MacConkey Agar Media Research Articles

Extended culture methods among orthopaedic surgery patients and recovery of various microorganisms in a tertiary care hospital

Introduction: Extended culture has made a marked tremendous impact on orthopaedic post operative treatment due to prolonged incubation period. Aim: To identify various organisms isolated from different surgical procedures in Orthopaedic department and their rapidity of growth of various microorganisms isolated along with their sensitivity patterns. Materials and method: The prospective study was conducted in the department of Microbiology, KIMS, Secunderabad over a period of November 2015 to October 2016. A total of 104 samples were received from orthopaedic department during different orthopaedic surgical procedures. The received specimens were inoculated onto 5% sheep blood agar, chocolate agar, MacConkey agar and fluid thioglycolate medium and incubated at 37˚C, 5% CO2 for 40-48 hours and read daily for the growth. If no growth was observed, subcultures were made from fluid thioglycolateand observed daily till 7 days. Positive growths processed in Vitek 2 compact system (Biomerieux). The data collected was analyzed using Microsoft Excel. UK Standard for microbiology Investigations Investigation for orthopaedic implants associated infections (NICE accredited guidelines 2016) are the basis of the study. Results: A total of 104 patients were investigated in the present study, out of which 60 (57.6%) were culture positive. 44 (42.30%) cultures were sterile after 7 days of incubation. The orthopaedic surgical procedures were 52 Open fractures with Open reduction internal fixation (50%), 10 Septic arthritis (9.61%), 7 Sinovitis (6.7%), 7 Discharging sinus (6.7%) and 29 others (27.8%). Among all culture positives (n=60), 38 Gram negative organisms (61.66%) were in majority as compared to 23 Gram positive microorganisms (38.33%). Among them, 17 E.coli (28.3%), 8 Enterococcus (13.33%), 7 MSSA (6.7%), 5 Pseudomonas (4.8%) were the predominant pathogen. Conclusion: As 15(25%) microorganism showed prolonged incubation (48hrs to 7 days) period, which could be missed in conventional cuture method (24-48hrs). The outcome of all culture positive patient were good, 95% successfully treated.

Dermatopathy Caused by Enterobacter aerogenes and Pseudomonas aeruginosa in Boa constrictor amarali

Background: Bacterial diseases are the main cause of the high mortality rates of snakes, especially those caused by gramnegative agents. However, studies on dermatopathy caused by these bacterial agents in snakes are scarce; and no reports have been found on Enterobacter aerogenes as causative agent of dermatopathy in snake species. Thus, the objective of this study was to describe the clinical signs, and lesion evolution of a dermatopathy in a male snake (Boa constrictor amarali) specimen of approximately seven years old; and to describe the isolation and identification of the Enterobacter aerogenes and Pseudomonas aeruginosa agents involved in the cause of this disease.Case: The Boa constrictor amarali evaluated presented blackened cutaneous lesions in the dorsal, snout-vent and tail regions; and well-defined subcutaneous nodules of 2.0-3.0 cm diameter, with soft consistency, reddish color, cutaneous flaccidity, and areas of scale ulceration in the dorsolateral region. The clinical evaluation of the animal showed dehydration signs and pale mucous membranes. The blackened lesions were subjected to mycological analysis - after procedure of deep scale scraping - which showed presence of septate hyphae. The nodule was punctured for microbiological and biochemical analysis. The sample was collected with a sterilized alginate cotton tip swab, and was stored in a plastic tube containing a semi-solid Stuart transport medium, for microbiological analysis. Then, this sample was incubated in a bacteriological oven at 37°C for 24 h. Typical colonies of Pseudomonas and Enterobacter grew on MacConkey agar medium; these bacteria were identified by the colony morphology and their typical odor. The colonies grown in MacConkey agar were also identified through biochemical tests in the mediums: Phenol red, Lysine, Phenylalanine, Citrate, Urea and SIM (Sulfide, Indole, Motility). The results of these tests were able to confirm and identify the P. aeruginosa and E. aerogenes species. The animal died within 36 h, before the identification of the causative agents of the disease, thus, no pharmacological interference was possible.Discussion: Immunodepression, malnutrition, and temperatures and humidity outside the animal thermal comfort zone, are predisposing factors for the development of bacterial diseases in reptiles. Little information about pathogen agents affecting Boa constrictor specimens in their native area is available; however, captive snakes are subject to a wide variety of diseases - most of which caused or intensified by the captivity conditions. Among the bacteria involved in reptile diseases, few are primary causative agents. In general, clinical bacterial infections tend to be secondary to viral infections. The bacterial agents found in this study are commonly described in scientific literature with location in the oral cavity, differently from the results found in this study. Moreover, the bacterium E. aerogenes has not yet been described in other studies as a causative agent of dermatopathy. Reptiles are considered reservoirs of important zoonotic microorganisms, such as P. aeruginosa, which can be transmitted by fecal contact, bites and wounds. However, the little information on P. aeruginosa in captive reptiles indicates the need for further studies to establish its zoonotic potential. A most adequate management conditions for the snake species could have decreased the severity of the lesions. The occurrence of P. aeruginosa and E. aerogenes found in this work may alert professionals for future clinical suspicions and adequate therapeutic management.

