Ten-year assessment of a cancer fast-track programme to connect primary care with oncology: reducing time from initial symptoms to diagnosis and treatment initiation.

BACKGROUND: Cancer is the second leading cause of mortality worldwide. Integrating different levels of care by implementing screening programmes, extending diagnostic tools and applying therapeutic advances may increase survival. We implemented a cancer fast-track programme (CFP) to shorten the time between suspected cancer symptoms, diagnosis and therapy initiation. PATIENTS AND METHODS: Descriptive data were collected from the 10 years since the CFP was implemented (2009-2019) at the Clinico-Malvarrosa Health Department in Valencia, Spain. General practitioners (GPs), an oncology coordinator and 11 specialists designed guidelines for GP patient referral to the CFP, including criteria for breast, digestive, gynaecological, lung, urological, dermatological, head and neck, and soft tissue cancers. Patients with enlarged lymph nodes and constitutional symptoms were also considered. On identifying patients with suspected cancer, GPs sent a case proposal to the oncology coordinator. If criteria were met, an appointment was quickly made with the patient. We analysed the timeline of each stage of the process. RESULTS: A total of 4493 suspected cancer cases were submitted to the CFP, of whom 4019 were seen by the corresponding specialist. Cancer was confirmed in 1098 (27.3%) patients: breast cancer in 33%, urological cancers in 22%, gastrointestinal cancer in 19% and lung cancer in 15%. The median time from submission to cancer testing was 11 days, and diagnosis was reached in a median of 19 days. Treatment was started at a median of 34 days from diagnosis. CONCLUSIONS: The findings of this study show that the interval from GP patient referral to specialist testing, cancer diagnosis and start of therapy can be reduced. Implementation of the CFP enabled most patients to begin curative intended treatment, and required only minimal resources in our setting.

Using multi-criteria risk ranking methodology to select case studies for a generic risk assessment framework for exotic disease incursion and spread through Europe.

We present a novel approach of using the multi-criteria pathogen prioritisation methodology as a basis for selecting the most appropriate case studies for a generic risk assessment framework. The approach uses selective criteria to rank exotic animal health pathogens according to the likelihood of introduction and the impact of an outbreak if it occurred in the European Union (EU). Pathogens were evaluated based on their impact on production at the EU level and international trade. A subsequent analysis included criteria of relevance to quantitative risk assessment case study selection, such as the availability of data for parameterisation, the need for further research and the desire for the case studies to cover different routes of transmission. The framework demonstrated is flexible with the ability to adjust both the criteria and their weightings to the user's requirements. A web based tool has been developed using the RStudio shiny apps software, to facilitate this.

Atención primaria en los estudios médicos de pregrado / Primary care in undergraduate medical studies

La atención de salud debe ser ambulatoria, y la hospitalización solo cuando sea indispensable, la causa son la urbanización y el aumento de la expectativa de vida. La tendencia comienza en 1945 en Inglaterra, la Declaración de Alma Ata en 1978 de OMS. En Latino América 1975 y en Venezuela en 1979. Se creó la Especialidad Médica MEDICINA FAMILIAR, el primer posgrado fue en el Centro Ambulatorio de Caricuao del IVSS por Carmen Cedraro de Carpio y Rafael Anselmi 1982. La OPS dirigió la ejecución y el desarrollo. Describimos la estructura curricular Medicina Familiar. En 16 países estudiados hay docencia de APS en pregrado. Hoy debe haber un Médico General-Familiar por cada 500 familias o 2 500 personas. Necesitamos 12 000 médicos con posgrado en Medicina Familiar. Son las conclusiones y recomendaciones: 1. La Atención Primaria de Salud (APS) es el eje en la conservación de la salud, la prevención y el tratamiento. 2. La APS debe junto con la Historia Médica Electrónica ser enseñada en el Pregrado de la Facultad Medicina. 3. Creen la OPS y la OMS, que debe ser desempañada por los Municipios junto con la Educación Preescolar artículo 178 aparte 5 de la Constitución Nacional. 4. La Academia Nacional de Medicina de Venezuela debe participarlo a: 1. Las Facultades y Escuelas de Medicina, 2. Sociedades Científicas de la Especialidades Médicas, y 3. Autoridades Gremiales Colegios de Médicos y Federación Médica (AU)

Non-invasive short-term assessment of retinoids effects on human skin in vivo using multiphoton microscopy.

