Characterization of the O/ME-SA/Ind-2001d foot-and-mouth disease virus epidemic recorded in the Maghreb during 2014-2015.

The O/ME-SA/Ind-2001d has been the main foot-and-mouth disease virus (FMDV) lineage responsible for FMD epidemics outside the Indian subcontinent from 2013 to 2017. In 2014, outbreaks caused by this FMDV lineage were reported in Maghreb, where it was initially detected in Algeria and Tunisia and later in Morocco. This was the first incursion of an FMDV type O of exotic origin in the Maghreb region after 14 years of absence. In this study, we report analyses of both VP1 and whole-genome sequences (WGSs) generated from 22 isolates collected in Algeria and Tunisia between 2014 and 2015. All the WGSs analysed showed a minimum pairwise identity of 98.9% at the nucleotide level and 99% at the amino acid level (FMDV coding region). All Tunisian sequences shared a single putative common ancestor closely related to FMDV strains circulating in Libya during 2013. Whereas sequences from Algeria suggest the country experienced two virus introductions. The first introduction is represented by strains circulating in 2014 which are closely related to those from Tunisia, the second one, of which the origin is more uncertain, includes strains collected in Algeria in 2015 that gave origin to the 2015 outbreak reported in Morocco. Overall, our results demonstrated that a unique introduction of O/Ind-2001d FMDV occurred in Maghreb through Tunisia presumably in 2014, and from then the virus spread into Algeria and later into Morocco.

Wide recognition of Culex pipiens and lack of detection of Culex torrentium through biomolecular differentiation of mosquitoes in the Emilia-Romagna region, Northern Italy.

The Culex pipiens complex includes species with reported differences in vector competence for arthropod-borne viruses, many of which are of significant importance to human health such as the West Nile virus and the Sindbis virus. This group of mosquitoes is difficult to distinguish morphologically; particularly as adult females. In Europe, the two species of the complex, Culex pipiens Linnaeus 1758 and Culex torrentium Martini 1925, are often found sympatrically. With the aim to characterize the presence and spread of both species in the Emilia-Romagna region, Northern Italy, mosquitoes of the complex - collected during the West Nile virus surveillance plans - were tested by multiplex real-time PCR for the detection of the two species Cx. pipiens and Cx. torrentium. A total of 24 165 mosquitoes, collected between 2012 and 2014 from 105 sites, and sorted in 204 pools, were tested. All tested pools were found to be composed of Cx. pipiens, whereas Cx. torrentium was not detected. These results indicate a likely absence of Cx. torrentium mosquitoes within the surveyed territory, whereas Cx. pipiens is widely distributed in the area mentioned. This is in line with previous reports, which describe a pre-alpine distribution of Cx. torrentium in Italy.

Human and entomological surveillance of Toscana virus in the Emilia-Romagna region, Italy, 2010 to 2012.

Toscana virus (TOSV), transmitted by phlebotomine sandflies, is recognised as one of the most important causes of viral meningitis in summer in Mediterranean countries. A surveillance plan based on both human and entomological surveys was started in 2010 in the Emilia-Romagna region, Italy. Clinical samples from patients with neurological manifestations were collected during 2010 to 2012. The surveillance protocol was improved during these years, allowing the detection of 65 human infections. Most of these infections were recorded in hilly areas, where sandflies reach the highest density. Entomological sampling around the homes of the patients resulted in a low number of captured sandflies, while later sampling in a hilly area with high number of human cases (n=21) resulted in a larger number of captured sandflies. Using this approach, 25,653 sandflies were sampled, of which there were 21,157 females, which were sorted into 287 pools. TOSV RNA was detected by real-time PCR in 33 of the pools. The results highlighted the role of Phlebotomus perfiliewi as the main vector of TOSV and a potential link between vector density and virus circulation. This integrated system shows that an interdisciplinary approach improves the sensitiveness and effectiveness of health surveillance.

The experience of West Nile virus integrated surveillance system in the Emilia-Romagna region: five years of implementation, Italy, 2009 to 2013.

