New insights into the role of ticks in African swine fever epidemiology.

African swine fever (ASF), one of the most important diseases of swine, is present in many African countries, as well as in eastern Europe, Russia and Sardinia. It is caused by a complex virus, ASF virus (ASFV), for which neither vaccine nor treatment is available. ASFV affects swine of all breeds and ages, and also replicates in soft ticks of the genus Ornithodoros, facilitating ASFV persistence and reocurrence of disease. Depending on the involvement of these ticks, and the presence or not of sylvatic asymptomatic animals, several epidemiological cycles have been identified. The disease persists in East and southern African countries in a sylvatic cycle between O. porcinus (of the O. moubata species complex) and common warthogs. In some countries a domestic pig-tick cycle exists, whereas in other regions, notably West Africa, the role of soft ticks has not been demonstrated, and ASFV is transmitted between domestic pigs in the absence of tick vectors. Even in several East and Central African countries which have the sylvatic or domestic cycle, the majority of outbreaks are not associated with ticks or wild suids. In Europe, O. erraticus was detected and identified as a crucial vector for ASF maintenance in outdoor pig production on the Iberian Peninsula. However, in most parts of Europe, there is a lack of information about the distribution and role of Ornithodoros ticks in ASF persistence, particularly in eastern regions. This article reviews ASF epidemiology and its main characteristics, with a special focus on the distribution and role of soft ticks in ASF persistence in different settings. Information abouttick detection, control measures and future directions for research is also included.

African swine fever diagnosis update.

African swine fever (ASF) is one of the most complex livestock diseases. The significant losses that it causes, coupled with the lack of a vaccine against ASF virus and the possible resemblance with other swine hemorrhagic diseases, make early detection and laboratory diagnosis essential for controlling and managing the disease. All the techniques currently used to diagnose ASF have been fully validated showing high sensitivity and specificity to detect both antigen and antibodies against all 22 known genotypes; and enable the correct diagnosis of ASF in all possible epidemiological situations. Because no vaccine is available, the presence of antibodies always indicates previous infection, and serological diagnosis must always be performed in parallel with antigen detection to increase the sensitivity and specificity of the analyses. Recent developments in ASF diagnosis, specifically the new field diagnostic tests, have improved and facilitated the likelihood of ASF early detection, essential to fighting the disease.

Metabolomic changes in lactating multiparous naturally MAP-infected Holstein-Friesian dairy cows suggest changes in mitochondrial energy pathways.

Mycobacterium avium subspecies paratuberculosis (MAP) is the causative organism of Johne's Disease, a chronic intestinal infection of ruminants. Infected cows begin shedding MAP within the asymptomatic, subclinical stage of infection before clinical signs, such as weight loss, diarrhoea and reduced milk yields develop within the clinical stages of disease. Herein, we examine the milk metabolomic profiles of naturally MAP-infected Holstein-Friesian cows. The study used biobanked milk samples which were collected 73.4 ± 3.79 (early lactation) and 143 ± 3.79 (mean ± SE) (mid-lactation) days post-calving from 5 MAP-infected and 5 control multiparous cows. The milk metabolome was assessed using flow infusion electrospray high-resolution mass spectrometry (FIE-HRMS) for sensitive, non-targeted metabolite fingerprinting. Metabolite fingerprinting assessments using partial least squares discriminate analyses (PLS-DA) indicated that lactation stage was a larger source of variation than MAP status. Examining each lactation stage separately for changes associated to MAP-infection status identified 45 metabolites, 33 in early lactation and 12 in mid-lactation, but only 6 metabolites were targeted in both stages of lactation. Pathway enrichment analysis suggested that MAP affected the malate-aspartate shuffle during early lactation. Pearson's correlation analysis indicated relationships between milk lactose concentrations in mid-lactation and 6 metabolites that were tentatively linked to MAP-infection status. The targeted metabolites were suggestive of wider changes in the bioenergetic metabolism that appear to be an acceleration of the effects of progressing lactation in healthy cows. Additionally, milk lactose concentrations suggest that MAP reduces the availability of lactose derivatives.

Impact of AQP3 inducer treatment on cultured human keratinocytes, ex vivo human skin and volunteers.

