Species diversity and molecular insights into phlebotomine sand flies in Sardinia (Italy)-an endemic region for leishmaniasis.

This study provides updated information on the distribution of the phlebotomine sand fly species and their genetic characterization in Sardinia, a Mediterranean island where leishmaniasis is endemic. From April to November 2017, sand flies were trapped in five different capture sites using sticky traps and light traps as collection methods, operated nearby sheep, poultry, cat, and dog shelters. Phlebotomine specimens (n = 513) collected were morphologically and molecularly identified as Phlebotomus perniciosus (249/513, 48.5%), Phlebotomus perfiliewi (236/513; 46%), and Sergentomyia minuta (28/513, 5.5%). Sand flies were collected from the second half of May to October confirming the well-defined seasonal activity, which peaks in August in Sardinia. Overall, correlation analyses indicated a significant positive association between the monthly number of sand flies collected and the mean temperature (r = 0.88, rho = 0.87, and tau = 0.69, P < 0.05), while there was non-significant, moderately negative correlation between the monthly number of sand flies collected and the monthly mean relative humidity and wind (r = - 0.22, rho = 0.02, and tau = 0.04, P > 0.05). This study provides the first data on the molecular characterization of phlebotomine sand flies in this region and confirms the presence of three sand fly species. Molecular results suggest that the morphological features used for analysis represent synapomorphic-derived characters which are shared among descendant taxa and the common ancestor.

Gongylonema infection of wild mammals in Japan and Sardinia (Italy).

The gullet worms, classical Gongylonema pulchrum and newly differentiated Gongylonema nepalensis, are prevalent in various mammals in Japan and Sardinia, Italy, respectively. The former species is cosmopolitan in distribution, dwelling in the mucosa of the upper digestive tract of a variety of domestic and wild mammals, and also humans. At present, the geographical distribution of G. nepalensis is known in Nepal and Sardinia, with the nematode having been recorded from the oesophagus of water buffaloes (Nepal), cattle, sheep, goats and wild mouflon (Sardinia). To clarify their natural transmission cycles among domestic and wild mammals, the present study analysed the ribosomal RNA gene (rDNA) and mitochondrial cytochrome c oxidase subunit 1 gene (cox1) of worms of various origins: G. pulchrum worms from sika deer, wild boars, Japanese macaques, and feral alien Reeves's muntjacs in Japan, and G. nepalensis worms from a red fox and a wild boar in Sardinia. Although the internal transcribed spacer (ITS) regions of rDNA and partial cox1 nucleotide sequences of G. pulchrum from native wild mammals in Japan were distinct from those of the worms in cattle, the worms from feral alien Reeves's muntjacs showed the cattle-type ITS genotype and cox1 cattle-I and II haplotypes. The rDNA and cox1 nucleotide sequences of G. nepalensis from a red fox in Sardinia were almost identical to those of the worms from domestic and wild ruminants on the island. The ecological interaction between domestic and wild mammals and their susceptibility to different Gongylonema spp. must be considered when trying to elucidate this spirurid's transmission dynamics in nature.

Simultaneous infection by four feline lungworm species and implications for the diagnosis.

Besides Aelurostrongylus abstrusus, other parasites belonging to the superfamily Metastrongyloidea, namely Oslerus rostratus, Troglostrongylus brevior and to the family Trichuridae, i.e. Eucoleus aerophilus (syn. Capillaria aerophila), have also been reported as agents of respiratory infection in domestic cats. A case of simultaneous infection by four feline lungworm species in Sardinia is herein described. An adult female cat (Felis silvestris catus), road-killed in the southeast part of Sardinia (municipality of Villacidro, province of Cagliari), Italy, was referred to the Laboratory of Parasitology of the Veterinary Teaching Hospital in Sassari. At necropsy, the lungs were examined and dissected under a stereomicroscope for the presence of parasites, and first-stage larvae (L1) of broncho-pulmonary nematodes were searched for in a faecal sample using the Baermann method. Parasites collected in the lungs were morphologically identified as A. abstrusus, E. aerophilus, and O. rostratus. In addition to the above species, L1s of Troglostrongylus spp. were detected at coproscopy but no adult specimen was found in the lungs. The morphological identification was confirmed by the molecular amplification and sequencing of cox1 mitochondrial gene, 18S and ITS2 ribosomal DNA. This finding stands as the first simultaneous infection by four feline lungworm species in the same animal, and as the first report of O. rostratus and E. aerophilus in Sardinia.