Determination of Prevalence of Urinary Tract Infection Among the Pregnant Women with Lower Abdominal Pain

The present study was aimed to determine the prevalence of urinary tract infection among the pregnant women with lower abdominal pain and its aetiological micro-organism. Cross sectional study was done at Mount meru hospital laboratory, Arusha, Tanzania. 225 pregnant women attending an antenatal clinic in Levolosi hospital (Arusha, Tazania) were enrolled. To diagnose urinary tract infection in the enrolled participants, mid stream urine was collected and culture on Macconkey agar media and blood agar media. Urine analysis was done using dipstick test, urine microscopy and biochemical tests. The prevalence of urinary tract infection was found to be 31.6%. High incidence of infection was found in 33-40 years age group (41.6%). The incidence of infection was high in the third trimester of pregnancy compared to first and second trimester. The prevalence of infection is more in participants who had past history of infection. The bacterial pathogens isolated include E.coli (40.8%), Staphylococcus species (30.0%), Klebsiella species (14.1%), Proteus species (11.3%), Citrobacter species (1.4%) and Enterobacter species (1.4%). Diagnosis of urinary tract infection in pregnant women during all antenatal visits should be considered a vital care in the community. This helps to keep away from complications in pregnancy at an early stage.

Isolation and Antimicrobial Susceptibility Pattern of Urinary Escherichia Coli in Dhaka Medical College Hospital, Bangladesh

Background: Escherichia coli is the major bacterial pathogen being isolated and reported from urine, globally. For these uropathogens the therapeutic management becomes limited, so knowledge about the antimicrobial resistance pattern of these agents at a specific area may help the doctors to choose correct treatment regimen. The aim of the current study was to detect the common uropathogens from urine and the antimicrobial susceptibility pattern of Escherichia coli in Dhaka Medical College Hospital (DMCH).Methods: Samples were collected over a period of 12 months from July 2011 to June 2012 from the patients of DMCH irrespective of age and sex. Samples were cultured in blood agar and MacConkey agar media and organisms were identified by different biochemical tests such as oxidase test, reaction in MIU and simmon's citrate media and different sugar fermentation tests. Data were analyzed using Microsoft Excel (2007).Results: From total 300 urine samples, 177 (59%) Gram-positive and Gram-negative bacteria were isolated. Among them, Esch. coli was the most predominant 112 (63.28%) organism followed by Enterobacter spp. (11.87%). The prevalence of Esch. coli was significantly higher in females (63.39%) of younger age group than in males (36.61%). Esch. coli was highly sensitive to imipenem (100%) followed by ceftriaxone (62.50%), ceftazidime (55.36%) and azitromycin (35.72%) respectively. Low sensitivity patterns were found against doxycycline (13.39%), co-trimoxazole (16.07%), amoxiclav (21.43%) and ciprofloxacin (23.22%) respectively.Conclusion: The result of this study provides insight into the high proportion of multidrug resistant Escherichia coli and creates a need to report them routinely in laboratories.Anwer Khan Modern Medical College Journal Vol. 7, No. 1: Jan 2016, P 40-44

Prevalence and antibiogram of ESBL producing gram negative bacilli isolated from urine in Dhaka Medical College Hospital, Bangladesh

Extended spectrum ß-lactamase (ESBL) producing strains are resistant to a wide variety of common antimicrobials and become a major clinical concern worldwide that has complicated treatment strategies. The current study has been carried out to detect ESBL producing gram negative bacilli with their antimicrobial susceptibility pattern from urine collected over a period of 12 months from July 2011 to June 2012 from Dhaka Medical College Hospital. Samples were cultured in blood agar and MacConkey's agar media and organisms were identified by different biochemical tests such as oxidase test, reaction in MIU and simmon's citrate media and different sugar fermentation tests. ESBL producers were detected by double-disk synergy test (DDST). From total of 300 urine samples, 157 (52.33%) gram negative bacilli causing UTI were isolated and most of them were Escherichia coli (71.34%) followed by Enterobacter aerogenes (13.38%). Among the isolates, 45 (28.66%) ESBL producers were detected. The highest of ESBL producer was observed in Escherichia coli (32.14%) followed by Klebsiella pneumoniae (28.57%), Enterobacter aerogenes (23.80%) and Acinetobacter spp (20%). ESBL producers were significantly more resistant to ciprofloxacin, gentamycin, doxycycline, amoxiclav and co-trimoxazole than non-ESBL producers. All the ESBL producing strains were sensitive to imipenem. The result of this study provides insight into the high proportion of highly resistant ESBL producing organisms and more effective strategies are needed to control the spread of these resistant organisms.Bangladesh J Med Microbiol 2015; 9 (1): 17-21

