Protozoan parasites and free-living amoebae contamination in organic leafy green vegetables and strawberries from Spain.

In this study, the presence of Acanthamoeba spp., Blastocystis sp., Cryptosporidium spp., Cyclospora cayetanensis, Entamoeba histolytica, Giardia sp., Toxoplasma gondii and Vermamoeba vermiformis was assessed in organic leafy green vegetables (lettuce, spinach, cabbage) and fruits (strawberry), which are usually consumed raw. A total of 110 organic samples were collected in Valencia (Spain). Protozoa were concentrated before detection by immunofluorescence (Cryptosporidium spp. and Giardia sp.) or real-time qPCR (Acanthamoeba spp., Blastocystis sp., C. cayetanensis, E. histolytica, T. gondii and V. vermiformis). The most abundant protozoa in organic vegetables and berry fruits were Acanthamoeba (65.5%), followed by T. gondii (37.2%), V. vermiformis (17.3%), C. cayetanensis (12.7%), Cryptosporidium spp. (6.8%), Blastocystis sp. (1.8%) and Giardia sp. (1.7%). E. histolytica was not found in any of the organic samples. Thus, results showed that consumers can be exposed to protozoan parasites by consuming organic vegetables and berry fruits. This is the first report in Spain describing the presence of the protozoan pathogens Acanthamoeba spp., Blastocystis sp., C. cayetanensis, T. gondii and V. vermiformis, Cryptosporidium spp. and Giardia sp. in organic fresh produce. The results of this research will help determine the risk of foodborne protozoan parasites on organic leafy greens and strawberries that are available at local markets.

Impact of mobilization in patients with short-term mechanical circulatory support such as Levitronix® CentriMag as a bridge to heart transplantation.

INTRODUCTION: Despite the benefits of mobilisation in the critical patient, the evidence in patients with Levitronix® CentriMag as a bridge to heart transplantation (HT) is scarce. The objective of this study is to analyze the impact of mobility on these patients. METHODS: Retrospective observational study of patients who received a HT with Levitronix® CentriMag admitted between 2010 and 2019 to a tertiary hospital. Degree of mobility and nutritional status were assessed at the time of HT. Outcomes including infections, length of hospital admission and mortality were evaluated. RESULTS: 27 patients were included and divided in two groups according to degree of mobility (22 with low mobility and 5 with high mobility). 90-day survival after HT was 63.6% in patients with low mobility and 80% in high mobility group; no statistically significant differences were observed. No differences were observed regarding ICU discharge after HT at 30 days. Nevertheless, lower albumin levels were observed in low mobility group (24,5 g/L (IQR: 23-30) vs 33 g/L (IQR: 26-36); p = .029). Invasive mechanical ventilation (IMV) post HT was longer in patients with low mobility (p = .014). There were no significant differences in appearance of pressure ulcers, or post HT infections among mobility groups. CONCLUSIONS: Patients with high mobility had a shorter time of IMV and a better nutritional status. No complications were observed associated to mobility. No differences were observed between the degree of mobility and 90-day mortality, ICU stay or post HT adverse events.

Simultaneous detection of less frequent waterborne parasitic protozoa in reused wastewater using amplicon sequencing and qPCR techniques.

Waterborne parasitic protozoa (WPP) infections have a worldwide distribution and are a source for epidemic and endemic human diseases. Although a variety of protozoa are commonly detected in wastewater and cited as causative agents of outbreaks, effluents from wastewater treatment plants (WWTPs) used for irrigation can contain other pathogenic protozoa that are not currently being controlled. The lack of control on a routine basis using rapid and sensitive methods to detect these parasites in water may keep them under-recognized. This study focused on using molecular tools, 18 S rRNA amplicon-based sequencing and qPCR, to characterize WPP distribution in wastewater samples from urban WWTPs used for irrigation. A total of eight wastewater samples (from secondary and tertiary disinfection treatment effluents) were collected. Potentially pathogenic protozoa identified by 18 S rRNA sequencing and/or qPCR in the analyzed samples included Acanthamoeba spp., Blastocystis sp., Entamoeba coli, Entamoeba dispar, Entamoeba hartmanni, Giardia intestinalis assemblage A and Toxoplasma gondii Positive results by qPCR were in non-quantifiable levels. Blastocystis sp. was the most represented protozoa among the sequences retrieved from the amplicon sequencing. Blastocystis ST1 and ST2 were the most abundant subtypes among the obtained OTUs. Moreover, Blastocystis sp. ST3, ST4, ST6 and ST8 were also detected, although in lower abundances. Results of this study showed that WWTP effluents used for irrigation can provide a source of WPP.

