Detection of biofilm among uropathogenic Escherichia coli and its correlation with antibiotic resistance pattern.

BACKGROUND: Escherichia coli accounts for 70%-95% of urinary tract infections (UTIs). UTI is a serious health problem with respect to antibiotic resistance and biofilms formation being the prime cause for the antibiotic resistance. Biofilm can restrict the diffusion of substances and binding of antimicrobials. In this context, the present study is aimed to perform in vitro detection of biofilm formation among E. coli strains isolated from urine and to correlate their susceptibility pattern with biofilm formation. MATERIALS AND METHODS: A total of 100 E. coli strains isolated from patients suffering from UTI were included in the study. The identification of E. coli was performed by colony morphology, Gram staining, and standard biochemical tests. The detection of biofilm was carried out by Congo Red Agar (CRA) method, tube method (TM), and tissue culture plate (TCP) method. Antimicrobial sensitivity testing was performed by Kirby-Bauer disc diffusion method on Muller-Hinton agar plate. RESULTS: Of the 100 E. coli strains, 49 (49%) and 51 (51%) were from catheterized and noncatheterized patients, respectively. Biofilm production was positive by CRA, TM, and TCP method were 49 (49%), 55 (55%), and 69 (69%), respectively. Biofilm producers showed maximum resistance to co-trimoxazole (73.9%), gentamicin (94.2%), and imipenem (11.6%) when compared to nonbiofilm producers. Significant association was seen between resistance to antibiotic and biofilm formation with a P = 0.01 (<0.05). CONCLUSION: A greater understanding of biofilm detection in E. coli will help in the development of newer and more effective treatment. The detection of biofilm formation and antibiotic susceptibility pattern helps in choosing the correct antibiotic therapy.

Whole-Genome Sequence of Brucella melitensis CIIMS-BH-2, a Biovar 2 Strain Isolated from Human Blood.

Brucella species are the etiological agent of brucellosis in humans and animals. Here, we report the whole-genome sequence of Brucella melitensis strain CIIMS-BH-2, belonging to biovar 2. The draft assembly of CIIMS-BH-2 is 3.31 Mb in size, with 57.2% G+C content.

Protean clinical manifestations and diagnostic challenges of human brucellosis in adults: 16 years' experience in an endemic area.

A prospective study was carried out to elucidate the clinical, epidemiological and laboratory features of human brucellosis. A total of 26 948 blood samples (from adults aged 15 years and above) were screened for serological evidence of brucellosis over a period of 16 years. The slide agglutination/Rose Bengal plate agglutination test gave positive results in 517 patients, of which 509 had detectable titres by the standard tube agglutination test (SAT). The diagnosis of brucellosis was documented in 495 (1.8 %) patients based on diagnostic titres (> or = 1 : 160, 490 cases) and rising titres from insignificant titres (four cases) by serology and for one case by blood-culture isolation alone. Blood cultures were carried out in 345 cases, of which 191 cases (55.3 %) yielded Brucella melitensis. In 77/79 cases undertaken for follow up, there was a steady fall in 2-mercaptoethanol (2ME) agglutination titres along with clinical improvement (P < 0.01). SAT titres remained detectable in most cases for a longer period in spite of an effective antimicrobial therapy and clinical recovery. A substantial number of patients (84.2 %) presented with fever, this being the only complaint in 51.1 % of the cases. Complications were present in 8.8 % of the patients (arthritis excluded): this included the unusual complications of hydrocele (two cases), Stevens-Johnson syndrome (one case) and urinary tract infection (one case). Brucella agglutinins were demonstrated in synovial, testicular, hydrocele and cerebrospinal fluids. There was no clinical suspicion of brucellosis in 439 cases (88.7 %) and the diagnosis was made only by routine serology. A two-drug regimen for 42-84 days with a follow-up 2ME test resulted in lower levels of relapse. These results suggest that, in endemic areas of the world, it should be mandatory to screen routinely for brucellosis due to protean clinical manifestations.

Brucellosis in Occupationally Exposed Groups.

