A bacteriological study of hospital beds before and after disinfection with phenolic disinfectant

In hospitals, one of the ways to control microbial contamination is by disinfecting the furniture used by patients. This study's main objective was to evaluate the microbiological condition of hospital mattresses before and after such disinfection, in order to identify bacteria that are epidemiologically important in nosocomial infection, such as Staphylococcus aureus and Pseudomonas aeruginosa. RODAC plates with two different culture media were used to collect specimens. Patient beds were selected according to previously established criteria, and surface areas on the mattresses were chosen at random. From the total of 1 040 plate cultures from 52 mattresses, positive results were obtained from 500 of them (48.1%), 263 before disinfection and 237 after disinfection. Considering the selectivity of the culture media, the positivity rate was high. There were high prevalences of S. aureus both before and after mattress disinfection. The study results suggest that the usual disinfection procedures, instead of diminishing the number of microbes, merely displace them from one part of the mattress to another, and the number of microorganisms remains the same.

Health and disease status of Thai pangas, Pangasius hypophthalmus cultured in rural ponds of Mymensingh, Bangladesh

Thai pangas, Pangasius hypophthalmus is one of the important aquaculture species in Bangladesh. Over the last few years spectacular development has been taking place in Thai pangas farming in Mymensingh district. Due to availability of easy breeding and culture techniques as well as quick return, more and more people are converting their rice fields into pangas farms overnight. The present study was carried out to examine health and disease status of Thai pangas mainly through clinical, histopathological and bacteriological techniques. In addition, for collecting primary data on disease and health status of Thai pangas and the resultant socioeconomic impacts on rural households, questionnaire interview and participatory rural appraisal tools were used with selected farming households in three upazilas of Mymensingh district. The most prevalent diseases as reported by the farmers were red spot, followed by anal protrusion, tail and fin rot, pop eye, dropsy and gill rot. Other conditions like cotton wool type lesion, ulceration and white spot were reported but with lower incidence. Four isolates of Aeromonas hydrophila were recovered from kidney and lesion of diseased fish. Hemorrhage over the body especially near mouth and caudal region was noticed in the fishes associated with aeromonad infection. Internally, kidney, liver and spleen became swollen and enlarged. The isolates varied with their pathogenicity. All the four isolates were sensitive to Nitrofurantoin, Cotrimoxazole and Tetracycline but were resistant to Amoxycilline. An attempt was made to treat diseased fish with extracts from neem leaf, garlic and turmeric. Recovery of infection was monitored through mortality and histopathology. General histopathological changes of different organs were also studied. Extract from neem (Azadirachta indica) leaf gave better result. Telangiectasis, lamellar hypertrophy and hyperplasia hemorrhage, lamellar fusion, necrosis of lamellar epithelial cells, presence of parasites and their cysts were the major pathology of gills. Hemorrhagic lesion, pyknotic nuclei and melanomacrophage centers (MMC) were found in the liver of fish. Major pathologies in kidney of fish included presence of MMC, necrotic and ruptured kidney tubules, severe haemopoietic necrosis, and hemorrhage. The economic loss due to disease in Thai pangas farming was estimated from the difference between expected production and actual production. On an average, Thai pangas farmers of Mymensingh incur a loss of Tk. 23,104/ha/cycle due to fish disease (3.6% of expected total production). The loss, however, varied with location and size of farms, type of farmers and management practices. The study also highlighted fish health management related problems and recommended further work for the development of user-friendly farmer-oriented fish health management packages.

WHO guidelines for antimicrobial treatment in children admitted to hospital in an area of intense Plasmodium falciparum transmission: prospective study.

