The polyanionic substances lambda and kappa carrageenan, pentosan polysulfate, fucoidan, dextran sulfate and heparin were investigated for their inhibitory effect on the replication of African swine fever virus (ASFV) in vitro. Lambda carrageenan was the most efficacious with a selectivity index, as based on the ratio of the 50% cytotoxic concentration to the 50% antiviral effective concentration, of 120, followed by pentosan polysulfate with 30, kappa carrageenan 13.3 and fucoidan 10. Dextran sulfate and heparin were almost inactive. In general, the substances had low toxicity for Vero cells. The studies with radiolabeled ASF virions suggest that the sulfated polysaccharides inhibit virus adsorption. Inhibition of virus replication was found for all the polysaccharides only when the substances were present during virus adsorption, with the exception of lambda and kappa carrageenan, which were also inhibitory when added immediately after the adsorption period.

The European Commission requested EFSA to compare the reliability of wild boar density estimates across the EU and to provide guidance to improve data collection methods. Currently, the only EU-wide available data are hunting data. Their collection methods should be harmonised to be comparable and to improve predictive models for wild boar density. These models could be validated by more precise density data, collected at local level e.g. by camera trapping. Based on practical and theoretical considerations, it is currently not possible to establish wild boar density thresholds that do not allow sustaining African swine fever (ASF). There are many drivers determining if ASF can be sustained or not, including heterogeneous population structures and human-mediated spread and there are still unknowns on the importance of different transmission modes in the epidemiology. Based on extensive literature reviews and observations from affected Member States, the efficacy of different wild boar population reduction and separation methods is evaluated. Different wild boar management strategies at different stages of the epidemic are suggested. Preventive measures to reduce and stabilise wild boar density, before ASF introduction, will be beneficial both in reducing the probability of exposure of the population to ASF and the efforts needed for potential emergency actions (i.e. less carcass removal) if an ASF incursion were to occur. Passive surveillance is the most effective and efficient method of surveillance for early detection of ASF in free areas. Following focal ASF introduction, the wild boar populations should be kept undisturbed for a short period (e.g. hunting ban on all species, leave crops unharvested to provide food and shelter within the affected area) and drastic reduction of the wild boar population may be performed only ahead of the ASF advance front, in the free populations. Following the decline in the epidemic, as demonstrated through passive surveillance, active population management should be reconsidered.

Keywords: African swine fever; passive surveillance; population density; population density threshold; population reduction; population separation; wild boar.

The US livestock safety strongly depends on its capacity to prevent the introduction of Transboundary Animal Diseases (TADs). Therefore, accurate and updated information on the location and origin of those potential TADs risks is essential, so preventive measures as market restrictions can be put on place. The objective of the present study was to evaluate the current risk of African swine fever (ASF) and Classical swine fever (CSF) introduction into the US through the legal importations of live pigs and swine products using a quantitative approach that could be later applied to other risks. Four quantitative stochastic risk assessment models were developed to estimate the monthly probabilities of ASF and CSF release into the US, and the exposure of susceptible populations (domestic and feral swine) to these introductions at state level. The results suggest a low annual probability of either ASF or CSF introduction into the US, by any of the analyzed pathways (5.5*10-3). Being the probability of introduction through legal imports of live pigs (1.8*10-3 for ASF, and 2.5*10-3 for CSF) higher than the risk of legally imported swine products (8.90*10-4 for ASF, and 1.56*10-3 for CSF). This could be caused due to the low probability of exposure associated with this type of commodity (products). The risk of feral pigs accessing to swine products discarded in landfills was slightly higher than the potential exposure of domestic pigs through swill feeding. The identification of the months at highest risk, the origin of the higher risk imports, and the location of the US states most vulnerable to those introductions (Iowa, Minnesota and Wisconsin for live swine and California, Florida and Texas for swine products), is valuable information that would help to design prevention, risk-mitigation and early-detection strategies that would help to minimize the catastrophic consequences of potential ASF/CSF introductions into the US.