Predicting West Nile virus (WNV) circulation and the risk of WNV epidemics is difficult due to complex interactions of multiple factors involved. Surveillance systems that timely detect virus activity in targeted areas, and allow evidence-based risk assessments may therefore be necessary. Since 2009, a system integrating environmental (mosquitoes and birds) and human surveillance has been implemented and progressively improved in the Emilia-Romagna region, Italy. The objective is to increase knowledge of WNV circulation and to reduce the probability of virus transmission via blood, tissue and organ donation. As of 2013, the system has shown highly satisfactory results in terms of early detection capacity (the environmental surveillance component allowed detection of WNV circulation 3­4 weeks before human cases of West Nile neuroinvasive disease (WNND) occurred), sensitivity (capacity to detect virus circulation even at the enzootic level) and area specificity (capacity to indicate the spatial distribution of the risk for WNND). Strong correlations were observed between the vector index values and the number of human WNND cases registered at the province level. Taking into consideration two scenarios of surveillance, the first with environmental surveillance and the second without, the total costs for the period from 2009 to 2013 were reduced when environmental surveillance was considered (EUR 2.093 million for the first scenario vs EUR 2.560 million for the second). Environmental surveillance helped to reduce costs by enabling a more targeted blood unit testing strategy. The inclusion of environmental surveillance also increased the efficiency of detecting infected blood units and further allowed evidence-based adoption of preventative public health measures.

Mosquito surveillance in northwestern Italy to monitor the occurrence of tropical vector-borne diseases.

Mosquito-borne arboviruses (MBV) represent an important health problem, causing diseases and deaths both in human and animals mainly in tropical and subtropical countries. In recent years, they have emerged also in temperate regions where they have caused epidemics. Of mounting concern among public health authorities in Europe are zoonotic mosquito-borne viruses belonging to the Flavivirus genus. The aim of this study was to carry out active surveillance on mosquitoes in two regions of northwestern Italy (Liguria and Piedmont) to gain a better knowledge of the mosquito populations by identifying potential vectors of arboviruses and to investigate arbovirus infection. A network of 61 CO2 CDC traps was placed in the study area; sampling was conducted from May to October 2011. A total of 46,677 mosquitoes was collected, identified to species level, and classified according to their vector competence. Mosquitoes collected from 16 traps, selected according to risk-based factors, were tested by biomolecular analysis to detect flavivirus infection. This study highlights the importance of entomological surveillance in northwestern Italy because most of the mosquitoes collected were found to have high vector competence. Moreover, the risk-based virological surveillance allowed to detect the presence of mosquito flavivirus RNA, phylogenetically closely related to the MMV Spanish isolate, in three pools and USUV RNA in one pool in new areas where it has not been reported previously. The availability of continuous data on mosquito populations provides invaluable information for use in cases of an epidemic emergency. Maintenance of this integrated system for the next years will provide stronger data that can inform the design of a risk-based surveillance for the early detection of the occurrence of outbreaks of tropical MBDs.

Carbon monoxide, cigarettes and family doctors.

AIMS AND BACKGROUND: General practitioners could play a key role in preventive programs against tobacco-related diseases. However, they seldom take action in the office even with minimal advice counselling. Such behaviour might reflect the lack of academic teaching and the lack of practice with motivational and dependence questionnaires, considered basic tools to help smokers to quit successfully. The study was aimed to investigate the awareness of a sample of Italian family doctors as regards tobacco epidemiology and smoking cessation strategies. METHODS: A total of 428 family doctors were administered a questionnaire with a set of questions on their personal smoking habits and on personal initiatives in the office towards smokers. Another set of questions regarded their knowledge on tobacco issues, with special attention to carbon monoxide, which is widely perceived as a very dangerous poison and works as a motivational tool on smokers and adolescents. Carbon monoxide measurement was carried out on all participants to obtain objective data on smoking and to show the feasibility of the test. RESULTS: The percentage of self-reported current smokers among general practitioners was 24%, with a high prevalence of ex-smokers (46%), and 29% of never smokers. Family doctors were more keen to counsel adolescents than adults about tobacco, and they were very interested in continuing medical education on the issue. The doctors who took part in our study showed a surprising limited knowledge of all the issues associated with smoking cessation and prevention such as epidemiology, cigarette characteristics, success rate of smoking cessation programs, Fagerström's tolerance questionnaire, safety of nicotine replacement therapy and the knowledge of carbon monoxide as a product of cigarette smoke. CONCLUSIONS: The scenario depicted by our survey underscores the necessity to improve the knowledge and performance of primary care physicians on tobacco-related issues in order to implement primary and secondary prevention in clinical practice.

Endothelin-A blockade attenuates systemic and renal hemodynamic effects of L-NAME in humans.