BACKGROUND: The occlusive patch test developed for assessing topical retinoids activity in human skin has been extended as a short-term screening protocol for anti-ageing agents. In this model, biopsies are performed at the end of the occlusion period for morphological and immuno-histochemistry analysis. Multiphoton microscopy is a recent non-invasive imaging technique that combined with image processing tools allows the in vivo quantification of human skin modifications. OBJECTIVE: To validate with gold standards of anti-ageing that are retinoids, the relevance of multiphoton microscopy for kinetic and quantitative assessment in this model. METHODS: Twenty women, aged 50-65 years, were enrolled. Retinol 0.3% (RO) and Retinoic acid 0.025% (RA) were applied to the dorsal photo-damaged side of their forearm under occlusive patches for 12 days. A patch alone was applied to a third area as control. Evaluation was performed at day D0, D12 (end of treatment), D18 and D32 using multiphoton microscopy. Epidermal thickness, normalized area of the dermal-epidermal junction (DEJ) and melanin density were estimated using 3D image processing tools. RESULTS: Main significant results are: Epidermal thickening at D12, D18 and D32 with RO and at D12, D18 with RA vs. baseline and vs. CONTROL: Increased DEJ undulation at D32 with RO and at D12 with RA vs. baseline and vs. CONTROL: Decreased melanin content with RO (at D12 and D18 vs. baseline and at D32 vs. baseline and vs. control) and with RA (at D12 vs. baseline). CONCLUSIONS: This study shows that multiphoton microscopy associated to specific 3D image processing tools allows cutaneous effects induced by topical retinoids in this in vivo model to be non-invasively detected, quantified and followed over time. This innovative approach could be applied to the evaluation of other active compounds.

Treatment of childhood acute lymphoblastic leukemia in central America: a lower-middle income countries experience.

BACKGROUND: Five Asociación de Hemato-Oncología de Centroamérica (AHOPCA) countries have used an adapted BFM-based protocol for childhood acute lymphoblastic leukemia (ALL). PROCEDURE: In the AHOPCA-ALL 2008 protocol, patients were stratified by age, white blood cell count, immunophenotype, central nervous system involvement, day 8 prednisone response, and morphologic bone marrow response to induction therapy. Patients at Standard Risk (SR) received a three-drug induction regimen, a reinduction phase, and maintenance with protracted intrathecal therapy. Those at Intermediate (IR) and High Risk (HR) received, in addition, daunorubicin during induction therapy, a consolidation phase and two or three reinduction phases respectively. RESULTS: From August 2008 through July 2012, 1,313 patients were enrolled: 353 in SR, 548 in IR, 412 in HR. During induction therapy, 3.0% of patients died, 2.7% abandoned treatment, 1.1% had resistant ALL, and 93.2% achieved morphological complete remission (CR). Deaths and abandonment in first CR occurred in 2.7% and in 7.0% of patients, respectively. The relapse rate at a median observation time of 2.1 years was 15.0%. At 3 years, the event-free survival (EFS) and overall survival (OS), with abandonment considered as an event, were 59.4% (SE 1.7) and 68.2% (SE 1.6). Three-year EFS was 68.5% (SE 3.0), 62.1% (SE 2.6), and 47.8% (SE 3.2) for SR, IR, and HR groups. Adolescents had a significantly higher relapse rate (P = 0.001). CONCLUSIONS: This experience shows that common international studies are feasible in lower-middle income countries. Toxic deaths, abandonment of treatment, and relapses remain major obstacles to the successful treatment. Alternative treatment strategies may be beneficial.

Assessment of phenylurea herbicides sorption on various Mediterranean soils affected by irrigation with wastewater.

The retention values of two herbicides, chlorotoluron and isoproturon, in five Mediterranean soils were assessed by two different approaches, a dynamic method, using a batch technique (BT) and a static method, using a soil saturated paste (SP). The SP method led in all cases to lower herbicide sorption when compared with BT, although pesticide distribution constants from both methods were linearly related for the set of used soils (R(2)⩾0.99) showing that both methods similarly reflected the behaviour of the different soils. Low-quality water, evaluated by employing recycled urban wastewater, did not modify herbicide sorption when compared with high quality water, in any soil and with any method.

Association of FcgR2a, but not FcgR3a, with inflammatory bowel diseases across three Caucasian populations.

