Testing the efficiency of capture methods for questing Hyalomma lusitanicum (Acari: Ixodidae), a vector of Crimean-Congo hemorrhagic fever virus.

Available methods to census exophilic tick populations have limitations in estimating true population size due to their inability to capture a high proportion of the actual tick population. We currently ignore the efficacy of these methods to capture questing Hyalomma spp. ticks, vectors of the Crimean-Congo hemorrhagic fever virus. To address the need of accurately estimating questing densities of Hyalomma spp., we designed a field experiment to test the efficacy of blanket dragging, blanket flagging, CO2-baited traps, and an ad hoc designed method, absolute surface counts, in capturing adult Hyalomma lusitanicum ticks from known numbers of preset fluorescent-marked ticks. The experiment was designed in 2 stages to estimate the point (1-day sampling) and cumulative (3-day serial sampling) efficacy of the methods under varying sampling effort and habitat. Tick survival, host interference, and weather effects on efficacy were controlled for in multiple regression models. There was high variability in method efficacy for capturing ticks, which was also modulated by effort, habitat, tick density, hosts, and soil temperature. The most effective method was absolute surface counts for both point estimates (39%) and cumulative efficacy (83%). CO2-baited traps reached a maximum efficacy of 37%, while blanket dragging and blanket flagging captured a maximum of the 8% of the marked ticks. Our results reveal the strengths and weaknesses of the different tick capture methods applied to adult H. lusitanicum and lay the groundwork for more accurate inferences about the true size of exophilic tick populations.

Insights into the spatiotemporal dynamics of West Nile virus transmission in emerging scenarios.

The incidence of West Nile fever (WNF) is highly variable in emerging areas, making it difficult to identify risk periods. Using clinical case records has important biases in understanding the transmission dynamics of West Nile virus (WNV) because asymptomatic infections are frequent. However, estimating virus exposure in sentinel species could help achieve this goal at varying spatiotemporal scales. To identify the determinants of inter-annual variation in WNV transmission rates, we designed a 15-year longitudinal seroepidemiological study (2005-2020) in five environmentally diverse areas of southwestern Spain. We modeled individual annual area-dependent exposure risk based on potential environmental and host predictors using generalized linear mixed models. Further, we analyzed the weight of predictors on exposure probability by variance partitioning of the model components. The analysis of 2418 wild ungulate sera (1168 red deer - Cervus elaphus - and 1250 Eurasian wild boar - Sus scrofa) with a highly sensitive commercial blocking ELISA identified an average seroprevalence of 24.9% (95% confidence interval (CI): 23.2-26.7%). Antibody prevalence was slightly higher in wild boar (27.5%; CI: 25.1-30.1%) than in deer (22.2%; CI: 19.8-24.7%). We observed a spatial trend in exposure, with higher frequency in the southernmost areas and a slight, although area-dependent, increasing temporal trend. Host-related predictors were important drivers of exposure risk. The environmental predictor with the highest weight was annual cumulative precipitation, while temperature variations were also relevant but with less weight. We observed a coincidence of spatiotemporal changes in exposure with the notification of WNF outbreaks in horses and humans. That indicates the usefulness of wild ungulates as sentinels for WNV transmission and as models to understand its spatiotemporal dynamics. These results will allow the development of more accurate predictive models of spatiotemporal variations in transmission risk that can inform health authorities to take appropriate action.

Risk Factors for Exposure of Wild Birds to West Nile Virus in A Gradient of Wildlife-Livestock Interaction.

West Nile virus (WNV) transmission rate is shaped by the interaction between virus reservoirs and vectors, which may be maximized in farm environments. Based on this hypothesis, we screened for WNV in wild birds in three scenarios with decreasing gradient of interaction with horses: (i) the farm (A1); (ii) the neighborhood (A2); and (iii) a wild area (A3). We captured wild birds and analyzed their sera for WNV antibodies by blocking ELISA and micro-virus neutralization test. Flavivirus infections were tested with generic and specific PCR protocols. We parameterized linear mixed models with predictors (bird abundance and diversity, vector abundance, vector host abundance, and weather quantities) to identify Flavivirus spp. and WNV exposure risk factors. We detected a low rate of Flavivirus infections by PCR (0.8%) and 6.9% of the birds were seropositive by ELISA. Exposure to Flavivirus spp. was higher in A1 (9%) than in A2 and A3 (5.6% and 5.8%, respectively). Bird diversity was the most relevant predictor of exposure risk and passerines dominated the on-farm bird community. Our results suggest that measures deterring the use of the farm by passerines should be implemented because the environmental favorability of continental Mediterranean environments for WNV is increasing and more outbreaks are expected.