Eight Na-repleted volunteers underwent 3 separate 90-minute infusions of either N(G)-nitro-L-arginine methyl ester (L-NAME) 3.0 mg. kg(-1). min(-1) or endothelin-A receptor (ET-A) blocker BQ-123 (BQ) 0.125 nmol. kg(-1). min(-1) or both. Mean arterial pressure (MAP), glomerular filtration rate (GFR), renal blood flow (RBF), renal vascular resistances (RVR), and sodium excretion rate (UNaV) were measured at baseline (b) and from 0 to 45 minutes (period 1) and 45 to 90 minutes (period 2) of infusion. BQ alone had no effect. GFR declined by 4.9% (P<0.001 versus b) in period 1, to 9.9% (P<0. 001) in period 2 with L-NAME, and by 3.3% (P<0.01) to 6.6% (P<0.001) with L-NAME plus BQ (P=NS between L-NAME and L-NAME plus BQ). UNaV fell equally with L-NAME or L-NAME plus BQ. MAP rose significantly in period 2 with L-NAME (6.9%; P<0.001) but not with coinfused BQ (2. 1%; P=NA versus b, P=0.005 versus L-NAME alone). RBF declined by 12. 2% (P<0.001) to 18.3% (P<0.001) with L-NAME and by 4.6% (P<0.005) to 8.2% (P<0.001) with L-NAME plus BQ. These changes were smaller with L-NAME plus BQ (P<0.05 in period 1 and P<0.02 in period 2). Blunted changes were also seen for RVR (P<0.005 in period 1 and P<0.001 in period 2 between L-NAME alone and L-NAME plus BQ). These findings show that systemic and renal vasoconstriction due to L-NAME are attenuated by BQ, which suggests that an interaction between endogenous nitric oxide production and ET-A activity participates in the maintenance of baseline systemic and renal vascular tone in humans.

Regional sympathetic activity in pre-hypertensive phase of spontaneously hypertensive rats.

Imbalances in central and peripheral sympathetic nervous system (SNS) activity have been observed in essential and experimental hypertension. This study was carried out in order to evaluate SNS activity in two distinct tissue types of spontaneously hypertensive rats (SHR), compared to Wistar-Kyoto normotensive (WKY) rats, in the pre-hypertensive phase (4-5 weeks of age). Interstitial concentrations of norepinephrine (NE) and other catecholamines were measured by microdialysis in striated muscle, whose tone is controlled by baroreflexes, and in the subcutaneous adipose tissue where sympathetic output mainly controls metabolism. Two groups of SHR and WKY male rats were studied, aged 4-5 weeks, with a mean body weight of 92 and 86 g respectively. Systolic blood pressure (SBP, tail-cuff) values were 113 mm Hg (SD +/- 6.2) in SHR and 108 mm Hg (SD +/- 7.3) in WKY rats (p=0.28,t test). Two microdialysis probes were positioned in the subcutaneous fatty tissue and in the striated muscle of the parascapular region and perfused with Ringers' solution. The dialysate was collected every thirty minutes for 3 hours and analyzed in HPLC-ED to determine the content of NE and other catecholamines. Interstitial levels of NE were higher in SH than in WKY rats in both tissues. Mean NE values from subcutaneous adipose tissue in 4-5 week-old SHR were 1162 +/- 193 pg/ml compared to 496 + 188 pg/ml in WKY rats (p<0.001, t test). Muscle tissue NE levels in SHR were 1241 +/- 337 pg/ml vs. 521 +/- 138 pg/ml in WKY rats (p<0.001, t test). Plasma NE concentrations (279 +/- 61 pg/ml in SHR vs 246 + 69 pg/ml in WKY P = 0.65, t test) were not significantly different between the two strains at this young age. These findings suggest SNS hyperactivity in young SHR, though still normotensive, possibly dissociated from regional components of regulation (baroreceptor control in striated muscle and metabolic control in subcutaneous adipose tissue).

Use of Fourier shape descriptors to improve the reproducibility of echographic measurements of arterial intima-media thickness.