One of the main functions of the skin is to protect the organism against environmental threats, such as thermal stress. Aquaporin-3 (AQP3) facilitates water and glycerol transport across cell membranes and therefore regulates osmotic balance in different situations of stress. This mechanism seems to be particularly important for the resistance of different organisms to cold stress. Consequently, we were interested in investigating the effect of cold and osmotic stress on AQP3 expression in normal human keratinocytes. We developed a new active ingredient to stimulate aquaporins in skin and demonstrated the partial restoration of AQP3 expression in keratinocytes transfected with AQP3 siRNA. Moreover, we examined the effect of cold stress on cell morphology and the impact of a pre-treatment with the active ingredient. Our results indicated that induction of AQP3 helped maintain a correct organization of the actin cytoskeleton, preserving cell morphology and preventing cells from rounding. Immunofluorescent staining revealed cytoplasmic localization of AQP3 and its translocation to the cell membrane following osmotic stress. Histological ex vivo studies of skin under different conditions, such as cold environment and tape-stripping, indicated that increase in AQP3 expression appears to be involved in skin protection and showed that the pattern of AQP3 expression was more enhanced in the active ingredient-treated samples. In vivo confocal microscopy by Vivascope showed a generally healthier appearance of the skin in the treated areas. These results attest to the potential value of the active ingredient in optimizing environmental stress resistance and protecting the skin from stratum corneum damage.

The control of chlorophyll catabolism and the status of yellowing as a biomarker of leaf senescence.

The pathway of chlorophyll catabolism during leaf senescence is known in a fair amount of biochemical and cell biological detail. In the last few years, genes encoding a number of the catabolic enzymes have been characterized, including the key ring-opening activities, phaeophorbide a oxygenase (PaO) and red chlorophyll catabolite reductase (RCCR). Recently, a gene that modulates disassembly of chlorophyll-protein complexes and activation of pigment ring-opening has been isolated by comparative mapping in monocot species, positional cloning exploiting rice genomics resources and functional testing in Arabidopsis. The corresponding gene in pea has been identified as Mendel's I locus (green/yellow cotyledons). Mutations in this and other chlorophyll catabolic genes have significant consequences, both for the course of leaf senescence and senescence-like stress responses, notably hypersensitivity to pathogen challenge. Loss of chlorophyll can occur via routes other than the PaO/RCCR pathway, resulting in changes that superficially resemble senescence. Such 'pseudosenescence' responses tend to be pathological rather than physiological and may differ from senescence in fundamental aspects of biochemistry and regulation.

Why is African swine fever still present in Sardinia?

African swine fever (ASF) is an infectious disease of swine that has been present in Sardinia since 1978. Soon after introduction of the disease, several control and eradication programmes were established with limited success. Some researchers attributed the persistence of the disease in central and eastern areas to certain socio-economic factors, the existence of some local and traditional farming practices (i.e., unregistered free-ranging pigs known as brado animals) and the high density of wild boar in the region. In the past, scarcity of swine data in Sardinia complicated the evaluation and study of ASF on the island. More complete, accurate and reliable information on pig farms has become available as a result of the most recent eradication programmes. Here, we perform statistical modelling based on these data and the known distribution of domestic pig and wild boar to identify the main risk factors that have caused ASF persistence in Sardinia. Our results categorized, identified and quantified nine significant risk factors, six of which have not been previously described. The most significant factors were the number of medium-sized farms, the presence of brado animals and the combination of estimated wild boar density and mean altitude above sea level. Based on these factors, we identified regions in eastern and central Sardinia to be at greatest risk of ASF persistence; these regions are also where the disease has traditionally been endemic. Based on these risk factors, we propose specific control measures aimed at mitigating such risks and eradicating ASF from the island.

Understanding African Swine Fever infection dynamics in Sardinia using a spatially explicit transmission model in domestic pig farms.