Design, Development and Validation of a Knee Brace to Standardize the US Imaging Evaluation of Knee Osteoarthritis.

OBJECTIVE: A repeatable and reliable follow-up of knee injuries would be desirable to prevent delayed diagnosis and to monitor the efficacy of the applied treatment over time. Ultrasound (US) techniques are an attractive option to this purpose, since they are safe, low-cost and non-invasive. However, its use in the clinical practice is limited by the high dependency on the operator's experience. Hence, the objective of this study is to provide a standardization of the US image acquisition process for knee osteoarthritis (OA) allowing an extended clinical use of US technologies in this domain. METHODS: Clinical specifications were provided by expert musculoskeletal radiologists thus identifying the subject poses and the US probe positions needed to evaluate the cartilage structure, signs of synovitis and joint effusion. Such considerations were used to derive the technical requirements needed for the development of a wearable brace equipped with specific openings to guide the correct placement of the probe. The feasibility of the developed wearable brace was tested on three healthy volunteers, which were asked to acquire informative US images, similar to the reference images performed by the musculoskeletal radiologist. RESULTS: Thanks to the knee brace, the untrained subjects were able to self-acquire informative B-mode images comparable to the corresponding images acquired by an expert clinician. DISCUSSION/CONCLUSION: The use of a knee brace intended for knee OA US diagnosis demonstrated the possibility to standardize the acquisition protocol and make its application achievable also for untrained subjects, representing a key step toward tele-ultrasonography.

Comparative analysis of polyphenolic profiles and antioxidant and antimicrobial activities of tunisian pome fruit pulp and peel aqueous acetone extracts.

Pome trees, apple, pear, and quince, are classified into the subfamily Pomoideae, belonging to the Rosaceae family. Their autumnal fruits are consumed worldwide in different forms, that is, fresh or transformed into jams, jelly, juices, etc. Their well-established beneficial properties to human health were found mainly related to their phenolic content. Pulp and peel aqueous acetone extracts obtained from Tunisian fruits at commercial maturity were comparatively evaluated for their phenolic profiles and antioxidant and antimicrobial potentials. The phenolic compounds present in the extracts were identified and quantified using RP-HPLC-DAD and ESI-MS techniques. Significant differences in the chromatographic profiles among these fruits, as well as between pulp and peel extracts of each fruit, were observed. Quince, followed by 'Red Delicious', peel extracts showed the highest phenolic content (160.33 and 110.90 mg/100 g of fresh weight). The stronger inhibitory effect on DPPH radicals corresponded to those obtained from peel materials. A comparative analysis of the antimicrobial potential against a range of microorganism strains was also carried out. Staphylococcus aureus, Pseudomonas aeruginosa, and Bacillus cereus were the most sensitive to the active extracts. Among the examined phenolic extracts, 'Red Delicious' and quince peels showed the highest effects for inhibiting bacteria growth. Minimum inhibitory and bactericide concentrations ranged from 10(2) to 10(4) microg of polyphenol/mL. Red skin apple and quince peels could be of great interest as important antioxidant and antimicrobial polyphenol sources.

In vitro interaction between ochratoxin A and different strains of Saccharomyces cerevisiae and Kloeckera apiculata.

The interaction of ochratoxin A (OTA) and 20 yeast strains of Saccharomyces cerevisiae and Kloeckera apiculata during alcoholic fermentation was studied. Levels of OTA were determined in the fermentation liquid and in the yeast cells solid using a high-performance liquid chromatography system with a fluorescence detector. Yeast cells do not adsorb OTA, and for all yeasts, OTA levels did not affect the alcoholic fermentation. Some yeast strains reduced levels of OTA, whereas other strains did not show any effect demonstrating that OTA level reduction is not a genus species characteristic but a strain trait.

First record of Gongylonema nepalensis in domestic and wild ruminants in Europe.

