical site infections in a tertiary care hospital of North India

PURPOSE A prospective study was conducted to assess the role of coryneform bacteria in surgical site infections among obstetric and gynecological patients undergoing surgery. MATERIALS AND METHODS The surgery was graded according to the degree of contamination, and surgical site infections (SSIs) were classified as superficial or deep. Pus samples were collected from SSIs according to rigorous aseptic precautions, and the quality of specimens was assessed by Q-score. A detailed clinical and treatment history was elicited from all patients. The samples were processed using standard protocols. Coryneform bacteria were considered significant pathogens only if they fulfilled rigorous clinical and microbiological criteria. Antibiotic susceptibility testing was performed using the Kirby-Bauer method according to the CLSI guidelines. RESULTS In total, 127 patients developed SSIs among 882 postoperative patients. Of these, 89 (70.1%) were culture positive: 40 (44.9%) were Gram-positive cocci, 27 (30.3%) were coryneform, and 22 (24.7%) were Gram-negative bacilli. All coryneform-infected patients had fever and post-operative wound dehiscence leading to a prolonged hospital stay. The most commonly isolated organism was Staphylococcus aureus (33.7%), followed by Corynebacterium amycolatum (11.2%), Escherichia coli (8.9%), Citrobacter spp. (7.8%) and coagulase-negative Staphylococci (6.7%). In our study, 45.5% were ESBL producers, 18.2% were Amp C producers, and 40% were MRSA. All the coryneform bacteria were multidrug resistant, and 51.8% of isolates were sensitive to only gatifloxacin and vancomycin. Symptomatic improvement was observed in all coryneform-infected patients after the administration of appropriate therapy. CONCLUSION Coryneform bacteria appear to be emerging as significant nosocomial surgical site pathogens. The high level of multidrug resistance observed in coryneform bacteria in our study is cause for alarm.

Prevalence of leptospira in acute hepatitis syndrome and assessment of IL-8 and TNF-alpha level in leptospiral hepatitis.

Abstract To study the prevalence of leptospira in acute hepatitis syndrome and to assess interleukin (IL)-8 and tumour necrosis factor (TNF)-alpha levels in the pathogenesis of hepatitis due to leptospiral infection. Two hundred and forty-seven consecutive cases with symptoms of acute hepatitis and 30 healthy controls were enrolled in the study and detailed clinical history was elicited from them. Enzyme-linked immunosorbent assays (ELISAs) for HAV, HBV, HCV and HEV were performed to rule out common viral aetiology of hepatitis. IgM antibodies to leptospira were detected by ELISA. IL-8 and TNF-alpha levels were estimated in leptospira-positive cases and healthy controls by ELISA. Out of 247 cases of acute hepatitis, 46 (18·62%) were observed to be positive for IgM antibodies for leptospira. The mean age of these patients was 31·99±0·28 years (25 males and 21 females; M/F ratio: 1·19∶1). The mean ALT, AST and ASP were raised in the majority of patients. IL-8 was found to be elevated (130·81 pg/ml) in a large majority of cases 41/46, 89·1% (P<0·001). Patients with more severe symptoms were associated with higher levels of IL-8. One mortality was observed due to leptospira. Unpredictably, TNF-alpha level was largely suppressed (45·63 pg/ml) in most of the leptospira-positive patients in comparison with healthy controls. Leptospira-induced hepatitis should be actively looked for in patients negative for A–E viral hepatitis. IL-8 appears to play an important role in the pathogenesis of leptospiral hepatitis. High TNF-alpha should alert clinicians for aggressive in hospital management of patients.

Screening children with severe short stature for celiac disease using tissue transglutaminase

ObjectiveThis prospective study was planned with the aim to determine the prevalence of anti-tissue transglutaminase in children and adolescents with severe short stature (<-3 SD) attending the Pediatric outpatient, inpatient and/or the Pediatric Gastroenterology clinic.MethodsAll children in age group of 1–18 years having height less than -3 SD for their age and sex, were included. For each child age and sex matched healthy control (height more than -2 SD) was taken. The included subjects (study & control group) were subjected to anti tissue transglutaminase (tTG) (IgA) antibody assay estimation.ResultsOf the 112 cases, 23 were tTG positive, giving a prevalence of 20.5% for seropositivity among cases of short stature while all the controls were seronegative for tTG. All the 23 had tTG values above 40 U/ml and 11 had values above 100 U/ml. On univariate analysis we found that the presence of chronic diarrhea (OR = 2.55, 95%CI − 1.08–5.98), bulky stools (OR = 3.03, 95%CI − 1.52–6.05), hemoglobin < 7 gm/dl (OR = 3.12, 95%CI − 1.55–6.29) and more severe short stature (<-4 SD) (OR = 0.41, 95%CI − 0.17–0.95) had significant association with the tTG positivity. On logistic regression analysis in all cases, hemoglobin < 7gm/dl (OR = 0.090, 95%CI = 0.024–0.342) and bulky stools (OR=0.212, 95%CI = 0.069–0.649) were significantly associated with tTG positivity.ConclusionMore than one fifth of all severe short stature are seropositive for tTG and the chances of seropositivity increases if severe anemia and bulky stool are also associated.

Typhidot M and Diazo test vis-à-vis blood culture and Widal test in the early diagnosis of typhoid fever in children in a resource poor setting

OBJECTIVE Typhoid fever is a major public health problem. A test which is simple, reliable and can be carried out in small laboratories is the need of the hour. We prospectively evaluated typhidot M and Diazo tests vis-à-vis blood culture and Widal test in children. METHODS Patients aged 6 months to 12 years, having fever of more than four days duration with clinical suspicion of typhoid fever were enrolled. Patients in whom other diagnosis was made served as control. The tests under scrutiny were validated against blood culture and then all the four tests were evaluated among patients who presented in the first week of illness. RESULTS Blood culture was positive in only 27.3% of the cases. Among these culture positive cases, typhidot M test had the highest sensitivity, specificity, PPV and NPV of 90% (95% CI = 74.4-96.5), 100% (95% CI = 90.1-100), 100% (95% CI = 87.5-100), and 92.1% (95% CI = 79.2-97.3) respectively. Diazo test ranked next with sensitivity, specificity, PPV and NPV of 86.7% (95% CI = 70.3-94.7), 85.7% (95% CI = 70.6-93.7), 83.9% (95% CI = 67.4-92.9), 88.2% (95% CI = 73.4-95.3) respectively. Among clinically suspected typhoid cases, the overall sensitivity, of blood culture, Widal, typhidot M, Diazo was 27.3% (95% CI = 19.8- 36.3), 64.6% (95% CI = 55.3-72.9), 89.1% (95% CI = 81.9-93.7), 80.9% (95% CI = 72.6-87.2) respectively. In the first week of illness, typhidot M showed the best sensitivity [86.2% (95% CI = 69.4-94.5)] followed by Diazo [79% (95% CI = 61.6-90.2)], Widal [41.4% (95% CI = 25.5-59.3)] and blood culture [31% (95% CI = 17.3-49.2)]. CONCLUSION Both Typhidot M and Diazo are good screening tests for the diagnosis of typhoid fever. Typhidot M is superior to Diazo but the latter is more suitable to resource poor settings being economic and easy to perform.