Co-occurrence of the cephalosporinase cepA and carbapenemase cfiA genes in a Bacteroides fragilis division II strain, an unexpected finding.

BACKGROUND: Bacteroides fragilis, an anaerobic gut bacterium and opportunistic pathogen, comprises two genetically divergent groups (or divisions) at the species level. Differences exist both in the core and accessory genomes and the beta-lactamase genes, with the cephalosporinase gene cepA represented only in division I and the carbapenemase gene cfiA only in division II. METHODS: Multidrug resistance in a clinical B. fragilis strain was examined by whole-genome sequencing. RESULTS: Strain CNM20200260 carried the antimicrobial resistance genes cepA, cfiA2, ant(6'), erm(F), mef(En2), est(T), tet(Q) and cat(A), along with 82-Phe mutation in gyrA (together with 47 amino acid changes in gyrA/B and parC/parE). bexA/B and other efflux pump genes were also observed. None of the detected insertion sequences was located upstream of cfiA2. The genome-based taxonomy coefficients (average nucleotide identity, DNA-DNA hybridization similarity and difference in genomic G + C%) with respect to genomes of the strains of B. fragilis division II and the novel species Bacteroides hominis (both cfiA-positive) met the criteria for CNM20200260 to belong to either species (>95%, >70% and <1%, respectively). No such similarity was seen with type strain NCTC 9343 or the representative genome FDAARGOS 1225 of B. fragilis (division I, cfiA-negative). Strain CNM20200260 harboured four out of nine Kyoto Encyclopedia of Genes and Genomes orthologues defined for division I and one of two defined for division II. CONCLUSIONS: This is the first description of the co-occurrence of cepA and cfiA in a Bacteroides strain, confirming the complexity of the taxonomy of this species.

Macroparasites of allis shad (Alosa alosa) and twaite shad (Alosa fallax) of the Western Iberian Peninsula Rivers: ecological, phylogenetic and zoonotic insights.

Samples of anadromous Alosa alosa (Clupeidae) (n = 163) and Alosa fallax (Clupeidae) (n = 223), caught in Western Iberian Peninsula Rivers from 2008 to 2013, were examined for buccal, branchial and internal macroparasites, which were identified using morphological and molecular methods. Alosa alosa were infected with Anisakis simplex s.s., Anisakis pegreffii, Hysterothylacium aduncum, Rhadinorhynchus pristis, Mazocraes alosae, Hemiurus appendiculatus, Ceratothoa italica and an unidentified ergasilid copepod. Ceratothoa italica represents a new host record for A. alosa. Alosa fallax were infected with A. simplex s.s., A. pegreffii, H. aduncum, H. appendiculatus, Clavellisa emarginata and an unidentified cymothoid isopod. This is the first report of C. italica, C. emarginata and M. alosae in the Iberian Peninsula. The phylogenetic positions of M. alosae, H. appendiculatus and C. emarginata were assessed using 18S and 28S ribosomal RNA (rRNA); our contributions provide a better understanding of the phylogenetic relationships within their groups. Qualitative and quantitative differences in the parasite faunas of these two shad species are consistent with different feeding strategies. The results provide information about host migration behaviour and transmission pathways through diet during the marine trophic phase of the shad's life cycle and their roles as paratenic or final hosts and transporters of parasites between seawater and freshwater environments. The zoonotic parasites A. simplex s.s. and A. pegreffii pose a risk for consumers or riverine mammals (e.g. European otter). The use of parasites as biological tags for shad stocks in Western Iberian Rivers could be a useful approach in multidisciplinary studies concerning fish stock delimitation and characterization.

Anisakis infection in allis shad, Alosa alosa (Linnaeus, 1758), and twaite shad, Alosa fallax (Lacépède, 1803), from Western Iberian Peninsula Rivers: zoonotic and ecological implications.

Spawning individuals of allis shad, Alosa alosa (Linnaeus, 1758), and twaite shad, Alosa fallax (Lacépède, 1803), were sampled from three rivers on the Atlantic coast of the Iberian Peninsula (Ulla, Minho, Mondego) during 2008 to 2013 to assess the presence of the zoonotic marine parasite Anisakis spp. larvae. The results revealed that both shad species were infected by third-larval stage Anisakis simplex s.s. and Anisakis pegreffii. The latter is reported in mixed infections in both shad species of Western Iberian Peninsula for the first time. In A. alosa, the prevalence of Anisakis infection can reach 100%, while in A. fallax, prevalence was up to 83%. Infected individuals of the former species also often contain much higher number of parasites in theirs internal organs and flesh: from 1 to 1138 Anisakis spp. larvae as compared to 1 to 121 larvae, respectively. In general, numbers of A. pegreffii were higher than those of A. simplex s.s. Our results suggest that in the marine environment of the Western Iberian Peninsula, both anadromous shad species act as paratenic hosts for A. simplex s.s. and A. pegreffii, thus widening the distribution of the infective nematode larvae from the marine to the freshwater ecosystem. This finding is of great epidemiological relevance for wildlife managers and consumers, considering the zoonotic and gastroallergic threats posed of these parasites.