INTRODUCTION: In India, high incidence of human brucellosis may be expected, as the conditions conducive for human brucellosis exist. Limited studies have been undertaken on human brucellosis especially in occupationally-exposed groups. AIM: To estimate prevalence of anti-brucellar antibodies, evaluate the clinical manifestations, risk factors and Knowledge, Attitude and Practices (KAP) levels about brucellosis among occupationally exposed groups. MATERIALS AND METHODS: Blood samples were collected from 2337 occupationally exposed individuals. The serum samples were screened for the presence of anti-brucellar antibodies by Rose Bengal Plate Test (RBPT), Serum Agglutination Test (SAT) and 2-Mercaptoethanol test (2-ME). Clinical manifestations, risk factors and KAP levels were evaluated by personal interview using a structured questionnaire. RESULTS: Seroprevalence of brucellosis by RBPT, SAT and 2-ME test was 9.46%, 4.45% and 3.64 % respectively. Clinical symptoms resembling brucellosis were seen in 91 subjects. The major risk factors were animal exposure in veterinarians and abattoirs, both animal exposure and raw milk ingestion in farmers and shepherds, exposure to raw milk and its ingestion in dairy workers and exposure to Brucella culture in laboratory workers. Except laboratory workers, few veterinarians and dairy workers none had heard about brucellosis. KAP levels regarding brucellosis were too poor in all the groups except laboratory workers. CONCLUSION: Brucellosis most of the times was missed or misdiagnosed. Regular screenings for brucellosis and awareness programmes to increase KAP levels are necessary to control brucellosis in occupationally exposed groups.

Klebsiella Pneumoniae in Septicemic Neonates with Special Reference to Extended Spectrum ß-lactamase, AmpC, Metallo ß-lactamase Production and Multiple Drug Resistance in Tertiary Care Hospital.

BACKGROUND: ß-lactamases viz., extended spectrum ß-lactamase (ESBL), AmpC, and metallo ß-lactamase (MBL) production in Klebsiella pneumoniae has led to a serious concern about septicemic neonates in Neonatal Intensive Care Units due to high resistance against commonly used antimicrobials. PURPOSE: To study the prevalence of ESBL, AmpC, and MBL production in K. pneumoniae isolates in neonatal septicemia, to check antimicrobial susceptibility to various drugs including tigecycline; and to assess burden of multiple drug resistance (MDR). MATERIALS AND METHODS: Total 24 clinical isolates of K. pneumoniae isolated from 318 blood samples of suspected cases of neonatal septicemia were studied. Isolates were screened for ESBL, AmpC, and MBL production by Clinical and Laboratory Standards Institute (CLSI) disk method, AmpC cefoxitin screen, and imipenem, meropenem, ceftazidime disk screen respectively; and confirmation was done by CLSI phenotypic disk confirmatory test, AmpC sterile disk method, and imipenem ethylenediamine tetracetic acid double disk synergy test respectively. Antimicrobial susceptibility was determined by Kirby-Bauer's disk diffusion method. Efficacy of tigecycline was evaluated using United States Food and Drug Administration guidelines. RESULTS: Of the 24 K. pneumoniae isolates, co-production of AmpC + MBL was found in more number of isolates (67%) (P < 0.0001) compared to single enzyme production (ESBL and MBL 8% both, AmpC 12.5%). Rate of resistance for penicillins and cephalosporins was highest. Susceptibility was more for imipenem, co-trimoxazole, and meropenem. Nonsusceptibility to tigecycline was low (21%). A total of 23 (96%) isolates were MDR. CONCLUSIONS: Routine detection of ESBL, AmpC, and MBL is required in laboratories. Carbapenems should be kept as a last resort drugs. Trend of tigecycline susceptibility has been noted in the study. Continued monitoring of susceptibility pattern is necessary to detect true burden of resistance for proper management.

Serological, Clinical, and Epidemiological Profile of Human Brucellosis in Rural India.