BACKGROUND: A major source of error in the longitudinal assessment of the intima-media thickness (IMT) is the difficulty in retrieving the same echographic view of the vessel. OBJECTIVE: To present a method for increasing the reproducibility of IMT measurements by ultrasound in large arteries. METHOD: The Fourier descriptor is a well-known means of describing an object's shape. By means of the discrete Fourier transform (DFT), the shape was represented in a frequency domain; the computational advantages of the DFT then permitted a measure of unlikeness between different shapes (the 'distance' measure; DM) to be defined and used as a criterion for reproducing the contour. When the sonographer compared successive images of a complex vascular segment, like the carotid bifurcation, the identity of the echographic cut was deduced from the identity of the vessel's contour. The best match of the baseline image was the view that minimized the contour DM. RESULTS: Preliminary studies in the carotid artery bifurcations of eight subjects showed that the DM responds to systematic variations in the ultrasound interrogation angle and reveals minimal changes in transducer position. Duplicate scans of 12 subjects were performed by three sonographers with different strategies for acquisition of the same images: a low DM was associated with a low difference in pairs of IMT measurements. Data were classified into two groups (normal or borderline vessels with a pooled mean IMT of 0.62 mm and overtly thickened segments with a pooled mean IMT of 1.31 mm). When minimization of the DM was the criterion for the acquisition of replicate scans, the mean absolute difference of paired data for the mean IMT of the distal common carotid artery was 0.03 +/- 0.02 mm for the first group and 0.06 +/- 0.03 mm for the second group. This is a significant reduction in comparison with non-quantitative alternative criteria for image reproduction. For the maximum IMT of the same segments the mean absolute differences were 0.07 +/- 0.03 and 0.13 +/- 0.06 mm in the first and second groups, respectively. CONCLUSION: This method can be applied to the serial assessment of single atherosclerotic segments. The computational time is negligible. By reducing the scatter in sequential IMT data, longitudinal investigations (e.g. of the results of antihypertensive therapy) with shorter durations and smaller sample groups may be rendered feasible.

[The relationship between regional sympathetic activity and the onset of arterial hypertension in spontaneously hypertensive rats]. / Relazione fra attività simpatica distrettuale e sviluppo di ipertensione arteriosa in ratti spontaneamente ipertesi.

Several studies on spontaneously hypertensive rats (SHR) have demonstrated increased activity of the sympathetic nervous system (SNS). Using microdialysis, we have observed a greater release of norepinephrine (NE) into the interstitia of striated muscle, than that observed in control Wistar-Kyoto (WKY) rats in the prehypertensive phase. We confirmed these results in the subcutaneous adipose tissue where the sympathetic output controls metabolism. This study was carried out in order to evaluate SNS activity in two district tissue types conducted during both the prehypertensive phase (4-5 weeks of age) and the established hypertensive phase (15-16 weeks of age). Interstitial concentrations of NE were measured by microdialysis in striated muscle and subcutaneous adipose tissue. Two groups of rats were studied. Each group was made up of 8 subjects, SHR and WKY, males of 4-5 weeks of age with a mean body weight of 80 and 75 g respectively. Arterial systolic pressure (tail-cuff) values were 106 mmHg (standard deviation +/-8.2) in SHR and 101 mmHg (standard deviation +/-6.9) in WKY rats (NS). Two microdialysis probes were positioned in the subcutaneous fatty tissue and in the striated muscle of the parascapular region and perfused with Ringers' solution. The dialysate was collected every 30 min for 150 min and analyzed in high-performance liquid chromatography-every day. The content of NE and other catecholamines was determined. The same animals in both groups were reevaluated at 15-16 weeks of age. The mean body weight at this time was 246 g for the SHR and 289 g for the WKY rats. Arterial systolic pressure was 161 mmHg (standard deviation +/-13.3) and 108 mmHg (standard deviation +/-15.6) respectively (p < 0.01, Student's t test). Interstitial levels of NE were higher in SHR than in WKY rats in both tissues examined in the prehypertensive phase and in the established hypertensive phase. Mean NE values from subcutaneous adipose tissue in 4-5 week-old SHR were 1362.1 +/- 181.3 pg/ml compared to 479.0 +/- 162.3 pg/ml in WKY rats (p < 0.001, Student's t test). Muscle tissue NE levels in SHR were 1292.7 +/- 319.1 vs 536.3 +/- 146.7 pg/ml in WKY rats (p < 0.001, Student's t test). Values from the same rats at 15-16 weeks of age were 1405.0 +/- 148.3 pg/ml in SHR compared to 501.6 +/- 131.2 pg/ml in fatty tissue from WKY rats and 1893.7 +/- 214.6 vs 502.0 +/- 118.8 pg/ml in muscle tissue from the respective groups (p < 0.001, Student's t test). Significant differences (p < 0.01, Student's t test) were also observed in mean NE values in striated muscle tissue during the developing phase of hypertension. These findings document SNS hyperactivity in SHR when compared to WKY normotensive controls. This increase in SNS activity was observed in both the prehypertensive phase and in the established hypertensive phase indicating a complete disassociation from regional components of regulation (baroreceptor control and metabolic control), at least in the prehypertensive phase. These results may suggest as alteration in primitive sympathetic central outflow. Higher interstitial NE concentrations in the muscle tissue from SHR during the hypertensive phase compared to levels of young animals that are still normotensive, reveal an interesting pathophysiological aspect for the development of arterial hypertension.

Serological diagnosis of feline immunodeficiency virus infection using recombinant transmembrane glycoprotein.