The altered Schaedler flora (ASF) is a model microbial community with both in vivo and in vitro relevance. Here we provide the first characterization of the ASF community in vitro, independent of a murine host. We compared the functional genetic content of the ASF to wild murine metagenomes and found that the ASF functionally represents wild microbiomes better than random consortia of similar taxonomic composition. We developed a chemically defined medium that supported growth of seven of the eight ASF members. To elucidate the metabolic capabilities of these ASF species-including potential for interactions such as cross-feeding-we performed a spent media screen and analyzed the results through dynamic growth measurements and non-targeted metabolic profiling. We found that cross-feeding is relatively rare (32 of 3570 possible cases), but is enriched between Clostridium ASFASFASF dynamics and spatial distribution in vivo, to design pre- and probiotics that modulate relative abundances of ASF members, and will be essential for validating computational models of ASF metabolism. Well-characterized, experimentally tractable microbial communities enable research that can translate into more effective microbiome-targeted therapies to improve human health.

About half of Ethiopians belong to the Orthodox Tewahedo religion. Annually, more than 200 days are dedicated to religious fasting, which includes abstaining from all types of food, animal source foods, and water. However, the association of fasting with undernutrition remains unknown in Ethiopia. Therefore, dietary pattern and nutritional status of lactating women during lent fasting and non-fasting periods were studied, and predictor variables for maternal underweight were identified. To achieve this, lactating mothers in lent fasting (N = 572) and non-fasting (N = 522) periods participated from rural Tigray, Northern Ethiopia. Average minimum diet diversity (MDD-W) was computed from two 24-h recalls, and nutritional status was assessed using body mass index (BMI). Binary logistic regression was used to identify potential predictors of maternal underweight. Wilcoxon signed-rank (WSRT) and McNemar's tests were used for comparison of the two periods. The prevalence of underweight in fasting mothers was 50.6%. In the multivariate logistic regression model, younger age, sickness in the last four weeks preceding the survey, fasting during pregnancy, lactation periods, grandfathers' as household decision makers, previous aid experience, non-improved water source, and not owning chicken were positively associated with maternal underweight. In WSRT, there was no significant (p>> 0.05) difference on maternal body weight and BMI between periods. The average number of meals, diet diversity, and animal source foods (ASFs), consumption scores were significantly increased in non-fasting compared to fasting periods in both fasting and non-fasting mothers (p < 0.001, p < 0.05, and p < 0.001, respectively). Consumption of dark green leafy vegetables was higher in the fasting period (11%) than non-fasting (3.6%), in the study population. As a conclusion, Ethiopian Orthodox fasting negatively affected maternal nutritional status and dietary pattern in rural Tigray, Northern Ethiopia. To reduce maternal malnutrition in Ethiopia, existing multi-sectoral nutrition intervention strategies, should include religious institutions in a sustainable manner.

The study reports the development of a polymerase cross-linking spiral reaction (PCLSR) for the detection of African swine fever virus (ASFV) DNA in blood collected from infected pigs and wild boars. The method uses 3 specifically designed primers. Two outer-spiral primers comprising of 3' sequences complementary to ASFV p72 gene sequence and 5'end sequences complementary to exogenous gene of black widow alpha-latrotoxin as well as additional ASFV specific cross-linking primer. The method is specific exclusively to ASFV DNA without cross-reactions with cDNA of classical swine fever virus (CSFV), porcine reproductive respiratory syndrome (PRRSV) or porcine epidemic diarrhea virus (PEDV). The sensitivity of this technique reached 7.2 × 102 copies per μl-1 of plasmid containing p72 gene. The PCLSR was conducted at 65 °C creating cross-linked complex structures. The results of PCLSR were visualized using SYBR Green I dye, gel electrophoresis while the reaction progress was traced using real-time PCR system that resulted in registration of fluorescent curves and melting peaks at 85.3 °C. The developed PCLSR was examined using blood or tissue samples collected from selected 17 ASF cases from infected wild boars and 3 outbreaks in pigs. Further tests have been also conducted using 55 tissue samples from 23 outbreaks and 22 cases. These results showed that PCLSR might be further used for preliminary and cost-effective detection and surveillance of ASFV.

African swine fever virus (ASFV) is a major threat for porcine production that has been slowly spreading in Eastern Europe since its first appearance in the Caucasus in 2007. ASFV enters the cell by endocytosis and gains access to the cytosol to start replication from late endosomes and multivesicular bodies. Cholesterol associated with low-density lipoproteins entering the cell by endocytosis also follows a trafficking pathway similar to that of ASFV. Here we show that cholesterol plays an essential role in the establishment of infection as the virus traffics through the endocytic pathway. In contrast to the case for other DNA viruses, such as vaccinia virus or adenovirus 5, cholesterol efflux from endosomes is required for ASFV release/entry to the cytosol. Accumulation of cholesterol in endosomes impairs fusion, resulting in retention of virions inside endosomes. ASFV also remodels intracellular cholesterol by increasing its cellular uptake and redistributes free cholesterol to viral replication sites. Our analysis reveals that ASFV manipulates cholesterol dynamics to ensure an appropriate lipid flux to establish productive infection.

