Pestivirus. Based on partial sequences, three genotypes can be distinguished that do not, however, directly correlate with virulence. Depending on both virus and host factors, a wide range of clinical syndromes can be observed and thus, laboratory confirmation is mandatory. To this means, both direct and indirect methods are utilized with an increasing degree of commercialization. Both infections in domestic pigs and wild boar are of great relevance; and wild boars are a reservoir host transmitting the virus sporadically also to pig farms. Control strategies for epidemic outbreaks in free countries are mainly based on classical intervention measures; i.e., quarantine and strict culling of affected herds. In these countries, vaccination is only an emergency option. However, live vaccines are used for controlling the disease in endemically infected regions in Asia, Eastern Europe, the Americas, and some African countries. Here, we will provide a concise, updated review on virus properties, clinical signs and pathology, epidemiology, pathogenesis and immune responses, diagnosis and vaccination possibilities.Classical swine fever (CSF) remains one of the most important transboundary viral diseases of swine worldwide. The causative agent is CSF virus, a small, enveloped RNA virus of the genus  Classical swine fever (CSF) is one of the most important viral diseases of domestic pigs and wild boar. It has tremendous impact on animal health and pig industry and is therefore notifiable to the World Organization for Animal Health (OIE) . After iA binding legal framework exists for the surveillance and control in most countries. Integral parts of the control measures are timely and reliable diagnosis, stamping out of infected herds, establishment of restriction zones, movement restrictions, and tracing of possible contacts. Prophylactic vaccination and other treatments are often also strictly prohibited. However, in Europe, where affected wild boar populations were shown to be an important reservoir for the virus, and acted as a source for reintroduction into the domestic pig population ,3, emergDesign of control measures and risk assessment depends on the knowledge of factors that influence disease dynamics and epidemiology. For this purpose, the presented review aims at providing an updated overview on the disease and the underlying mechanisms but also control and diagnostic options. Classical swine fever virus (CSFV) belongs to the genus Pestivirus within the Flaviviridae family [Bovine viral diarrhea virus 1 and 2 (BVDV-1 and -2), Border disease virus (BDV) and a growing number of unclassified and so-called atypical pestiviruses, from giraffe-virus over HoBi-like viruses to recently discovered Bungowannah virus and atypical porcine pestivirus [e family . Other mstivirus ,11,12,13rns [pro, p7, NS2-3, NS2, NS3, NS4A, NS4B, NS5A, and NS5B. The latter have various functions in the viral replication, e.g., NS5B represents the RNA-dependent RNA polymerase [The enveloped viral particles consist of four structural proteins, namely the core protein (C), and envelope glycoproteins E1, E2, and Erns ,16,17,18rns ,20,21,22lymerase  and NS3 lymerase ,25.rns encoding region [Virus replication takes place in the cytoplasm after receptor mediated endocytosis and does normally not lead to a cytopathic effect in cell culture  . A putatg region , namely rns are necessary for viral attachment [rns which interacts with glycosaminoglycans [pro is cleaved from the polyprotein [rns, E1 and E2, E2 and p7 as well as NS2-3. NS2-3 is then partially processed through the autocatalytic cysteine protease activity of NS2 into NS2 and NS3. In this way NS2 generates its own C-terminal ending [In any case, glycoproteins E2 and Etachment ,10, and oglycans . For recoglycans . After foglycans ,14,15. Toglycans ,17,18. Toglycans . Initialyprotein ,21. Subsl ending ,23. The l ending . While rl ending . Virion l ending ,27.The survival of CSFV under different ambient conditions varies considerably and is influenced especially by the temperature but also by the matrix in which it is found. Generally, survival times are higher under cold, moist and protein rich conditions . The depFor animal disease control, survival in excretions (left in the pen or stored as slurry) and stability in meat products are crucial parameters. For CSFV in excretions, survival times were demonstrated that range from a few days at room temperature to several weeks at 5 °C . If temppro, C, E1, and Erns [Classical swine fever virus strains can be divided into three genotypes with three to four sub-genotypes. The most recently added sub-genotype 1.4 was only very recently described for CSFV strains from Cuba. These strains had so far been placed into sub-genotype 1.2 but formed a distinct cluster when compared based on longer genome fragments, e.g., full-length E2, Nand Erns . Furtherand Erns ,45,46,47and Erns . Up to very recently, phylogenetic studies were mainly based on two short fragments, namely a 150 nucleotide (nt) fragment of the 5′NTR and a 190 nt fragment of the E2 encoding region . MoreoveThe distribution of genotypes shows a distinct geographical pattern ,56: WherEuropean CSFV sequences were collected and made available through the semi-public CSF-database (DB) at the EU and OIE reference laboratory for CSF in Hannover, Germany ,93,94,95pro [rns [The search for virulence markers indicated a role of the Npro , the E2 pro , the ribpro [rns ,99, and pro [rns . Furtherpro [rns ,104,105.pro [rns ,106. Invpro [rns . There wpro [rns .Classical swine fever can be divided into the following forms of the disease: an acute , a chronic and a persistent course, which usually requires infection during pregnancy . In geneIn acute-lethal courses, death usually occurs 2–4 weeks after CSFV infection. Mortality can reach up to 100% from 10 to 30 days depending on the age of the animal and the virulence of the virus strain ,110,111.Chronic course occurs when an infected pig is not able to mount an adequate immune response. In general, only non-specific clinical signs are observed in infected animals like remittent fever, depression, wasting and diffuse dermatitis see . It is aWhen infection occurs during pregnancy, the virus can also infect the fetus in the womb due to its ability to pass the placental barrier which in turn might lead to persistent infection in the piglets. While the sows often show only mild clinical signs, an infection depending on the stage of gestation, leads to absorption or mummification of the fetuses and to abortions or stillbirth ,122,123.The pathological findings  depend oAs mentioned above, clinical signs of CSFV infections can vary considerably from peracute deaths to unapparent courses depending on virulence of the virus strain involved and different (partly unknown) host factors . UnspeciInfection with CSFV is followed by primary replication in the tonsils and subsequently spread to surrounding lymphoid tissues . The virEspecially in the acute-lethal course, CSF is accompanied by severe lymphopenia and resulting immunosuppression as well as granulocytopenia ,145,146.rns and the non-structural protein NS3 [Despite the immunopathogenesis of most CSF-related lesions, pigs recovering from CSFV infection mount an effective immune response with E2-specific antibodies detectable after 10–14 days. The E2 antibodies are able to neutralize CSFV in vitro and induce protective immune responses ,155. Thetein NS3 ,157. Immtein NS3 ,159,160.tein NS3 ,162,163.Suidae family, particularly domestic pigs (Sus scrofa domesticus) and European wild boar (Sus scrofa scrofa) [Phacochoerus africanus) and bushpigs (Potamochoerus larvatus) was recently demonstrated [Susceptible hosts are different members of the  scrofa) ,164. Mornstrated .Classical swine fever virus can be transmitted both horizontally and vertically. Horizontal transmission takes places through direct or indirect contact between infected and susceptible pigs. Important indirect routes include feeding of virus contaminated garbage/swill and mechanical transmission via contact to humans or agricultural and veterinary equipment . AerogenUpon contact, infection usually occurs through the oronasal route, or less frequently via conjunctiva, mucus membranes, skin abrasions, insemination, and the use of contaminated instruments ,172,173.Classical swine fever affected wild boar populations can serve as reservoir of the virus and present a constant risk for domestic pigs. Fritzemeier et al.  could shOver the last decades, a decreasing virulence was observed for the CSFV strains involved in many outbreaks among wild boar and domestic pigs. In Europe, the most prevalent genotype 2.3 strains showed moderate virulence with a highly age-dependent clinical picture and rather unspecific clinical pictures in older animals (see above). These strains showed potential to establish endemicity in affected wild boar populations rather than showing the self-limiting behavior of the historical highly virulent CSFV strains. It was discussed whether these strains are somewhat the ideally adapted variants of CSFV for long-term perpetuation in wildlife . In endemically affected countries with official but imperfect vaccination, circulation of less virulent CSFV strains is often masked by partial protection. In combination with management and biosecurity issues , the virus is maintained over prolonged periods in the domestic pig population. Rapid and reliable diagnosis is of utmost importance for the timely implementation of control measures against CSF. On the international level, laboratory methods as well as sampling and shipping guidelines can be found in the OIE Manual of Diagnostic Tests and Vaccines for Terrestrial Animals and the respective EU Diagnostic Manual (European Commission Decision 2002/106/EC). rns) is usually at least comparable with virus isolation, most CSF specific assays lack sensitivity [For CSFV, primary detection is performed using well established real-time reverse transcription polymerase chain reaction (RT-qPCR) systems ,182,183,sitivity . Serologsitivity .rns [Reliable DIVA  assays are needed when using DIVA vaccines. Commercially available tests that can accompany both E2 subunit vaccines and chimeric vaccines such as “CP7_E2alf”, target the detection of antibodies directed against glycoprotein Erns ,198,199.rns , anotherrns : Moreoverns . Due to the increased sensitivity of diagnostic tools , vaccine virus detections are quite common in oral vaccination campaigns of wild boar and vaccination programs of domestic pigs. For this reason, different RT-qPCR systems have been developed and tested, these allow differentiation between vaccine and field viruses (genetic DIVA) ,207,208.Sampling can be the bottleneck of swine fever diagnosis, especially in the case of wild boar, but also in remote areas. For this reason, alternative sampling strategies and sample matrices have been tested for CSF (often combined with African swine fever sampling) especially for wildlife specimens and under rural conditions ,211,212.Highly efficacious and safe live-attenuated CSF vaccines have existed for decades . The und® Pesti, MSD Animal Health, Unterschleißheim, Germany). These vaccines are safe and were shown to provide clinical protection and limit the spread of CSF [In addition, these vaccines were also adapted to a bait format for oral immunization of wild boar ,215,216 d of CSF ,234,235.d of CSF ,236 and d of CSF . Due to d of CSF . Among td of CSF . In 2014d of CSF ,256,257.Oral emergency vaccination of wild boar with baits has proven to be a potent tool to control the disease in wildlife and to safeguard domestic pigs . For thi