We developed an antibody detection enzyme-linked immunosorbent assay (ELISA) using recombinant surface (SU), transmembrane (TM) and capsid (CA) antigens of feline immunodeficiency virus (FIV) expressed in Escherichia coli. The three antigens were tested with sera collected from experimentally infected cats in order to follow the course of seroconversion and of the antibody levels throughout the infection. An early and marked increase of TM antibodies was observed. Antibodies to TM were demonstrated at high levels throughout the observation period. The immune response to SU and to CA was less pronounced and in some cats the level of antibodies to SU and CA tended to decline 6 months after infection. In addition, 413 FIV negative and positive cat sera were tested in order to define for each antigen the diagnostic sensitivity, specificity and efficiency. TM showed the highest diagnostic sensitivity (98%) while its specificity was 97%. Its diagnostic efficiency of 97% was better than that of SU and CA and exceeded that of tests utilizing conventionally grown and gradient purified FIV. Therefore, recombinant TM can be considered a very important antigen for FIV ELISA testing. An interesting perspective is offered in the combination of TM with other recombinant antigens in a dot assay form.

Hepatic uptake and biliary secretion of bile acids in the perfused rat liver.

Hepatic uptake and biliary secretion have been evaluated in the isolated perfused rat liver for cholic, chenodeoxycholic, ursodeoxycholic acid, both free and taurine-conjugated; the physicochemical properties of the bile acids have also been calculated and related to these experimental parameters. Cholic acid disappearance rate from the perfusate was the fastest, followed by that of ursodeoxycholic and chenodeoxycholic; it was also faster for taurine-conjugated bile acids than for their respective unconjugated forms. The recovery in bile was higher for conjugated than for unconjugated bile acids, and among each class, was higher for cholic than for chenodeoxycholic and ursodeoxycholic. The hepatic uptake correlated negatively (r = -0.99) with the bile acid lipophilicity, while the biliary secretion correlated with the solubility of the molecules. These results show the effect of the physicochemical properties of BA on their hepatic handling, at the physiological concentration of BA in the portal blood.

[Generalized histoplasmosis due to Histoplasma duboisii with mediastino-pulmonary infection. Cure after 15 months of treatment with ketoconazole]. / Histoplasmose généralisée à Histoplasma duboisii avec atteinte médiastino-pulmonaire. Guérison après 15 mois de traitement par kétoconazole.

We report a case of a young burkinabian, who presented with the disseminated african form of histoplasmosis, Histoplasma duboisii. There was pulmonary mediastinal disease with superficial lymphadenopathy and also hepatic involvement. The diagnosis was achieved by a histological examination of the peripheral glands and micrological examination of the expectorate. A clinical cure (despite the radiological persistence of enlarged but stable left hilar glands) with the follow up 10 months was achieved after 15 months of treatment with ketoconazole in a dose which was increased to 10 mg/kg/day.

Pharmacokinetics of ursodeoxycholic acid in rat.

The pharmacokinetic behaviour and metabolism of ursodeoxycholic acid (UDCA) have been studied in the rat. After oral administration of both 3H-labelled (4 muCi/kg body wt) and unlabelled (20 mg) UDCA, UDCA appeared in serum almost entirely in conjugated form (taurine conjugated); UDCA was present in bile mostly as taurine conjugated; the more relevant metabolite is 3 alpha,6 alpha, 7 beta-trihydroxycholanoic acid which represents 10% of the total bile acid pool. UDCA increased bile flow and selectively decreased biliary cholesterol secretion, while phospholipid secretion was unaffected. Faecal UDCA excretion was 15-20% while the urinary extraction was 1.5% during 24 h. The data show that UDCA, when administered in high dose, is promptly secreted into bile almost entirely metabolized to tauroursodeoxycholic acid, where it (1) desaturates the cholesterol in bile, (2) exerts choleretic properties.

[Cutaneous leishmaniasis in Algeria (author's transl)]. / La leishmaniose cutanée en Algérie.

Report of 27 cases and comments on their epidemiological, clinical, immunological features and on the question of their treatment and prophylaxis.

[Staphylococcal septicemias in Algiers (report of 14 cases) (author's transl)]. / Les septicémies à staphylocoques à Alger. A propose de 14 observations.

Staphyococcal septicemia is rare (0.19 p. 100 of admittances). The origin of this kind of infection could be located in all cases out of the sanitary units, and in 12 out of the 14 cases, patients under 30 years old were concerned. The main disorders were in pulmonary tract, kidneys and heart. A fatal issue occured 6 times due in most cases to a delayed admittance. A good study of sensitivity to antibiotics and a treatment associating several antibiotics are necessary.