Initial Impact of Perinatal Loss on Mothers and Their Partners.

(1) Background: Perinatal Loss affects one in ten women worldwide. It is known to have a deep impact on the physical and psychological wellbeing of the mother. Moreover, there is a lack of information in regard to gender differences. The role of culture, environment, personal characteristics, and gender is yet to be determined in most reports; (2) Objective and Methods: Our aim is to study the initial impact of perinatal losses in an unselected sample of couples, focusing on gender differences. We conducted a longitudinal prospective study with 29 mothers and 17 fathers. Upon discharge from the hospital, they filled out the Edinburgh Postnatal Depression Scale (EPDS), among others. After one-month post-loss, they performed the EPDS and the Short Version of the Perinatal Grief Scale. We used descriptive statistics for the sample and non-parametric tests for the comparison of gender; (3) Results: We found no gender differences in initial depressive symptoms, nor in depressive symptoms, perinatal grief symptoms, or grief level (total scores or complicated grief) one month after the loss; (4) Conclusions: we need to better understand the psychological evolution of couples in cases of perinatal loss without falling into preconceived ideas about the influence of gender.

The Common Mosquito (Culex pipiens) Does Not Seem to Be a Competent Vector for Hepatitis E Virus Genotype 3.

An experimental infection approach was used to estimate the competence of the common mosquito, Culex pipiens, for hepatitis E virus replication and transmission, using an isolate of hepatitis E virus genotype 3 of human origin in varying infectious doses. The experimental approach was carried out in biosafety level 2 conditions on three batches of 120 Cx. pipiens females, each using an artificial feeding system containing the virus in aliquots of fresh avian blood. Mosquitoes from each batch were collected 1, 7, 14, and 21 days post-infection (dpi) and dissected. The proboscis was subjected to forced excretion of saliva to estimate potential virus transmission. HEV RNA presence in abdomen, thorax, and saliva samples was analyzed by PCR at the selected post-infection times. HEV RNA was detected in the abdomens of Cx. pipiens females collected 1 dpi in the two experimentally-infected batches, but not in the saliva or thorax. None of the samples collected 7-21 dpi were positive. Our results show that Cx. pipiens is not a competent vector for HEV, at least for zoonotic genotype 3.

The spatial pattern of human exposure to Crimean-Congo haemorrhagic fever virus is not consistent with red deer-based risk predictions.

The objective of this study was to evaluate the spatial risk of exposure to Crimean-Congo haemorrhagic fever virus (CCHFV) infection of healthy blood donors in an enzootic region with a predicted risk gradient based on a virus-animal interaction risk model. We designed a cross-sectional study to test if the exposure pattern of the human population to CCHFV spatially matches the predicted risk. We randomly selected 1384 donors from different risk gradients and analyzed their sera searching for CCHFV antibodies. None of the selected blood donors showed exposure to CCHFV. This study shows that exposure risk spatial patterns, as predicted from animal-tick-virus models, does not necessarily match the pattern of human-infected tick interactions leading to CCHFV infection and CCHF cases, at least in a region of predicted moderate infection risk. The findings suggest that future studies should bear the potential drivers of tick-human encounter rates into account to more accurately predict risks.

Confirmed presence of aedes (rusticoidus) refiki Medschid, 1928 in a continental dry Mediterranean peri-urban environment in south-central Spain.