Most studies of infectious diseases in East African cattle have concentrated on gastro-intestinal parasites and vector-borne diseases. As a result, relatively little is known about viral diseases, except for those that are clinically symptomatic or which affect international trade such as foot and mouth disease, bluetongue and epizootic haemorrhagic disease. Here, we investigate the seroprevalence, distribution and relationship between the viruses involved in respiratory disease, infectious bovine rhinotracheitis virus (IBR), bovine parainfluenza virus Type 3 (PIV3) and bovine viral diarrhoea virus (BVDV) in East African Shorthorn Zebu calves. These viruses contribute to the bovine respiratory disease complex (BRD) which is responsible for major economic losses in cattle from intensive farming systems as a result of pneumonia. We found that calves experience similar risks of infection for IBR, PIV3, and BVDV with a seroprevalence of 20.9%, 20.1% and 19.8% respectively. We confirm that positive associations exist between IBR, PIV3 and BVDV; being seropositive for any one of these three viruses means that an individual is more likely to be seropositive for the other two viruses than expected by chance. •IBR, PIV3 and BVDV are circulating in East Africa.•Calves experience similar risks of infection for IBR, PIV3, and BVDV with a seroprevalence of 20.9%, 20.1% and 19.8%. As a result, relatively little is known about viral diseases, except for those that are clinically symptomatic or which affect international trade such as foot and mouth disease, bluetongue and epizootic haemorrhagic disease . Yet, inThere are many factors which contribute to the BRD, including stress, management practices and biological agents . VirusesThe purpose of this study is to improve current knowledge on the seroprevalence and distribution of IBR, PIV3, and BVDV in cattle in sub-Saharan Africa by estimating the seroprevalence to these viruses in East African shorthorn zebu calves from Western Kenya using data gathered by the Infectious Diseases of East Africa Livestock (IDEAL) project. We aim to quantify the associations between these viruses within this study population.22.1The study reported here uses data gathered as part of the Infectious Diseases of East Africa Livestock (IDEAL) project, the design of which has previously been reported by Calves were selected using a stratified two-stage random cluster study design. In the first stage, 20 sublocations  were selected from 5 agro-ecological zones, in a radius of 45 km from the town of Busia for logistical reasons. This area is dominated by smallholder mixed crop-livestock production systems, with an average farm size of 2 ha and about 5 cattle per farm . During 2.2At the recruitment visit, the calf's owner completed a questionnaire. This questionnaire collected environmental information about the farm, other livestock, water sources and animal husbandry practices. Calf locations were geo-referenced using hand-held GPS devices . During the study calves were weighed (measured to the nearest 0.5 kg) and the dam's girth was measured (measured in cm) and the calves had biological samples taken for laboratory analysis.2.32.3.1Jugular vein blood samples were collected in plain Vacutainer™  tubes. Blood samples were allowed to clot, the serum was recovered and aliquots were stored at − 20 °C, until serological analysis could take place.Serological samples taken from calves at 51 weeks of age were screened using SVANOVIR kits obtained from Svanova Biotech AB  to identify antibodies specific to the following viruses: a) infectious bovine rhinotracheitis (IBR); b) bovine viral diarrhoea virus (BVDV) antibody and c) bovine parainfluenza virus type 3 (PIV3). In addition, the sera were screened for the presence of BVDV antigens using kits obtained from IDEXX . All the kits (apart from the BVDV antigen kit) were designed to detect virus specific IgG antibodies in serum using a procedure based upon a solid phase indirect Enzyme Linked Immunosorbent Assay (Indirect ELISA). All assays were performed and analysed according to the manufacturer's instructions using single wells. There is no evidence for cross-reactivity between the tests.corr). To calculate the ODcorr, values in wells coated with a particular antigen are corrected by subtracting the OD values of corresponding wells containing the control antigen (ODtest antigen − ODcontrol = ODcorr). The Percent Positivity (PP) values are calculated as follows:The SVANOVIR ELISA tests are based on the measurement of corrected optical density , which was calculated according to the manufacturer's instructions for the test.To convert the continuous PP (or S/P%) values into a serostatus which is a binary seropositive/seronegative outcome, the PP (or S/P%) values were interpreted according to the manufacturer's cut-offs specified in the appendix. Justification for grouping the inconclusive IBR results with the IBR seropositive individuals is presented in the appendix. Therefore, seropositive/seronegative means antibodies to the antigen of interest are detectable/not detectable in the blood of the host, whereas seroconversion is the movement from a seronegative to a seropositive state.2.42.4.1epicalc package in R v.2.15.2  at 51 weeks of age for each virus, and is included in the models as a fixed effect (β). Sublocation  is included in the model as a random effect (b) to account for the study design and environmental similarity between calves clustered into each sublocation.The association between the three viruses was assessed using generalised linear mixed models (GLMM) fitted with a logit link function and binomial errors and a Laplace approximation to the maximum likelihood estimation in R v.2.15.2 using the  package . We inveA separate model was constructed for each virus. Viruses were said to be co-distributed if they occurred in an individual more often than expected by chance. The same patterns of association were examined using continuous measures of PP values instead of serostatus see the . Interacp < 0.2 in the univariate analysis (listed in the Appendix), which may affect respiratory pathogens or contact between calves, were added into each of the models. Backwards stepwise selection was then used to remove variables to produce the most parsimonious model.Following construction of the virus-only model, environmental confounders associated with the virus with a By using a two stage statistical approach it was possible to examine whether the associations between being seropositive for the different viruses was due to coinfection or if the associations were being driven by a joint underlying correlation such as a shared environmental factor or calf characteristic.33.1The seroprevalence at 51 weeks was similar for each virus; IBR had an adjusted seroprevalence of 20.9% whilst PIV3 and BVDV antibodies had adjusted seroprevalences of 20.1% and 19.8%, respectively . We obse3.2The virus-only GLMMs indicated that IBR, PIV3 and BVDV were co-distributed . There wInclusion of environmental confounders in the virus-only models did not affect the relationship observed between the serostatus of the three viruses; the most parsimonious model for all three viruses was the one which excluded all environmental variation . Even in4This study aimed to describe the prevalence and association of viruses involved in the bovine respiratory disease complex (BRD) in Western Kenya. We have shown that IBR, PIV3 and BVDV all have an estimated seroprevalence of around 20%. The observed seroprevalence of IBR is within the range (16%–54%) that Cross-reactivity may occur between a virus and its related viruses. For example, BVDV is cross-reactive with other pestiviruses such as Classical Swine Fever and Border Disease Virus of sheep; IBR is cross-reactive with four herpesviruses from other animals including goats and buffalo; and PIV3 cross-reacts with human strains of the virus . HoweverIn accordance with numerous other epidemiological studies, but in a previously unstudied setting, this analysis has found that IBR, PIV3 and BVDV are associated . InclusiTo conclude, this study shows that the viruses IBR, PIV3 and BVDV are co-circulating in East African shorthorn zebu calves in Western Kenya. We identified positive associations occurring between IBR, PIV3 and BVDV in a previously unstudied setting. Further studies are needed to identify the long-term impact of these viruses on cattle productivity and their interactions with other parasites.The IDEAL project received approval by the University of Edinburgh Ethics Committee (reference number OS 03–06), and the Animal Care and Use Committee of the International Livestock Research Institute. All participating farmers gave informed consent in their native language prior to recruiting their animals into the study.