Multiple identification of most important waterborne protozoa in surface water used for irrigation purposes by 18S rRNA amplicon-based metagenomics.

Understanding waterborne protozoan parasites (WPPs) diversity has important implications in public health. In this study, we evaluated a NGS-based method as a detection approach to identify simultaneously most important WPPs using 18S rRNA high-throughput sequencing. A set of primers to target the V4 18S rRNA region of WPPs such as Cryptosporidium spp., Giardia sp., Blastocystis sp., Entamoeba spp, Toxoplasma sp. and free-living amoebae (FLA) was designed. In order to optimize PCR conditions before sequencing, both a mock community with a defined composition of representative WPPs and a real water sample inoculated with specific WPPs DNA were prepared. Using the method proposed in this study, we have detected the presence of Giardia intestinalis, Acanthamoeba castellanii, Toxoplasma gondii, Entamoeba histolytica and Blastocystis sp. at species level in real irrigation water samples. Our results showed that untreated surface irrigation water in open fields can provide an important source of WPPs. Therefore, the methodology proposed in this study can establish a basis for an accurate and effective diagnostic of WPPs to provide a better understanding of the risk associated to irrigation water.

Standard and new faecal indicators and pathogens in sewage treatment plants, microbiological parameters for improving the control of reclaimed water.

This study involved collaboration between three centres with expertise in viruses, bacteria and protozoa. The focus of the research was the study of the dissemination and removal of pathogens and faecal indicators in two sewage treatment plants (STP1 and STP2) using tertiary treatments. Samples were collected over a period of five months through the sewage treatment processes. Analysis of the samples revealed that the plants were not efficient at removing the faecal indicators and pathogens tested during the study. From entry point (raw sewage) to effluent level (tertiary treatment effluent water), the experimental results showed that the reduction ratios of human adenoviruses were 1.2 log10 in STP1 and 1.9 log10 in STP2. Whereas for Giardia spp. and Cryptosporidium spp. the reduction ratios were 2.3 log10 for both pathogens in STP1, and 3.0 and 1.7 log10 in STP2, respectively. Furthermore, the presence of faecal indicators and pathogens at different sampling points was evaluated revealing that the tested pathogens were present in reclaimed water. Human adenovirus and Arcobacter spp. showed positive results in infectivity assays for most of the tertiary effluent water samples that comply with current legislation in Spain. The pathogens detected must be evaluated using a risk assessment model, which will be essential for the development of improved guidelines for the re-use of reclaimed water.

LNA probes in a real-time TaqMan PCR assay for genotyping of Giardia duodenalis in wastewaters.

AIMS: This study describes an approach for genotyping Giardia cysts obtained from wastewater treatment plants (WTPs) in Spain using real-time PCR (qPCR) in combination with immunomagnetic beads. METHODS AND RESULTS: A 50-cycle amplification of a 74-bp fragment of the Giardia beta-giardin gene was adopted from a previous qPCR method. Additionally, two locked nucleic acid (LNA) probes were designed (LNA P434 P1 for assemblage A and LNA P434 H3 for assemblage B). All 16 wastewater samples analysed were positive with the immunofluorescence assay (IFA). Assemblage A was detected in all WTP samples using primer-LNA probe P434 P1 set. Giardia duodenalis identification was confirmed by PCR-RFLP analysis and sequencing of the beta-giardin gene in the water samples found positive by IFA and qPCR. Among the 16 assemblage A isolates that were sequenced, two subtypes were identified; 11 corresponded to the A2 subgenotype, whereas three corresponded to the subgenotype A3. A mixture of subgenotypes was found in the remaining two isolates. CONCLUSIONS: The newly developed qPCR assays were able to discern G. duodenalis assemblages A and B in wastewater. SIGNIFICANCE AND IMPACT OF THE STUDY: The real-time PCR assays provided a rapid method for detection and one-step genotyping of G. duodenalis from wastewater samples, and its application would contribute to understanding the distribution and abundance of G. duodenalis assemblages A and B in wastewater.

New investigational drugs for the management of acute heart failure syndromes.

Acute heart failure syndromes (AHFS) enclose a broad spectrum of conditions with different clinical presentations, heart failure history, pathophysiology, prognosis and treatment. AHFS represent a major public health problem because of their high prevalence, high rates of mortality and readmissions and significant healthcare costs, and a therapeutic challenge for the clinicians because management strategies vary markedly. Traditionally used drugs for the treatment of AHFS, including diuretics, vasodilators and positive inotropics, improve clinical signs and symptoms as well as hemodynamics, but present important limitations, as they fail to reduce and may even increase in-hospital and postdischarge mortality, especially in patients with coronary artery disease. Thus, we need new pharmacological agents to not only improve signs and symptoms and cardiac performance, but also improve both short- and long-term outcomes (hospitalizations/survival). In the last decade, significant efforts have been made to identify new therapeutic targets involved in the genesis/progression of AHFS and to develop new therapeutic strategies that may safely improve outcomes. As a result, several new families of drugs have been developed and are currently studied in experimental models and in Phase II and III clinical trials, in an attempt to define their efficacy and safety profiles as well as their precise role in the treatment of AHFS patients. This review firstly analyzes the main clinical applications and limitations of conventional drugs, and then focuses on the mechanisms of action and effects of recently approved drugs and of new investigational agents on signs, symptoms, hemodynamics and outcomes in AHFS patients.