BACKGROUND: Brucellosis is an important but neglected zoonotic disease in India. Due to frequent animal contact, high prevalence of this disease, though expected in rural population, has not been much studied. AIM: The study was carried out to determine serological, clinical, and epidemiological profile including associated risk factors for human brucellosis in rural India. MATERIALS AND METHODS: In this cross-sectional study, serum samples from 1,733 individuals residing in rural areas were screened for the presence of anti-brucellar antibodies by Rose Bengal Plate test (RBPT), Serum Agglutination test (SAT), and 2-Mercaptoethanol test (2-ME). Clinical symptoms, epidemiological data including risk factors and knowledge about brucellosis were evaluated by personal interview using a structured questionnaire. RESULTS: Of the 1,733 individuals, 998 had direct contact with animals, whereas 735 had no direct contact. The overall positivity rates by RBPT, SAT, and 2-ME test were 10.50% (182), 7.32% (127), and 5.88% (102), respectively. Clinical symptoms resembling brucellosis were seen in 151 (8.71%) subjects. Animal contact especially during milking, parturition/abortion was the major risk factor, followed by raw milk ingestion. None of the participant knew about brucellosis. CONCLUSION: Regular surveillance of the disease with awareness programs emphasizing prevention and control are needed.

Evaluation of conventional castaneda and lysis centrifugation blood culture techniques for diagnosis of human brucellosis.

We investigated the role of the lysis centrifugation blood culture technique over the conventional Castaneda technique for the diagnosis of human brucellosis. The lysis centrifugation technique has been found to be more sensitive in both acute (20% higher sensitivity; P < 0.00001) and chronic (40% higher sensitivity; P = 0.087) forms of brucellosis. The major advantage of lysis centrifugation was in the mean detection time, which was only 2.4 days in acute and 2.7 days in chronic cases, with 103 out of 110 (93.6%) and 17 out of 20 (85%) cultures from acute and chronic brucellosis, respectively, detected before the conventional culture was positive. Our results confirmed the potential usefulness of the lysis technique in diagnosis and institution of appropriate antibiotic therapy.

Childhood brucellosis--a microbiological, epidemiological and clinical study.

A total of 5726 blood specimens (from children aged 14 years and younger) were studied for the serological evidence of brucellosis. Ninety-three (1.6 per cent) showed diagnostic agglutinin titres with a geometric mean titre of 403 (SD +/- 547). Forty-three (59.7 per cent) blood specimens yielded the growth of Brucella melitensis. Thirty-nine patients (41.93 per cent) were shepherds, who constituted the major occupational group affected in the present series. More than 60 per cent of the patients had a history of both consumption of fresh goat's milk and close animal contact. The habit of consuming fresh goat's milk to obtain relief from chronic ailments was noted in nine patients. Seventy-three (78.49 per cent) were males and 20 (21.51 per cent) were females, with a male to female ratio of 3:1. The disease occurred mainly in the school age group (mean age 10.3 years). All the patients had an acute history of less than 2 months. Forty-nine (52.68 per cent) patients presented with persistent fever, 19 (20.43 per cent) with joint pain, and the rest with a combination of fever and joint pain with and without low backache, fever being the commonest complaint. One case presented with involuntary movements of limbs alone and the other with burning feet only. Pityriasis alba was the consistent physical finding, with fever in the majority of the patients. The major joint found to be involved was the knee (52.77 per cent). The synovial fluid obtained from the knee joint of five patients demonstrated Brucella agglutinins and also three grew B. melitensis. Eight patients presented with complications that included skin lesions (3), carditis (2), neurobrucellosis such as chorea (1), peripheral neuritis (1), and meningitis (1). Brucella melitensis biotype 1 was successfully isolated from the papular eruption of one out of three cases who presented with skin lesions. To our knowledge this is the fourth confirmed isolation of B. melitensis from skin lesions with brucellosis, reported in the literature. The cerebrospinal fluid obtained from the meningitis patient was positive for B. agglutinins. To our knowledge chorea of brucellar origin appears to be the first case reported in the literature. In 15 cases (16.13 per cent) brucellosis was suspected clinically whereas 78 (83.87 per cent) cases, only serological evidence of brucellosis confirmed the diagnosis. None of the cases relapsed. In our experience an initial combination therapy with a three-drug regimen followed by a two-drug regimen for a minimum of 6 weeks has been found to be effective in the prevention of a relapse.