The gullet worm, Gongylonema pulchrum, is cosmopolitan in distribution, infecting a variety of mammals including domestic and wild ruminants. Gongylonema nepalensis recently collected from the esophageal epithelium of water buffaloes in Nepal was separated from G. pulchrum based on its distinctly shorter left spicule relative to body length and unique nucleotide sequences of the ribosomal RNA gene (rDNA) and the mitochondrial cytochrome c oxidase subunit 1 gene (cox-1). During meat inspections at four abattoirs on Sardinia Island, Italy, 25 Gongylonema worms were collected from one each individual of cattle (n=8), sheep (n=7), goats (n=4), and mouflon (Ovis aries musimon; n=6), and characterized morphologically and genetically. Intriguingly, all of the collected worms from these ruminants were G. nepalensis, exhibiting comparable body lengths to G. pulchrum in cattle from other regions but with significantly shorter left spicules like G. nepalensis (less than 20.9% of the entire body length in contrast to 21.8-65.6%, the reported proportion of G. pulchrum). Furthermore, the rDNA nucleotide sequences of these worms from different ruminant species on Sardinia Island were almost identical to each other and to Nepalese G. nepalensis isolates. With the exception of one worm from a sheep (displaying a single nucleotide substitution), the 369-bp cox-1 nucleotide sequences of all the Sardinian G. nepalensis isolates from the different host sources were absolutely identical, forming a clade with Nepalese G. nepalensis isolates and not G. pulchrum isolates. The present study reveals that G. nepalensis is not a local parasite in the Indian subcontinent (Nepal), but instead has a certain geographical distribution in Europe and takes several ruminant species as a definitive host.

Trichinella britovi from domestic to wild animals of Sardinia, Italy.

The zoonotic nematode Trichinella britovi has been documented in animals and/or humans of the Mediterranean islands of Corsica and Sardinia since 2004. From 2005 to 2007 in the Sardinia island, several surveys had shown that T. britovi was circulating among backyard and free-ranging pigs reared in the Orgosolo municipality but all attempts had failed to detect this parasite in wild susceptible animals. The aim of the present work was to investigate the circulation of T. britovi in pigs and wildlife of the Orgosolo municipality, and of surrounding municipalities and provinces in the 2010-2014 slaughtering/hunting seasons. The results show that the T. britovi circulation was still restricted to the Orgosolo municipality with a prevalence of 2.6% in free-ranging pigs and 0.2% in backyard pigs but, for the first time, this parasite was detected also in 0.4% of wild boar, and 27.6% of red foxes. No infection was detected in backyard pigs, wild boar, and red foxes of the other municipalities and provinces. Since 1978, African swine fever is endemic in Sardinia and foci of this virus are still active in the investigated areas favoring cannibalism and, consequently, the T. britovi transmission, due to the high mortality rate caused by this virus. This is the first documented report on the transmission of T. britovi between the domestic and the sylvatic cycle. The health authority of the island must provide a service to dispose animal carcasses and offal, stamping out illegal free-ranging pigs, and train hunters and pig owners to manage waste and by-products according to the EU regulations.

Clinical experience with gabapentin in patients with bipolar or schizoaffective disorder: results of an open-label study.

BACKGROUND: This study was carried out to evaluate the efficacy, tolerability, and safety of gabapentin as an adjunctive treatment in patients with bipolar or schizoaffective disorder with manic or hypomanic symptoms. METHOD: Twenty-five patients fulfilling DSM-IV diagnostic criteria for bipolar I disorder or schizoaffective disorder underwent a 16-week, open-trial treatment with gabapentin. Symptom severity was measured using the Clinical Global Impressions scale (CGI) and the Brief Psychiatric Rating Scale (BPRS). Baseline scores and final scores were compared by using the Student t test and the Friedman range variance analysis. RESULTS: Twenty-two patients (88%) completed the 16 weeks of treatment with gabapentin; 19 (76%) had a positive response as measured by changes in CGI and BPRS scores. The mean dose was 1440 mg/day. The only side effect observed was oversedation, which decreased with continuing treatment. CONCLUSION: Gabapentin was effective in the treatment of mania and hypomania in patients with bipolar and schizoaffective disorders. If confirmed in controlled studies, these findings suggest that gabapentin represents a well-tolerated, rapidly acting antimanic agent.