African swine fever virus (ASFV) has been endemic in Sardinia since 1978, resulting in severe losses for local pig producers and creating important problems for the island's veterinary authorities. This study used a spatially explicit stochastic transmission model followed by two regression models to investigate the dynamics of ASFV spread amongst domestic pig farms, to identify geographic areas at highest risk and determine the role of different susceptible pig populations (registered domestic pigs, non-registered domestic pigs [brado] and wild boar) in ASF occurrence. We simulated transmission within and between farms using an adapted version of the previously described model known as Be-FAST. Results from the model revealed a generally low diffusion of ASF in Sardinia, with only 24% of the simulations resulting in disease spread, and for each simulated outbreak on average only four farms and 66 pigs were affected. Overall, local spread (indirect transmission between farms within a 2 km radius through fomites) was the most common route of transmission, being responsible for 98.6% of secondary cases. The risk of ASF occurrence for each domestic pig farm was estimated from the spread model results and integrated in two regression models together with available data for brado and wild boar populations. There was a significant association between the density of all three populations (domestic pigs, brado, and wild boar) and ASF occurrence in Sardinia. The most significant risk factors were the high densities of brado (OR = 2.2) and wild boar (OR = 2.1). The results of both analyses demonstrated that ASF epidemiology and infection dynamics in Sardinia create a complex and multifactorial disease situation, where all susceptible populations play an important role. To stop ASF transmission in Sardinia, three main factors (improving biosecurity on domestic pig farms, eliminating brado practices and better management of wild boars) need to be addressed.

Serological Surveillance and Direct Field Searching Reaffirm the Absence of Ornithodoros Erraticus Ticks Role in African Swine Fever Cycle in Sardinia.

African swine fever (ASF), one of the most important diseases of swine, has been endemic in the Italian island of Sardinia for more than 35 years. During these decades, several strategies and eradication efforts have been implemented in the island with limited success. Strong climatic and ecological similarities exist between Sardinia and one area of the Iberian Peninsula where Ornithodoros erraticus ticks were involved in the persistence of ASF from 1960 to 1995. This fact leads to the hypothesis that, potentially, Ornithodoros ticks could be also involved in the ASF cycle in Sardinia, thus accounting for some of the reoccurring ASF outbreaks in this island. Initial efforts aimed at detection of Ornithodoros ticks in Sardinia were performed during the 1970s/1980s with no positive results. Accordingly, the absence of Ornithodoros ticks in Sardinia has been generally accepted. However, since a new and reinforced ASF eradication programme has been recently launched in Sardinia, it is essential to clarify the presence and role of these soft ticks in the epizootiology of ASF in this island. For that purpose, 1767 porcine serum samples collected from all around the island (1261 from domestic and 506 from wild boar) were analysed by ELISA for antibodies to salivary antigens of Ornithodoros erraticus. In addition, Ornithodoros ticks were directly searched in a number of pig premises that have suitable habitats for these ticks and were located in areas repeatedly affected by ASF. Only one serum sample resulted positive in the serological assay, and no Ornithodoros ticks were collected in none of the premises. These results indicate that these soft tick species are not involved in the epizootic cycle of ASF in Sardinia and highlight the importance of controlling other risk factors still present in the island for effectively eradicate the disease.

Thirty-Five-Year Presence of African Swine Fever in Sardinia: History, Evolution and Risk Factors for Disease Maintenance.

Despite the implementation of control efforts and funds to fight against the disease, African swine fever (ASF) has been present in Sardinia since 1978. It has caused serious problems for both the industrial pig sector and the regional authorities in Sardinia, as well as the economy of Italy and the European Union, which annually supports the costly eradication programme. During this time, ASF has persisted, especially in the central-east part of Sardinia where almost 75% of the total outbreaks are concentrated. The Sardinian pig sector is clearly divided into two categories based on the specialization and industrialization of production: industrial farms, which represents only 1.8% of the farms in the island and non-professional holdings, which are comprised of small producers (90% of pig holdings have <15 pigs) and apply little to no biosecurity measures. Additionally, illegally raised pigs are still bred in free-ranging systems in certain isolated parts of the island, despite strict regulations. The illegal raising of pigs, along with other high-risk management practices (e.g., use of communal areas) are likely the primary reasons for endemic persistence of the virus in this area. The compensation provided to the farmers, and other aspects of the eradication programme have also negatively influenced eradication efforts, indicating that socio-cultural and economic factors play an important role in the epidemiology of ASF on the island. The aim of this study was to comprehensively review the evolution of the 35-year presence of ASF in Sardinia, including control measures, and the environmental and socio-economic factors that may have contributed to disease endemicity on the island. The present review highlights the need for a coordinated programme that considers these socio-economic and environmental factors and includes an assessment of new cost-effective control strategies and diagnostic tools for effectively controlling ASF in Sardinia.