Haematophagous feeding of newly metamorphosed European sea lampreys Petromyzon marinus on strictly freshwater species.

Approximately 6% of the freshwater living northern straight-mouth nase Pseudochondrostoma duriense in two Spanish rivers had attached post-metamorphic sea lamprey Petromyzon marinus. Minimum prey size was 16·1 cm fork length and 56·3 g mass. The condition factor of attacked P. duriense was 16% lower than that of unattacked conspecifics.

Brain abscesses caused by anaerobic microorganisms: a systematic review.

The objective of this study was to perform a systematic review of the characteristics, causative microorganisms and outcome of brain abscesses caused by anaerobic bacteria over the past 25 years. We reviewed studies on brain abscesses which included infection due to anaerobic microorganisms published between 1998 and 2022. We excluded reports with polymicrobial infections (more than 2 anaerobic bacteria isolated) and those that do not provide enough information to make comparisons, the reports with only one case of brain abscess due to anaerobes, as well as those focused on an only anaerobic bacterium. Also, we have excluded the cases in pediatric population. We searched the scientific literature through the Cochrane Library, EMBASE and PubMed/MEDLINE databases for studies of this condition. We finally included 28 studies with 6,167 patients, of which 715 (11.5%) were cases caused by anaerobic bacteria. There was a male predominance (70%) and mean age of 40.3 years. Most infections were monomicrobial (59.4%). The most common anaerobic microorganisms isolated were Bacteroides spp (43.4%) and Gram-positive anaerobic cocci (35.1%). Cases of brain abscesses caused by anaerobic bacteria were most frequent in Asia and Europe. The source of infection most frequent was otogenic in 84.6% followed by a neurosurgery procedure infection in 23% of patients. The main symptom observed was headache in 95.6% of patients followed by fever (69.5%). Surgical treatment was performed in 48 % of patients and the percentage of patients in whom antibiotic treatment was applied range 88.8% to 100%. The main limitation of this review is the non-inclusion of studies published before of 1998 in which MALDI-TOF MS system had not been introduced in the majority of laboratories for routine identification. The patient rate with isolation of anaerobic bacteria in brain abscesses is low, but these data could be underestimated mainly due to the fastidious nature of these microorganisms and the difficulties in the identification of some anaerobes.

Therapy-related myelodysplastic syndromes deserve specific diagnostic sub-classification and risk-stratification-an approach to classification of patients with t-MDS.

In the current World Health Organization (WHO)-classification, therapy-related myelodysplastic syndromes (t-MDS) are categorized together with therapy-related acute myeloid leukemia (AML) and t-myelodysplastic/myeloproliferative neoplasms into one subgroup independent of morphologic or prognostic features. Analyzing data of 2087 t-MDS patients from different international MDS groups to evaluate classification and prognostication tools we found that applying the WHO classification for p-MDS successfully predicts time to transformation and survival (both p < 0.001). The results regarding carefully reviewed cytogenetic data, classifications, and prognostic scores confirmed that t-MDS are similarly heterogeneous as p-MDS and therefore deserve the same careful differentiation regarding risk. As reference, these results were compared with 4593 primary MDS (p-MDS) patients represented in the International Working Group for Prognosis in MDS database (IWG-PM). Although a less favorable clinical outcome occurred in each t-MDS subset compared with p-MDS subgroups, FAB and WHO-classification, IPSS-R, and WPSS-R separated t-MDS patients into differing risk groups effectively, indicating that all established risk factors for p-MDS maintained relevance in t-MDS, with cytogenetic features having enhanced predictive power. These data strongly argue to classify t-MDS as a separate entity distinct from other WHO-classified t-myeloid neoplasms, which would enhance treatment decisions and facilitate the inclusion of t-MDS patients into clinical studies.

Update on the Diagnosis of Sexually Transmitted Infections. / Actualización en el diagnóstico de las infecciones de transmisión sexual.

Sexually transmitted infections (STIs) are one of the most frequent and universal Public Health problems. Health professionals should be aware of the possibility of STIs due to their high morbidity and the presence of sequelae. The delay in the diagnosis is one of the factors that justifies the difficulty to infections control. Diagnostic tests allow the introduction of aetiological treatment and also lead to treating symptomatic and asymptomatic patients more effectively, as well as to interrupt the epidemiological transmission chain without delay. In this review we have made an update of the main existing diagnostic methods for the more important STIs.