BACKGROUND: The Fc receptors II and III (FcgR2a, and FcgR3a) play a crucial role in the regulation of the immune response. The FcgR2a*519GG and FcgR3a*559CC genotypes have been associated with several autoimmune diseases including systemic lupus erythematosus, rheumatoid arthritis, nephritis, and possibly to type I diabetes, and celiac disease. In a large multicenter, two-stage study of 6570 people, we tested whether the FcgR2a and FcgR3a genes were also involved in inflammatory bowel disease (IBD), which includes Crohn's disease (CD) and ulcerative colitis (UC). METHODS: We genotyped the FcgR2a*A519G and FcgR3a*A559C functional variants in 4205 IBD patients in six well-phenotyped Caucasian IBD cohorts and 2365 ethnically matched controls recruited from the Netherlands, Spain, and New Zealand. RESULTS: In the initial Dutch study we found a significant association of FcgR2a genotypes with IBD (P-genotype = 0.02); while the FcgR2a*519GG was more common in controls (23%) than in IBD patients (18%; odds ratio [OR] = 0.75; 95% confidence interval [CI] 0.61-0.92; P = 0.004). This association was corroborated by a combined analysis across all the study populations (Mantel-Haenszel [MH] OR = 0.84; 0.74-0.95; P = 0.005) in the next stage. The Fcgr2a*GG genotype was associated with both UC (MH-OR = 0.84; 0.72-0.97; P = 0.01) and CD (MH-OR = 0.84; 0.73-0.97; P = 0.01), suggesting that this genotype confers a protective effect against IBD. There was no association of FcgR3a*A559C genotypes with IBD, CD, or UC in any of the three studied populations. CONCLUSIONS: The FcgR2a*519G functional variant was associated with IBD and reduced susceptibility to UC and to CD in Caucasians. There was no association between FcgR3a*5A559C and IBD, CD or UC.

The feasibility of developing a risk assessment for the impact of climate change on the emergence of Crimean-Congo haemorrhagic fever in livestock in Europe: a review.

Crimean-Congo haemorrhagic fever virus (CCHFV) is one of the most widespread of all medically important arboviruses with ticks of the Hyalomma spp. serving as the main vectors. Infection of livestock by CCHFV serves as a route of exposure to humans, as a reservoir of disease and as a route of importation. This study discusses the pathways and data requirements for a qualitative risk assessment for the emergence of CCHFV in livestock in Europe. A risk map approach is proposed based on layers that include the potential routes of release (e.g. by migrating birds carrying infected ticks) together with the main components for exposure, namely the distributions of the tick vectors, the small vertebrate host reservoirs and the livestock. A layer on landscape fragmentation serves as a surrogate for proximity of livestock to the tick cycle. Although the impact of climate change on the emergence of CCHF is not clear, comparing the distribution of risk factors in each layer currently with those predicted in the 2080s with climate change can be used to speculate how potential high-risk areas may shift. According to the risk pathway, transstadial and/or transovarial transmission in the tick vector are crucial for CCHFV spread. Vector competence and tick vector switching, however, remain critical factors for CCHFV colonization of new regions in Europe. The species of migratory bird is also an important consideration in the release assessment with greater abundance and biodiversity of ground-dwelling birds in southern Europe than in northern Europe.

Ochratoxin A exposure assessment of the inhabitants of Lisbon during winter 2007/2008 through bread and urine analysis.

A survey on the occurrence of ochratoxin A (OTA) in 41 bread samples was carried out in the Portuguese capital, Lisbon. Maize (5) and wheat bread (36) and 43 representative urine samples from the Lisbon region were assayed for OTA levels using immunoaffinity column cleanup (IAC) and HPLC with fluorimetric detection (LC­FD). The percentage of OTA-positive samples was slightly higher for maize bread (80%) than wheat bread (70.8%), although, due to its higher consumption, the latter contributes more to OTA exposure, featuring a higher estimated daily intake (EDI). In the urine samples analyzed, both female and male residents displayed similarly high levels of OTA frequency and average contamination. In summary, OTA is a food contaminant of concern and may constitute a hazard for public health through consumption of cereal-based products.

An assessment of the effect of data partitioning on the performance of modelling algorithms for habitat suitability for ticks.

