Integrative transcriptome and proteome analyses define marked differences between Neospora caninum isolates throughout the tachyzoite lytic cycle.

Neospora caninum is one of the main causes of transmissible abortion in cattle. Intraspecific variations in virulence have been widely shown among N. caninum isolates. However, the molecular basis governing such variability have not been elucidated to date. In this study label free LC-MS/MS was used to investigate proteome differences between the high virulence isolate Nc-Spain7 and the low virulence isolate Nc-Spain1H throughout the tachyzoite lytic cycle. The results showed greater differences in the abundance of proteins at invasion and egress with 77 and 62 proteins, respectively. During parasite replication, only 19 proteins were differentially abundant between isolates. The microneme protein repertoire involved in parasite invasion and egress was more abundant in the Nc-Spain1H isolate, which displays a lower invasion rate. Rhoptry and dense granule proteins, proteins related to metabolism and stress responses also showed differential abundances between isolates. Comparative RNA-Seq analyses during tachyzoite egress were also performed, revealing an expression profile of genes associated with the bradyzoite stage in the low virulence Nc-Spain1H isolate. The differences in proteome and RNA expression profiles between these two isolates reveal interesting insights into likely mechanisms involved in specific phenotypic traits and virulence in N. caninum. SIGNIFICANCE: The molecular basis that governs biological variability in N. caninum and the pathogenesis of neosporosis has not been well-established yet. This is the first study in which high throughput technology of LC-MS/MS and RNA-Seq is used to investigate differences in the proteome and transcriptome between two well-characterized isolates. Both isolates displayed different proteomes throughout the lytic cycle and the transcriptomes also showed marked variations but were inconsistent with the proteome results. However, both datasets identified a pre-bradyzoite status of the low virulence isolate Nc-Spain1H. This study reveals interesting insights into likely mechanisms involved in virulence in N. caninum and shed light on a subset of proteins that are potentially involved in the pathogenesis of this parasite.

Vaccines for bovine neosporosis: current status and key aspects for development.

Bovine neosporosis is a worldwide concern due to its global distribution and great economic impact. Reproductive failure in cattle due to abortion leads to major economic losses associated with the disease. Currently, there is no treatment or vaccine available against abortion or transmission caused by Neospora caninum infection in cattle. However, vaccination is considered the best measure of control against bovine neosporosis. Several host and parasite factors can influence the dynamics of the infection in bovines. Moreover, the availability of well-defined infection models is a key factor for the evaluation of vaccine candidates. However, working with cattle is not easy due to difficult handling, facilities and costs, and therefore, 'more affordable' models could be used for screening of promising vaccines to establish proof of concept. So far, live-attenuated vaccines have shown good efficacy against exogenous transplacental transmission; however, they have relevant disadvantages and associated risks, which render inactivated or subunit vaccines the best way forward. The identification of novel potential targets and vaccines, and the application of innovative vaccine technologies in harmonized experimental animal models, will accelerate the development of an effective vaccine against bovine neosporosis.

A longitudinal study of verotoxin-producing Escherichia coli in two dairy goat herds.

A longitudinal study was conducted on two dairy farms to investigate the pattern of shedding of verotoxin-producing Escherichia coli (VTEC) in goats. Faecal samples were taken from 20 goat kids once weekly during the first 4 weeks of life and then once every month for the next 5 months of life, and from 18 replacement animals and 15 adults once every month for 12 months. The proportion of samples containing VTEC was higher for replacement animals and adults (85.7% and 78.7%, respectively) than for goat kids (25.4%). About 90% of the VTEC colonies isolated from healthy goats belonged to five serogroups (O33, O76, O126, O146 and O166) but the most frequent serogroups of these isolates, except one, were different in the two herds studied. E. coli O157:H7 was found in three goat kids on only one occasion. None of the VTEC isolates, except the three E. coli O157:H7 isolates, was eae-positive. The patterns of shedding of VTEC in goat kids were variable, but, in contrast, most of the replacement animals and adults were persistent VTEC shedders. Our results show that isolates of VTEC O33, O76, O126, O146 and O166 are adapted for colonising the intestine of goats but that, in contrast, infection with VTEC O157:H7 in goats seems to be transient.

Prevalence and associated factors for gallstone disease: results of a population survey in Spain.