Background: Soldiers’ perception of leadership during military deployment has gained research attention as a potentially modifiable factor to buffer against the development of postdeployment post-traumatic stress disorder (PTSD). Within nonmilitary research, the organizational justice (OJ) framework, i.e. distributive justice, procedural justice (PJ) and interactional justice (IJ), has been found to relate to mental health outcomes. Aspects of OJ may, therefore, be protective against PTSD.Objectives: We examined the prospective relationship between aspects of OJ, namely the perceptions of PJ and IJ by subordinate soldiers without leadership obligations in relationship to immediate superiors and PTSD.Method: Participants were soldiers (n = 245) deployed to Helmand Province in Afghanistan in 2009. Logistic regression procedures were used. The primary analysis measured PTSD cases using the Structured Clinical Interview for DSM-IV-TR Axis-I Disorder (SCID) 2½ years after homecoming. PJ/IJ was measured during deployment with a 6-item composite measure ranging from 0 to 12. Supplementary primary analyses were performed with PJ/IJ measured before and immediately after deployment. A secondary PJ/IJ analysis also tested against four postdeployment measures with the Post-Traumatic Stress Disorder Checklist Civilian (PCL-C) dichotomized at screening symptom levels.Results: Higher levels of perceived PJ/IJ for soldiers without leadership obligations during deployment had a prospective relation  with PTSD on the SCID 2½ years after homecoming after adjustment for factors including predeployment PTSD symptoms, trauma and combat exposure, and state affectivity. Similar results were found by measuring PJ/IJ before  but not immediately after homecoming . A relationship with PTSD symptoms at the screening level at the four measurements of PCL-C was found, but only when predeployment PTSD symptoms were not controlled for.Conclusions: These results suggest that PJ/IJ exercised by superiors in relation to military deployments may protect subordinate soldiers against the development of postdeployment PTSD. OJ refers to employees’ perceptions of being treated fairly and rewarded for their efforts by the organization and the leadership they work for. Early research on OJ focused mainly on the fairness of distribution of rewards under the term ‘distributive justice’, while later research found that the fairness of ‘procedural justice’ (PJ) in organizations is also important for the perception of justice and fairness. Additional theoretical developments also separated the leadership’s enactment of procedures into the term ‘interactional justice’ (IJ)  military personnel’s judgements of procedural and interactional justice (PJ/IJ) during deployment as enacted by their immediate superior and (b) PTSD identified with the Structured Clinical Interview for DSM-IV-TR Axis-I Disorder (SCID) or by self-reported symptoms exceeding a screening cut-off on the Post-Traumatic Stress Disorder Checklist Civilian scale (PCL-C).2.2.1.Before, during and after a 6-month deployment to Helmand Province in Afghanistan in 2009, Danish soldiers from the International Security Assistance Force (ISAF) were asked to complete several questionnaires concerning their background and their experiences during and possible reactions to the deployment. The questionnaire data were obtained at a Danish military camp approximately 1–2 months before deployment, at the base or in Afghan airports approximately three months into the deployment, and 1–3 weeks after homecoming, at standard homecoming meetings. Two to three months after their homecoming, the questionnaire data were obtained at military camps or via mail for those now civilians. At 7–8 months, all were contacted via mail, and two cinema tickets were given for their participation. At the time, 2½–3 years after homecoming, soldiers were invited via mail to answer a questionnaire and participate in an SCID, for which they received travel reimbursement and a gift certificate . Participants were continuously informed that the data were being gathered for research purposes only. An overview of the data can be found in Supplemental data Table S1.The SCID interviews were performed by six graduate psychology students who went through an intensive training programme and a certification course. The interrater reliability of videorecorded SCIDs was tested after one month and showed a .73 for the questions in the SCID module, but there was full agreement on the overall PTSD diagnoses  and negative affectivity (α = 0.80), both ranging from 10 to 50, from the Positive And Negative Affect Schedule  through RStudio (v. 0.99.491). Descriptive data on the distribution of predictors and additional background information were produced, along with a zero-order correlation between all variables used in the regression models.For the primary analysis, we used a logistic regression model with incremental adjustment. The ‘basic model’ contained PJ/IJ and predictors related to trauma exposure during this deployment or during additional deployments. The ‘main model’ included the risk/resilience variables age, gender, predeployment PTSD symptoms, the two scales on state affectivity during deployment and the additional trauma exposure before SCID. A ‘predeployment depressive model’ further included baseline depressive symptom levels, to account for the effect of possible reverse causation. Supplementary primary analyses were conducted to test relationships of the PJ/IJ and PANAS measures taken 1–2 months before and 1–3 weeks after with the SCID.The secondary analysis included the predictors of the ‘main model’ in models with the PCL-C scales dichotomized at the screening cut-off as an outcome, to test the relation to self-reported PTSD symptoms at the four postdeployment measures. The indicators of additional trauma exposure were excluded since these were first measured together with the last PCL-C outcome. In line with previous research conducted on the data , the SCID identified n = 22 cases and n = 223 non-cases of PTSD. The distribution of cases by the variables used in the primary analysis and additional background information are presented in Of the participants  and perceptions of PJ/IJ  maintained a significant relationship with PTSD on the SCID. Including the predeployment depressive symptoms into the ‘Predeployment depressive model’ did not overall change the significant relationships with number of traumatic events  or PJ/IJ . Positive affectivity during deployment, measured with the PANAS, however, bordered on having a significant relation to a higher risk of PTSD on the SCID in this model .The supplementary primary analysis of the relation of PJ/IJ and PANAS with PTSD on the SCID, throughout the deployment cycle, showed that a better perception of PJ/IJ 1–2 months before deployment  was also a significant predictor for lower odds of PTSD. This was, however, not the case for the measure 1–3 weeks after deployment  (results not shown).3.2.The secondary analysis introduced predictors of the main model into four ‘screening models’, to test the relation with having screening levels of PTSD symptoms measured with the PCL-C at four measures postdeployment . Better perceptions of PJ/IJ bordered on having a significant relation to PTSD symptom levels at 1–3 weeks and 2–3 months after deployment, but not at 7–8 months or 2½–3 years. A more negative affectivity during deployment showed a significant relation to screening levels of PTSD on the PCL-C at 1–3 weeks and 7–8 months, but not at 2–3 months or 2½–3 years. Finally, a consistently significant relationship of higher predeployment PTSD symptoms with PTSD at screening levels was found, in the range of OR 1.08–1.12, but the outcome at 7–8 months only bordered on significance. None of the models showed significant relationships to any of the other risk/resilience factors. Differences in the zero-order correlation and the regression model prediction by the predeployment PCL-C measure of the SCID PTSD and the screening levels of PTSD symptoms, even when obtained at the same time point, could indicate overcontrol.When excluding predeployment PTSD symptoms, measured with the same scale, the PJ/IJ measure was found to predict PTSD at screening levels, at 1–3 weeks  and 2–3 months  postdeployment .4.The present multiwave cohort study examined the prospective relationship between PJ/IJ in relation to immediate superiors, as perceived by soldiers deployed into a combat zone in Helmand Province in Afghanistan in 2009, and PTSD. The PTSD outcomes were diagnosis with the SCID approximately 2½ years after homecoming and reporting a screening level of self-reported PTSD symptoms at four different time points after homecoming. Whilst studies of OJ or soldiers’ perceptions of leadership in relation to PTSD in military settings are not new, this is, to our knowledge, the first study to examine the prospective relationships between soldiers’ perception of aspects of OJ during deployment to a war zone and postdeployment PTSD.4.1.In line with our hypothesis, the result showed that experiencing higher levels of PJ/IJ in relation to an immediate superior during military deployment is related to lower odds of meeting the criteria for PTSD on the SCID 2½–3 years after homecoming. This relation was found while accounting for gender and age, predeployment PTSD and depressive symptom scores, perceived levels of general traumatic exposure and combat exposure during deployment, and positive and negative state affectivity while judging PJ/IJ. Further analysis suggested that such a relation also existed when PJ/IJ was measured before deployment. Measures of PJ/IJ were overall stable across the deployment cycle, even though some variation must be expected given the major changes that occur at home compared to that in the war zone (data not shown). However, we did not find the perceptions of PJ/IJ immediately after deployment to relate to PTSD on the SCID. While this could pertain to the measurement point at 1–3 week after homecoming, these results may also be explained by the theory of conservation of resources. This suggests that positive and negative aspects of leadership spiral over time into more or fewer resources and result in resilience or vulnerability Hobfoll, . We do fFurther, the secondary analyses of the relation between PJ/IJ during deployment and screening levels of PTSD symptoms at the four postdeployment time points showed some of the same tendencies. However, a significant relation to PJ/IJ only occurred when we excluded the predeployment PCL-C as a predictor, given the possibility of overcontrol. We can therefore not make a firm conclusion about the relation to screening levels of self-reported PTSD symptoms.Overall, the results of the relationship between PJ/IJ and PTSD do suggest that experience of lower or higher levels of PJ/IJ in relation to an immediate superior, throughout deployment into an active combat zone, may be a genuine risk/resilience factor for postdeployment PTSD diagnoses, for soldiers without their own leadership obligations.The ‘alarm-system perspective’ from the OJ literature may provide a valid theoretical explanation for the relationship found, since perception of PJ/IJ may act as a substitute for real information on dangers, which may thus calm down, or give rise to, further negative reactions for the deployed soldier. An alternative explanation for the relationship may, however, relate to the individual’s relations with the group. Group cohesion and support have been extensively studied in relation to the risk of postdeployment mental health problems  and the few cases of PTSD based on the SCID (n = 22) should be acknowledged. This restricted the possibilities of inferring results from possible predictors beyond the traumatic exposure–OJ–PTSD relationship, such as interactions between measures like PJ/IJ and trauma exposures. Second, the current study is based on a secondary analysis of data obtained for general research purposes, for which reason the questions used for the PJ/IJ measure were not originally included for this purpose and why the scale validation using the Rasch procedures was conducted in this population only. Furthermore, it was not possible to investigate the aspect of distributive justice. Thus, while the PJ/IJ measure did show fine properties, future studies on OJ in a deployed military environment should therefore also consider using other tailored questionnaires, such as that of Olsen et al. (Several limitations of the study must also be acknowledged. First, the small sample size (n et al. . Third, 5.In summary, this study examined the longitudinal relationship between perceptions of PJ/IJ in relation to immediate superiors and the risk of developing PTSD after deployment amongst Danish soldiers without their own leadership obligations, operating in Helmand Province in Afghanistan in 2009. As hypothesized, we found an association between higher levels of PJ/IJ during deployment and lower odds of developing PTSD 2½–3 years after deployment. This association was found after controlling for factors such as predeployment PTSD symptoms, positive and negative state affectivity, experiences of traumatic events and combat exposure during deployment, and traumatic events after homecoming. These findings suggest that experiencing higher levels of PJ/IJ in relation to immediate superiors during deployment may independently protect soldiers deployed into an active war zone against developing postdeployment PTSD. We further suggest that soldiers’ relationships with immediate superiors during deployment may be understood within the OJ framework. Modifying factors that lead to better perceptions of PJ/IJ in relation to an immediate superior may thus possibly aid in the prevention of PTSD cases. However, given the limitations of the study, these conclusions must be considered provisional.