A comparison of the performance of five modelling methods using presence/absence (generalized additive models, discriminant analysis) or presence-only (genetic algorithm for rule-set prediction, ecological niche factor analysis, Gower distance) data for modelling the distribution of the tick species Boophilus decoloratus (Koch, 1844) (Acarina: Ixodidae) at a continental scale (Africa) using climate data was conducted. This work explicitly addressed the usefulness of clustering using the normalized difference vegetation index (NDVI) to split original records and build partial models for each region (cluster) as a method of improving model performance. Models without clustering have a consistently lower performance (as measured by sensitivity and area under the curve [AUC]), although presence/absence models perform better than presence-only models. Two cluster-related variables, namely, prevalence (commonness of tick records in the cluster) and marginality (the relative position of the climate niche occupied by the tick in relation to that available in the cluster) greatly affect the performance of each model (P < 0.05). Both sensitivity and AUC are better for NDVI-derived clusters where the tick is more prevalent or its marginality is low. However, the total size of the cluster or its fragmentation (measured by Shannon's evenness index) did not affect the performance of models. Models derived separately for each cluster produced the best output but resulted in a patchy distribution of predicted occurrence. The use of such a method together with weighting procedures based on prevalence and marginality as derived from populations at each cluster produced a slightly lower predictive performance but a better estimation of the continental distribution of the tick. Therefore, cluster-derived models are able to effectively capture restricting conditions for different tick populations at a regional level. It is concluded that data partitioning is a powerful method with which to describe the climate niche of populations of a tick species, as adapted to local conditions. The use of this methodology greatly improves the performance of climate suitability models.

Design of a monitoring network and assessment of the pollution on the Lerma river and its tributaries by wastewaters disposal.

While the 2005 progress report of the United Nations Millennium Development Goals stresses out the need of a dramatic increase in investment to meet the sanitation target in the third world, it is important to anticipate about some parallel negative impacts that may have this optimistic programme (extension of sewer networks without sufficient treatment works). Research was initiated on Lerma River (Mexico), subjected to many rejects disposal, to design a monitoring network and evaluate the impact of wastewaters on its water quality. The discharges was inventorized, geo-positioned with a GPS and mapped, while the physico-chemical characteristics of the river water, its tributaries and main rejects were evaluated. Microtox system was used as an additional screening tool. Along the 60 km of the High Course of Lerma River (HCLR), 51 discharges, with a diameter or width larger than 0.3 m (including 7 small tributaries) were identified. Based on the inventory, a monitoring network of 21 sampling stations in the river and 13 in the important discharges (>2 m) was proposed. A great similitude was found between the average characteristics of the discharges and the river itself, in both the wet and dry seasons. Oxygen was found exhausted (<0.5 mg/L) almost all along the high course of the river, with COD and TDS average levels of 390 and 1980 mg/L in the dry season, against 150 and 400 mg/L in the wet season. In the dry season, almost all the sites along the river revealed some toxicity to the bacteria test species (2.9 to 150 TU, with an average of 27 TU). Same septic conditions and toxicity levels were observed in many of the discharges. Four of the six evaluated tributaries, as well as the lagoon (origin of the river), were relatively in better conditions (2 to 8 mg/L D.O., TU<1) than for the Lerma, acting as diluents and renewal of the HCLR flow rate. The river was shown to be quite a main sewer collector. The high surface water contamination by untreated wastewaters that is depicted in this research should be taken into account in the Millennium Goals strategies, by promoting treatment plan works simultaneously, when sewer networks in the third world would extend.

Use of a three-day estimated food record, a 72-hour recall and a food-frequency questionnaire for dietary assessment in a Mediterranean Spanish population.

AIM: To assess the validity and the short-term reproducibility of a semi-quantitative, self-administrated food frequency questionnaire and a structured 72-hour recall in a Mediterranean Spanish population. METHODS: 44 free-living volunteers participated in the study. Macronutrient, vitamin and mineral intake, recorded on the food frequency questionnaire and the structured 72-h recall were compared with intakes derived from a three-day food record (reference method). Validity of the dietary assessment methods was further assessed by comparing urinary nitrogen, plasma vitamin C, plasma beta-carotene and whole blood glutathione peroxidase activity levels with the corresponding nutrient intakes from the questionnaires. The food frequency questionnaire and the 72-h recall were administered twice to assess the short term reproducibility. RESULTS: Pearson's correlation coefficients between urinary nitrogen, plasma vitamin C, plasma beta-carotene and whole blood glutathione peroxidase activity levels and the reported nitrogen, vitamin C, beta-carotene and selenium intakes were 0.26, 0.53, 0.17, 0.26 for the food frequency questionnaire; 0.41, 0.09, 0.34, 0.42 for the structured 72-h recall and 0.50, 0.54, 0.44, 0.38 for the three-day food record, respectively. The short term reproducibility of analysed nutrient intake showed average intra-class correlation coefficients of 0.91 and 0.69 for the food frequency questionnaire and the structured 72-h recall irrespectively. CONCLUSION: The food frequency questionnaire and the structured 72-h recall provide valid estimates of nutrient intake and could be used for dietary assessments.