We conducted a cross-sectional survey to determine the prevalence of gallstone disease (gallstone or cholecystectomy) in a random sample of the adult population of Guadalajara, Spain. The sample stratified by age and sex was drawn from the municipal census. Stratum sample sizes were proportional to population sizes and to the expected prevalence rates calculated through a meta-analysis of the European literature. The screening protocol included a gallbladder ultrasonography, a questionnaire on personal and family history, a physical examination, and a blood sample for biochemical determinations. The response rate was 61.2%. The overall prevalence of gallstone disease was 9.7% (95% CI, 7.3-12.0). Prevalence was higher (but not statistically significant) in women (11.5%; 95% CI, 8.2-14.7) than in men (7.8%; 95% CI, 4.6-11.1). After controlling for confounding by multiple logistic regression, increasing age, body mass index, dyspeptic symptoms, smoking habit, and use of hypolipidemic drugs were positively associated with gallstone disease. Total serum cholesterol and alcohol consumption were negatively associated.

Spleen abscess caused by Eikenella corrodens.

A case is reported of splenic abscess due to Eikenella corrodens, a gram-negative rod which is found as part of normal flora in human mucous surfaces. A 64-year-old man presented with fever, chills, anorexia and abdominal pain. Abdominal ultrasound examination showed a perisplenic fluid collection which was considered to be either blood or a subcapsular spleen abscess. The presence of a splenic abscess was later confirmed during surgery and a splenectomy was performed. Splenic purulent material and blood cultures yielded Eikenella corrodens. The patient received cefotaxime for 19 days and was discharged asymptomatic.

[Complications in diabetic patients on dialysis: experiences in 10 years of treatment using 3 technics]. / Complicaciones de los pacientes diabéticos en diálisis: experiencia en diez años de tratamiento con tres técnicas.

In treated diabetic patients with terminal renal failure we have examined the complications arising from the dialysis technique used. 55 patients were followed up for 1.279 months. Out of the latter, 24 were on hemodialysis (HD) 33 on continuous ambulatory peritoneal dialysis (CAPD) and 9 on intermittent peritoneal dialysis (IPD). Intraperitoneal insulin in patients on CAPD best controlled glycemia. Antihypertensive therapy was not necessary in 86%, 66%, and 46% of the patients on CAPD, IPD and HD respectively (CAPD vs HD, p less than 0.001). Acute myocardial infarction was more common in patients on HD (33%) (p less than 0.05). 22% of the patients on IPD had limb amputation (IPD vs CAPD-HD, p less than 0.05). The type of dialysis method used does not seem to influence the course of the retinopathy. The neuropathy behaved in a similar manner in those on CAPD and on HD when followed up for two years. Vascular complications were frequent in those on HD with one thrombotic event every 9.7 months/patient and a new vascular access every 7.4 months/patient. Peritonitis was the main complication of CAPD and IPD which was found more frequent in the former and longer hospitalization in the latter. The election of the dialysis technique in diabetic patients can directly influence the glycemia levels and blood pressure reading, as well as heart complications and those due to the method used. However they seem to influence less the neuropathy, retinopathy and peripheral vasculopathy.

[Morbidity and mortality of diabetic patients on dialysis in a 10-year program: value of combined treatment]. / Morbilidad y mortalidad de pacientes diabéticos en diálisis en diez años de programa: validez del tratamiento combinado.

Morbidity and mortality derived from the employed techniques after ten year experience in treating patients afflicted with end-stage diabetic nephropathy by means of dialysis are evaluated. Hemodialysis (HD) was applied to 24 patients, continuous ambulatory peritoneal dialysis (OCPD) was employed in 33 patients and intermittent peritoneal dialysis (IPD) in nine patients. Cumulated experience with each technique was 529, 644, and 107 months, respectively. Ten patients were treated with a combination of two or three techniques. Hospitalization rate in patients receiving CAPD was 32 days/year, peritonitis was the most frequent condition leading to admittance (54.8%), followed by cardiovascular alterations (14.4%) and gastrointestinal complications (10.5%). On hemodialyzed patients, hospitalization rate was 24.7 days/year and complications derived from vascular access were the most frequent admittance cause (34%), followed by cardiovascular complications (20.4%) and hypertension (11.3%). Peritonitis (45%), vascular complications (15%) and metabolic impairment (15%) were the most frequent causes of admittance in IPD patients; hospitalization rate was 88 days/year. Actuarial survival in patients on CAPD was 92% during first year, 79% during second, and 64% during third and fourth years. On hemodialyzed patients, 90% of patients survived during first year and 80%, 48%, and 36% during second, third, and fourth years, respectively. On IPD, survival during first year was 63% and 12% at 18 months. By combinating two or the three methods, survival was 100% at 12 months and 88% at 4 years. Although CAPD as isolated technique may offer to diabetic patients a longer survival and HD a lower hospitalization rate, combined treatment (HD-CAPD-IPD) may provide a survival matchable to that achieved in renal transplantation.