Neumonía cavitada por tifus murino / No disponible

No disponible

Assessment of toxicity of a glyphosate-based formulation using bacterial systems in lake water.

A new Aeromonas bioassay is described to assess the potential harmful effects of the glyphosate-based herbicide, Roundup, in the Albufera lake, a protected area near Valencia. Viability markers as membrane integrity, culturability and beta-galactosidase production of Aeromonas caviae were studied to determine the influence of the herbicide in the bacterial cells. Data from the multifactor analysis of variance test showed no significant differences (P>0.05) between A. caviae counts of viability markers at the studied concentrations (0, 50 and 100 mg l-1 of glyphosate). The effects of Roundup on microbial biota present in the lake were assessed by measuring the number of indigenous mesophilic Aeromonas in presence of different amounts of the herbicide at 0, 50 and 100 mg l-1 of glyphosate. In samples containing 50 and 100 mg l-1 of glyphosate a significant (P<0.05) increase in Aeromonas spp. counts and accompanying flora was observed. The acute toxicity of Roundup and of Roundup diluted with Albufera lake water to Microtox luminescent bacterium (Vibrio fischeri) also was determined. The EC50 values obtained were 36.4 mg l-1 and 64.0 mgl-1 of glyphosate respectively. The acidity (pH 4.5) of the herbicide formulation was the responsible of the observed toxicity.

[Uncommon etiology of gastrointestinal bleeding: duodenal metastases from renal cell carcinoma]. / Causa poco habitual de hemorragia digestiva alta: metástasis duodenal de adenocarcinoma renal.

Because of its unpredictable behavior, renal cell carcinoma is one of the most controversial neoplasms. On the one hand, patients frequently show metastases at diagnosis because of its slight manifestations, while on the other, the neoplasm can remain stable after nephrectomy and can then metastasize many years later. When this happens, the metastases usually involve more than 2 organs. The most frequent sites of metastases are the lung and lymph nodes, followed by the bones and liver, while duodenal involvement is rare. Indeed, intestinal metastases are found in only 2% of autopsies and of these, renal cell carcinoma metastases account for 7.1%. We present a case of a solitary late recurrence presenting as upper gastrointestinal bleeding 19 years after nephrectomy for clear cell renal carcinoma.

Causa poco habitual de hemorragia digestiva alta: metástasis duodenal de adenocarcinoma renal / Uncommon etiology of gastrointestinal bleeding: duodenal metastases from renal cell carcinoma

El adenocarcinoma renal es una de las neoplasias más controvertidas debido a su comportamiento impredecible. En muchas ocasiones los pacientes presentan enfermedad metastásica en el momento del diagnóstico dada su escasa expresividad clínica, y en otras, tras una nefrectomía, el tumor puede permanecer estable y metastatizar años después. Cuando la enfermedad metastásica ocurre, suele ser múltiple y los lugares afectados con mayor frecuencia son el pulmón, los ganglios, el hueso y el hígado, mientras que la afectación intestinal es inusual. De hecho, la afectación intestinal como zona de implantación de metástasis es infrecuente y representa sólo el 2% de las autopsias. De este pequeño porcentaje, el 7,1% corresponde a metástasis de adenocarcinoma renal. Presentamos un caso de metástasis única duodenal que se manifestó como hemorragia digestiva alta al cabo de 19 años de una nefrectomía por carcinoma renal de células claras

Because of its unpredictable behavior, renal cell carcinomais one of the most controversial neoplasms. On the one hand, patients frequently show metastases at diagnosis because of its slight manifestations, while on the other, the neoplasm can remain stable after nephrectomy and can then metastasize many years later. When this happens, the metastases usually involve more than 2 organs. The most frequent sites of metastases are the lung and lymph nodes, followed by the bones and liver, while duodenal involvement is rare. Indeed, Intestinal metastases are found in only 2% of autopsies and of these, renal cell carcinoma metastases account for 7.1%. We present a case of a solitary late recurrence presenting as upper gastrointestinal bleeding 19 years after nephrectomy for clear cell renal carcinoma

[Wilson's disease with severe neurological manifestations: response to trientine plus zinc therapy]. / Enfermedad de Wilson con afectación neurológica grave: respuesta al tratamiento combinado con trientina y acetato de cinc.