In-vitro Antimicrobial Susceptibility Pattern of Isolates from Urine in Butembo, Democratic Republic of the Congo

Objective: This study aimed to determine the isolates from urine cultures at patients suspected with urinary tract infection and their in-vitro susceptibility pattern to antimicrobials at the Central Laboratory of Research of the “Universite Catholique du Graben” (UCG) in Butembo. Methodology: This was a cross-sectional study adopting a descriptive approach, conducted from January, 2015 to December, 2016. Six hundred and seventeen patients were screened. Freshly voided mid- stream urine sample were taken according to standard method. Isolation of the microbial agents of urinary tract infections was done on blood agar and MacConkey agar media. The culture was repeated in contrast with significant bacteria and urinalysis if the isolate at the first culture is known non-pathogenic. The antimicrobial susceptibility testing was done by using disk diffusion (Kirby Bauer's) technique. Results: Out of the 617 patients screened, the culture was positive in 77.3% (477 cases). The most isolated bacteria from urine culture were Staphylococcus aureus (47%), Streptococcus spp (12%), Escherischia coli (10.9%), Moraxella spp (7.5%) and Bacillus spp (2.3%). Antibiotics which were sensible to three or more isolates were ciprofloxacin, cefuroxime, cefotaxime and vancomycin. It was also observed that all the bacterial species have a Multiple Antibiotic Resistance Index greater than 0.2. Conclusion: Continuous monitoring of antimicrobial susceptibility pattern of bacterial isolates implicated in urinary tract infections is needed seeing the high Multiple Antibiotics Resistance Indices of all samples isolates. And this should be prior to antibiotic prescription in order to ensure an optimal, desired and cost-effectiveness treatment. For empiric treatment of urinary tract infections in Butembo, we suggested ciprofloxacin, cefuroxime and cefotaxime as the first line antibiotics of choice.

Port-site infections by nontuberculous mycobacterium: A retrospective clinico-microbiological study.

Port-site infection (PSI) is a prevailing, chronic, nagging, treatment refractory complication of laparoscopic surgery (LS). It neutralizes the advantages of minimally invasive surgery and increases morbidity, treatment cost of patient, leading to loss of confidence on operating surgeon. PSIs are preventable with appropriate preoperative, intraoperative, and postoperative measures. Atypical mycobacterium is most commonly associated with nonhealing postlaparoscopic wound infections, causing outbreaks or sporadic cases worldwide. We retrospectively studied the occurrence of nontuberculous mycobacterium (NTM) from PSIs following LS that did not respond to antibiotics used for pyogenic infections and having sterile routine aerobic cultures and their antimicrobial susceptibility pattern to guide proper management. The study was done in a tertiary care hospital of Eastern India over a 1-year period which included PSI cases with delayed onset not responding to antibiotics, following different types of LS. Pus/discharge from 32 patients was collected and examined for isolation and identification of the causative agents. Gram stain and Ziehl-Neelsen staining methods were used for direct examination. Culture media included blood agar, Robertson's cooked meat broth, MacConkey agar, and Lowenstein-Jensen medium. Isolates from the cases were identified using biochemical tests or molecular methods and studied the antimicrobial susceptibility pattern by the standard microbiologic procedures. Mycobacterium abscessus (13) and Mycobacterium fortuitum (2) were isolated from 15 serosanguinous drainage obtained from 32 cases by routine microbiological techniques. All isolates analyzed for antimicrobial susceptibility pattern were highly sensitive to clarithromycin (93.3%), amikacin (93.3%), and imipenem (80%) but were variable to ciprofloxacin, ofloxacin, and linezolid. Our present study shows frequent association of NTM with laparoscopic port-site nonhealing chronic infection or wound dehiscence. Although direct microscopy can give us a clue to diagnosis, culture isolation is required for speciation and antimicrobial susceptibility testing, which helps formulate therapeutic regimen.