Rotavirus-associated medical visits and hospitalizations in South America: a prospective study at three large sentinel hospitals.

BACKGROUND: Knowledge of the impact of rotavirus-associated disease on the health care systems of South America can aid in defining strategies for diagnosis, management and prevention. Up to date information on the impact of rotavirus disease in South America is scarce. AIM: To determine prospectively the impact of rotavirus disease as a cause of medical visits and hospitalizations at three large sentinel pediatric hospitals in Argentina, Chile and Venezuela. METHODS: A 2-year prospective surveillance for rotavirus-associated medical visits and hospitalizations was conducted during 1997 through 1998 at three large sentinel public hospitals, one each in Argentina, Chile and Venezuela. A common surveillance protocol was implemented at the three sites, and a representative number of nonbloody diarrhea stool samples from children <36 months of age were tested for rotavirus by enzyme-linked immunosorbent assay. RESULTS: For our target age group, acute diarrhea-associated medical visits/hospitalizations represented 41%/2%, 5%/6% and 9%/13% of all medical visits/all hospitalizations at the Argentinean, Chilean and Venezuelan sites, respectively (P < 0.001 for difference among the three sites). Rotavirus detection rates among a total of 5,801/1,256 medical visit/hospitalization diarrhea stool samples tested were 39%/71% in Argentina, 34%/47% in Chile and 29%/38% in Venezuela (P < 0.01 by chi square for difference among the three sites). Rotavirus was associated with a mean of 1.5, 1.8 and 3% of total medical visits and 1.6, 2.8 and 5% of hospitalizations among children <36 months of age at the Argentinean, Chilean and Venezuelan sites, respectively. Seasonality was evident for medical visits at all three sites (although less striking in Chile) with peak activity occurring between November and May. Rotavirus-associated hospitalizations had a marked peak in Venezuela, represented largely by short stays, but not in Argentina and Chile. CONCLUSIONS: Rotavirus was a significant cause of medical visits at all three sentinel sites. Rotavirus caused less hospitalizations than previously reported in Argentina and Chile. On the basis of our findings we estimate that approximately 106,000/ 21,000, 48,000/8,000 and 98,000/31,000 rotavirus-associated medical visits/hospitalizations occur yearly in Argentina, Chile and Venezuela, respectively.

[Thallium-201 scintigraphy and dobutamine echocardiography in the assessment of myocardial viability]. / Gammagrafía con talio-201 y ecocardiografía con dobutamina en la valoración de viabilidad miocárdica.

BACKGROUND: The possibility of differentiating viable from non-viable tissue among patients with severe coronary artery disease and severe left ventricular impairment entails relevant clinical and therapeutic implications since it may influence the indication of patient revascularization. To evaluate the presence of myocardial viability two techniques are available in the clinical setting: echocardiography with intravenous infusion of dobutamine and scintigraphy with myocardial perfusion with thalliem-201 by means of single-photon emission tomography. OBJECTIVE: To compare prospectively the value of these techniques for detecting viable myocardium. MATERIAL AND METHODS: Thirty-five patients with severe coronary disease and severe left ventricular dysfunction were included in the study. All patients underwent an echocardiogram using incremental doses of dobutamine, from 5 up to 40 micrograms/kg/min in three-minute periods. For thallium-201 scintigraphy the rest redistribution protocol with delayed images at 4 hours was used. The criteria for detecting viability were: a) for thallium-201, the presence of redistribution in delayed images and normal uptake at rest, and b) for dobutamine echocardiography, a sustained improvement in regional motion, biphasic response, and worsening. RESULTS: By considering the segmental improvement post-revascularization as "gold standard" of viability, the statistically significant variables in a logistic regression model and, therefore, predictors of segmental functional recovery were the biphasic response and the sustained response for dobutamine echocardiography and normal uptake at rest and redistribution in the delayed images for thallium-201. Taken together, the result was significant for the biphasic response of dobutamine echocardiography. CONCLUSIONS: The biphasic response with dobutamine echocardiography is the echocardiographic pattern that best predicts the functional recovery of the ischemic myocardium. A normal uptake and redistribution at four hours is the only scintigraphic pattern that can predict functional improvement. Of both patterns, the biphasic response is the best predictor of the functional recovery of the dysfunctional myocardium.

Validation of flow cytometry for assessment of viability and acrosomal integrity of dog spermatozoa and for evaluation of different methods of cryopreservation.