OBJECTIVE

Since its appearance in the Caucasus in 2007, African swine fever (ASF) has been spreading westwards to neighboring European countries, threatening porcine production. Due to the lack of an effective vaccine, ASF control relies on early diagnosis and widespread culling of infected animals. We investigated early stages of ASFV infection to identify potential cellular targets for therapeutic intervention against ASF. The virus enters the cell by endocytosis, and soon thereafter, viral decapsidation occurs in the acid pH of late endosomes. We found that ASFV infection requires and reorganizes the cellular lipid cholesterol. ASFV requires cholesterol to exit the endosome to gain access to the cytoplasm to establish productive replication. Our results indicate that there is a differential requirement for cholesterol efflux for vaccinia virus or adenovirus 5 compared to ASFV.

African swine fever (ASF) is believed to have evolved in eastern and southern Africa in a sylvatic cycle between common warthogs (Phacochoerus africanus) and argasid ticks of the Ornithodoros moubata complex that live in their burrows. The involvement of warthogs and possibly other wild suids in the maintenance of ASF virus means that the infection cannot be eradicated from Africa, but only prevented and controlled in domestic pig populations. Historically, outbreaks of ASF in domestic pigs in Africa were almost invariably linked to the presence of warthogs, but subsequent investigations of the disease in pigs revealed the presence of another cycle involving domestic pigs and ticks, with a third cycle becoming apparent when the disease expanded into West Africa where the sylvatic cycle is not present. The increase in ASF outbreaks that has accompanied the exponential growth of the African pig population over the last three decades has heralded a shift in the epidemiology of ASF in Africa, and the growing importance of the pig husbandry and trade in the maintenance and spread of ASF. This review, which focuses on the ASF situation between 1989 and 2017, suggests a minor role for wild suids compared with the domestic cycle, driven by socio-economic factors that determine the ability of producers to implement the control measures needed for better management of ASF in Africa.

Injury to lens epithelial cells (LECs) leads to epithelial-mesenchymal transition (EMT) with resultant fibrosis. The tropomyosin (Tpm) family of cytoskeleton proteins is involved in regulating and stabilizing actin microfilaments. Aberrant expression of Tpms leads to abnormal morphological changes with disintegration of epithelial integrity. The EMT of LECs has been proposed as a major cause of posterior capsule opacification (PCO) after cataract surgery. Using in vivo rodent PCO and human cataractous LECs, we demonstrated that the aberrant expression of rat Tpm and human Tpm1α/2β suggested their association in remodelling of the actin cytoskeleton during EMT of LECs. Expression analysis from abnormally growing LECs after lens extraction revealed elevated expression of α-smooth muscle actin (α-SMA), a marker for EMT. Importantly, these cells displayed increased expression of Tpm1α/2β following EMT/PCO formation. Expression of Tpm1α/2β was up-regulated in LECs isolated from cataractous lenses of Shumiya Cataract Rats (SCRs), compared with non-cataractous lenses. Also, LECs from human patients with nuclear cataract and anterior subcapsular fibrosis (ASF) displayed significantly increased expression of Tpm2β mRNA, suggesting that similar signalling invokes the expression of these molecules in LECs of cataractous SCR and human lenses. EMT was observed in LECs overexpressed with Tpm1α/2β, as evidenced by increased expression of α-SMA. These conditions were correlated with remodelling of actin filaments, possibly leading to EMT/PCO and ASF. The present findings may help clarify the condition of the actin cytoskeleton during morphogenetic EMT, and may contribute to development of Tpm-based inhibitors for postponing PCO and cataractogenesis.

This report represents a case study of an outbreak of African swine fever (ASF) that took place in the autonomic region of Extramadura, Badajoz Province, during the month of August, 1992, and aims to describe the Spanish ASF eradication programme as carried out in the field at producer level. Extensive husbandry methods used in the management of Iberian pigs in ASF endemic areas of Spain (about 4% of Spanish territory in 4/52 provinces) makes eradication particularly difficult. Since there is no vaccine available for ASF, the identification and slaughter of carrier animals is crucial to the control of the disease. A case of ASF, which caused one secondary outbreak, was diagnosed on 26 August 1992 and is described in terms of case history, laboratory examination, disease eradication measures and epidemiological observations. Additional evidence is presented in an attempt to evaluate and gain an understanding of the long-term risk of carrier animals, wild European pigs, vector ticks and biting flies as a cause of the persistence in endemic areas of recrudescence of ASF in areas from which the disease was previously eradicated.