Molecular Study of Quinolone Resistance in Klebsiella pneumoniae and Citrobacter freundii Isolates

A total of 97 burns swab collected from patients treated in burns unit of Al-Sader Hospital, Al-Najaf Province, during the period five months (1/6/2014 to 1/11/2014). Only 75/97(77.31%) were positive bacterial culture on MacConkey agar medium compared with 22/97(22.68%) specimens gave negative bacterial growth. The results of microscopic, morphology culture, biochemical test and Vitek-2 system revealed that 23/75(30.66%) isolates, were obtained as following 21/75(28%) Klebsiella pneumoniae ssp pneumoniae and 2/75(2.66%) Citrobacter freundii were recovered from 75 positive specimens obtained aseptically from admitted patients in the burn unit. Out of these, 15/23 (65.21%) were isolated from female patients and 8/23 (34.78 %) from male patients.Antimicrobial susceptibility were done using disk diffusion method and minimal inhibitory concentration (MIC) strip test. Results showed high degree resistance to most antibiotic under study and all tested isolates were at least resistant to three or more of antibiotic classes and then consider as multidrug resistant isolates (MDR) , At same time, ertapenem revealed maximum effectiveness against K. pneumoniae and C. freundii isolates with resistance rate reached to (28.57%) and (0%) respectively. While cloxacillin, oxacillin, and carbenicillin were offered less effective among antibiotic through resistance 100%. 14(66.66%) of K. pneumoniae isolates gave high level of resistance to nalidixic acid and ciprofloxacin with MIC ≥ 256 µg/ml and MIC ≥32 µg/ml respectively compared with 1(50%) in C. freundii isolates and these were considered as highly resistance. The PCR data showed that aac(6’)-Ib –cr gene high prevalence 15(71.42%) in K. pneumoniae isolates compared to 1(50%) in C. freundii no.2. Also qnr B gene was positive in 10(47.61%) K. pneumoniae, while only 1 (50%) of C. freundii isolates was harbored qnr B gene. While qnr, qnr C and qnr D genes did not detect in this study.

Prevalence and Antibiogram of Microbial Agents Causing Nosocomial Urinary Tract Infection in Surgical Ward of Dhaka Medical College Hospital

Background: Nosocomial infections pose substantial risk to patients receiving care in hospitals. In Bangladesh, this problem is aggravated by inadequate infection control due to poor hygiene, resource and structural constraints and lack of awareness regarding nosocomial infections.Objective: We carried out this study to determine the prevalence of different microorganisms from urine in surgery ward and antimicrobial susceptibility pattern against various antibiotics.Materials and Methods: This cross sectional study was carried out in Department of Microbiology, Dhaka Medical College, Dhaka over a period of 12 months from July 2011 to June 2012. A total of 52 urine specimens were collected from catheterized patients admitted in general surgery ward of Dhaka Medical College Hospital (DMCH) and incubated in blood agar, MacConkey agar media and the isolates were identified by different biochemical tests – oxidase test and reaction in MIU (motility indole urease) and Simmon’s citrate and TSI (triple sugar iron) media. ESBL producers were detected by double-disk synergy test (DDST).Results: Bacteria were isolated from 35 specimens and Escherichia coli was the commonest isolate (23, 65.71%) followed by Pseudomonas aeruginosa 6 (17.14%), Klebsiella pneumoniae 3 (8.57%), Acinetobacter baumannii 2 (5.72%) and Proteus vulgaris 1 (2.86%) respectively. Among the isolates, 10 (28.57%) ESBL producers were detected and the highest ESBL production was observed in Escherichia coli (8, 22.85%) followed by Klebsiella pneumoniae 1 (2.86%) and Pseudomonas aeruginosa 1 (2.86%). The isolates were resistant to most of the commonly used antimicrobial agents.Conclusion: The emergence of multi-drug resistant (MDR) bacteria poses a difficult task for physicians who have limited therapeutic options. However, the high rate of nosocomial infections and multi-resistant pathogens necessitate urgent comprehensive interventions of infection control.J Enam Med Col 2016; 6(2): 75-79

Changing trends in the epidemiology ofVibrio choleraein an outbreak of 2013 in Solapur, Maharashtra