BACKGROUND: The 'snow-melt mosquito' aedes (rusticoidus) refiki is a rare species with a wide distribution in Europe that is usually defined as an aggressive mosquito for mammals, including humans. During a mosquito survey in a peri-urban area in south-central mainland Spain, adult Ae. refiki females were captured and identified by morphological traits. The presence of this species of mosquito has never been molecularly confirmed under continental dry Mediterranean climatic influence with scarce number of days with snow on soil. The aim of this study was to confirm by amplification and sequencing of mitochondrial cytochrome c oxidase subunit I (COI) and internal transcribed spacer 2 (ITS2) region. RESULTS: We also successfully amplified and typed the species molecularly by COI and ITS2 regions. The peri-urban area where Ae. refiki was found contrasts with the reported cold, humid and snowy environments required by the species to breed. CONCLUSIONS: This finding suggests that the species is already adapted to continental dry Mediterranean environments, questioning whether it is a truly stenotopic species of cold snowy environments.

Red deer reveal spatial risks of Crimean-Congo haemorrhagic fever virus infection.

Crimean-Congo haemorrhagic fever virus (CCHFV) continues to cause new human cases in Iberia while its spatial distribution and ecological determinants remain unknown. The virus remains active in a silent tick-animal cycle to which animals contribute maintaining the tick populations and the virus itself. Wild ungulates, in particular red deer, are essential hosts for Hyalomma ticks in Iberia, which are the principal competent vector of CCHFV. Red deer could be an excellent model to understand the ecological determinants of CCHFV as well as to predict infection risks for humans because it is large, gregarious, abundant and the principal host for Hyalomma lusitanicum. We designed a cross-sectional study, analysed the presence of CCHFV antibodies in 1444 deer from 82 populations, and statistically modelled exposure risk with host and environmental predictors. The best-fitted statistical model was projected for peninsular Spain to map infection risks. Fifty out of 82 deer populations were seropositive, with individual population prevalence as high as 88%. The highest prevalence of exposure to CCHFV occurred in the southwest of the Iberian Peninsula. Climate and ungulate abundance were the most influential predictors of the risk of exposure to the virus. The highest risk regions were those where H. lusitanicum is most abundant. Eight of the nine primary human cases occurred in or bordering these regions, demonstrating that the model predicts human infection risk accurately. A recent human case of CCHF occurred in northwestern Spain, a region that the model predicted as low risk, pointing out that it needs improvement to capture all determinants of the CCHFV infection risk. In this study, we have been able to identify the main ecological determinants of CCHFV, and we have also managed to create an accurate model to assess the risk of CCHFV infection.

A Duplex Quantitative Real-Time Reverse Transcription-PCR for Simultaneous Detection and Differentiation of Flaviviruses of the Japanese Encephalitis and Ntaya Serocomplexes in Birds.

High impact, mosquito-borne flaviviruses such as West Nile virus (WNV), Usutu virus (USUV), Japanese encephalitis virus (JEV), Tembusu virus (TMUV), and Bagaza/Israel turkey meningoencephalomyelitis virus (BAGV/ITV) are emerging in different areas of the world. These viruses belong to the Japanese encephalitis (JE) serocomplex (JEV, WNV, and USUV) and the Ntaya serocomplex (TMUV and BAGV/ITV). Notably, they share transmission route (mosquito bite) and reservoir host type (wild birds), and some of them co-circulate in the same areas, infecting overlapping mosquito and avian population. This may simplify epidemiological surveillance, since it allows the detection of different infections targeting the same population, but also represents a challenge, as the diagnostic tools applied need to detect the whole range of flaviviruses surveyed, and correctly differentiate between these closely related pathogens. To this aim, a duplex real-time RT-PCR (dRRT-PCR) method has been developed for the simultaneous and differential detection of JE and Ntaya flavivirus serocomplexes. The method has been standardized and evaluated by analyzing a panel of 49 flaviviral and non-flaviviral isolates, and clinical samples of different bird species obtained from experimental infections or from the field, proving its value for virus detection in apparently healthy or suspicious animals. This new dRRT-PCR technique is a reliable, specific and highly sensitive tool for rapid detection and differentiation of JE and Ntaya flavivirus groups in either domestic or wild animals. This novel method can be implemented in animal virology diagnostic laboratories as screening tool in routine surveillance and in the event of bird encephalitis emergence.

Prospective evaluation of the uterocervical angle as a predictor of spontaneous preterm birth.