In patients with Wilson's disease and neurological manifestations, treatment with D-penicillamine can cause worsening of neurological symptoms, usually in the first few weeks of treatment. Because the neurological damage can be severe and irreversible, the use of D-penicillamine is controversial, and several authors believe that it should be avoided. Studies of the use of ammonium tetrathiomolybdate as an alternative chelating agent for the initial treatment of neurologic Wilson's disease are still in the experimental phase. Published experience on the simultaneous use of trientine, another chelating agent, and zinc, which blocks intestinal absorption of copper, is promising but limited. We present the case of a 17 year-old boy with severe neurologic Wilson's disease that had first presented six years previously. The patient showed a complete recovery after six months of treatment with a combination of trientine and zinc acetate.

Quantitative determination of total coliforms and Escherichia coli in marine waters with chromogenic and fluorogenic media.

This study compared the performance of LMX(R) broth (LMX), Chromocult Coliform(R) agar (CC) and Chromocult Coliform agar plus cefsulodin (10 microg ml-1) (CC-CFS), with standard methods multiple tube fermentation (MTF), for the enumeration of total coliforms and Escherichia coli from marine recreational waters. LMX and CC are two media designed to concurrently detect total coliform (TC) bacteria and E. coli by the specific action of beta-galactosidase (total coliforms) and beta-glucuronidase (E. coli). Overall results for the TC test showed that LMX, CC and MTF recovered 2.63, 1.95 and 1.90 times as many TCs as CC-CFS, respectively. Data from the multiple range test showed significant differences (P < 0.05) between TC counts on CC-CFS and LMX. The traditional MTF was less sensitive for E. coli enumeration. However, there was no statistically significant differences between LMX, CC, CC-CFS and the MTF method for E. coli enumeration. Background interference was reduced on CC-CFS and the counts obtained reflected more accurately the number of TCs. Therefore, the contribution of beta-galactosidase positive, non coliform bacteria (Aeromonas spp. and Vibrio spp.) to TC counts should not be neglected.

Comparison and recovery of Escherichia coli and thermotolerant coliforms in water with a chromogenic medium incubated at 41 and 44.5 degrees C.

This study compared the performance of a commercial chromogenic medium, CHROMagarECC (CECC), and CECC supplemented with sodium pyruvate (CECCP) with the membrane filtration lauryl sulfate-based medium (mLSA) for enumeration of Escherichia coli and non-E. coli thermotolerant coliforms (KEC). To establish that we could recover the maximum KEC and E. coli population, we compared two incubation temperature regimens, 41 and 44.5 degrees C. Statistical analysis by the Fisher test of data did not demonstrate any statistically significant differences (P = 0.05) in the enumeration of E. coli for the different media (CECC and CECCP) and incubation temperatures. Variance analysis of data performed on KEC counts showed significant differences (P = 0.01) between KEC counts at 41 and 44.5 degrees C on both CECC and CECCP. Analysis of variance demonstrated statistically significant differences (P = 0.05) in the enumeration of total thermotolerant coliforms (TTCs) on CECC and CECCP compared with mLSA. Target colonies were confirmed to be E. coli at a rate of 91.5% and KEC of likely fecal origin at a rate of 77.4% when using CECCP incubated at 41 degrees C. The results of this study showed that CECCP agar incubated at 41 degrees C is efficient for the simultaneous enumeration of E. coli and KEC from river and marine waters.

A rapid procedure for the isolation of plasmid DNA from environmental bacteria.

The INSTA-MINI-PREP method, a rapid protocol for plasmid DNA extraction, was originally developed to prepare plasmid DNA from 1 to 3 ml miniprep Escherichia coli cultures. Direct extraction of plasmid DNA is achieved by a two-phase solution which is separated by centrifugation in the presence of the INSTA-PREP gel barrier material. This method has been successfully tested on various environmental Salmonella strains, although it was not suitable for Pseudomonas aeruginosa and enterococci strains. The INSTA-MINI-PREP method is a new alternative procedure to screen plasmid contents of Salmonella and E. coli strains rapidly and easily.

Differential susceptibility of aeromonads and coliforms to cefsulodin.

Volumn 62, no. 6, p. 1885, Abstract, line 3: "For 102 of 129 Aeromonas isolates" should read "For 102 of 130 Aeromonas isolates." Page 1887, column 1, lines 16 and 17: "and for 13 of 21 coliform isolates" should read "and for 13 of 94 coliform isolates." [This corrects the article on p. 1885 in vol. 62.].