The utility of the polymerase chain reaction assay for aetiologic definition of unspecified bacterial meningitis cases

Most patients with acute suppurative meningitis are otherwise healthy individuals with regard to immune mechanisms against invasive bacterial disease. This medical emergency is among the most dramatic and potentially ravaging diseases that affect humans, particularly young children. The illness often strikes suddenly, and can either result in death or leave the survivors with significant neurological dysfunctions. The demonstration of a bacterial aetiology is necessary for decisions regarding treatment and prophylaxis. Conventional bacteriological methods frequently fail to identify an agent, as a result of administration of antibiotics or delayed lumbar punctures. We investigated the major aetiologic sources of unspecified bacterial meningitis cases (G00.9, ISCD-10) by polymerase chain reaction (PCR)-based identification of Neisseria meningitidis (crgA), Streptococcus pneumoniae (ply) and Haemophilus influenzae (bexA) in cerebrospinal fluid samples. The multiplex PCR detected N. meningitidis in 92%, S. pneumoniae in 4% and H. influenzae in 1% of the 192 clinical samples assayed; 3% were negative for all three DNA targets. Bacterial DNA detection was found to be a valuable adjunct to enhance bacterial meningitis surveillance when the yield of specimens by culture is reduced. The implementation of PCR assays as a diagnostic procedure in Public Health Laboratories is perceived to be a significant advance in the investigation of bacterial meningitis.

On the etiology of an outbreak of winter dysentery in dairy cows in Brazil

Winter dysentery (WD) is a seasonal infectious disease described worldwide that causes a marked decrease in milk production in dairy cows. In the Northern hemisphere, where the disease is classically recognized, bovine coronavirus (BCoV) has been assigned as a major etiologic agent of the disease. Nonetheless, in the Southern hemisphere, an in-deep etiological survey on WD cases had not been carried out. This study aimed to survey for BCoV by nested-RT-PCR, rotavirus by polyacrylamide gel electrophoresis (PAGE) and ELISA, bacteria by classical bacteriological methods and PCR for virulence factors and parasites by sugar flotation test on fecal samples of 21 cows from a farm during an outbreak of WD in São Paulo state, Southeastern Brazil. BCoV was detected in all 21 samples, while rotavirus was detected in two symptomatic cows. Escherichia coli, Yersinia intermedia, Providencia rustigianii Proteus penneri, Klebsiella terrigena and Enterobacter aglomerans were detected in samples from both asymptomatic and healthy cows in different associations. The study of E. coli virulence factors revealed that the strains isolated were all apathogenic. Cysts of Eimeria sp. and eggs of Strongyloidea were detected at low numbers in four of the symptomatic cows, with one co-infestation. These results suggest BCoV as the main etiologic agent of the cases of WD in Brazil, a conclusion that, with the clinical and epidemiological patterns of the disease studied herein, match those already described elsewhere. These findings give basis to the development of preventive measures and contribute to the understanding of the etiology of WD.

‘Studies on Bacterial Indicators and Pathogen Vibrio parahaemolyticus in Cochin Backwater

Microbiological studies on the incidence, behaviour, activity and ecological implications of marine micro~organisms, particularly microbial pathogens in coastal waters and estuaries exhibit the increasing concern and awareness of environmental impacts on health and wealth. Marine microbiologists have been active in investigating on the distribution, kinds of organisms and their activity in the environment. However, informations on the effect of environment on the ecology or on the distribution (spatial/temporal) of microbial comunity and competition among groups inhabiting the ecosystem are sparE§L Estuarine environment are complex with respect to diversity of habitats, variation in physicochemical parameters and contamination by terrestrial bacterial species. Being the organisms of‘public health significance, ecological studies on total coliforms, faecal coliforms, faecal streptococci, §. ggli and X. parahaemolyticus have great relevance as studies of these types would provide a wealth of information to environmentalists and to fishery industry. In order to evalé%e the status, role and significance of potentially hazardous bacterial species in natural environment it is necessary to monitor the ecology of such organisms systematically in relation to physico-chemical parameters

