Vaginal microbial communities from synchronized heifers and cows with reproductive disorders.

AIM: To evaluate changes in the resident microbial population in the cranial vaginal mucosa induced by a progesterone-releasing intravaginal device (PRID) compared to the vaginal microbiota of cows with reproductive disorders. METHODS AND RESULTS: Vaginal discharge was evaluated by clinical examination and a Vaginitis Diagnosis Score was performed by exfoliative cytology. All samples classified as positive and some classified as negative by clinical evaluation were later diagnosed as positive for vaginitis by cytological analysis. Bacterial diversity profiles were performed by PCR-DGGE and clustered according to the reproductive health status of the specimens, revealing a correspondence between the structures of the communities in the vagina and the clinical profile. Representative bands from each group were sequenced and identified as Ruminococcus sp., Dialister sp., Escherichia sp./Shigella sp., Virgibacillus sp., Campylobacter sp., Helcoccoccus sp., Staphylococcus sp., Bacillus sp., Actinopolymorpha sp., Exiguobacterium sp., Haemophilus sp./Histophilus sp., Aeribacillus sp., Porphyromonas sp., Lactobacillus sp. and Clostridium sp. CONCLUSION: Our results contribute to the knowledge of the vaginal microbiome in synchronized heifers showing positive or negative clinical vaginitis. SIGNIFICANCE AND IMPACT OF THE STUDY: This study contributes to the understanding of a dynamic vaginal colonization by bacterial consortiums during the synchronization with a widely used PRID protocol. Also, the results reveal the presence of well-known metritis-related pathogens as well as emerging uterine opportunistic pathogens. The provided information will allow to carry out further studies to elucidate functional roles of these native micro-organisms in the bovine reproductive tract.

Survival and beneficial properties of lactic acid bacteria from raniculture subjected to freeze-drying and storage.

AIM: To evaluate the effect of freeze-drying and storage conditions on the viability and beneficial properties of lactic acid bacteria (LAB) for raniculture. METHODS AND RESULTS: Lactococcus lactis CRL 1584, L. lactis CRL 1827, Lactococcus garvieae CRL 1828 and Lactobacillus plantarum CRL 1606 viability under different conditions was studied. 10% lactose and 5% skim milk + 5% lactose were excellent lyoprotectants, but 5% skim milk + 5% lactose and whey protein concentrated (WPC) or WPC + sugars were the lower cost lyoprotective options. The effect of temperature depended on both lyoprotectants and storage time. Thus, for Lactococcus, skim milk, skim milk + sucrose and WPC + sucrose were selected for lyophilization and storage at 4°C and skim milk + lactose for 25°C. For Lact. plantarum CRL 1606, the best lyoprotectants for lyophilization and storage at 4°C were milk + sugars and WPS + sucrose and, at 25°C, skim milk + sucrose. CONCLUSIONS: Lactic acid bacteria viability after freeze-drying was strain-specific and depended on the lyoprotectant used. Highest viability was obtained when stored at 4°C, and the beneficial properties remained stable for 18 months independently of storage temperature. SIGNIFICANCE AND IMPACT OF THE STUDY: The studies reported for the first time in this work are of primary interest to obtain dried bacteria to be included in beneficial products for raniculture.

Probiotic properties of vaginal lactic acid bacteria to prevent metritis in cattle.

AIMS: The isolation of bovine vaginal lactic acid bacteria (LAB) and the screening of their beneficial properties to select those that could be used as probiotics in the prevention of bovine metritis were performed. METHODS AND RESULTS: Out of 76 Lactobacillus sp. and seven Streptococcus sp. strains, a small number showed high- and medium hydrophobicity when the microbial adhesion to hydrocarbons method (MATH) was applied. In the agar plate diffusion test, a large number of strains inhibited vaginal bovine Escherichia coli 99/14 and human E. coli. This inhibition was due to acid. Only a few strains inhibited Actinomyces pyogenes 96/393, a pathogen isolated from bovine metritis. This inhibition remained after neutralization. The taxonomic identification of the selected strains was carried out by an amplified ribosomal DNA restriction analysis (ARDRA). Most of the strains were identified as Lactobacillus fermentum, a few as Lactobacillus gasseri and one as Lactobacillus rhamnosus. CONCLUSIONS: Bovine vaginal lactobacilli strains have differential surface properties. The strains selected are capable of inhibiting specific metritis pathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results can be applied for future studies to design a probiotic product to prevent metritis in dairy postpartum cows.