Nephila clavipes, known for its sexual size dimorphism, is abundant and widespread in the New World. The first annotated genome of orb-weaver spiders, exploring N. clavipes, has recently been reported. The study, focused primarily on the diversity of silk specific genes, shed light into the complex evolutionary history of spiders. Furthermore, a robust transcriptome analysis provided a massive resource for N. clavipes RNA survey. Here, I present evidence of viral sequences corresponding to the first 10 extant virus species associated to N. clavipes and indeed, nephilids. The putatively new species are linked to ssRNA positive-strand viruses, such as Picornavirales, and to ssRNA negative-strand and dsRNA viruses. In addition, I detected sequence data of new strains of two recently reported arthropod viruses, which complemented and extended the corresponding sequence references. The identified viruses appear to be complete, potentially functional, and presenting the typical architecture and consistent viral domains. The intrinsic nature of the detected sequences and their absence in the recently generated genome assembly, suggest that they correspond to bona fide RNA virus sequences. The available RNA data allowed for the first time to address a tissue/organ specific analysis of virus loads/presence in spiders, suggesting a complex spatial and differential distribution of the tentative viruses, encompassing the spider brain and also silk and venom glands. Until recently, the virus landscape associated to spiders remained elusive. The discovered viruses described here provide only a fragmented glimpse of the potential magnitude of the Aranea virosphere. Future studies should focus not only on complementing and expanding these findings, but also on addressing the potential ecological role of these viruses, which might influence the biology of these outstanding arthropod species.The golden orb-weaver spider  Neoscona nautica, Parasteatoda tepidariorum, Plexippus setipes, Pirata sp., and an unidentified Araneae. The viruses were assigned to the Mononegavirales order, two Nairovirus like and a Plebovirus like , an Orthomyxoviridae like and a new genus of non-segmented circular RNA viruses: Chuvirus. Remarkably, employing the same RNA library Shi et al.  and phylogenetic analyses suggested they form a new clade within the Picornavirales order. The preceding studies have inaugurated Araneae virology by assessing the RNA landscape of only a dozen spiders. There are ca. 45.7 k spider species described  collected from four female individuals. These 1.53 × 109 reads corresponding to an assembly of 1.5 million transcripts, which represent the most extensive deposited RNA assembled data of any organism at the NCBI TSA database, were used as input for the objective of this study: The first identification and characterization of potential RNA viruses associated to N. clavipes.The golden orb-weaver N. clavipes, the RNA data from Babb et al. -depleted, 100-bp paired-end reads. In sum, a total of 1,848,260,474 raw RNA reads were quality controlled and filtered, and the curated 1,531,402,748 reads were de novo assembled using Trinity (rel_2.25.13) yielding 1,507,505 unique strand-specific transcripts. These 1.53 × 109 reads corresponding to an assembly of 1.5 million transcripts from Babb et al.  using as probe the complete predicted non-redundant viral proteins in a local server. Significant hits were explored by hand, and redundant contigs discarded. Potential virus genome segments sequences were curated by iterative mapping of reads using Bowtie 2 v2.3.2 http://bowtie-bio.sourceforge.net/bowtie2/index.shtml. To identify/rule out additional segments of no homology to the closely associated viruses I used diverse in silico approaches based on RNA levels by the sequencing depth of the transcript, predicted gene product structure, domain architecture, or conserved genome termini, and significant co-expression levels with the remaining viral segments. Potential open reading frames (ORF) were predicted by ORFfinder. Translated putative proteins were blasted against the non-redundant protein sequences NR database and best hits were retrieved. Predicted proteins were subjected to a domain-based Blast search against the Conserved Domain Database (CDD) v3.16 https://www.ncbi.nlm.nih.gov/Structure/cdd/cdd.shtml and integrated with SMART http://smart.embl-heidelberg.de/, Pfam http://pfam.xfam.org/ and PROSITE http://prosite.expasy.org/ to characterize the functional domains. Secondary protein structure was predicted with Garnier http://emboss.sourceforge.net/apps/release/6.6/emboss/apps/garnier.html, signal and membrane cues were assessed with SingnalP v4.1 http://www.cbs.dtu.dk/services/SignalP/ and prediction of transmembrane topology and signal peptides by Phobius http://www.ebi.ac.uk/Tools/pfa/phobius/. RNA secondary prediction was performed with the mfold Web Server http://unafold.rna.albany.edu/?q=mfold/rna-folding-form. Eventually, sequence similarity levels were visualized using the Circoletto tool with standard parameters http://tools.bat.infspire.org/circoletto/ Potential panhandle structures derived from partially complementary virus sequence termini were assessed with the RNAcofold web server http://rna.tbi.univie.ac.at/cgi-bin/RNAWebSuite/RNAcofold.cgi. Ribosomal frameshifting events were predicted using the KnotInFrame tool https://bibiserv.cebitec.uni-bielefeld.de/knotinframe. The potential functions of the ORFs products were predicted by the annotated data and similarity with known viral proteins. Virus RNA levels were calculated with Cufflinks http://cole-trapnell-lab.github.io/cufflinks/ or alternatively with the Geneious suite 8.1.9 (Biomatters Inc.) as Fragments Per Kilobase of virus transcript per Million mapped reads (FPKM) based on million non-rRNA host transcriptome reads that map to the host genome assembly NepCla1.0. These normalized values avoid potential inconsistencies associated to the success of rRNA capture (RNA depleted libraries), the presence of other viruses and the diluting effects of variable non-host levels of reads. Tentative virus detections were contrasted on the N. clavipes NepCla1.0 genome assembly  by BLASTN searches  and inspected by hand to rule out the identification of transcripts associated to putative integrated viruses or endogenous viral-like elements (EVE). In addition, the complete DNA derived raw read collection of Babb et al. In order to identify putative RNA virus associated with b et al.  were intb et al.  were useb et al.  was mappN. clavipes.Based on sequence similarity to best hits, sequence alignments, predicted proteins, and domains, and phylogenetic comparisons to reported species based in MAFFT and FastTree, I found evidence of 10 diverse new virus species and three new strains of reported virus species associated to Picornavirales order  is predicted to have a 10,198 nt long genome, presenting a single ORF between coordinates 761–10,147, encoding a putative 357.963 kDa and 3,128 aa long polyprotein. Domain prediction based on InterproScan, NCBI-CD database v3.15, THHM, PHOBIUS, SMART, Pfam, PROSITE and Garnier resulted in the identification of diverse motifs associated to Picornavirales coat and replicase proteins (RP) , P. tepidariorum (3), P. setipes (3), Pirata sp. (1), and eight unrecognized individuals (Araneae sp.)  shares the proposed Picorna-Calici ssRNA(+) super clade with NcV1, but presents a different genome organization and domain architecture  and of Wuhan spider virus 6 , both of unknown function. ORF 4  encodes a 29.7 kDa, 251 aa long protein, with a central coiled-coil region, and of unknown function. Although it is evident that NcPV1 and NcPV2 share several conserved domains, probably suggesting a common origin, their divergent spatial arrangement and their low sequence identity might be interpreted as an separation followed by recombination and reorganization of the putative proteins on the respective viruses. NcPV2 is similar to WSV6 . WSV6 was also recently identified from a pooled sample library of individuals of diverse spiders, and has been provisionally assigned to a Picorna-calici superclade. Its specific host spider remains unidentified.N. clavipes. NcPV3 is predicted to have a 9,141 nt long genome, presenting a single ORF between coordinates 680–8,914, encoding a putative 312.010 kDa and 2,744 aa long polyprotein presenting several Picornavirales associated domains . WBPV was released to GenBank last year by the University of Washington Virology NGS group, and identified in a bat RNA library, but there is no related literature available. Interestingly, NcPV3 RP shares 41% similarity  to Hubei Tetragnatha maxillosa virus 3, which was reported by Shi et al. (T. maxillosa (Tetragnathidae) RNA library. This spider is a member of the Araneoidea superfamily, which includes the Araneidae family, thus it is the closest spider to N. clavipes in which a virus was identified.An additional picorna-like virus could be associated to s Figure . The 5′  Figures . Blastp i et al. , and ideN. clavipes picorna-like virus 4 (NcPV4) shares the genome architecture of NcPV2. NcPV4 presents an 11,237 nt RNA genome enclosing four putative ORFs. NcPV4 ORF1 encodes a 2,533 aa long replicase protein . NcPV4 ORF3 encodes a 202 aa protein similar to WSV4 hypothetical protein 3, of unknown function . The 360 aa protein encoded in ORF4 shares no significant similarity to known proteins. NcPV1-NcPV3 and NcPv2-NcPV4 not only share genome organization, but also presents higher levels of similarity at their respective replicases, and sequence identity is significantly higher in equilocal regions suggesting a shared protein architecture . NcPV2 and NcPV4 appear to be more closely related to a distinctive phylogroup of unclassified Picornavirales, which diverges branching between the Dicistroviridae and the Marnaviridae families of viruses. This phylogroup is enriched with Myriapoda derived viruses, however among NcPV2 and NcPV4 distinctive subgroups there are several spider derived viruses, such as WSV4, WSV5, and WSV6. There appears to be some distinguishing cues in the replicase sequences of arthropod picorna-like viruses, which is reflected in the particular clustering correlated with evolutionary history of putative hosts in phylogenetic trees. It is worth noting that these unclassified picorna-like viruses have been identified in broad non-targeted transcriptomic studies, and the biological and ecological implications of the virus presence remains unclear. The associated literature is exceptionally limited; thus until future studies explore the potential impact of these viruses on their host, it might be prudent not to speculate on their biology. Distant virus species of this order have been studied in more detail, such as Acute bee paralysis virus  or Sacbrood Virus  which are reported to have drastic effects on their bee host, resulting in larvae death and sudden colony collapse  is predicted to have a 10,569 nt long ssRNA (+) strand genome, presenting two partially overlapping ORFs, and three additional ORFs. ORF1  encodes a putative replicase protein (RP) of 260.4 kDa and 2,266 aa long  corresponding to RNA 2 of the bisegmented Chronic bee paralysis virus (CBPV). To date, the