OBJECTIVES: To assess the performance of WHO's "Guidelines for care at the first-referral level in developing countries" in an area of intense malaria transmission and identify bacterial infections in children with and without malaria. DESIGN: Prospective study. SETTING: District hospital in Muheza, northeast Tanzania. PARTICIPANTS: Children aged 2 months to 13 years admitted to hospital for febrile illness. MAIN OUTCOME MEASURES: Sensitivity and specificity of WHO guidelines in diagnosing invasive bacterial disease; susceptibility of isolated organisms to recommended antimicrobials. RESULTS: Over one year, 3639 children were enrolled and 184 (5.1%) died; 2195 (60.3%) were blood slide positive for Plasmodium falciparum, 341 (9.4%) had invasive bacterial disease, and 142 (3.9%) were seropositive for HIV. The prevalence of invasive bacterial disease was lower in slide positive children (100/2195, 4.6%) than in slide negative children (241/1444, 16.7%). Non-typhi Salmonella was the most frequently isolated organism (52/100 (52%) of organisms in slide positive children and 108/241 (45%) in slide negative children). Mortality among children with invasive bacterial disease was significantly higher (58/341, 17%) than in children without invasive bacterial disease (126/3298, 3.8%) (P<0.001), and this was true regardless of the presence of P falciparum parasitaemia. The sensitivity and specificity of WHO criteria in identifying invasive bacterial disease in slide positive children were 60.0% (95% confidence interval 58.0% to 62.1%) and 53.5% (51.4% to 55.6%), compared with 70.5% (68.2% to 72.9%) and 48.1% (45.6% to 50.7%) in slide negative children. In children with WHO criteria for invasive bacterial disease, only 99/211(47%) of isolated organisms were susceptible to the first recommended antimicrobial agent. CONCLUSIONS: In an area exposed to high transmission of malaria, current WHO guidelines failed to identify almost a third of children with invasive bacterial disease, and more than half of the organisms isolated were not susceptible to currently recommended antimicrobials. Improved diagnosis and treatment of invasive bacterial disease are needed to reduce childhood mortality.

Clinical microbiologists facing an anthrax alert.

Microbiological war and terrorist attacks are made to weaken populations by transmitting pathogenic and epidemic microorganisms. These bacteria or viruses are often difficult to diagnose. Anthrax alerts following September 2001 showed that most clinical microbiology laboratories were not adequately prepared, using obsolete diagnostic methods or being too slow to use accurate tools when facing a major threat. Following this period, most microbiology laboratories were prepared for bioterrorism alerts, in order to provide accurate and rapid results, although such events are rare and unexpected. In this review, we describe the organization and preparedness of our clinical microbiology laboratory regarding bioterrorism risk, although its main task is to perform routine diagnostic microbiology tests. To illustrate the difficulties, we briefly describe an anthrax alert.

Which anatomical sites should be sampled for screening of methicillin-resistant Staphylococcus aureus carriage by culture or by rapid PCR test?

The nose is the anatomical site usually recommended for methicillin-resistant Staphylococcus aureus (MRSA) screening. Other sites are also recommended, but are more controversial. We showed that the sensitivities of MRSA detection from nasal swabs alone were 48% and 62% by culture or by rapid PCR test, respectively. These percentages increased to 79% and 92% with the addition of groin swabs, and to 96% and 99% with the addition of groin and throat swabs. In conclusion, neither by culture nor by rapid PCR test is nose sampling alone sufficient for MRSA detection. Additional anatomical sites should include at least the groin and throat.

Influence of age on the reactivity of the BANA test among Brazilian children.

Three hundred and twenty samples of subgingival plaque were obtained from 80 caucasian girls, ranging from 10 to 13 years of age. The samples were analyzed to verify the influence of age upon colonization of the gingival sulcus by microorganisms potentially pathogenic to the periodontal tissues. The gingival and plaque status were evaluated through the gingival index (GI) and plaque index (PlI) and the microflora was assessed by the enzymatic method benzoyl-arginine-naphthylamide (BANA). The results of the BANA test were positive for 62.50% of the tested individuals and 40% of the examined sites. The influence of age was statistically significant on BANA reactivity, and the number of positive sites was greater at 11 (57.5%) than at 12 years (28.8%).

Paracoccidioides brasiliensis antigen batches from the same isolate show immunological and biochemical differences

We investigated the occurrence of antigenic and biochemical variability among Paracoccidioides brasiliensis antigen batches prepared according to the same protocol. Initially (experiment 1), we analyzed two antigen lots of two human isolates (Bt1 and Bt2), cultured in two media (PYG: bactopeptone, yeast extract, glucose; MMM: McVeigh and Morton medium) in SDS-PAGE and in two immunological tests (immunodiffusion-ID and footpad swelling test-FPT). Afterwards (experiment 2), we compared the antigenic profile of three antigen hatches from three human isolates (Bt1, Bt2 and Bt3) by two-dimensional immunoelectrophoresis (2 D-IEP) against a reference system for P. brasiliensis antigens. In experiment 1, there were important intra- and inter-strain antigenic differences between batches of the fungal isolates cultured on both media. The block titration of the antigen batches for the immunological tests revealed correlation between protein concentration and biological activity in ID and no correlation in FPT. In experiment 2, the reference system for P brasiliensis showed 26 antigen peaks. There were important differences between batches prepared from the same isolate and between batches from different isolates. Our data suggested the occurrence of instability in the synthesis of antigenic components by a same P. brasiliensis isolate, under controlled incubation conditions.