This study assesses the relation between hypothesized risk factors and African swine fever virus (ASFV) distribution in Sardinia (Italy) after the beginning of the eradication program in 1993, using a Bayesian multivariable logistic regression mixed model. Results indicate that the probability of ASFV occurrence in Sardinia was associated to particular socio-cultural, productive and economical factors found in the region, particularly to large number of confined (i.e., closed) farms (most of them backyard), high road density, high mean altitude, large number of open fattening farms, and large number of pigs per commune. Conversely, large proportion of open farms with at least one census and large proportion of open farms per commune, were found to be protective factors for ASFV. Results suggest that basic preventive and control strategies, such as yearly census or registration of the pigs per farm and better control of the public lands where pigs are usually raised, together with endanced effords of outreach and communication with pig producers should help in the success of the eradication program for ASF in the Island. Methods and results presented here will inform decision making to better control and eradicate ASF in Sardinia and in all those areas with similar management and epidemiological conditions.

BACKGROUND

Acute myeloid leukemia (AML) is the most common type of blood cancer in adults. Emerging evidence is establishing a connection between AML and aberrant alternative splicing of pre-mRNA, which may result from aberrant expression of splicing factors, the mediators of splicing reactions.

METHODS

Using quantitative real-time polymerase chain reaction, we measured mRNA expression of 7 splicing factors belonging to the serine/arginine-rich (SR) protein family, SRSF1 (SF2/ASF), SRSF2 (SC35), SRSF3 (SRp20), SRSF4 (SRp75), SRSF5 (SRp40), SRSF6 (SRp55), and SRSF7 (9G8), and 1 non-SR factor, heterogeneous nuclear ribonucleoprotein A1 (HNRNPA1), in peripheral blood mononuclear cells of 26 patients with newly diagnosed AML and 26 healthy controls. In addition, the relationship between splicing factors and the mRNA splicing patterns of the caspase-8 gene (CASP8) was investigated.

RESULTS

Compared to healthy controls, the expression of splicing factors was obviously aberrant in newly diagnosed AML patients. The expression of SRSF1, SRSF3 and SRSF4 mRNAs was significantly decreased. Moreover, a significant correlation was observed between several splicing factors and caspase-8 pre-mRNA splicing in AML patients, but not in control subjects.

CONCLUSIONS

These data suggest that aberrant expression of splicing factors in AML may potentially connect with abnormal expression of oncogenes and be useful for early diagnosis, prognosis, and therapy of AML.

African swine fever (ASF), caused by African swine fever virus (ASFV), is a devastating infectious disease of domestic pigs and wild boars, and has tremendous negative socioeconomic impact on the swine industry and food security worldwide. It is characterized as a notifiable disease by World Organisation for Animal Health (OIE). No effective vaccine or treatment against ASF has so far been available. Early detection and rapid diagnosis are of potential significance to control the spread of ASF. Recombinase-based isothermal amplification assay, recombinase polymerase amplification (RPA) developed by TwistDx (Cambridge, United Kingdom) or recombinase-aided amplification (RAA) by Qitian (Wuxi, China), is becoming a molecular tool for the rapid, specific, and cost-effective identification of multiple pathogens. In this study, we aim to investigate if RPA/RAA can be a potential candidate for on-site, rapid and primary detection of ASFV. A panel of 152 clinical samples previously well-characterized by OIE-recommended qPCR was enrolled in this study, including 20 weak positive (Ct value ≥ 30) samples. This panel was consisted of different types, such as EDTA-blood, spleen, lung, lymph node, kidney, tonsil, liver, brain. We evaluated two recombinase-based isothermal amplification assays, RPA or RAA, by targeting the ASFV B646L gene (p72), and validated the clinical performance in comparison with OIE real-time PCR. Our result showed that the analytical sensitivity of RPA and RAA was as 93.4 and 53.6 copies per reaction, respectively at 95% probability in 16 min, at 39°C. They were universally specific for all 24 genotypes of ASFV and no cross reaction to other pathogens including Classical swine fever virus (CSV), Foot-and-mouth disease virus (FMDV), Pseudorabies virus, Porcine circovirus 2 (PCV2), Porcine Reproductive and respiratory syndrome virus (PPRSV). The results on detection of various kinds of clinical samples indicated an excellent diagnostic agreement between RPA, RAA and OIE real-time PCR method, with the kappa value of 0.960 and 0.973, respectively. Compared to real-time PCR, the specificity of both RPA and RAA was 100% (94.40% ∼ 100%, 95% CI), while the sensitivity was 96.59% (90.36% ∼ 99.29%, 95% CI) and 97.73% (92.03% ∼ 99.72%, 95% CI), respectively. Our data demonstrate that the developed recombinase-based amplification assay (RPA/RAA), promisingly equipped with field-deployable instruments, offers a sensitive and specific platform for the rapid and reliable detection of ASFV, especially in the resource-limited settings for the purpose of screening and surveillance of ASF.