The aims of the present study were: (i) to validate the accuracy of flow cytometry for assessment of viability and acrosomal status of canine spermatozoa; and (ii) to evaluate the cryopreservation protocols currently used for dog spermatozoa using flow cytometry. Data obtained by flow cytometry analysis of fresh dog spermatozoa stained with carboxyfluorescein diacetate (CFDA) and propidium iodide, or with fluorescein isothiocyanate (FITC)-conjugated Pisum sativum agglutinin (PSA) and propidium iodide, were compared with those obtained by microscopic evaluation. The results demonstrated that flow cytometry is a precise method for evaluating the viability and acrosomal status of fresh samples of dog semen. A new triple staining procedure, using carboxy-SNARF-1, propidium iodide and FITC-PSA, was developed and was an efficient method for evaluating the following aspects of cryopreservation protocols for dog spermatozoa: (i) addition of 0.5% (v/v) Equex STM paste to a Tris-egg yolk-based extender; (ii) dilution of the semen in one or two steps; (iii) freezing semen by placing 0.5 ml straws horizontally above liquid nitrogen in a styrofoam box or lowering them vertically into a liquid nitrogen tank; (iv) thawing semen at two different rates; (v) packaging semen at different sperm concentrations; and (vi) diluting semen at different rates after thawing. The highest sperm survival and longevity was obtained when Equex was present in the semen extender, the semen dilution was performed in two steps to obtain a concentration of 2.0 x 10(8) spermatozoa ml-1, the freezing was carried out using the styrofoam box, the straws were thawed at 70 degrees C for 8 s and the semen was diluted 1:4 after thawing.

Accuracy and cost-effectiveness of a new strategy to screen for celiac disease in children with Down syndrome.

OBJECTIVES: To investigate the best approach to screen for celiac disease (CD) in patients with Down syndrome (DS). STUDY DESIGN: One hundred thirty-seven children with DS were followed up longitudinally. CD screening was offered in 1994, 1996, and 1999 by determination of serum immunoglobulin A-anti-endomysium antibodies (AEA). The HLA-DQA1*0501/DQB1*02 allelic combination known to be strongly positively associated with CD was typed. All IgA-AEA-positive children were given the opportunity to undergo a small bowel biopsy: if villous atrophy was found, the diagnosis of CD was established. RESULTS: CD was diagnosed in 11 (8%) children: 8 in 1994 and 3 in 1996. All of them carried the HLA-DQ alleles associated with CD. The presence of symptoms was not useful in discriminating which children could have CD. CONCLUSIONS: Screening once in a lifetime is not enough to detect CD in patients with DS. We propose a new, accurate, and cost-sparing 2-step strategy for screening, based on selection of the individuals with potential CD by HLA-DQ typing and on longitudinal serologic CD screening in this selected group.

Lyme borreliosis habitat assessment.

Tick ecologists throughout Europe provided descriptions of Lyme borreliosis habitats according to a standardised format and data for 105 habitats in 16 countries were received. The data showed that high risk situations, as defined by the presence of large numbers of B. burgdorferi sensu lato-infected nymphal I. ricinus, occur in heterogeneous deciduous woodland, usually with a recreational function and with a diverse fauna, usually including deer. Large numbers of ticks occurred in some other habitats, but infection prevalence was usually low. The situation for adult I. ricinus was similar but less clearly defined. Tick infection rates were found to be lower in western Europe than in the east, and the infection rate in I. persulcatus, the most easterly vector species, was markedly higher than in I. ricinus. In the vast majority of habitats the infection rate in adult I. ricinus was greater than in nymphs. Larvae were rarely found to be infected.

Viability assessment of dog spermatozoa using flow cytometry.

The percentages of living and dead spermatozoa in fresh dog semen samples were assessed by means of a dual staining technique using carboxifluorescein diacetate (CFDA) and propidium iodide (PI). Two ejaculates were obtained from dogs, each ejaculate was divided into 4 aliquots, and different proportions of freeze-killed cells were added to each aliquot. Data obtained by flow cytometry analysis of each sample were compared with those obtained by the microscopic evaluation under epifluorescence illumination and by phase-contrast microscopy evaluation of the samples stained with eosin-nigrosin. Regression analysis was used to compare the 3 methods for membrane integrity assessment of canine spermatozoa, and high correlation coefficients were found between the flow cytometry procedure and the 2 microscopy techniques. The results from this study validate the use of flow cytometry as a precise method for assessing the viability of dog spermatozoa.