ssRNA(+) CBPV remains unclassified by the International Committee on Taxonomy of Viruses (ICTV), and only its RP sequence presents similarities with members of the Nodaviridae and Tombusviridae families. It has been suggested that CBPV might be the prototype species of a new family of positive single-stranded RNA viruses . ORF4 and ORF5 encode a 423 and 102 aa hypothetical proteins of unknown function. NcVV1 shares with LV 51.8% nt sequence identity at the RNA level, and 28.4% at the RP protein. RP based phylogenetic trees, suggest that NcVV1 is further related to Virgaviridae distant like viruses, forming distinctive clusters of invertebrate viruses, such as the mosquito derived Negevirus. NcVV1 forms a divergent phylogroup with the proposed LV   shares with NcVV1 a super clade of Virga-like viruses. However, the divergent NcVV2 clusters within a distinct group of ssRNA(+) viruses, some of them associated with nematodes, such as the recently reported Xinzhou nematode virus 1 (XzNV1) and Xingshan nematode virus 1 . ORF3  encodes a 27.3 kDa, 244 aa long protein, akin to the hypothetical protein 3 of XgNV1 .N. clavipes bunya-like virus (NcBV) is predicted to have a segmented genome. Sequence analyses resulted in the identification of two genome segments, tentatively assigned as genomes L and G. Genome segment L of NcBV is 7,365 nt long, presenting a single ORF between coordinates 56–7,285 nt (3′−5′orientation), encoding a putative 278.9 kDa and 2,409 aa long polyprotein  is predicted to have a multisegmented dsRNA genome. RNA segment 1 is 4,060 nt long, presenting a single ORF between coordinates 10–4,005 nt, encoding a putative 152.4 kDa, 1,331 aa long RP  sharing a 23% aa identity . Hidden Markov Models (HMM) searches hint that the NcRV1 RP is similar to the Hubei reo-like virus 10 (HrlV10) RdRP , a recently described dsRNA Reoviridae like virus found in a Odonata (Odonatoptera) RNA pooled library  to the putative minor capsid protein of Hubei reo-like virus 11. Reoviridae are multipartite viruses composed of ca. 10 dsRNA genome segments. Nevertheless, there are only two virus segments available for HrlV10 encoding a RP and a second RNA genome segment encoding a minor structural protein. Given the high divergence of NcRV1 with reported virus species, subsequent sequence analyses focused on co-expression levels of virus derived RNA levels, allowed to predict two additional tentative genome segments of NcRV1. Genome segment 3 of NcRV1 is 4,302 nt long, encoding a single 1,410 aa protein, similar to the hypothetical protein 4 of Hubei odonate virus 14 . Hov14 was identified in a dragonflies and damselflies pooled RNA library; thus its specific host remains unclear. Hov14 has been tentatively assigned to the reovirus-like superclade by Shi et al.  of the Reoviridae Banna virus (BAV-Seadornavirus). BAV has been isolated from ticks and from humans suffering from meningoencephalitis . Segments 1–9 of NcRV2 segment presented a single ORF encoding a set of proteins which shares significant similarity (30–74% at the aa level) with the cognate proteins of Bloomfield virus, a proposed Reovirus, yet unclassified, described recently to be derived from a pooled sample of diverse wild caught Drosophila sp. flies  with the hypothetical protein encoded in RNA 3 of Hov14. Structural and homology based annotation rendered only a few signatures that could envision the functions of the divergent gene products of NcRV2. Homology detection and structure prediction with the HMM tool HHPred suggest that the gene products of RNA 2 and RNA 3 could have a structural function and that RNA 6 might be involved in methylation. Future studies could expand the current limited sequences reference set associated to these cluster of viruses, which might lead, in turn, to the eventual identification of highly divergent potential virus segments that should not be ruled out this soon, based on absence of evidence. RPs based phylogenetic trees hint that NcRV viruses form two divergent clusters within the Reoviridae, supporting their potential assignment to two new clades of invertebrate reo-like viruses  presents a monopartite ssRNA(+) 7,569 nt long genome. NcAV presents two partially overlapping ORFs (ORF1a-b) followed by ORF2. Based on sequence analyses, ORF1a-b appear to encode a fusion protein of 1,756 aa generated by −1 ribosomal frameshifting (RF)  and Sindbis virus  (U_UUU_UUA). The heptanucleotide signal cue is supported by the presence of a highly stable H-type pseudoknot immediately downstream the slippery sequence , a proposed astro-like virus identified by Shi et al. (Nudaurelia capensis omega virus (NucOV), the type species of the Omegatetravirus genus of Alphatetraviridae. Interestingly, Alphatetravirus capsids present a T = 4 icosahedral symmetry whereas Astrovirus capsids are T = 3. NucOV coat protein assembles into a stable particle called the procapsid, which is 450 Å in diameter. Lowering the pH to 5.0 leads to a conformational change and maturation of the capsid, mediated by an autoproteolytic cleavage dependent on the presence of an Asn-570, which is at the cleavage site  tail share a conserved secondary structure made up of three stem loops. This structure, reminiscent of the IRES Picornavirales region, is involved in virus replication  binding to the 3′ UTR, which is required for Astrovirus replication  and bird (Avastrovirus) infecting Astroviridae. CP based trees add some complexity to the picture  Figure . Astrovie Figure . HHpred e Figure . ORF1b si et al. . Intrigus Figure . The 3′ e Figure . NcAV alN. clavipes were identified and are presented as Supplementary Data N. clavipes , the corresponding and expected segment RNAs were found as independent units, presenting expected UTRs, ORFs and predicted gene products and shared a dynamic pattern of presence/absence and co-expression levels in the diverse RNA libraries.In addition to the tentative new virus species detected, new strain of reported invertebrate virus associated to N. clavipes multiple RNA-seq data derived from 16 different tissue/organ/individual isolates  collected from four female individuals , replicates in the host's nervous tissue and induces a severe neuropathy. Interestingly the wasp transmits the virus to a coccinellid host, the Spotted lady beetle Coleomegilla maculate, in which virus replication induces an antiviral immune response that correlates with paralytic symptoms. This behavior manipulation of the coccinellid, characterized by tremors, gait disturbance and limited movements, facilitates parasitism and correlates with virus RNA levels in the cerebral ganglia . Data from Babb et al. (PRJNA356433 and BioSamples accessions SAMN06132062–SAMN06132080. All short-read sequencing data are deposited in the NCBI Short Read Archive (SRX2458083–SRX2458130), and transcriptome data are available at the Transcriptome Shotgun Assembly (TSA) under accession GFKT00000000.b et al.  are avaiThe author confirms being the sole contributor of this work and approved it for publication.The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Moreover, the perpendicular magnetic anisotropy is robust, with an anisotropy constant two orders of magnitude greater than the in-plane anisotropy of the perovskite/perovskite interface. The present work demonstrates the great potential of symmetry engineering in designing artificial materials on demand.Grouping different transition metal oxides together by interface engineering is an important route toward emergent phenomenon. While most of the previous works focused on the interface effects in perovskite/perovskite heterostructures, here we reported on a symmetry mismatch-driven spin reorientation toward perpendicular magnetic anisotropy in perovskite/brownmillerite heterostructures, which is scarcely seen in tensile perovskite/perovskite heterostructures. We show that alternately stacking perovskite La Complex oxide heterostructures exhibit multifunctional behaviour that could be used in a range of device applications. Here, the authors observe that reconstruction at oxide perovskite/brownmillerite interfaces leads to perpendicular magnetic spin orientation, with potential use in spintronic devices. Grouping different TMOs into a heterostructure will break the delicate balance between different interactions, resulting in unforeseen effects5. To get an atomic level control of interface quality, usually the TMOs with similar crystal symmetry are chosen to compose artificial structures via interface engineering. Due to the excellent match in crystal symmetry and atomic configurations, the perovskite/perovskite (P/P) heterostructures have been intensively studied18. As revealed by Gibert et al.18, the charge transfer from LaMnO3 to LaNiO3 makes the originally paramagnetic LaNiO3 interfacial layer FM, yielding a magnetic pining to neighboring LaMnO3. As shown by Liao et al.19, transferring octahedron rotation from NdGaO3 to La2/3Sr1/3MnO3 (LSMO) caused an in-plane switching of the easy axis of LSMO by an angle of 90°; titling octahedron has modified the hopping rate of the eg electrons of Mn ions along the a- and b-axes, thus the magnetic anisotropy. As reported by Kan et al.20, introducing a Ca0.5Sr0.5TiO3 buffer layer to tune the network of the RuO6 octahedra leads to a lateral 45° rotation of the easy axis of SrRuO3.Transition metal oxides (TMOs) are known for their exotic properties stemming from strongly competitive mechanisms, such as ferromagnetic (FM) double exchange versus antiferromagnetic (AFM) superexchange, charge ordering vs. electron delocalization, electronic homogeneity vs. phase separation21, so far most works on interface engineering focused on the P/P type heterostructure20, a special heterostructure. In fact, there are many oxides that have similar structural framework and topotactic relationship as perovskite oxide but different atomic/electronic configurations. A typical example is the brownmillerite oxide with alternately stacked oxygen octahedra and tetrahedra30. Brownmillerite oxides have atomically ordered one-dimensional vacancy channel, showing advantages as ionic conductors, oxygen separation membranes, and catalyzers. In particular, some of them own the AFM order, providing a space for spin engineering with FM oxides. Because of the different symmetries of the two constituents, the perovskite/brownmillerite (P/B) heterostructure is expected to undergo considerable interface reconstruction, exhibiting various physical behaviors. Unfortunately, works on the effects of interlayer coupling in this kind of heterostructures are scarce, although several heterostructures have been fabricated31, probably due to the difficulty to obtain high quality P/B interfaces. Here we show an atomic level-controlled fabrication of the P/B type LSMO/LaCoO2.5 (LCO) interfaces. The present work demonstrated the great potential of symmetry engineering in the exploration for emergent phenomena in magnetic complex oxides. Its application to, as an example, the LSMO/LCO multilayers has produced a strong perpendicular magnetic anisotropy (PMA), which has been in hot pursuit of spintronics, of the LSMO layer, which is otherwise of easy plane; the maximal perpendicular energy is ~ 1.3 J cm−3, which is more than one order of magnitude higher