Methods of experimental induction of periapical inflammation. Microbiological and radiographic evaluation

Aim: To evaluate the influence of coronal filling and apical perforation on the induction of periapical inflammation. Methodology: Fifty-eight root canals in the teeth of dogs were divided into four groups. Groups I and II: root canals were exposed for 180 days; groups III and IV: root canals were exposed for 7 days and then the access cavity filled for 53 days. The root apices of groups I and III were perforated after the coronal opening, whilst those of groups II and IV remained intact. Standard radiographs were taken before and after the experimental periods. Digital images of the radiographs were created and then analysed by three examiners. After induction of periapical inflammation, the root canal contents were collected using paper points. Microbiologic evaluation of the type of microorganism was carried out by culture in different growth media. The radiographic and microbiologic data were statistically analysed using ANOVA at a 5% significance level. Results: There were a greater total number of microorganisms in groups I and II (P < 0.05). The number of anaerobes was greater than the number of aerobes (P < 0.05). The size of the periapical radiolucencies were not significantly different between the experimental groups. Conclusions: The different methods analysed induced similar areas of periapical radiolucency in dogs with predominantly anaerobic bacteria. However, the time required for induction was less when the method with coronal filling was used. © 2005 International Endodontic Journal.

In vitro effect of low-level laser therapy on typical oral microbial biofilms

The aim of this study was to evaluate the effect of specific parameters of low-level laser therapy (LLLT) on biofilms formed by Streptococcus mutans, Candida albicans or an association of both species. Single and dual-species biofilms - SSB and DSB - were exposed to laser doses of 5, 10 or 20 J/cm 2 from a near infrared InGaAsP diode laser prototype (LASERTable; 780 ± 3 nm, 0.04 W). After irradiation, the analysis of biobilm viability (MTT assay), biofilm growth (cfu/mL) and cell morphology (SEM) showed that LLLT reduced cell viability as well as the growth of biofilms. The response of S. mutans (SSB) to irradiation was similar for all laser doses and the biofilm growth was dose dependent. However, when associated with C. albicans (DSB), S. mutans was resistant to LLLT. For C. albicans, the association with S. mutans (DSB) caused a significant decrease in biofilm growth in a dose-dependent fashion. The morphology of the microorganisms in the SSB was not altered by LLLT, while the association of microbial species (DSB) promoted a reduction in the formation of C. albicans hyphae. LLLT had an inhibitory effect on the microorganisms, and this capacity can be altered according to the interactions between different microbial species.

Recovery of mutans streptococci on MSB, SB-20 and SB-20M agar media

Objective: The recovery of mutans streptococci in saliva and dental biofilm samples depends, in part, on the culture medium used. In this study, we compared (i) the culture media Sucrose-Bacitracin agar (SB-20), Modified SB-20 (SB-20M) and Mitis Salivarius Bacitracin agar (MSB) in the count of colony forming units (cfu) of mutans streptococci and (ii) in the morphological and biochemical differentiation between Streptococcus mutans and Streptococcus sobrinus. Design: Samples of non-stimulated saliva from 20 children were plated on SB-20, SB-20M and MSB, and incubated in microaerophilia at 37 °C for 72 h. Identification of microorganisms was based on analysis of colony morphology under stereomicroscopy. The biochemical identification of colonies was done by biochemical tests using sugar fermentation, resistance to bacitracin and hydrogen peroxide production. Results: There was no significant difference (p > 0.05) in the number of cfu of mutans streptococci recovered on SB-20 and SB-20M agar. Comparing the media, SB-20 and SB-20M yielded a larger number of mutans streptococci colonies (p < 0.05) and were more effective than MSB in the identification of S. sobrinus (p < 0.05), but not of S. mutans (p > 0.05). Conclusion: There was no significant difference between SB-20 and SB-20M culture media in the count of mutans streptococci, demonstrating that the replacement of sucrose by coarse granular cane sugar did not alter the efficacy of the medium. Compared with MSB, SB-20 and SB-20M allowed counting a larger number of mutans streptococci colonies and a more effective morphological identification of S. sobrinus. © 2012 Elsevier Ltd.

Análise comparativa de métodos de diagnóstico para linfadenite caseosa em ovinos sintomáticos e assintomáticos

Pós-graduação em Ciência Animal - FMVA

Influence of temperature of incubation and type of growth medium on pigmentation in Serratia marcescens.