Alphitobius diaperinus spp como veiculador de Clostridium perfringens em granjas avícolas do interior paulista - Brasil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Clostridium perfringens em rações e águas fornecidos a frangos de corte em granjas avícolas do interior paulista: Brasil

Através de métodos bacteriológicos convencionais, avaliou-se a contaminação por Clostridium perfringens na ração e água utilizadas na alimentação e dessedentação de frangos de corte em diferentes regiões avícolas do interior paulista. C. perfringens esteve presente em 42 e 30% das amostras de ração e águas analisadas, respectivamente. As médias das contagens foram 6,7 x 10-2UFC mL para as amostras de água e 3,69 x 10-2UFC g para as de rações. As altas freqüências e contagens de C. perfringens verificadas nas rações e nas águas podem estar associadas à falta de higiene geral na manipulação e armazenamento dos mesmos. Sugere-se o monitoramento periódico da presença de C. perfringens nestas fontes, com a finalidade de evitar tal patógeno, em vista que o mesmo pode causar um surto de enterite necrótica levando, assim a grandes prejuízos na produção avícola.

Isolamento, identificação, avaliação da capacidade de adesão in vitro e efeito imunomodulatório de lactobacillus isolados de aves

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Avaliacão das técnicas de cultivo microbiológico e soroaglutinação rápida em cartão com e se 2-Mercaptoetanol da Brucelose canina

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Salmonella spp. e Clostridium perfringens em farinhas de origem animal utilizadas na fabricação de rações e avaliação de aditivo na inibição de patógeno

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Salmonella spp. in meat-type quails (Coturnix coturnix coturnix) in the State of São Paulo, Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Pathogens in animal meal and the use of Salmex (R) in the elimination of Clostridium perfringens

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Isothermal loop-mediated amplification (LAMP) for diagnosis of contagious bovine pleuro-pneumonia.

BACKGROUND Contagious Bovine Pleuropneumonia (CBPP) is the most important chronic pulmonary disease of cattle on the African continent causing severe economic losses. The disease, caused by infection with Mycoplasma mycoides subsp. mycoides is transmitted by animal contact and develops slowly into a chronic form preventing an early clinical diagnosis. Because available vaccines confer a low protection rate and short-lived immunity, the rapid diagnosis of infected animals combined with traditional curbing measures is seen as the best way to control the disease. While traditional labour-intensive bacteriological methods for the detection of M. mycoides subsp. mycoides have been replaced by molecular genetic techniques in the last two decades, these latter approaches require well-equipped laboratories and specialized personnel for the diagnosis. This is a handicap in areas where CBPP is endemic and early diagnosis is essential. RESULTS We present a rapid, sensitive and specific diagnostic tool for M. mycoides subsp. mycoides detection based on isothermal loop-mediated amplification (LAMP) that is applicable to field conditions. The primer set developed is highly specific and sensitive enough to diagnose clinical cases without prior cultivation of the organism. The LAMP assay detects M. mycoides subsp. mycoides DNA directly from crude samples of pulmonary/pleural fluids and serum/plasma within an hour using a simple dilution protocol. A photometric detection of LAMP products allows the real-time visualisation of the amplification curve and the application of a melting curve/re-association analysis presents a means of quality assurance based on the predetermined strand-inherent temperature profile supporting the diagnosis. CONCLUSION The CBPP LAMP developed in a robust kit format can be run on a battery-driven mobile device to rapidly detect M. mycoides subsp. mycoides infections from clinical or post mortem samples. The stringent innate quality control allows a conclusive on-site diagnosis of CBPP such as during farm or slaughter house inspections.