Background: Cholera, one of the Notifiable Diseases is known to cause serious epidemics and pandemics in developing countries like India. An outbreak had occurred in 2013 in Solapur and the isolates revealed certain change in the trends of the epidemiological markers. Methods: A total of 30 cases presented with gastroenteritis during the months of July and August of 2013. 30 stool specimens were cultured on Blood agar, Mac Conkey agar and TCBS medium and enriched on alkaline peptone water broth. Identification of the isolates was done by standard Biochemical and Serological Agglutination tests. Antibiotic susceptibility testing was done using Kirby Bauer disc diffusion method, according to CLSI guidelines. Phage typing was done at the National Institute of Cholera and Enteric Diseases, Kolkata. Results: Out of the 30 stool specimen cultured, 14 strains of V.cholerae biotype El Tor serotype Ogawa was isolated. Phage Typing could be done on 6 isolates. All the strains identified belonged to T-2 phage type, according to Basu and Mukherjee scheme. According to the new scheme, two isolates belonged to T-27 phage type, two belonged to T-3 phage type, while one each belonged to T-6 and T-12 phage types. Most of the strains isolated in 2013 epidemic were sensitive to tetracycline, cotrimoxazole, and ciprofloxacin and resistant to ampicillin and gentamicin. Interpretation and Conclusion: Due to the changing epidemiological markers of V.cholerae such as Phage type and Antibiogram, all the epidemics must be thoroughly investigated and the isolates must be subjected to complete identification and Antibiotic Susceptibility Testing. Keywords: Antibiogram, Basu and Mukherjee scheme, epidemiological markers, phage typing, Vibrio cholerae

Current Trends of Urinary Pathogens and their Antimicrobial Susceptibility Pattern in a Tertiary Care Hospital

Introduction: Urinary Tract Infection (UTI) is one of the most common bacterial diseases worldwide that can present as asymptomatic or symptomatic characterized by a wide range of symptoms from mild irritative voiding to bacteremia, sepsis or even death. Increase in resistance of urinary pathogens to conventional antimicrobial agents is gaining the attention of many microbiologists worldwide in respect to treatment of urinary tract infection. Since the pattern of bacterial resistance is continuously changing, the monitoring of antibiotic susceptibility pattern becomes more important.Objective: The objective of this study was to update the distribution of current urinary pathogens and to find out their antimicrobial susceptibility pattern.Methods: A Cross sectional descriptive study was conducted at Border Guard Hospital (BGB Hospital) Peelkhana, Dhaka during the period of February 2013 to September 2013. A total of three hundred ninety urine samples were collected from patients with suspected UTI. All the samples were clean catch mid stream urine. Urine samples were cultured on MacConkey agar medium and blood agar medium. Colony counts yielding single type of bacterial growth of 105 CFU/ml were deemed significant.Results: Out of 390 samples, 49(12.56%) samples were culture positive. The isolated organisms were identified by conventional methods and were subjected to determine antimicrobial susceptibility testing by Kirby Bauer’s disc diffusion method. Escherichia coli (E.coli) was found as the most prevalent isolates 31(63.26%) followed by klebsiella spp 06(12.24%), Enterobacter spp o4(8.17%), Pseudomonas aeruginosa 04(8.17%), Staphylococcus aureus (Staph aureus) 02(4.08%) and Enterococcus spp 02(4.08%). Eight antimicrobial agents namely Amoxycillin, Co-trimoxazole, Levofloxacin, Nitrofurantoin, Gentamicin, Ceftriaxone, Imipenem and Amikacin were used for antimicrobial susceptibility testing. The most effective drug found against urinary isolates was Imipenem (95.91%), followed by Amikacin (81.63%) and Ceftriaxone (69.38%). None of the drug found was 100% effective against urinary pathogens. E.coli was found to be fairly sensitive to Ceftriaxone, Gentamicin and Nitrofurantoin.Conclusion: Urinary tract infection remains one of the most common bacterial infections. Antimicrobial drug resistance is increasing among urinary pathogens. This study updated the current occurrence of urinary pathogens and their antibiotic sensitivity pattern. We recommend periodical determination of antimicrobial sensitivity pattern of urinary pathogens in a particular hospital or area.Journal of Armed Forces Medical College Bangladesh Vol.10(2) 2014

Development of an improved selective medium for the detection of Shigella spp.

We developed an improved selective medium (HEX medium, modified Hektoen Enteric agar medium) for the detection of Shigella spp. from foods. Conventional culture media for Shigella spp. are neither specific nor sensitive. HEX medium contains lactose, sucrose, d-xylose, and salicin as a differentiation marker. The concentration of bile salts No.3 was reduced to a level of 0.3% which completely inhibited tested gram positive bacteria. All Shigella spp. tested (Shigella flexneri, Shigella dysenteriae, Shigella boydii, and Shigella sonnei) produced green colonies on HEX medium, while Hafnia alvei, found to be false positive for Shigella on HE agar, appeared as differentiable orange colonies on HEX medium. A total of 300 uninoculated and inoculated food samples were used to evaluate the specificity and sensitivity of HEX medium. The specificity was 13.50, 38.50, and 83.50 (%) on MacConkey agar, HE agar, and HEX medium, respectively. The sensitivity was 76.00, and 84.00 (%) for S. flexneri and 80.00, and 92.00 (%) for S. sonnei on HE agar and HEX medium, respectively. HEX medium had superior specificity and sensitivity compared to HE agar. Therefore HEX medium can be an appropriate selective and differential medium for detection of Shigella spp. from foods.