Detection of African Swine Fever Antibodies in Experimental and Field Samples from the Russian Federation: Implications for Control.

African swine fever (ASF) re-entered in Europe in 2007 by Georgia rapidly affecting neighbouring countries. Since then, ASF has caused severe problems to the Russian Federation (RF) and spread to Northern and Western regions, including Ukraine (2012 and 2014) and Belarus (2013). At the beginning of 2014, dead wild boars were found in Lithuania and Poland. Several outbreaks have been later notified in the European Union(EU), affecting domestic pigs and wild boar of Latvia, Lithuania and Poland, and also wild boar in Estonia, causing major problems for the EU pig sector. Some studies have been performed with this ASFV isolate, revealing that it belongs to genotype II and causes an acute form of the disease. However, few data are available about the presence of antibodies in field and experimental samples from the affected area. This study analysed samples from experimental infections with ASFV isolated from the RF in 2013 (74 sera and 3 tissue exudates), and field samples from the RF from 2013 to 2014 (266 samples, including 32 and 7 tissue exudates from domestic pigs and wild boar, respectively). All samples were tested by a commercial ELISA and, some of them (79), also by immunochromatographic tests. Positive and doubtful samples were confirmed by immunoblotting test. Positive results were found in experimental and field samples, which confirm the presence of antibodies against ASFV in the RF. Antibodies were detected in animals inoculated with three different ASFV isolates, with some differences found among them. Only a small percentage of field samples was positive for ASF antibodies (3.7%), in agreement with other observations that reported a high virulence for the ASFV isolates in the area. These results confirm the potential presence of survivor animals that should be considered in affected areas to help design effective control and eradication plans against ASF.

An update on the epidemiology and pathology of African swine fever.

African swine fever (ASF) is one of the most important infectious diseases of swine and has major negative consequences for affected countries. ASF is present in many sub-Saharan countries, Sardinia and several countries of eastern and central Europe, where its continuous spread has the swine industry on heightened alert. ASF is a complex disease for which no vaccine or treatment is available, so its control is based on early detection and rapid control of spread. For a robust and reliable early detection programme it is essential to be able to recognize the clinical signs and pathological changes of ASF, keeping in mind that in most cases the first introductions don't show high mortality nor characteristic clinical signs or lesions, but fever and some hemorrhagic lymph nodes. Knowledge of the main characteristics of this infection, including its current distribution and routes of transmission, is also essential for preventing and controlling ASF. This review addresses each of these topics and aims to update knowledge of the disease in order to improve early detection of ASF in the field and allow implementation of public health programmes.

Assessing the Risk of African Swine Fever Introduction into the European Union by Wild Boar.

The presence of African swine fever (ASF) in the Caucasus region and Russian Federation has increased concerns that wild boars may introduce the ASF virus into the European Union (EU). This study describes a semi-quantitative approach for evaluating the risk of ASF introduction into the EU by wild boar movements based on the following risk estimators: the susceptible population of (1) wild boars and (2) domestic pigs in the country of origin; the outbreak density in (3) wild boars and (4) domestic pigs in the countries of origin, the (5) suitable habitat for wild boars along the EU border; and the distance between the EU border and the nearest ASF outbreak in (6) wild boars or (7) domestic pigs. Sensitivity analysis was performed to identify the most influential risk estimators. The highest risk was found to be concentrated in Finland, Romania, Latvia and Poland, and wild boar habitat and outbreak density were the two most important risk estimators. Animal health authorities in at-risk countries should be aware of these risk estimators and should communicate closely with wild boar hunters and pig farmers to rapidly detect and control ASF.

Efficiency of light utilization of Chlamydomonas reinhardtii under medium-duration light/dark cycles.