Typing and antimicrobial susceptibility of 134 Neisseria gonorrhoeae strains from Southern Spain.

OBJECTIVE: Last guidelines have recommended the introduction of dual antimicrobial therapy in order to avoid treatment failure. In the present report, the susceptibility to some antibiotics was evaluated, and the typing of Neisseria gonorrhoeae strains was performed. METHODS: Gonococcal isolates were tested for susceptibility according to the recommendations of both CLSI and EUCAST. A total of 134 isolates were typed by the NG-MAST technique. RESULTS: Seventy-two different N. gonorrhoeae types were found, and the most frequent types obtained were ST 1407, ST 14958, ST 7192, ST 13251 and ST 5405. If CLSI/EUCAST criteria were applied, a ST 9807 type was found nonsusceptible to ceftriaxone and cefixime (MIC 0.5 mg/L), and a ST 12800 type was found nonsusceptible only to cefixime (MIC 0.25 mg/L). When only EUCAST breakpoints were taken into account, three strains were also resistant to cefixime (MIC 0.25 mg/L) and three isolates were resistant to ceftriaxone (MIC 0.19, 0.16 and 0.25 mg/L, respectively). The majority of strains were resistant to ciprofloxacin (68.6%), and all N. gonorrhoeae strains were susceptible to spectinomycin; 9.7% of isolates were resistant to azithromycin. CONCLUSIONS: Molecular typing may be a useful tool to predict antimicrobial resistance. High rates of resistance to penicillin, tetracycline and ciprofloxacin were found in this area. It is highly recommended to carry out antimicrobial susceptibility in all gonorrhoea cases and to identify treatment failures to verify emerging resistance.

Whole-blastocyst culture followed by laser drilling technology enhances the efficiency of inner cell mass isolation and embryonic stem cell derivation from good- and poor-quality mouse embryos: new insights for derivation of human embryonic stem cell lines.

The optimization of human embryonic stem (hES) cell line derivation methods is challenging because many worldwide laboratories have neither access to spare human embryos nor ethical approval for using supernumerary human embryos for hES cell derivation purposes. Additionally, studies performed directly on human embryos imply a waste of precious human biological material. In this study, we developed a new strategy based on the combination of whole-blastocyst culture followed by laser drilling destruction of the trophoectoderm for improving the efficiency of inner cell mass (ICM) isolation and ES cell derivation using murine embryos. Embryos were divided into good- and poor-quality embryos. We demonstrate that the efficiency of both ICM isolation and ES cell derivation using this strategy is significantly superior to whole-blastocyst culture or laser drilling technology itself. Regardless of the ICM isolation method, the ES cell establishment depends on a feeder cell growth surface. Importantly, this combined methodology can be successfully applied to poor-quality blastocysts that otherwise would not be suitable for laser drilling itself nor immunosurgery in an attempt to derive ES cell lines due to the inability to distinguish the ICM. The ES cell lines derived by this combined method were characterized and shown to maintain a typical morphology, undifferentiated phenotype, and in vitro and in vivo three germ layer differentiation potential. Finally, all ES cell lines established using either technology acquired an aneuploid karyotype after extended culture periods, suggesting that the method used for ES cell derivation does not seem to influence the karyotype of the ES cells after extended culture. This methodology may open up new avenues for further improvements for the derivation of hES cells, the majority of which are derived from frozen, poor-quality human embryos.

Clinical and microbiologic characteristics of pleuro-pulmonary infection due to Streptococcus intermedius.

The clinical and microbiological characteristics of pleuro-pulmonary infection (PPI) caused by Streptococcus intermedius is described, including 6 cases in the literature and 9 cases handled at the present centre. Out of the 15 patients, 12 were male; mean age at diagnosis was 62.06 ± 15 years. Twelve had risk factors for S. intermedius infection such as alcoholism in 5 (35.7%) patients, periodontal disease in 3 (24.6%) cases, chronic obstructive pulmonary disease in 3 (24.6%), and diabetes mellitus in 2 (14.2%). Cough was present in 12 (80%) patients and chest pain and dyspnea in 9 (60%). The mean diagnosis interval was 34 days. The diagnosis was obtained from pleural fluid aspirate in 13 (86.6%) cases and from biopsy/tissue samples in 2. The most frequently antimicrobials used for treatment were ceftriaxone + levofloxacin. Ten patients cured with a combination of medical and surgical treatment and 2 patients died as a consequence of infection. The incidence of PPI caused by S. intermedius is increasing in our health area; drainage along with antibiotic therapy is recommended for treatment.

Clinical and microbiological findings of septic arthritis caused by Haemophilus parainfluenzae.