Isolation and identification of Bacteria and their drug susceptibility pattern in children to establish the correlation between childhood Septicemia with C-reactive protein (CRP) levels

Context: Neonatal sepsis is one of the most important causes of morbidity and mortality and C-reactive protein (CRP) an excellent biomarker has significant diagnostic and prognostic value for the treatment of septicemia patient. Objectives: To isolate and identify viable pathogens from blood culture and their antibiogram and to correlate CRP levels with septicemic child. Materials and Methods: A total of 273 cases among which 233 were clinically suspected septicemia cases and 40 were healthy controls in age group 0 day to 15 years were selected from United Hospital Ltd., Dhaka Bangladesh. Blood culture was analyzed by the instrument BACTEC 9120 series. CRP was measured from blood serum by the auto biochemical analyzer OLYMPUS AU 640 followed by immuno-turbidimetric method. The organisms were isolated by inoculation on blood agar and MacConkey agar media. Identification of the organisms was done by colony morphology, gram staining and biochemical tests. Sensitivity of isolates was done against antimicrobial agents by disc diffusing method. Results: Blood samples in total 233 cases of suspected septicemia in children were studied between the ages of 0 day to 15 years. Culture proven septicemia 39 (16.74%), probable septicemia 136 (58.37%) and nonsepticemic febrile patients 58 (24.90%) were found. The highest rate of blood culture positivity found among 5-10 yrs. age group (25.64%). Salmonella typhi (41.03%) was the most common infective agent. The rate of blood culture positivity was significantly higher (p<0.001) among patients without antimicrobial therapy (23.74%) than those in patients with antimicrobial therapy (6.38%). S. typhi were 75% sensitive to Ceftriaxone while azithromycin showed high rate resistance (85.71%). E. coli and Klebsiella pneumoniae were highly sensitive (100%) to imipenem but E. coli resistant (100%) to amikacin, amoxyclavonic acid ciprofloxacin, gentamicin, Cefepime and netilmicin. Resistant (100%) to gentamicin was observed from K. pneumoniae. Mean CRP values (mg/l) of blood culture proven septicemia group, probable septicemia group, non-septicemic febrile group and control group were 70.42, 34.05, 3.08 and 0.98 respectively. Both proven septicemia and suspected septicemia cases showed CRP concentration above the cut-off value (>6 mg/l) and p value significant (p<0.001). Statistically significant difference (p<0.001) was found when mean CRP level of proven septicemia group, probable septicemia group and non-septicemic febrile group each compared with control group. CRP concentration were significantly (p < 0.001) different among three study group. Conclusion: Most of the gram negative bacteria isolated from blood culture showed resistance to commonly used antibiotics. The predominant infective isolate was Salmonella typhi. In this study, CRP level is high (cut-off value 6 mg/l) both in proven and probable septicemia group. CRP may have a good biomarker tools in diagnostic and prognostic value. Investigation of blood culture should be done before antimicrobial therapy, DOI: http://dx.doi.org/10.3329/jbs.v21i0.22524 J. bio-sci. 21: 99-108, 2013

Antimicrobial sensitivity pattern of bacterial pathogens in urinary tract infections in South Delhi, India.

Seventy-four bacterial proven cases of urinary tract infections were studied, and identified by Mac Conkey agar and blood agar medium separately; all the isolates were subjected to antimicrobial sensitivity testing by Stokes technique. Ninty-six percent of total isolated organisms were found to be gram negative while remaining 4% were gram positive. Among gram negatives, E. coli and gram positive S. aureus were the most prevalent organisms. The percentage of gram negative isolates were as follows, E. coli (79.7%) followed by Klebsiella (9.5%), Pseudomonas, Acinetobacter were (2.7% each), Proteus constituted (1.4%). and among gram positive S. aureus (4%). The antibiotic resistance of identified organisms was carried out by disc-diffusion method with commercially available disc of thirteen antibiotics having different mode of actions such as inhibition of cell wall synthesis, membrane permeability alternatives, inhibition of protein synthesis and DNA synthesis inhibitors. Gram negatives showed more resistance to these antibiotics as compared to gram positive organisms. The most effective antibiotic for gram negative UTI isolates is amikacin showing 63% efficacy followed by Cefotaxime 55% efficacy, Amoxicillin and Ciprofloxacin with (49% each) efficacy. Among gram positives, Chloramphenicol, Co-trimoxazole, Gentamicin, Amikacin, Ciprofloxacin and Cefotaxime are most effective with (66.6% each) efficacy, then Ampicillin, Amoxicillin, Tetracycline and norfloxacin with (33.3% each) efficacy.