The light regime inside a photobioreactor is characterized by a light gradient with full (sun)light at the light-exposed surface and darkness in the interior of the bioreactor. Consequently, depending on the mixing characteristics, algae will be exposed to certain light/dark cycles. In this study the green alga Chlamydomonas reinhardtii was cultivated under five different light regimes: (1) continuous illumination; (2) a square-wave light/dark cycle with a light fraction (epsilon) of 0.5 and a duration (t(c)) of 6.1 s; (3) epsilon=0.5, t(c)=14.5 s; (4) epsilon=0.5, t(c)=24.3 s and (5) epsilon=0.8, t(c)=15.2 s. The biomass yield on light energy, protein per photons, decreased under light/dark cycles (epsilon=0. 5) in comparison to continuous light (CL), from 0.207 (CL) to 0.117-0.153 g mol(-1) (epsilon=0.5). Concomitantly, the maximal specific photosynthetic activity, oxygen production per protein, decreased from 0.94 (CL) to 0.64-0.66 g g(-1) h(-1) (epsilon=0.5). Also the quantum yield of photochemistry, yield of the conversion of light energy into chemical energy, decreased from 0.47 (CL) to 0. 23 (epsilon=0.5, t(c)=24.3 s). Apparently, C. reinhardtii is not able to maintain a high photosynthetic capacity under medium-duration light/dark cycles and since specific light absorption did not change, light utilization efficiency decreased in comparison to continuous illumination.

Waist circumference values in Spanish children--gender related differences.

OBJECTIVE: To obtain reference values of the waist circumference in Spanish children, and to investigate their dependence on age and gender. DESIGN: Cross-sectional study. SETTING: General school-age population. SUBJECTS: A representative sample of the schools in Zaragoza, Spain, was drawn from seven schools. The population selected comprised 1728 children with ages ranging from 6.0-14.9y. Of the original sample, 368 children (21.29%) were excluded because of chronic diseases or refusal. Finally, 1360 children and adolescents: 701 boys and 659 girls, were studied. INTERVENTIONS: Waist and hip circumferences were measured with an unelastic tape. RESULTS: Waist circumference tended to be higher in males than in females and this difference was significant after 11.5y. In general, hip circumference was higher in females than in males (statistically significant differences at 7.5, 10.5, 12.5 and 13.5 y). In general, percentile values of waist circumference were higher in males than in females, especially after 12.5 y. Difference between males and females on percentile 95 at 14.5 y was 7.6 cm. Hip was greater than waist in both sexes, and the two curves run nearly parallel in males. In females, while hip enlarges continuously, waist shows the reverse tendency between 11.5 and 14.5 y. CONCLUSIONS: Waist circumference showed higher values in boys than in girls, especially after 11.5 y, and waist values increase with age both in males and females. These findings justify the use of age and gender specific reference standards.

[Plasma concentrations of fibronectin in septic, hypovolemic, and cardiogenic shock. Prognostic usefulness of their variation]. / Concentraciones plasmáticas de fibronectina en el shock séptico, hipovolémico y cardiogénico. Utilidad pronóstica de sus variaciones.

BACKGROUND: Fibronectin is a protein with opsonic capacity. Its plasma level is diminished in septic shock. The aim of the present was to study its behavior in other types of shock. METHODS: A prospective study of 60 patients in shock (septic, hypovolemic and cardiogenic) was carried out. Serial plasma levels of fibronectin were determined in these patients over a period of 72 hours. The values of the cases which evolved favorably were compared with the values of those which did not. RESULTS: In all the cases, the plasma levels of fibronectin were diminished within a minimum of approximately 12 hours with a tendency to recovery of initial values being observed within 72 hours, except in cases in which the evolution of the patient was poor. In these cases fibronectin values remained diminished in such statistical significant. CONCLUSIONS: Plasma levels of fibronectin behave similarly in the three types of shock studied (septic, hypovolemic and cardiogenic). Serial determination of fibronectin provides a good prognostic index in patients with shock.

[Effect of age, nutritional status and tumor extension on various immunologic parameters in cancer patients]. / Influencia de la edad, el estado nutricional y la extensión tumoral sobre diversos parámetros inmunitarios en pacientes neoplásicos.

We studied cellular immune (total lymphocytes, lymphocytary populations and delay hypersensitivity skin tests) and humoral (immunoglobulins and complement) response, as well as nutritional status (anthropometric and biochemical para meters), antitumoral treatment. The effect of tumoral extension, nutritional status and age on immunity was assessed. We have demonstrated a poor relationship between the cellular immunological response and the extension of neoplasia, a moderate effect of age and a significant impact of nutritional status.

Epidemiology of African swine fever virus.