INTRODUCTION: The uterocervical angle has been proposed as an ultrasound marker to predict spontaneous preterm birth; however, the studies that provided this evidence were retrospective and their results heterogeneous. This study aimed to assess the ability of the uterocervical angle to predict spontaneous preterm birth before 34 and 37 weeks of gestation. MATERIAL AND METHODS: A prospective cohort study with singleton pregnancies between 19.0 and 22.6 weeks of gestation. Uterocervical angle and cervical length were measured by transvaginal ultrasound. Maternal history and pregnancy data were recorded. Delivery data were subsequently collected. RESULTS: The final analysis included 1453 singleton pregnancies. Spontaneous preterm birth before 37 weeks occurred in 52 cases (3.6%) and before 34 weeks in 17 (1.2%). For the prediction of spontaneous preterm birth before 34 weeks, the area under the curve for the uterocervical angle was 0.67 (95% CI 0.54-0.79) and the detection rates were 5.9% and 23.5% for fixed false-positive rates of 5% and 10%, respectively. For the prediction of spontaneous preterm birth before 37 weeks, the area under the curve was 0.58 (95% CI 0.50-0.67) and the detection rates were 5.8% and 18% for fixed false-positive rates of 5% and 10%, respectively. Combined predictive models were studied. To predict spontaneous preterm birth before 34 weeks, the best model was provided by a combination of uterocervical angle and cervical length (area under the curve 0.72; 95% CI 0.58-0.86). The detection rates of this model were 35.3% and 41.2% for fixed false-positive rates of 5% and 10%, respectively. To predict spontaneous preterm birth before 37 weeks of gestation, the best model was provided by a combination of uterocervical angle, cervical length, and previous history of spontaneous preterm birth (area under the curve 0.64; 95% CI 0.55-0.72). The detection rates of this model were 15.4% and 30.8% for fixed false-positive rates of 5% and 10%, respectively. Obese women and those with a history of cesarean section had a wider uterocervical angle. CONCLUSIONS: The uterocervical angle, measured mid-trimester, is a poor predictor of spontaneous preterm birth.

Dynamics of Aujeszky's disease virus infection in wild boar in enzootic scenarios.

Aujeszky's disease (AD) virus is enzootic in Iberian wild boar, thus posing a threat to the official eradication of AD on extensive domestic pig farms in Spain. Understanding the dynamics and drivers of ADV infection in wild boar will help prevent viral transmission at the wild boar-pig interface. This study analyses the dynamics of ADV infection in wild boar and tests relevant hypotheses in order to identify drivers of ADV infection dynamics. Wild boar sera (N = 971) and oropharyngeal tonsils (TN, N = 549) collected over 11 consecutive years in south-western Spain were tested for ADV antibodies and DNA, respectively. We tested the hypotheses that population immunity modulates the risk of ADV infection (H1 ) and that detecting ADV DNA in TN is a good proxy of the annual ADV infection pressure (H2 ). This was done by building logistic regression models that were subsequently employed to test the influence of a series of host population and host individual factors-including predictors of ADV immunity in the population-on the annual risk of new ADV infections and on the presence of ADV DNA in TN. The premise of H1 was that there would be a negative association between the proportion of ADV antibody-positive wild boar in a given year and the risk of ADV infection of naïve individuals. There was, however, a positive association, and H1 was, therefore, rejected. If detecting ADV in TN had been a good indicator of ADV infection pressure, a positive association with the proportion of ADV antibody-positive wild boar would have been found. However, this was not the case and H2 was also rejected. We confirmed that ADV infection is a dynamic phenomenon. The risk of infection with ADV can change considerably between consecutive years, and these changes are positively associated with the proportion of infected wild boar in the population.

Prevalence of hepatic steatosis as assessed by controlled attenuation parameter (CAP) in subjects with metabolic risk factors in primary care. A population-based study.