Staphylococcus aureus small colony variants in diabetic foot infections.

BackgroundStaphylococcus aureus (S. aureus) is one of the major pathogens causing chronic infections. The ability of S. aureus to acquire resistance to a diverse range of antimicrobial compounds results in limited treatment options, particularly in methicillin-resistant S. aureus (MRSA). A mechanism by which S. aureus develops reduced susceptibility to antimicrobials is through the formation of small colony variants (SCVs). Infections by SCVs of S. aureus are an upcoming problem due to difficulties in laboratory diagnosis and resistance to antimicrobial therapy.MethodsA prospective study was performed on 120 patients diagnosed with both type 2 diabetes mellitus and infected diabetic foot ulcers. The study was carried out from July 2012 to December 2013 in Hospital General de Mexico. The samples were cultured in blood agar, mannitol salt agar, and MacConkey agar media, and incubated at 37°C in aerobic conditions.ResultsWe describe the first known cases of diabetic foot infections caused by MRSA-SCVs in patients diagnosed with type 2 diabetes mellitus and infected diabetic foot ulcers. In all of our cases, the patients had not received any form of gentamicin therapy.ConclusionsThe antibiotic therapy commonly used in diabetic patients with infected diabetic foot ulcers fails in the case of MRSA-SCVs because the intracellular location protects S. aureus-SCVs from the host's defenses and also helps them resist antibiotics. The cases studied in this article add to the spectrum of persistent and relapsing infections attributed to MRSA-SCVs and emphasizes that these variants may also play a relevant role in diabetic foot infections.

Surgical Site Infections Due to Rapidly Growing Mycobacteria in Puducherry, India

Rapidly growing Mycobacteria are increasingly recognized, nowadays as an important pathogen that can cause wide range of clinical syndromes in humans. We herein describe unrelated cases of surgical site infection caused by Rapidly growing Mycobacteria (RGM), seen during a period of 12 months. Nineteen patients underwent operations by different surgical teams located in diverse sections of Tamil Nadu, Pondicherry, Karnataka, India. All patients presented with painful, draining subcutaneous nodules at the infection sites. Purulent material specimens were sent to the microbiology laboratory. Gram stain and Ziehl-Neelsen staining methods were used for direct examination. Culture media included blood agar, chocolate agar, MacConkey agar, Sabourauds agar and Lowenstein-Jensen medium for Mycobacteria. Isolated microorganisms were identified and further tested for antimicrobial susceptibility by standard microbiologic procedures. Mycobacterium fortuitum and M.chelonae were isolated from the purulent drainage obtained from wounds by routine microbiological techniques from all the specimens. All isolates analyzed for antimicrobial susceptibility pattern were sensitive to clarithromycin, linezolid and amikacin but were variable to ciprofloxacin, rifampicin and tobramycin. Our case series highlights that a high level of clinical suspicion should be maintained for patients presenting with protracted soft tissue lesions with a history of trauma or surgery as these infections not only cause physical but also emotional distress that affects both the patients and the surgeon.

Shiga toxin-producing Escherichia coli isolated from lettuce samples in tehran, iran.

Background:During the last decade, the prevalence of foodborne diseases due to contaminated food as well as the outbreaks of diseases due to Shiga toxin-producing Escherichia coli (STEC) strains has increased.Objectives:The aim of this study was to evaluate the prevalence and antibiotic resistance pattern of STEC strains in lettuce samples. Since lettuce is used as a raw vegetable in salads, the rates of infections caused by this vegetable are high.Materials and Methods:A total of 100 samples collected from Tehran, Iran, were transported to the laboratory, homogenized by a stomacher in E. coli broth containing cefixime, and cultured on MacConkey agar medium. Their DNA was extracted by boiling method and polymerase chain reaction (PCR) was performed, using five primers targeting the stx1, stx2, fliCh7, rbfO157, and eaeA genes. Susceptibility testing against ampicillin, imipenem, cephalosporin, tetracycline, aminoglycosides, chloramphenicol and quinolones was performed using disk diffusion method.Results:Eight samples were positive for presence of STEC strains, three contained stx1, five contained stx2, and one sample was positive for presence of both rbfO157 and fliCh7. They were susceptible to all the antibiotics except for ampicillin and tetracycline.Conclusions:This study indicated the contamination of lettuce by STEC strains and its possible role as the source of infection. Resistance to both tetracycline and ampicillin may be considered as an emergency alarm for a multidrug resistance of STEC strains.