than that achieved via the conventional approaches such as magnetoelastic coupling (from 0.01 to 0.1 J cm−3)35 and magnetocrystalline anisotropy (~ 0.018 J cm−3)32. This large PMA stems from the symmetry mismatch of the MnO6 and CoO4 layers at the LSMO/LCO interface, which results in cooperative distortions of the interfacial oxygen polyhedra as evidenced by high-resolution lattice structure analysis and density functional theory (DFT) calculations. Moreover, the symmetry break at the P/B interface is also expected to bring about emergent phenomena associated with directional electronic/ionic transport, two-dimensional electric polarization, and two-dimensional magnetism, etc., thus opening a promising avenue for the exploration for new concepts of physics and materials.Although hetero-phases have attracted increasing attention5 superlattices (SLs) were grown on TiO2-terminated (001)-SrTiO3 (STO) single crystal substrates via pulsed laser deposition following the procedures described in the Methods section. Figure 26, which own a modulation structure with a periodicity of 4a0 along c axis, where a0 is the lattice constant of the perovskite unit cell. Indeed, the typical feature of the brownmillerite structure is identified from the HAADF image. As shown in Fig. 2.5 phase, shake up satellite peaks of the Co2+ ions are also detected by X-ray photoelectron microscopy (XPS) for the LSMO/LCO multilayers  is ~ 4.2 Å at interface layer and ~ 3.8 Å elsewhere (red rhombuses). These experimental results are well reproduced by DFT calculations  as a function of the layer thickness of LCO (tLCO). Fitting the cSL-tLCO relation to a simple equation of cSL = 4cLSMO/(4 + tLCO) + tLCOcLCO/(4 + tLCO) gives rise to the out-of-plane lattice parameters of cLSMO = 3.855 Å and cLCO = 3.780 Å. Both cLSMO and cLCO are obviously smaller than the in-plane lattice constant (3.905 Å), i.e., the LSMO and LCO unit cells are tensely strained , we prepared a series of LSMO/LCO SLs with a number of repetitions of five. Figure d TMOs. To highlight the distinct effect of interlayer coupling, we performed a comparison investigation of the bare LSMO film and the LSMO/LCO heterostructure. Figure M) of the bare LSMO film and the [LSMO(4 nm)/LCO(3 nm)]5 SLs, respectively, as functions of temperature (T)  along film plane : the magnetic moment takes the maximal value at low temperatures, decreases smoothly upon warming when T is well below 320 K, and rapidly close to 320 K, the Curie temperature of LSMO. In contrast, perpendicular fields induce substantially low magnetic moments unless it is high enough. Clearly, the easy axis of the bare LSMO film is in film plane.The above results unambiguously show that the interface in our heterostructures is P/B type, totally different from the P/P interface. This is expected to have an impact on physical properties of the heterostructure. Indeed, we observed a symmetry mismatch-driven spin reorientation, which leads to a strong PMA that is not seen in the P/P heterostructure of 3M–T relations of the SLs. Take the data recorded with a field of 0.2 T as an example. Unexpectedly, the in-plane magnetic moment is low at low temperatures and smoothly increases upon warming until a local maximum is reached. After that, a rapid decrease appears. On the contrary, the out-of-plane magnetic moment is substantially high, larger by a factor of 2.6 than the in-plane one at 10 K. Increase in T causes a slow decrease of it but its superiority remains obvious up to ~ 200 K. Apparently, the spins of the P/B heterostructure prefer to align perpendicularly, rather than laterally like the bare LSMO film  is calculated. Depicting M as a function of H at fixed temperatures, we obtained a series of M–H curves based on the data in Fig. M–H loops). Figure M-H relations at 10 K, obtained with perpendicular and parallel fields, respectively. As shown, a saturation state is established in a perpendicular field of 0.2 T, whereas it is not reached in a parallel field close to 3 T. The energy required to laterally orientate the magnetic moments can be calculated from the area encircled by the two M–H curves in Fig. Ktot = 5.4 × 106 erg cm−3. The positive sign of Ktot implies PMA. Surprisingly, we obtained the PMA in tensile LSMO layer by sandwiching it between two LCO layers. For the SLs, the magnetic contributions could mainly come from LSMO, as no obvious magnetic transition is detected at the TC of LCO (~ 75 K)37. Following the similar procedure, we obtained the Ktot at high temperatures. Figure Ktot as a function of temperature. It is maximal at low temperature, slowly decreases upon warming, and changes its sign at ~ 220 K. Above 220 K, PMA is no longer supported and the easy plane state prevails. For comparison, the Ktot of the plain LSMO film is also shown in Fig. Ktot of the SLs could be even larger than that shown here, as it has to overcome the intrinsic lateral spin preference of the LSMO itself. With this consideration, the actual Ktot of the SLs could be Ktot(SLs) – Ktot(LSMO), as shown by the dashed line in Fig. 7 erg cm−3 at 10 K.To get a quantitative description of magnetic anisotropy, anisotropy constant /LSMO(tLSMO)/LCO(6 nm) trilayers, which own the main characters of the SLs but has a simple structure. The strongest PMA appears when tLSMO = 3 nm: the out-of-plane moment overwhelms the in-plane one in the whole temperature range investigated. However, in-plane component develops with the increase of the LSMO thickness and finally overtakes the out-of-plane one when tLSMO = 15 nm. To determine Ktot, M–T curves are also measured in higher fields for all trilayers /LSMO(6 nm)/LCO(tLCO) trilayers with varied LCO layer thickness. The most remarkable observation is the presence of a threshold LCO thickness for the PMA, i.e., the LCO–LSMO interlayer coupling takes effect only for thick enough LCO. As shown in Fig. M–T curves. The strongest effect occurs when tLCO = 6 nm for this series of trilayers. The corresponding Ktot is 4.5 × 106 erg cm−3 and the PMA weakens again when tLCO > 6 nm. This result shows that a too thin or a too thick LCO layer in the multilayers disfavors the PMA. Shown in Fig. Ktot – tLCO dependence. The threshold thickness for interlayer coupling is ~ 1.5 nm.To clarify the effect of LCO on interlayer coupling, we further investigated the magnetic behavior of the LCO. To highlight the effect of the P–B interface, we also conducted the computation for the 1/1-LSMO/LCO SLs , which is comparable to the experimentally obtained value (1.3 J cm−3). As a reference, the magnetic anisotropy of the P/P-type SLs (1/1-LSMO/LaCoO3) was also investigated and an easy axis in film plane is obtained Ba2Cu3O7/La2/3Ca1/3MnO3 interface, resulting in an orbital reconstruction)39. All these will cause a preferred occupation of the 40. According to the Bruno model42, easy axis prefers to take the direction of the orbital momentum. This explains the occurrence of the PMA and why the PMA prefers to appear at low temperatures.A further issue to be addressed is the physical origin of the PMA. According to the result of STEM analysis and theoretical calculations, the interfacial MnOers Fig.  and 3. Mons Fig. . This ef20. Due to the similarity of the oxygen polyhedra in two sides of the interface, however, the tuning effect is generally subtle: the typical anisotropy constant is in the order of 104 erg cm−3, as for the LSMO film on NdGaO3 19. Compared with P/P heterostructures, P/B is a promising combination. The A or B sublattice of these two oxides (ABO3 and ABO2.5) well matches each other, which allows epitaxial film growth, whereas the oxygen polyhedra symmetry at interface is completely different. This produces a great and unique interfacial reconstruction . An advantage of the present SLs over the typical metallic film for perpendicular magnetic tunnel junctions is that its PMA has a large tolerance of film thickness, occurring in the thickness range from 3 to 10 nm, rather than being limited to ~ 2 nm43. Moreover, noting the unique characters of the brownmillerite oxides as ionic conductors, catalyzers, and oxygen separation membranes, amazing effects29 in addition to PMA are expected for the P/B combinations. For example, the symmetry break at the P/B interface may bring about emergent phenomena associated with directional electronic/ionic transport, two-dimensional electric polarization and two-dimensional magnetism, etc., thus opening a promising avenue for the exploration for new concepts of physics and materials.As stated above, the PMA is important because of its irreplaceable role in high-density information storage, current-induced magnetic switching6 octahedra and CoO4 tetrahedra at the interface cooperatively relax in a manner that is unavailable for the P/P interface, resulting in unique orbital reconstructions, thus the PMA. The present work demonstrates the distinct effect arising from the interface between oxides of different symmetry. The principle proven here can be extended to other combinations of TMOs, substantially extending the space for the exploration of emergent phenomena.In summary, an atomic level controlled fabrication of the P/B-type LSMO/LCO heterostructure has been demonstrated and a symmetry mismatch-induced spin reorientation towards PMA has been observed even the heterostructure is in an in-plane tensile state. The easy axis can be deliberately tuned between parallel and perpendicular directions by altering interface effects. It is found that stacking perovskite and brownmillerite oxides alternately caused a strong and distinct interface reconstruction due to symmetry discontinuity at interface: neighboring MnO2-terminated (001)-STO single crystal substrates (3 × 5 × 0.5 mm3), using the technique of pulsed laser ablation. The fluence of the laser pulse was 2 J cm–2 and the repetition rate was 2 Hz . The substrate temperature was maintained at 700 °C (for LSMO) or 635 °C (for LCO), and the oxygen pressure . Magnetic measurements were conducted by a Quantum Designed Vibrating Sample Magnetometer (VSM-SQUID) in the temperature interval from 5 to 300 K and the magnetic field range up to 7 T.Surface morphology of the heterostructure was measured by atomic force microscope . Crystal structure of the films was determined by a Bruker X-ray diffractometer equipped with thin film accessories . Lattice images were recorded by a high-resolution STEM with double C46 using the generalized gradient approximation GGA-PBEsol functional47 for electronic exchange and correlation. A kinetic energy cutoff of 500 eV was used and the Brillouin zone was sampled with a 9 × 9 × 3 k-point grid in combination with a tetrahedron method. We performed the calculations including an on-site Coulomb repulsion term of U = 2 eV for both Mn and Co d electrons. We investigated various spin configurations, including FM, A-type AFM, and G-type AFM. As our LSMO/LCO SLs and LCO/LSMO/LCO interfaces have the same in-plane lattice constant with STO, we simulated the strain effect by fixing the in plane lattice constant to 3.905 Å unless specifically stated. The out-of-plane lattice constant was taken from experimental values deduced from the XRD and STEM data. All the internal atomic positions are optimized in the presence of the FM order. Moreover, we performed the calculations including spin-orbit coupling to describe magnetocrystalline anisotropy, which is expected to have the dominant role for the observed PMA.The DFT calculations were performed with the Vienna ab initio simulation package codeThe data that support the findings of this study are available from the corresponding author upon request.Supplementary InformationPeer Review File