Group A streptococcal bacteremia: outcome and prognostic factors.

In the last two decades, an increase in the incidence of invasive group A streptococcus (GAS) infections has been reported. The aim of this study was to determine the clinical and epidemiological characteristics and the natural history of GAS bacteremias at our hospital by performing a retrospective study of all cases of GAS bacteremia diagnosed at our University hospital from 1994 to 2003. We reported 42 cases of GAS bacteremia (27 men, mean age 42.3 +/- 31.6 years). None had more than one episode and four cases were nosocomial. The mean annual incidence rate was 1.01 cases per 100,000 population. An increase in the incidence but not in severity of GAS bacteremia was observed in the last 5-year period (p<0.001). The rates were highest in young children and the elderly and those with underlying medical conditions; 73.8% of patients had some underlying chronic illness, and the most relevant conditions included peripheral vascular disease and diabetes mellitus. Mortality was high and the worst outcome corresponded to elderly patients with streptococcal toxic shock syndrome (STSS). Thirty patients (71.4%) had a disruption in the integrity of the skin barrier, 14 (33.3%) were immunocompromised patients and 6 patients (14.3%) were intravenous drug users. A source of the bacteremia was noted in 38 patients (90.5%), with skin and soft tissue infection being the major portals of entry. Twelve patients (28.6%) fulfilled the STSS criteria. All strains were susceptible to penicillin and vancomycin. Resistance to erythromycin was 21.4% and to ciprofloxacin was 17.5%. The global mortality rate was 28.6%. Only STSS was significantly associated with increased mortality in the multivariate analysis.

Meningoencefalitis virales / Viral meningoencephalitis

Entre el 1§ de enero de 1983 y el 30 de septiembre de 1999 atendimos a 117 niños con diagnóstico de meningoencefalitis viral, en la Sección de Infectocontagiosos del Hospital Infantil Universitario "La Fe" (Valencia, España). En 27 casos fue posible averiguar el agente etiológico: 20 por herpesvirus (14 Herpes simplex, 4 Epstein-Barr y 2 citomegalovirus) y 7 por enterovirus, quedando los 90 restantes sin demostrar.Se comentan los aspectos clínicos, diagnósticos y terapéuticos. Ante la posibilidad, aún remota, de una etiología herpética, se recomienda iniciar el tratamiento con aciclovir, dada su eficacia y el mal pronóstico de esta meningoencefalitis.En general, la evolución de nuestros casos fue favorable, con pocas secuelas, sobre todo en forma de convulsiones controladas con tratamiento anticomicial. Sólo ha habido que lamentar un caso de muerte por meningoencefalitis herpética.

[Human immunodeficiency virus and resistance]. / Virus de la immunodeficiencia humana y resistencias.

It has been demonstrated that following prolonged treatment with reverse transcriptase inhibitors, especially with nucleoside analogs, resistant viral strains appear that show mutations in the nucleotides of the gene that codifies reverse transcriptase. These mutations cause changes in amino acids, which gradually increases resistance. In this study we evaluated the applicability af the LiPA HIV-1 RT test in determining the presence of mutations in this gene that can lead to resistance to antiretroviral treatments.Twenty-one samples were studied without previous knowledge of the antiviral treatments that the patients had received. No mutations in the HIV-1 stains from untreated patients were found. In the other patients who had undergone various treatment regimens, the results obtained With the LiPA HIV-1 RT made in possible to retrospectively identify the different nucleoside analogs they had been treated with. The most frequently found mutations were those that lead to a certain degree of resistance to AZT, followed by those related to resistance to ddl, which in effect in this order were the drugs used in the treatment of the patients. However, the LiPA HIV-1 RT test has certain limitations. It can only detect mutations that can be recognized by its sequences. As such, including sequences that would make it possible to study the possible genotypic variations in the gene that codifies the protease of HIV-1 would be helpful, given the importance of the protease inhibitors in clinical practice. In any case, direct sequencing continues to be standard method as it allows us to obtain full information on the genes that need to be studied.