Occurrence of Pasteurellosis and Newcastle Disease in indigenous chickens in Sirajgonj district

A study was carried out on indigenous layer chickens to know occurrence of Avian Pasteurellosis (AP) and Newcastle disease (ND) in Sirajgonj during the period from January/2012 to December/2013. The clinical signs showed before death was recorded by taking history and the birds were subjected to post mortem examination. In addition to the clinical and necropsy findings, ND was detected by Anigen® rapid antigen detection kit. The AP was confirmed by isolation and identification of Pasteurella (P.) multocida from liver, spleen and heart samples. The P. multocida was found to grow on nutrient broth, nutrient agar, blood agar and Eosin Methylene Blue agar where it produced whitish, opaque, round, flat, translucent colonies. It produced turbidity on nutrient broth. The organism was not found to grow on MacConkey agar media and did not cause hemolysis on blood agar media. The impression smear of liver and heart blood were stained by Gram’s staining, Leishman staining and Methylene blue staining techniques to detect P. multocida. P. multocida organism was also found to fermnt dextrose, lactose and mannitol with the production of only acid but did not ferment maltose and lactose. P. mutocida was found non-motile, indole positive and urease negative. On triple sugar iron test it produced H2S and fermented only glucose. It was found negative to both methyl red test and Voges Proskauer test. Out of 360 birds tested 59.72% (215) was AP positive and 40.28% was ND positive. Age, Sex, season and mortality due to other diseases were not considered.DOI: http://dx.doi.org/10.3329/bjvm.v11i2.19123 Bangl. J. Vet. Med. (2013). 11 (2): 97-105

Incidence of Bacteremia After Endoscopic Variceal Ligation of Oesophageal Varices in Cirrhotic Patients

Bacteremia and infectious sequelea are frequent complications after sclerotherapy. As cirrhotic patients are immune deficient, bacteremia occurring as a consequence of endoscopic variceal ligation (EVL) may develop systemic infection. The aim of the study was to see the incidence of bacteremia in cirrhotic patients after EVL. In this study 91 cirrhotic patients with grade II-III oesophageal varices were enrolled and 100 EVL sessions were done. Patients receiving antibiotics, having evidence of infection were excluded. After the completion of EVL, blood samples for culture and sensitivity were collected at 5minute and after 30minute and subculture done in Blood Agar and MacConkey Agar media. We followed the patients for a week. Of the one hundred cases there were 30 subjects in Child A, 48 in Child B and 13 in Child C class. Four patients (4%) had positive blood culture after EVL, three of Child A class and one of Child B class and non in Child C class. All three patients of Child A had culture positive in their 30minute sample and not in 5 minute sample. The Child B patient had culture positive in his 5minute sample and not in 30minute sample. The bacterium isolated was Coagulase negative Staphylococcus, which we considered it as contaminant. None of these patients developed any infectious sequelea. The incidence of bacteremia after EVL is yet to be ascertained. Post EVL antibiotic prophylaxis is not recommended. DOI: http://dx.doi.org/10.3329/medtoday.v24i1.14109 Medicine TODAY Vol.24(1) 2012 pp.20-22

Quicklime treatment and stirring of different poultry litter substrates for reducing pathogenic bacteria counts

Testing different management practices can help to identify conditions that decrease or even eliminate pathogenic bacteria in poultry litter. A trial was conducted to evaluate the effects of daily manual stirring (rotation of the litter with a pitchfork) for the first 14 d of a bird's life (WDR), in 3 types of poultry litter substrates and quicklime treatment (CaO) during layout time between flocks on pathogenic bacteria occurrence (cfu). A total of 216 male Cobb broilers were randomly allotted to 18 pens with new litter (experimental unit). A split-plot design, with 6 treatments allotted to the main plots, was used: 1) wood shavings (WS) + WDR, 2) WS without stirring up to 14 d (WODR), 3) rice hulls (RIH) + WDR, 4) RIH + WODR, 5) mixture of 50% RIH and WS + WDR, and 6) mixture of 50% RIH and WS + WODR. Two treatments were allotted to the subplots: 0 and 300 g of CaO•m(-2) litter. After depopulation, litter samples were collected, and CaO was incorporated into the litter in the designated half of each pen. The cfu from litter samples after 7 d of the quicklime treatment were counted on Chapman agar, brain heart infusion media, and MacConkey agar. The data were analyzed using ANOVA, and the means were compared by least squares means (P < 0.05). Neither the type of substrate nor the act of stirring affected the cfu. The incorporation of 300 g of CaO•m(-2) litter efficiently reduced the cfu observed on brain heart infusion, Chapman agar, and MacConkey agar media by 57.2, 66.9, and 92.1%, respectively, compared with control (6.4, 17.9, and 46.1%; P < 0.001). In conclusion, the incorporation of 300 g/m(-2) of quicklime in poultry litter reduces the cfu, regardless of the substrate and stirring performed.