In collaboration with state, tribal, local, and territorial health departments, CDC established the U.S. Zika Pregnancy Registry (USZPR) in early 2016 to monitor pregnant women with laboratory evidence of possible recent Zika virus infection and their infants.This report includes an analysis of completed pregnancies  in the 50 U.S. states and the District of Columbia (DC) with laboratory evidence of possible recent Zika virus infection reported to the USZPR from January 15 to December 27, 2016. Birth defects potentially associated with Zika virus infection during pregnancy include brain abnormalities and/or microcephaly, eye abnormalities, other consequences of central nervous system dysfunction, and neural tube defects and other early brain malformations.During the analysis period, 1,297 pregnant women in 44 states were reported to the USZPR. Zika virus–associated birth defects were reported for 51 (5%) of the 972 fetuses/infants from completed pregnancies with laboratory evidence of possible recent Zika virus infection ; the proportion was higher when restricted to pregnancies with laboratory-confirmed Zika virus infection . Birth defects were reported in 15% (95% CI = 8%–26%) of fetuses/infants of completed pregnancies with confirmed Zika virus infection in the first trimester. Among 895 liveborn infants from pregnancies with possible recent Zika virus infection, postnatal neuroimaging was reported for 221 (25%), and Zika virus testing of at least one infant specimen was reported for 585 (65%).These findings highlight why pregnant women should avoid Zika virus exposure. Because the full clinical spectrum of congenital Zika virus infection is not yet known, all infants born to women with laboratory evidence of possible recent Zika virus infection during pregnancy should receive postnatal neuroimaging and Zika virus testing in addition to a comprehensive newborn physical exam and hearing screen. Identification and follow-up care of infants born to women with laboratory evidence of possible recent Zika virus infection during pregnancy and infants with possible congenital Zika virus infection can ensure that appropriate clinical services are available. In response to the outbreak of Zika virus in the World Health Organization Region of the Americas and concerns about birth defects linked to Zika virus infection during pregnancy, CDC issued a travel notice on January 15, 2016, advising pregnant women to consider postponing travel to areas with active transmission of Zika virus. As part of the initial phase of the emergency response, CDC collaborated with state, tribal, local, and territorial health departments to establish the U.S. Zika Pregnancy Registry (USZPR) as an enhanced national surveillance system to monitor pregnancy and fetal/infant outcomes among pregnancies with laboratory evidence of possible recent Zika virus infection  recent Zika virus infection detected by a Zika virus RNA nucleic acid test  on any maternal, placental, or fetal/infant specimen or 2) detection of recent Zika virus infection or recent unspecified flavivirus infection by serologic tests on a maternal or infant specimen . Infants with positive or equivocal Zika virus IgM are included, provided a confirmatory PRNT has been performed on a maternal or infant specimen. The USZPR laboratory inclusion criteria are specified as “possible” recent Zika virus infection because the USZPR includes mother-infant pairs with serological evidence of a recent unspecified flavivirus infection, as well as those with laboratory-confirmed Zika virus infection.Analyses were done on both the overall completed pregnancies in the USZPR from the 50 U.S. states and DC and a subset of completed pregnancies that demonstrated confirmed recent Zika virus infection  women with symptoms or exposure in the first trimester onlyThe Zika virus–associated birth defects (henceforth referred to as “birth defects”) were analyzed in two mutually exclusive categories: 1) brain abnormalities and/or microcephaly regardless of the presence of additional birth defects, and 2) neural tube defects and other early brain malformations, eye abnormalities, and other consequences of central nervous system dysfunction, among fetuses and infants without evident brain abnormalities or microcephaly  was calculated, as well as the proportion who had laboratory testing for Zika virus reported on an infant specimen. CDC released updated Interim Guidance for the Evaluation and Management of Infants with Possible Congenital Zika Virus Infection in August 2016  of the 972 completed pregnancies with laboratory evidence of possible recent Zika virus infection. The proportion was higher among completed pregnancies with confirmed Zika virus infection . Among completed pregnancies with confirmed Zika virus infection, 217 of 250 (87%) tested positive by RT-PCR, including 24 pregnancies with a fetus or infant with birth defects.Birth defects were reported in similar proportions of fetuses/infants whose mothers did and did not report symptoms of Zika virus disease during pregnancy. Brain abnormalities and/or microcephaly were reported in 43 (84%) of 51 fetuses/infants with birth defects. Among pregnancies with confirmed Zika virus infection, brain abnormalities and/or microcephaly were reported in 18 (75%) of 24 fetuses/infants with birth defects. The 51 fetuses or infants with birth defects were from pregnancies with Zika virus exposure from the following 16 countries/territories with active Zika virus transmission: Barbados, Belize, Brazil, Cape Verde, Colombia, Dominican Republic, El Salvador, Guatemala, Guyana, Haiti, Honduras, Jamaica, Mexico, Puerto Rico, Republic of Marshall Islands, and Venezuela.Birth defects were reported in a higher proportion of fetuses or infants whose mothers were infected during the first trimester of pregnancy. Among 157 pregnancies in which women had symptom onset or exposure to Zika virus infection during the first trimester, 14 (9%) fetuses/infants had reported birth defects . When prAmong the 895 liveborn infants, postnatal neuroimaging results were reported to the USZPR for 221 (25%). Zika virus testing results of any specimen were reported for 585 (65%) infants; 94 (11%) of all 895 liveborn infants had positive Zika virus test results. Among the 45 liveborn infants with birth defects, 25 (56%) had positive infant Zika virus testing results reported, and 29 (64%) had postnatal neuroimaging reported to the USZPR . Among tThe number of pregnant women with laboratory evidence of possible recent Zika virus infection and the number of fetuses/infants with Zika virus–associated birth defects continues to increase in the United States. The proportion of fetuses and infants with birth defects among pregnancies with confirmed Zika virus infection at any time during pregnancy was more than 30 times higher than the baseline prevalence in the pre-Zika years, and a higher proportion of those with first trimester infections had birth defects  (These findings underscore the serious risk for birth defects posed by Zika virus infection during pregnancy and highlight why pregnant women should avoid Zika virus exposure and that all pregnant women should be screened for possible Zika virus exposure at every prenatal visit, with testing of pregnant women and infants in accordance with current guidance (• In 2016, a total of 1,297 pregnancies with possible recent Zika virus infection were reported to the U.S. Zika Pregnancy Registry from 44 states.• Approximately one in 10 pregnancies with laboratory-confirmed Zika virus infection resulted in a fetus or infant with Zika virus–associated birth defects.• The proportion of fetuses and infants with Zika virus–associated birth defects was highest among those with first trimester Zika virus infections.• Only 25% of infants from pregnancies with possible recent Zika virus infection reported receiving postnatal neuroimaging.• Identification and follow-up care of infants born to mothers with laboratory evidence of possible recent Zika virus infection during pregnancy and infants with congenital Zika virus infection can ensure that appropriate intervention services are available to affected infants.https://www.cdc.gov/vitalsigns/.• Additional information is available at


Viral encephalitis causes acute inflammation of the brain parenchyma and is a significant cause of human morbidity and mortality. Although Herpes Simplex encephalitis is the most frequent known cause of fatal sporadic encephalitis in humans, an increasingly wide range of viruses and other microbial pathogens are implicated. Up to 60% of cases of presumed viral encephalitis remain unexplained due to the failure of conventional laboratory techniques to detect an infectious agent. High-throughput DNA sequencing technologies have the potential to detect any microbial nucleic acid present in a biological specimen without any prior knowledge of the target sequence. While there remain challenges intrinsic to these technologies, they have great promise in virus discovery in unexplained encephalitis. Encephalitis refers to inflammation of the brain parenchyma that typically presents with a combination of features including fever, headache, clouding of consciousness, seizures, personality change, focal neurological deficits, and coma ,2. It isN-methyl-d-aspartate (NMDA) receptor [Forty to sixty percent of encephalitis cases are unexplained ,2,4,5. Ereceptor ,9.Viruses are an important, yet often poorly understood cause of encephalitis. A range of viral agents have been implicated . Herpes The most relevant means of neurological identification of a causative agent in encephalitis is by examination of the cerebrospinal fluid (CSF) or brain tissue obtained at either brain biopsy or autopsy. Methods employed include virus isolation, polymerase chain reaction (PCR), immunohistochemistry or electron microscopy, serology, and, more recently, high-throughput sequencing (HTS). Viral isolation is the classical method for diagnosis, but it is increasingly used in concert with PCR, which has become more accessible and can provide near immediate actionable data. Culture requires days to weeks and may fail in the event that the virus cannot be propagated in the cell line selected, whereas PCR requires only a few hours.The role of CSF PCR in the diagnosis of viral infections of the nervous system has been extensively reviewed . While tSerology can succeed where both culture and PCR fail; however, diagnosis may require the collection of samples at two different time points when IgM is negative, and cross-reactive antibodies can confound specificity, as during the early phase of investigation during the 1999 West Nile virus outbreak in North America when St. Louis encephalitis virus was inaccurately implicated ,14. FurtDiagnosis of an infection is best achieved by obtaining a specimen directly at the site of