Keywords: African swine fever virus; clinical validation; rapid isothermal amplification; recombinase aided amplification; recombinase polymerase amplification.

African swine fever (ASF) is a fatal disease caused by a virus in domestic pigs. In this study, a real-time loop-mediated isothermal amplification (LAMP) assay and visual LAMP assay were developed for the detection of African swine fever virus (ASFV). LAMP primers targeting the p10 gene of ASFV were designed, the LAMP reaction system was optimized with plasmid pUC57 containing the p10 gene sequence, and the specificities of the real-time LAMP and the visual assays were tested with the DNA or cDNA of pseudorabies virus (PRV), porcine circovirus type 2 (PCV2), classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine parvovirus (PPV) and ASFV, as well as the plasmid pUC57 containing the p10 gene sequence. The detection limits were determined using a serial dilution of plasmid pUC57 containing the p10 gene sequence. Our results showed that the LAMP assays could accurately and specifically detect ASFV with a detection limit of 30 copies per μl^-1 of pUC57 containing p10 gene sequence. In addition, the LAMP assays were further evaluated using various genotypes of ASFV strains. Furthermore, the LAMP assays are comparable with the well-established real-time PCR assay. This study provides promising solutions for facilitating preliminary and cost-effective surveillance for prevention and control of ASFV.

African swine fever (ASF) is an infectious and economically important disease of domestic pigs. There is no vaccine, and so reliable diagnosis is essential for control strategies. The performance of four recombinant ASF virus (ASFV) protein (pK205R, pB602L, p104R, and p54)-based enzyme-linked immunosorbent assays (ELISAs) was evaluated with European porcine field sera that had been established by Office International des Epizooties (OIE)-approved tests to be ASFV negative (n = 119) and ASFV positive (n = 80). The kappa values showed that there was almost perfect agreement between the results of the "gold standard" test (immunoblotting) and the results obtained by the p54-specific ELISA (kappa = 0.95; 95% confidence interval [CI], 0.90 to 0.99) and the pK205R-specific ELISA or the pB602L-specific ELISA (kappa = 0.92; 95% CI, 0.86 to 0.97). For the pA104R-specific ELISA, there was substantial to almost perfect agreement (kappa = 0.81; 95% CI, 0.72 to 0.89). Similar results were observed by the OIE-approved ELISA (kappa = 0.89; 95% CI, 0.82 to 0.95). Importantly, antibodies against these proteins were detectable early after infection of domestic pigs. Preliminary testing of 9 positive and 17 negative serum samples from pigs from West Africa showed identical results by the recombinant protein-based ELISA and the OIE-approved tests. In contrast, there was a high degree of specificity but a surprisingly a low level of sensitivity with 7 positive and 342 negative serum samples from pigs from East Africa. With poorly preserved sera, only the p104R-specific ELISA showed a significant reduction in sensitivity compared to that of the OIE-approved ELISA. Finally, these recombinant proteins also detected antibodies in the sera of the majority of infected warthogs. Thus, recombinant ASFV proteins p54, pB602L, and pK205R provide sensitive and specific targets for the detection of antibodies in European and West African domestic pigs and warthogs.