African swine fever virus used to occur primarily in Africa. There had been occasional incursions into Europe or America which apart from the endemic situation on the island of Sardinia always had been successfully controlled. But following an introduction of the virus in 2007, it now has expanded its geographical distribution into Caucasus and Eastern Europe where it has not been controlled, to date. African swine fever affects domestic and wild pig species, and can involve tick vectors. The ability of the virus to survive within a particular ecosystem is defined by the ecology of its wild host populations and the characteristics of livestock production systems, which influence host and vector species densities and interrelationships. African swine fever has high morbidity in naïve pig populations and can result in very high mortality. There is no vaccine or treatment available. Apart from stamping out and movement control, there are no control measures, thereby potentially resulting in extreme losses for producers. Prevention and control of the infection requires good understanding of its epidemiology, so that targeted measures can be instigated.

African swine fever: an epidemiological update.

African swine fever (ASF) is one of the most important swine diseases, mainly because of its significant sanitary and socioeconomic consequences. This review gives an update on the epidemiology of the disease and reviews key issues and strategies to improve control of the disease and promote its eradication. Several characteristics of ASF virus (ASFV) make its control and eradication difficult, including the absence of available vaccines, marked virus resistance in infected material and contaminated animal products, and a complex epidemiology and transmission involving tick reservoir virus interactions. The incidence of ASF has not only increased on the African continent over the last 15 years, so that it now affects West African countries, Mauritius and Madagascar, but it has also reached new areas, such as the Caucasus region in 2007. In fact, the rapid spread of the disease on the European continent and the uncontrolled situation in the Russian Federation places all countries at great risk as a result of intense global trade. The proximity of some affected areas to the European Union (EU) borders (<150 km) has increased concerns about the potential economic consequences of an ASF incursion into the EU pig sector. Establishing effective surveillance, control and eradication programmes that implicate all actors (veterinarians, farmers, and policy makers) is essential for controlling ASF. African swine fever -free countries should be aware of the potential risk of ASF incursion and implement risk reduction measures such as trade controls and other sanitary measures. This review will discuss lessons learnt so far about ASF control, current challenges to its control and future studies needed to support global efforts at prevention and control.

Monitoring of African swine fever in the wild boar population of the most recent endemic area of Spain.

Wild boars are natural hosts for African swine fever (ASF). The ASF virus (ASFV) can persist for long periods in the environment, such as in ticks and contaminated products, which may be sources of infection for wild boar populations. African swine fever was eradicated in domestic pig populations in Spain in 1995, after 35 years of significant effort. To determine whether ASFV can persist in wild boar hosts after it has been eradicated from domestic pigs and to study the role of wild boar in helping ASFV persist in the environment, we checked for the presence of ASFV in wild boars in Doñana National Park, one of the largest natural habitats of wild boar in Spain and one of the last areas where ASF was endemic prior its eradication. Samples from 158 animals collected between 2006 and 2010 were analysed using serological and nucleic acid-based diagnostic techniques recommended by the World Organization for Animal Health (OIE). None of the samples was found to be positive. These results confirm the absence of disease in wildlife in what was once one of the areas most affected by ASF in Spain, and they suggest that wild boars play a limited role in ASFV persistence. These results confirm that ASFV cannot persist in isolated wild boar populations for long periods of time without the interaction of other factors such as re-infection by contact with domestic pigs or by feeding on contaminated swill.

Quantitative risk assessment for the introduction of African swine fever virus into the European Union by legal import of live pigs.

The recent incursion and spread of African swine fever virus (ASFV) in the Russian Federation and Caucasus region, close to European Union (EU) borders, have increased the concerns regarding the probability of ASFV introduction into the EU. There are many potential routes of ASFV entry into EU, but here we specifically aimed to assess the probability of ASFV introduction by legal trade of pigs, which historically has been one of the most important ways of exotic diseases introduction into the EU. A stochastic model was used to estimate the monthly probability of ASFV introduction for each country of the EU. Results of this model suggest an annual probability for ASFV introduction in the whole EU by this way of 5.22*10(-3) , which approximately corresponds with one outbreak in 192years. The risk of ASFV introduction via live pigs was highest in Poland (69%), particularly during the months of November and December. As expected, Russian Federation is the country that most contributes to this risk, representing 68% of the overall annual risk. Methods and results presented here may be useful for informing risk-based surveillance and control programmes and, ultimately, for prevention and control of potential ASFV incursions into the EU.