OBJECTIVES: To report a case of septic arthritis due to H. parainfluenzae and to review the clinical and microbiological characteristics of published case patients. PATIENTS AND METHODS: Data was collected on age, sex, infection localization, underlying risk factors, symptom onset-diagnosis interval, analytical findings, microbiological diagnosis, treatment, outcome, and follow-up of the present patient (presenting with septic arthritis of the pubic symphysis due to H. parainfluenzae) and those identified in a literature analysis. RESULTS: Data of 18 patients, including 17 reported case patients, was collected. Mean age at presentation was 51±9 years. Underlying diseases for septic arthritis were recorded in 11 patients. The infection site was the knee in eight patients, hip and/or acromioclavicular joint in five. Pain was observed in 15 patients and fever in 10; the mean symptom onset-diagnosis interval was 9.4 days. Diagnosis was obtained from synovial fluid aspirate in 12 patients and from blood cultures in four. Susceptibility of H. parainfluenzae strains was reported in 12 cases. Eight patients were treated with cephalosporins and 10 with penicillins. A favorable outcome was observed in 13 patients. CONCLUSIONS: Septic arthritis caused by H. parainfluenzae is a rare entity that requires a high level of suspicion before application of laboratory methods for rapid diagnosis and treatment.

Pleural effusion due to Parvimonas micra. A case report and a literature review of 30 cases.

The clinical and microbiological characteristics of infections caused by Parvimonas micra is described, including 30 cases in the literature and a new case handled at the present centre. Out of the 31 patients, 18 were male; mean age at diagnosis was 65.1 ± 13.0 years. Infection site was the vertebral spine in 14 patients and joints and heart valves in 5 each one; pain was present in all patients with articular localization and in almost all patients with vertebral involvement. The diagnosis was obtained from fluid aspirate or drainage in 13 cases and blood cultures in 11. In 8 cases, molecular techniques were also applied. The most frequently used antimicrobials were clindamycin, penicillin, amoxicillin and ceftriaxone. The outcome was positive with the medical treatment in 28 patients. P. micra infections are uncommon and requires a high index of suspicion.

Streptococcus agalactiae: prevention and vaccine development.

Streptococcus agalactiae, group B Streptococcus (SGB), is the most important cause of morbi-mortality among newborn population, and an important pathogen among immunossupressed adult patients. Despite the advances in the treatment and prevention of neonatal infections as a consequence of implementation of national and international recommendations for prevention of infection, there are still some improvements for the final control of the disease. In this sense, the vaccination against SGB could be an effective measure for the prevention of disease in those cases where intrapartum prophylaxis is not useful and in adult patients with risk factors for invasive infection due to SGB. This review summarizes the efforts made until now in order to establish the control of the infection, and brings some information on the current state-of-the art of vaccines against SGB, in which different strategies in their design have been used.

Embryonic stem cell bank: a work proposal.

Human embryonic stem cells (hESCs) have an unlimited capacity to proliferate by a self-renewal process and can be differentiated in the three germ layers, opening doors to new clinical therapies to replace missing or damaged cells. The number of research groups and projects using human stem cells has increased largely in the last 5 yr. The creation of stem cell banks is another important step to support the advance of research in this field. Banks must be operated within the strict regulatory famework of good manufacturing practices and good laboratory practices that assure the highest quality standards and must implement a quality system that complies with international quality systems standards. It may also be appropriate to aim at an accreditation in order to assure correct laboratory practices at all times. Stem cell banks should receive the lines previously derived by other groups and hESCs should be provided for groups that justify their use in a research project previously approved by an ethical committee. The assays generally accepted as typical of hESCs together with the microbiological analysis should be performed in order to assure a consistent, reliable, and safe line for the researchers. In this article, the Andalusian Stem Cell Bank proposes a model of a stem cell banking process in order to create a flow diagram of hESC lines and, following the international initiatives in stem cells research, to achieve the full characterization of cells and a standardization of protocols that would simplify the hESCs culture.

Determinants of parasite drug resistance in human lymphatic filariasis.

Infection with filarial nematodes remains endemic in several countries worldwide and some of these infections are commonly associated with severe disease. The elimination of lymphatic filariasis relies on drug administration using the three drugs currently available for treatment: diethylcarbamazine, albendazole, and ivermectin. However, development of drug resistance is a reported phenomenon. The issue of resistance to antihelminthics used in humans has become increasing importance since the global program to eliminate lymphatic filariasis is implemented in larger population groups and the duration of the program is increasing. Recently, ivermectin resistance has been reported in Ghana, and widespread of resistance to benzimidazole (such as albendazole) is present because specific mutations in the gene encoding ß-tubulin have been associated with drug resistance. Moreover, it is well known that diethylcarbamazine susceptibility is not 100% for lymphatic filariasis treatment. A review of the mechanisms of resistance to these antihelminthics is necessary in order to optimize the treatment for human lymphatic filariasis.