The need for more sustainable production and consumption of animal source food (ASF) is central to the achievement of the sustainable development goals: within this context, wise use of land is a core challenge and concern. A key question in feeding the future world is: how much ASF should we eat? We demonstrate that livestock raised under the circular economy concept could provide a significant, nonnegligible part (9-23 g/per capita) of our daily protein needs (~50-60 g/per capita). This livestock then would not consume human-edible biomass, such as grains, but mainly convert leftovers from arable land and grass resources into valuable food, implying that production of livestock feed is largely decoupled from arable land. The availability of these biomass streams for livestock then determines the boundaries for livestock production and consumption. Under this concept, the competition for land for feed or food would be minimized and compared to no ASF, including some ASF in the human diet could free up about one quarter of global arable land. Our results also demonstrate that restricted growth in consumption of ASF in Africa and Asia would be feasible under these boundary conditions, while reductions in the rest of the world would be necessary to meet land use sustainability criteria. Managing this expansion and contraction of future consumption of ASF is essential for achieving sustainable nutrition security.

Stunting affects 160 million pre-school children globally with adverse life-long consequences. While work within nutritional science suggests that stunting in early childhood is associated with low intakes of animal-sourced foods (ASFs), this topic has received little attention from economists. We attempt to redress this omission through an analysis of 130,432 children aged 6-23 months from 49 countries. We document distinctive patterns of ASF consumption among children in different regions. We find evidence of strong associations between stunting and a generic ASF consumption indicator, as well as dairy, meat/fish, and egg consumption indicators, and evidence that consuming multiple ASFs is more advantageous than any single ASF. We explore why ASF consumption is low but also so variable across countries. Non-tradable ASFs (fresh milk, eggs) are a very expensive source of calories in low-income countries and caloric prices of these foods are strongly associated with children's consumption patterns. Other demand-side factors are also important, but the strong influence of prices implies an important role for agricultural policies-in production, marketing and trade-to improve the accessibility and affordability of ASFs in poorer countries.

Keywords: Animal-sourced foods; dietary diversity; fisheries; livestock; nutrition; stunting.

The comprehensive investigation reported herein provides compelling evidence that anion-π interactions are the main driving force in the formation of self-assembled Fe(II)-templated metallacycles with bptz [3,6-bis(2-pyridyl)-1,2,4,5-tetrazine] in high yields. It was demonstrated by X-ray crystallography, (1)H NMR, solution and solid-state MAS (19)F NMR spectroscopies, CV and MS studies that the anions [X](-) = [BF(4)](-), [ClO(4)](-) and the anions [Y](-) = [SbF(6)](-), [AsF(6)](-), [PF(6)](-) template molecular squares [Fe(4)(bptz)(4)(CH(3)CN)(8)][X](8) and pentagons [Fe(5)(bptz)(5)(CH(3)CN)(10)][Y](10), respectively. The X-ray structures of [{Fe(4)(bptz)(4)(CH(3)CN)(8)}⊂BF(4)][BF(4)](7) and [{Fe(5)(bptz)(5)(CH(3)CN)(10)}⊂2SbF(6)][SbF(6)](8) revealed that the [BF(4)](-) and [SbF(6)](-) anions occupy the π-acidic cavities, establishing close directional F···C(tetrazine) contacts with the tetrazine rings that are by ~0.4 Å shorter than the sum of the F···C van der Waals radii (ΣR(vdW) F···C = 3.17 Å). The number and strength of F···C(tetrazine) contacts are maximized; the F···C(tetrazine) distances and anion positioning versus the polygon opposing tetrazine rings are in agreement with DFT calculations for C(2)N(4)R(2)···[X](-)···C(2)N(4)R(2) (R = F, CN; [X](-) = [BF(4)](-), [PF(6)](-)). In unprecedented solid-state (19)F MAS NMR studies, the templating anions, engaged in anion-π interactions in the solid state, exhibit downfield chemical shifts Δδ((19)F) ≈ 3.5-4.0 ppm versus peripheral anions. NMR, CV, and MS studies also establish that the Fe(II) metallacycles remain intact in solution. Additionally, interconversion studies between the Fe(II) metallacycles in solution, monitored by (1)H NMR spectroscopy, underscore the remarkable stability of the metallapentacycles [Fe(5)(bptz)(5)(CH(3)CN)(10)][PF(6)](10) ≪ [Fe(5)(bptz)(5)(CH(3)CN)(10)][SbF(6)](10) < [Fe(5)(bptz)(5)(CH(3)CN)(10)][AsF(6)](10) versus [Fe(4)(bptz)(4)(CH(3)CN)(8)][BF(4)](8), given the inherent angle strain in five-membered rings. Finally, the low anion activation energies of encapsulation (ΔG(‡) ≈ 50 kJ/mol), determined from variable-temperature (19)F NMR studies for [Fe(5)(bptz)(5)(CH(3)CN)(10)][PF(6)](10) and [Zn(4)(bptz)(4)(CH(3)CN)(8)][BF(4)](8), confirm anion encapsulation in the π-acidic cavities by anion-π contacts (~20-70 kJ/mol).