disease. However, a brain biopsy is often a measure of last resort and, consequently, the specimen is rarely available. When accessible, the scarce amount of tissue material available greatly restricts the number of experiments that can be performed. In most cases, CSF is the only surrogate specimen available for the assessment of CNS disorders.In instances where CNS sampling is not possible or fails to provide clues to a causative agent, some clinicians and investigator survey blood, the oropharynx, feces, or urine. The identification of an agent in any sample type may lead to targeted assays of CSF or antibody tests, if seroconversion is consistent with acute infection.The advent of unbiased DNA sequencing technologies offers unprecedented opportunities to detect unexpected, previously unrecognized, or novel microbial pathogens in undiagnosed encephalitis. Unlike sequence-based methods that are limited to the detection of known or related agents, unbiased high-throughput sequencing (HTS) has the potential to detect any microbial nucleic acid present in a biological specimen without any prior knowledge of the target sequence. The ability of sample multiplexing using barcodes enables simultaneous sequencing of a large number of samples in a single run. This strategy fully exploits HTS systems by increasing throughput while reducing sequencing costs .Sample collection, transporting, handling, and storage conditions can profoundly affect the quality of the sample and thus sequencing results. The development of laboratory standard operating procedures is critical to ensure high quality input material.While HTS provides the most broad-based, unbiased approach for the detection of known and novel viral agents from biological samples, it also faces challenges related to sample preparation and data analysis that impede widespread use of the technique. The sample preparation protocols for HTS require the removal of host-derived sequences, which are present in the sample, in order to ensure that the sequence reads are of viral rather than host origin. Additionally, the massive amount of sequence information generated in HTS requires extensive support from bioinformatics experts, customized bioinformatic tools and powerful computer clusters to efficiently and rapidly analyze the data.The choice of a sample preparation protocol strictly depends on the nature of the biological specimen. Because host nucleic acid is present in great abundance as compared to virus genetic material in biological specimens, procedures such as the depletion of host background or the enrichment of viral genomes is critical to ensure the threshold detection of a viral signature. This is particularly true for complex clinical specimens such as tissue samples like brain . ConversViral nucleic acids may be enriched using either subtractive or positive selection methods. Host and non-viral templates can be reduced through filtration, ribosomal RNA subtraction, and nuclease treatment ,19,20,21The systematic analysis of extensive sequence data requires customized bioinformatic tools as well as powerful computers. Genomic footprints of viruses are detected using Basic Local Alignment Search Tool (BLAST) programs that compare sequences obtained by sequencing to those present in a reference database. This sequence-based approach relies on the concept that new viruses will share homology with known viruses.http://www.ncbi.nlm.nih.gov) using tBLASTx and BLASTx.The pipeline starts with the trimming of raw sequence reads to remove the adapter sequences from the reads. Sequences are then filtered to eliminate low-complexity reads and repetitive sequences. The dataset is further enriched for potential viral sequences by removing contaminating sequence reads derived from host and bacterial sequences, followed by the clustering of redundant sequences. The remaining sequences are further assembled into contiguous sequences and compared to the GenBank databases at the nucleotide (BLASTN) and to the non-redundant protein sequences (nr) database from NCBI  . The appThe system employs approximately 2 million biotinylated oligonucleotide probes designed to bind to the coding sequences of all viral taxa known to infect vertebrates at intervals of 50–100 nt. Libraries prepared from random primed cDNA are hybridized with the biotinylated VirCap probes. Streptavidin magnetic beads are added to trap biotinylated probes and their complementary library components. After magnetic capture and washing, nucleic acid is released from the beads and subjected to post-hybridization PCR prior to sequencing . DemultiA potential confounding factor is that VirCapSeq-VERT may not detect entirely new viruses; however, the likelihood of missing a virus that causes encephalitis is low because probe nucleic acid homology to target is not as stringent as PCR; the entire genomic sequence of each virus is accessible for capture and databases and probe libraries are continuously updated.The introduction of contaminating microbial sequences during the experimental process will lead to false-positive findings and erroneous data interpretation ,24,25,26bona fide microbial contaminations may be identified by systematically processing and sequencing a mock library preparation and a control sample in parallel with the experimental samples. Additionally, the use of a repository of contaminating sequences may help in some cases alleviate the effects of contamination by computationally excluding them from downstream analysis. Importantly, any microbial signature identified by HTS needs to further be validated in the original sample by specific PCR assays based on newly identified microbial genome sequences.The identification of A suggested flow chart for the laboratory investigation of unexplained viral encephalitis is shown in The presence of a signal provides an initial guide for therapy; however, CSF should be retested to ensure the validity of a finding. Cell counts and differential cell counts, as well as protein and glucose measures may be helpful; nonetheless, we proceed with comprehensive molecular analyses until a specific diagnosis is achieved, irrespective of whether the pattern is more consistent with bacterial or viral infection. Where PCR fails to implicate an agent, we move to high-throughput sequencing of CSF. High-throughput sequencing techniques have the potential to detect nucleic acids that represent viruses, bacteria, fungi, or parasites. However, the method is less sensitive than the analysis of libraries that target specific phyla. Accordingly, we typically sequence libraries prepared following 16S rRNA (bacteria) , internaUnbiased HTS has proven to be successful in detecting infectious agents in cases of encephalitis of unknown etiology, including a novel lymphocytic choriomeningitis virus-related arenavirus that caused transplant-associated encephalopathy and an astrovirus as the cause of fatal encephalitis in an immunocompromised patient ,31. ThesFurthermore, recent studies have identified both Parvovirus 4 (PARV4), an emerging virus identified recently in human blood and other tissues with no known associated disease, and a novel cyclovirus in CSF specimens from patients with acute central nervous system infections of unknown etiology ,37.This last point reinforces the notion that the identification of viral sequences in clinical material is not necessarily the same as showing a causal association between the virus and the neurological disease. Such an association is easiest to make when the infectious agent has been isolated at an appropriate site during the course of active disease. However, access to specimens such as brain biopsy is generally not available and the only surrogate specimen available for analysis is CSF. The possibility always exists that the novel virus identified may play no role in disease pathogenesis and represents an irrelevant co-infection. We must always consider and control for the possibility of sample contamination, a particular hazard with PCR . Nevertheless, identification of a virus in a case of presumed viral encephalitis would normally be construed as strong evidence of causation unless there was additionally strong evidence to suggest either accidental contamination during the procedure or an irrelevant co-existing infection.Since CSF is normally acellular and sterile, clear evidence of a microbial signature in CSF merits strong consideration as a candidate pathogen. Failure to detect a viral agent in a CNS disease such as viral encephalitis may reflect the absence of its molecular footprint at the time of sampling. Notwithstanding these caveats, investigating CNS infections of unknown etiology using unbiased HTS or VirCapSeq-VERT may identify the complete spectrum of causative agents associated with encephalitis. Further, early recognition of the causative agent of unexplained encephalitis cases could enable specific interventions that reduce illness and death and facilitate the recognition of outbreaks that threaten public health.Despite the progress that has been made with both classical virological and molecular biological technologies in identifying the numerous causes of viral encephalitis, there is an increasing need for better tools and new approaches for detecting both genuinely novel pathogens in such patients and determining new manifestations of known pathogens, including viruses. In evaluating the results of different studies, it is crucial to have uniform clinical and diagnostic criteria for establishing a definite diagnosis of viral encephalitis. There is not a standardized algorithm for the molecular investigation of unexplained encephalitis, though general guidelines for the investigation and management of suspected viral encephalitis are now available ,38. Prob


Culex neavei mosquitoes in South Africa. The first recognized infection in a human was in an African man with fever and rash in 1959 but was not reported until 1981.Usutu virus , named fIn 2001, Usutu virus emerged in Europe, when it was identified as the etiologic agent of bird—mainly blackbird—mortality. Retrospective analysis of archived tissue samples from wild bird deaths in the Tuscany region of Italy in 1996, however, revealed an earlier introduction of the virus to Europe. It was not thought to be associated with severe or fatal disease in humans until a neuroinvasive infection was reported to have occurred in an Italian woman in 2009.


We investigated dengue and chikungunya virus antibody seroprevalence in French Polynesia during 2014–2015. Dengue virus seroprevalence was ≈60% among schoolchildren and >83% among the general population; chikungunya virus seroprevalence was <3% before and 76% after Zika virus emergence (2013). Dengue virus herd immunity may affect Zika virus infection and pathogenesis. Flaviviridae, genus Flavivirus)  emerged there in 2013  in 2014  and the Fisher exact test. We set significance at p<0.05.We performed detection of DENV and chikungunya virus IgG on blood samples collected in 2014 by using a recombinant-antigen–based indirect ELISA  and schoolchildren (1% ± 1%) in 2014 corroborate previous findings for blood donors sampled during 2011–2013 (3%)  than Zika virus antibody seroprevalence for the general population in 2015 (22% ± 6%) (