Maximal amounts of prodigiosin were synthesized in either minimal or complete medium after incubation of cultures at 27 C for 7 days. Biosynthesis of prodigiosin began earlier and the range of temperature for formation was greater in complete medium. No prodigiosin was formed in either medium when cultures were incubated at 38 C; however, after a shift to 27 C, pigmentation ensued, provided the period of incubation at 38 C was not longer than 36 hr for minimal medium or 48 hr for complete medium. Washed, nonpigmented cells grown in either medium at 38 C for 72 hr could synthesize prodigiosin when suspended in saline at 27 C when casein hydrolysate was added. These suspensions produced less prodigiosin at a slower rate than did cultures growing in casein hydrolysate at 27 C without prior incubation at 38 C. Optimal concentration of casein hydrolysate for pigment formation by suspensions was 0.4%; optimal temperature was 27 C. Anaerobic incubation, shift back to 38 C, killing cells by heating, or chloramphenicol (25 mug/ml) inhibited pigmentation. Suspensions of washed cells forming pigment reached pH 8.0 to 8.3 rapidly and maintained this pH throughout incubation for 7 days. Measurements of viable count and of protein, plus other data, indicated that cellular multiplication did not occur in suspensions of washed cells during pigment formation. By this procedure utilizing a shift down in temperature, biosynthesis of prodigiosin by washed cells could be separated from multiplication of bacteria.

PCR-based assay using occult blood detection cards for detection of diarrheagenic Escherichia coli in specimens from U.S. travelers to Mexico with acute diarrhea.

Large field studies of travelers' diarrhea for multiple destinations are limited by the need to perform stool cultures on site in a timely manner. A method for the collection, transport, and storage of fecal specimens that does not require immediate processing and refrigeration and that is stable for months would be advantageous. This study was designed to determine if enterotoxigenic Escherichia coli (ETEC) and enteroaggregative E. coli (EAEC) DNA could be identified from cards that were processed for the evaluation of fecal occult blood. U.S. students traveling to Mexico during 2005 to 2007 were monitored for the occurrence of diarrheal illness. When ill, students provided a stool specimen for culture and occult blood by the standard methods. Cards then were stored at room temperature prior to DNA extraction. Fecal PCR was performed to identify ETEC and EAEC in DNA extracted from stools and from occult blood cards. Significantly more EAEC cases were identified by PCR that was performed on DNA that was extracted from cards (49%) or from frozen feces (40%) than from culture methods that used HEp-2 adherence assays (13%) (P < 0.001). Similarly, more ETEC cases were detected from card DNA (38%) than from fecal DNA (30%) or by culture that was followed by hybridization (10%) (P < 0.001). The sensitivity and specificity of the card test were 75 and 62%, respectively, compared to those for EAEC by culture and were 50 and 63%, respectively, compared to those for ETEC. DNA extracted from fecal cards that was used for the detection of occult blood is of use in identifying diarrheagenic E. coli.

Bacteriological investigations of prawn canneries 2. Incidence of Clostridium perfringens

Prawn processing factories of the three major fish processing centres of the West Coast of India, viz., Cochin, Mangalore and Calicut were surveyed to determine the occurrence of Clostridium perfringens in processing areas, and in processed products. Direct plating on Sulphite-polymyxin- sulphadiazine Agar and enrichment techniques were used. Samples of prawn, prawn guts, frozen prawns, canned prawns, water, ice, swab from utensils and soil from the factory premises were examined. Among a total of 461 samples examined, only 32 (6.9%) gave positive results. The incidence of C. perfringens was more in prawn guts (80%), followed by soil (50%), prawn (38%), ice (33.3%), frozen prawns (11%), swab (5.0%) and water (1.1%). No C. perfringens was isolated from canned prawns.

Computer vision techniques in seafood quality control

The application of computer vision based quality control has been slowly but steadily gaining importance mainly due to its speed in achieving results and also greatly due to its non- destnictive nature of testing. Besides, in food applications it also does not contribute to contamination. However, computer vision applications in quality control needs the application of an appropriate software for image analysis. Eventhough computer vision based quality control has several advantages, its application has limitations as to the type of work to be done, particularly so in the food industries. Selective applications, however, can be highly advantageous and very accurate.Computer vision based image analysis could be used in morphometric measurements of fish with the same accuracy as the existing conventional method. The method is non-destructive and non-contaminating thus providing anadvantage in seafood processing.The images could be stored in archives and retrieved at anytime to carry out morphometric studies for biologists.Computer vision and subsequent image analysis could be used in measurements of various food products to assess uniformity of size. One product namely cutlet and product ingredients namely coating materials such as bread crumbs and rava were selected for the study. Computer vision based image analysis was used in the measurements of length, width and area of cutlets. Also the width of coating materials like bread crumbs was measured.Computer imaging and subsequent image analysis can be very effectively used in quality evaluations of product ingredients in food processing. Measurement of width of coating materials could establish uniformity of particles or the lack of it. The application of image analysis in bacteriological work was also done