BACKGROUND

African swine fever (ASF) is a fatal, haemorrhagic disease of domestic pigs, that poses a serious threat to pig farmers and is currently endemic in domestic pigs in most of sub-Saharan Africa. To obtain insight into the factors related to ASF outbreaks at the farm-level, a longitudinal study was performed in one of the major pig producing areas in central Uganda. Potential risk factors associated with outbreaks of ASF were investigated including the possible presence of apparently healthy ASF-virus (ASFV) infected pigs, which could act as long-term carriers of the virus. Blood and serum were sampled from 715 pigs (241 farms) and 649 pigs (233 farms) to investigate presence of ASFV and antibodies, during the periods of June-October 2010 and March-June 2011, respectively. To determine the potential contribution of different risks to ASF spread, a questionnaire-based survey was administered to farmers to assess the association between ASF outbreaks during the study period and the risk factors.

RESULTS

Fifty-one (21 %) and 13 (5.6 %) farms reported an ASF outbreak on their farms in the previous one to two years and during the study period, respectively. The incidence rate for ASF prior to the study period was estimated at 14.1 per 100 pig farm-years and 5.6 per 100 pig farm-years during the study. Three pigs tested positive for ASFV using real-time PCR, but none tested positive for ASFV specific antibodies using two different commercial ELISA tests.

CONCLUSIONS

There was no evidence for existence of pigs that were long-term carriers for the virus based on the analysis of blood and serum as there were no seropositive pigs and the only three ASFV DNA positive pigs were acutely infected and were linked to outbreaks reported by farmers during the study. Potential ASF risk factors were present on both small and medium-scale pig farms, although small scale farms exhibited a higher proportion with multiple potential risk factors (like borrowing boars for sows mating, buying replacement from neighboring farms without ascertaining health status, etc) and did not implement any biosecurity measures. However, no risk factors were significantly associated with ASF reports during the study.

African swine fever (ASF) is a highly contagious, lethal and economically devastating haemorrhagic disease of domestic pigs. Insights into the dynamics and scale of virus transmission can be obtained from estimates of the basic reproduction number (R0). We estimate R0 for ASF virus in small holder, free-range pig production system in Gulu, Uganda. The estimation was based on data collected from outbreaks that affected 43 villages (out of the 289 villages with an overall pig population of 26,570) between April 2010 and November 2011. A total of 211 outbreaks met the criteria for inclusion in the study. Three methods were used, specifically; (i) GIS- based identification of the nearest infectious neighbour based on the Euclidean distance between outbreaks, (ii) epidemic doubling time, and (iii) a compartmental susceptible-infectious (SI) model. For implementation of the SI model, three approaches were used namely; curve fitting (CF), a linear regression model (LRM) and the SI/N proportion. The R0 estimates from the nearest infectious neighbour and epidemic doubling time methods were 3.24 and 1.63 respectively. Estimates from the SI-based method were 1.58 for the CF approach, 1.90 for the LRM, and 1.77 for the SI/N proportion. Since all these values were above one, they predict the observed persistence of the virus in the population. We hypothesize that the observed variation in the estimates is a consequence of the data used. Higher resolution and temporally better defined data would likely reduce this variation. This is the first estimate of R0 for ASFV in a free range smallholder pig keeping system in sub-Saharan Africa and highlights the requirement for more efficient application of available disease control measures.

BACKGROUND

African swine fever (ASF) is a contagious viral disease which can cause up to 100% mortality among domestic pigs leading to serious socio-economic impact on people's livelihoods. ASF is endemic in Uganda and there is paucity of information on the epidemiology of the disease. The major aim of this study was to determine the seroprevalence and prevalence of African swine fever virus (ASFV) in apparently healthy slaughter pigs at Wambizi slaughterhouse in Kampala city, Uganda. We also estimated the presence of ASFV antibodies and circulating viral antigens in pigs from selected districts of Uganda during targeted surveillance. We analysed 540 and 181 blood samples collected from slaughter pigs and pigs from targeted surveillance districts respectively.