BACKGROUND: Primary care is the ideal setting for early identification of patients with non-alcoholic fatty liver disease (NAFLD). NAFLD is a potentially progressive disease that may lead to cirrhosis and liver cancer but is frequently underrecognized because subjects at risk are often not evaluated. Controlled attenuation parameter (CAP) is a reliable method for non-invasive quantification of liver fat. It has the advantage of simultaneous measurement of liver stiffness (LS), an estimate of liver fibrosis. There is no information on CAP in subjects with risk factors from primary care. AIM: To investigate the prevalence of hepatic steatosis, as estimated by CAP, in subjects from the community with metabolic risk factors and correlate findings with clinical and biochemical characteristics and LS. PATIENTS AND METHODS: Population-based study of 215 subjects with metabolic risk factors without known liver disease identified randomly from a primary care center. A control group of 80 subjects matched by age and sex without metabolic risk factors was also studied. CAP and LS were assessed using Fibroscan. RESULTS: Subjects with risk factors had CAP values higher than those of control group (268±64 vs 243±49dB/m,p<0.001). Prevalence of severe steatosis (CAP> 280dB/m) in subjects with risk factors was 43%. In multivariate analysis, fatty liver index (FLI) and HOMA were independent predictive factors of severe steatosis. There was a direct correlation between CAP and FLI values (r = 0.52,p<0.001). Interestingly, prevalence of increased LS was 12.6% in the risk group vs 0% in the control group (p<0.001). Increased LS occurred predominantly in subjects with high CAP values. CONCLUSIONS: A high proportion of subjects with metabolic risk factors seen in primary care have severe steatosis. FLI could be used as a surrogate of CAP. Increased LS was found in a significant proportion of subjects with risk factors but not in control subjects.

The uterocervical angle and its relationship with preterm birth.

OBJECTIVES: The objective of this study is to determine whether the uterocervical angle (UCA) correlates with the risk of spontaneous preterm birth (sPTB) < 34 weeks and assess its interobserver variability. STUDY DESIGN: Case-control study of 275 women including 34 who started labor spontaneously and gave birth before 34 weeks of gestation (preterm group) and 241 who gave birth at term (control group). Images used to report cervical length were re-measured for UCA. RESULTS: Mean UCA in the second trimester was wider in the preterm group (105.16°) compared with the control group (94.53°), p = .015. The intraclass correlation coefficient was 0.821 (95% CI: 0.74-0.97) for masked interobserver variability, which implies correct agreement among UCA measurements. Mean UCA increased from the first to the second trimester (84.2° versus 94.5°; p = .019). CONCLUSIONS: Wider UCA in the second trimester is related to sPTB. UCA measurement is a reproducible technique. UCA appears to increase from the first to the second trimester. Prospective studies, with ultrasound examinations aimed at measuring UCA in vivo, are needed to accurately assess the characteristics of this marker and its potential as a predictor of sPTB in clinical practice.

Ultrasound examination at term for predicting the outcome of delivery in women with a previous cesarean section.

OBJECTIVE: To evaluate if ultrasound variables at term are associated with the mode of delivery in women with previous cesarean section (PCS). METHODS: This was a prospective study of singleton pregnant women who planned a trial of vaginal birth after cesarean delivery. Cervical length, posterior cervical angle, head-perineum distance, and estimated fetal weight were measured at 37-39 weeks of gestation. RESULTS: One hundred forty-four pregnancies were examined and vaginal delivery was achieved in 98 women (73%). Logistic regression analysis identified cervical length, head-perineum distance, age, previous vaginal delivery, previous cesarean for dystocia, and Bishop score as predictors of vaginal delivery. Combining ultrasound and clinical parameters, two models for risk scoring that differ in the variable Bishop score or cervical length were constructed. The AUC of these models was 0.867 and 0.855, respectively. CONCLUSIONS: In women with a PCS, measurement of cervical length and head-perineum distance at term is associated with the mode of delivery. A combination of clinical and sonographic parameters at term can predict the likelihood of vaginal delivery.

Decision-making criteria and indicators for water and sanitation projects in developing countries.