Economic aspects of a general vaccination against invasive disease caused by Haemophilus influenzae type b (Hib) via the experience of the Children's Hospital La Fe, Valencia, Spain.

With the aim of studying whether a general vaccination against invasive disease caused by Haemophilus influenzae type b (Hib) is economically profitable bearing in mind the efficacy and safety of the vaccine, its price and the global cost that this disease has in our area, a review is conducted of patients admitted due to invasive disease caused by Hib in the Children's Hospital La Fe, Valencia, born between 1984 and 1993. They total 100, 63 who have meningitis. In the 81 cases (56 with meningitis) born between 1984 and 1990 (years that can be regarded as "closed" since all the patients were younger than 5 years of age) the total cost has been calculated for hospitalization, care during the acute phase, care for the sequelae (6 severe and 7 mild) and death (5 cases). The mean annual cost of care can be calculated at 62 million pesetas, without making an economic valuation of the loss of life, and at 205 million pesetas taking this factor into account. The annual cost of vaccinating the 7000 babies under one year of age and falling within the Hospital's catchment area, on the basis of a vaccination pattern of three doses (at 2, 4 and 6 months) or four doses (at 2, 4, 6 and 15 months) would amount to 63 or 84 million pesetas, normal price to public (not covered by National Health Service), and 40 or 51 million pesetas if acquired by National Health Service. As a conclusion we can state that, even from the economic point of view, without quantifying the cost of the loss of life, a public general anti-Hib vaccination would be profitable in our area since it would mean an administration cost lower than that of the care required by patients. This is without taking into account the fact that emotional, family and social serious disturbances would also be avoided due to hospitalization, sequelae and deaths caused by a disease which is today perfectly preventable.

Risk/benefit in the treatment of children with imipenem-cilastatin for meningitis caused by penicillin-resistant pneumococcus.

A new case of pneumococcal meningitis in a child successfully treated with imipenem-cilastatin after failure of standard antibiotics is presented. Seizures, as the major adverse effect, were easily treated and no sequelae were later observed. The difficulty in distinguishing between seizures due to meningitis and those due to imipenem is discussed. We conclude that imipenem-cilastatin is a good alternative therapy in children's meningitis caused by pneumococcus resistant to standard antibiotics.

[Delay in childhood tuberculosis detection as a negative factor in the anti-tuberculosis struggle]. / El retraso en la detección de tuberculosis infantil como factor negativo en la lucha antituberculosa.

Childhood morbidity due to tuberculosis infection in Spain is not diminishing at the expected rate in relation with available knowledge and facilities. An important factor is the delay in diagnosis, which evaluation constitutes the objective of this publication. Said delay is due to several causes, patient does not consult on time (mean value between onset of symptoms and consult is 34.3 days), or physician does not think in tuberculosis as first diagnosis, which meant an added mean delay of 7.6 days more. These delays mean an important detriment for the patient, and one of the objectives in the anti-tuberculosis struggle is to shorten delays through information to the lay public and health professionals on the highly important current issue of tuberculosis.

[Fascioliasis in children. A study of 10 cases]. / Fasciolosis en los niños. Estudio de 10 casos.

These are the observation found in ten children with fascioliasis diagnosis at the Servicio de Parasitología, of the Instituto Nacional de Pediatría, from 1979 to 1990; six children in scholar age; one prescholar and three teenagers; only one was female. Four of them lined at the State of Mexico; three at Morelos; one at Puebla, another one at Oaxaca and the other one at Mexico City. In nine of the watercress eaten was confirmed. The most important clinic manifestations were: fever, weight lose, paleness, hepatomegaly, hiporexia, right hypochondrium pain and diarrhoea. Diagnosis was established by: counterimmunoelectrophoresis, indirect haemagglutination and there were found eggs by simple sedimentation, Ritchie's method and microscopic study of duodenal sample. Leukocytes counts were between 11,000 and 34,000/mm3. Eosinophils were to 77% with 24,430 totals. Only in three of them haemoglobin was found under 9 g/dL. All of them had hypergammaglobulinemia. In nine patients the alkaline phosphatase was found in higher levels; only in three of them, transaminase, oxalacetic and glutamic piruvic were found in higher levels. The most effective drug in the treatment was dehidroemetine.