RESULTS

The prevalence of ASFV in slaughter pigs was 52.96% (95% CI, 48.75-57.14) and 11.5% (95% CI, 9.06-14.45) by ELISA and PCR respectively. In surveillance districts, the proportion of ASFV positive pigs was 53.59% (95% CI, 46.33-60.71) and 0.55% (95% CI, 0.1-3.06) by ELISA and PCR respectively.

CONCLUSIONS

The study has found out a high seroprevalence of ASFV antibodies in apparently healthy slaughter pigs and also a high proportion of ASFV antibody seropositive pigs in surveyed districts in Uganda indicating exposure to ASFV. However, there was a lower prevalence of ASFV infection implying that there could be low virulent strains of ASFV circulating in domestic pigs in Uganda which requires further investigation.

The expression and role of RNA binding proteins (RBPs) controlling mRNA translation during tumor progression remains largely uncharacterized. Analysis by immunohistochemistry of the expression of hnRNP A1, hnRNPH, RBM9/FOX2, SRSF1/ASF/SF2, SRSF2/SC35, SRSF3/SRp20, SRSF7/9G8 in breast tumors shows that the expression of hnRNP A1, but not the other tested RBPs, is associated with metastatic relapse. Strikingly, hnRNP A1, a nuclear splicing regulator, is also present in the cytoplasm of tumor cells of a subset of patients displaying exceedingly worse prognosis. Expression of a cytoplasmic mutant of hnRNP A1 leads to increased translation of the mRNA encoding the tyrosine kinase receptor RON/MTS1R, known for its function in tumor dissemination, and increases cell migration in vitro. hnRNP A1 directly binds to the 5' untranslated region of the RON mRNA and activates its translation through G-quadruplex RNA secondary structures. The correlation between hnRNP A1 and RON tumoral expression suggests that these findings hold clinical relevance.

African swine fever (ASF) is a lethal haemorrhagic disease of domestic pigs for which there is no vaccine. Strains of the virus with reduced virulence can provide protection against related virulent strains of ASFV, but protection is not 100% and there are concerns about the safety profile of such viruses. However, they provide a useful tool for understanding the immune response to ASFV and previous studies using the low virulent isolate OUR T88/3 have shown that CD8+ cells are crucial for protection. In order to develop a vaccine that stimulates an effective anti-ASFV T-cell response we need to know which of the >150 viral proteins are recognized by the cellular immune response. Therefore, we used a gamma interferon ELIspot assay to screen for viral proteins recognized by lymphocytes from ASF-immune pigs using peptides corresponding to 133 proteins predicted to be encoded by OUR T88/3. Eighteen antigens that were recognized by ASFV-specific lymphocytes were then incorporated into adenovirus and MVA vectors, which were used in immunization and challenge experiments in pigs. We present a systematic characterization of the cellular immune response to this devastating disease and identify proteins capable of inducing ASFV-specific cellular and humoral immune responses in pigs. Pools of viral vectors expressing these genes did not protect animals from severe disease, but did reduce viremia in a proportion of pigs following ASFV challenge.

EFSA assisted four countries in the analysis of epidemiological data on African swine fever (ASF), collected until September 2017. The temporal analysis demonstrated that the average proportions of PCR and antibody-ELISA positive samples from the hunted wild boar remained below 3.9 and 6.6, respectively. A peak in the ASF incidence was observed 6 months after the first observed case, followed by a significant reduction of the number of cases and low levels of African swine fever virus (ASFV) circulation at the end of 38 months follow-up period at different spatial resolutions. The spatial analysis concluded that human-mediated spread of ASFV continues to play a critical role in the ASF epidemiology, despite all measures currently taken. 'Wild boar density', 'total road length' (as proxy for human activity) and 'average suitable wild boar habitat availability' were identified as predictors for the occurrence of ASF in Estonia by a Bayesian hierarchical model, whereas 'wild boar density' and 'density of pig farms' were predictors according to a generalised additive model. To evaluate the preventive strategies proposed in EFSA's Scientific Opinion (2015) to stop the spread of ASFV in the wild boar population, a simulation model, building on expert knowledge and literature was used. It was concluded that reduction of wild boar population and carcass removal to stop the spread of ASFV in the wild boar population are more effective when applied preventively in the infected area. Drastic depopulation, targeted hunting of female wild boar and carcass removal solely implemented as measures to control ASF in the wild boar population need to be implemented in a highly effective manner (at or beyond the limit of reported effectivity in wild boar management) to sustainably halt the spread of ASF.

Keywords: African swine fever; epidemiology; management; prevention; risk factors; wild boar.