Survey for papillomatous digital dermatitis in Australian dairy cattle

Objective To determine whether Treponema-associated papillomatous digital dermatitis (PDD) occurs in Australian dairy cattle. Design Mail-out questionnaire and histological and bacteriological examination of biopsy tissue from suspect PDD lesions. Procedure The questionnaire was mailed to 375 veterinarians to evaluate their knowledge of PDD, determine if they had observed the disease in Australian dairy cattle, and to request biopsy material from suspicious cases. Biopsies were examined for histological and bacteriological evidence of PDD, including for the presence of spirochaetes. Results Eighty-eight replies to the questionnaire were received (23.5%). Of 52 respondents who were aware of PDD as a possible cause of lameness, 26 reported observing the condition in Australian cattle. Of 32 respondents who were unaware of the condition, 6 reported observing lesions that might have been PDD. The majority of reports of PDD-like lesions came from the southern Australian states, the condition occurring during periods of high rainfall and proving responsive to topical or parenteral application of antimicrobials. Biopsies from five erosive lesions showed histological similarity to PDD whereas biopsies from five proliferative lesions were consistent with chronic inflammation, fibroma or cutaneous papilloma. The presence of spirochaetes was not demonstrated in any of the lesions by histological or bacteriological methods. Conclusion Anecdotal reports and analysis of biopsy material confirm that a condition similar to PDD does occur sporadically in dairy cattle in southern Australia. However, this condition has so far not been shown to be associated with the presence of spirochaetes in the lesions.

Standard methods of milk analysis, bacteriological and chemical.

Bibliography at end of each chapter except three.

Environmental impact on the bacteriological quality of domestic water supplies in Lagos, Nigeria

OBJECTIVE: To assess the impact of town planning, infrastructure, sanitation and rainfall on the bacteriological quality of domestic water supplies. METHODS: Water samples obtained from deep and shallow wells, boreholes and public taps were cultured to determine the most probable number of Escherichia coli and total coliform using the multiple tube technique. Presence of enteric pathogens was detected using selective and differential media. Samples were collected during both periods of heavy and low rainfall and from municipalities that are unique with respect to infrastructure planning, town planning and sanitation. RESULTS: Contamination of treated and pipe distributed water was related with distance of the collection point from a utility station. Faults in pipelines increased the rate of contamination (p<0.5) and this occurred mostly in densely populated areas with dilapidated infrastructure. Wastewater from drains was the main source of contamination of pipe-borne water. Shallow wells were more contaminated than deep wells and boreholes and contamination was higher during period of heavy rainfall (p<0.05). E. coli and enteric pathogens were isolated from contaminated supplies. CONCLUSIONS: Poor town planning, dilapidated infrastructure and indiscriminate siting of wells and boreholes contributed to the low bacteriological quality of domestic water supplies. Rainfall accentuated the impact.

Clinical and Bacteriological Survey of Diabetic Foot Infections in Lisbon

AIMS: An epidemiological survey of diabetic foot infections (DFIs) in Lisbon, stratifying the bacterial profile based on patient demographical data, diabetic foot characteristics (PEDIS classification), ulcer duration and antibiotic therapy. METHODS: A transversal observational multicenter study, with clinical data collection using a structured questionnaire and microbiological products (aspirates, biopsies or swabs collected using the Levine method) of clinically infected foot ulcers of patients with diabetes mellitus (DM). RESULTS: Forty-nine hospitalized and ambulatory patients were enrolled in this study, and 147 microbial isolates were cultured. Staphylococcus was the main genus identified, and methicillin-resistant Staphylococcus aureus (MRSA) was present in 24.5% of total cases. In the clinical samples collected from patients undergoing antibiotic therapy, 93% of the antibiotic regimens were considered inadequate based on the antibiotic susceptibility test results. The average duration of an ulcer with any isolated multi-drug resistant (MDR) organism was 29 days, and previous treatment with fluoroquinolones was statistically associated with multi-drug resistance. CONCLUSIONS: Staphylococcus aureus was the most common cause of DFIs in our area. Prevalence and precocity of MDR organisms, namely MRSA, were high and were probably related to previous indiscriminate antibiotic use. Clinicians should avoid fluoroquinolones and more frequently consider the use of empirical anti-MRSA therapy.