Water and sanitation projects for solving the needs of small communities in developing countries are complex in nature and involve complex decision-making, which must consider technical, socioeconomic and environmental dimensions. Multicriteria analysis (MCA) is a suitable decision-aid method that scores a finite number of options on the basis of a set of evaluation criteria. The main challenge in MCA is choosing the appropriate criteria and evaluation indicators to use for assessment. The aim of this paper is to present a comprehensive and wide list of criteria and evaluation indicators as a guideline in MCA of water and basic sanitation projects in small rural communities of developing countries. First, the paper details the general criteria to be considered in all the projects, which are classified in to 4 main groups: technical (e.g., local resources use, appropriate management); social (e.g., local community participation, overcoming discrimination of conflict); economic (e.g., low cost, employment of local staff) and environmental criteria (e.g., atmospheric emissions, water pollution). Then, it describes technical criteria to be additionally considered in each specific type of project: water supply (e.g., water needs, independent access), water treatment (e.g., constant resource availability, flexibility of use for different types of water) and basic sanitation projects (e.g., maximum number of people per system, effluent quality).

Síndrome de insensibilidad a los andrógenos como causa de amenorrea primaria / Androgen insensitivity syndrome as a cause of primary amenorrhea

El síndrome de insensibilidad a los andrógenos se caracteriza por la presencia de cariotipo 46,XY, fenotipo femenino y presencia de gónadas masculinas. Es la tercera causa más frecuente de amenorrea primaria, tras la disgenesia gonadal y la ausencia congénita de vagina (síndrome de Mayer-Rokitansky-Küster-Hauser). Se trata de una entidad de interés por su relevancia en la identificación sexual y por su posible asociación con tumores malignos de las gónadas masculinas, que hace necesario un correcto diagnóstico y tratamiento quirúrgico. En este artículo se describen dos casos de síndrome de insensibilidad a los andrógenos, con su estudio clínico-genético y tratamiento, así como su seguimiento (AU)

Androgen insensitivity syndrome is characterized by the presence of external female phenotype, 46,XY karyotype and intraabdominal testes. This syndrome is the third most frequent cause of primary amenorrhea, after gonadal dysgenesis and congenital absence of the vagina (Mayer-Rokitansky-Küster-Hauser syndrome). Androgen insensitivity syndrome is of interest due to its role in sexual identification and its possible association with malignant tumors of the male gonads, which require an accurate diagnosis and surgical treatment. We present two cases of androgen insensitivity syndrome. The results of the clinical and genetic examinations, as well as the treatment and follow-up of these two patients, are discussed (AU)

Psychrophilic anaerobic digestion of guinea pig manure in low-cost tubular digesters at high altitude.

Guinea pig is one of the most common livestock in rural communities of the Andes. The aim of this research was to study the anaerobic digestion of guinea pig manure in low-cost unheated tubular digesters at high altitude. To this end, the performance of two pilot digesters was monitored during 7 months; and two greenhouse designs were compared. In the dome roof digester the temperature and biogas production were significantly higher than in the shed roof digester. However, the biogas production rate was low (0.04 m(biogas)(3)m(digester)(-3) d(-1)), which is attributed to the low organic loading rate (0.6 kg(VS)m(digester)(-3)d(-1)) and temperature (23°C) of the system, among other factors. In a preliminary fertilization study, the potato yield per hectare was increased by 100% using the effluent as biofertilizer. Improving manure management techniques, increasing the organic loading rate and co digesting other substrates may be considered to enhance the process.

Multi-criteria analysis for improving strategic environmental assessment of water programmes. A case study in semi-arid region of Brazil.

Multi-criteria analysis (MCA) is a family of decision-making tools that can be used in strategic environmental assessment (SEA) procedures to ensure that environmental, social and economic aspects are integrated into the design of human development strategies and planning, in order to increase the contribution of the environment and natural resources to poverty reduction. The aim of this paper is to highlight the contribution of a particular multi-criteria technique, the analytic hierarchy process (AHP), in two stages of the SEA procedure applied to water programmes in developing countries: the comparison of alternatives and monitoring. This proposal was validated through its application to a case study in Brazilian semi-arid region. The objective was to select and subsequently monitor the most appropriate programme for safe water availability. On the basis of the SEA results, a project was identified and implemented with successful results. In terms of comparisons of alternatives, AHP meets the requirements of human development programme assessment, including the importance of simplicity, a multidisciplinary and flexible approach, and a focus on the beneficiaries' concerns. With respect to monitoring, the study shows that AHP contributes to SEA by identifying the most appropriate indicators, in order to control the impacts of a project.