Puerperal psychoses: a clinical case study with follow-up.

Thirty cases of post-partum psychotic disorders occurred between 1973 and 1987 and hospitalized at the Psychiatric Ward of Florence University were studied and followed up. A structured diagnostic interview was used, which explored DSM Ill-R diagnosis both for mood disorders and for psychotic features. The psychotic symptoms had started within 8 weeks of parturition in all cases. Only 36.7% of the patients showed no subsequent pathology after the puerperal symptoms. The diagnoses, both at the index episode and at the follow-up, revealed a great predominance of mood disorders and the absence of schizophrenia. The follow-up survey showed a greater proportion of bipolar disorders than it appeared at the puerperal onset of the disease. A high frequency of puerperal psychotic relapses has also occurred after subsequent deliveries during the follow-up period.

Gabapentin as a promising treatment for antipsychotic-induced movement disorders in schizoaffective and bipolar patients.

Improvement of antipsychotic-induced blepharospasm and involuntary oral-mandibulo movements was observed with the use of the anticonvulsant drug gabapentin among 14 of 16 affectively ill patients who had been exposed to maintenance neuroleptics of the conventional type. In many cases, the movement disorders of these patients had not responded to more standard measures such as clozapine. This finding permits a potential strategy for patients with treatment-emergent tardive dyskinesia, a well-known complication of extended conventional neuroleptic use. Gabapentin, whose mood stabilizing properties have been reported in several clinical reports, represents a more natural treatment in the setting of bipolar spectrum disorders.

Pesticide residues in grapes, wine, and their processing products.

In this review the results obtained in the 1990s from research on the behavior of pesticide residues on grapes, from treatment to harvest, and their fate in drying, wine-making, and alcoholic beverage processing are reported. The fungicide residues on grapes (cyproconazole, hexaconazole, kresoxim-methyl, myclobutanil, penconazole, tetraconazole, and triadimenol), the application rates of which were of a few tens of grams per hectare, were very low after treatment and were not detectable at harvest. Pyrimethanil residues were constant up to harvest, whereas fluazinam, cyprodinil, mepanipyrim, azoxystrobin, and fludioxonil showed different disappearance rates (t(1/2) = 4.3, 12, 12.8, 15.2, and 24 days, respectively). The decay rate of the organophosphorus insecticides was very fast with t(1/2) ranging between 0.97 and 3.84 days. The drying process determined a fruit concentration of 4 times. Despite this, the residue levels of benalaxyl, phosalone, metalaxyl, and procymidone on sun-dried grapes equalled those on the fresh grape, whereas they were higher for iprodione (1.6 times) and lower for vinclozolin and dimethoate (one-third and one-fifth, respectively). In the oven-drying process, benalaxyl, metalaxyl, and vinclozolin showed the same residue value in the fresh and dried fruit, whereas iprodione and procymidone resides were lower in raisins than in the fresh fruit. The wine-making process begins with the pressing of grapes. From this moment onward, because the pesticide on the grape surface comes into contact with the must, it is in a biphasic system, made up of a liquid phase (the must) and a solid phase (cake and lees), and will be apportioned between the two phases. The new fungicides have shown no effect on alcoholic or malolactic fermentation. In some cases the presence of pesticides has also stimulated the yeasts, especially Kloeckera apiculata, to produce more alcohol. After fermentation, pesticide residues in wine were always smaller than those on the grapes and in the must, except for those pesticides that did not have a preferential partition between liquid and solid phase (azoxystrobin, dimethoate, and pyrimethanil) and were present in wine at the same concentration as on the grapes. In some cases (mepanipyrim, fluazinam, and chlorpyrifos) no detectable residues were found in the wines at the end of fermentation. From a comparison of residues in wine obtained by vinification with and without skins, it can be seen that their values were generally not different. Among the clarifying substances commonly used in wine (bentonite, charcoal, gelatin, polyvinylpolypyrrolidone, potassium caseinate, and colloidal silicon dioxide), charcoal allowed the complete elimination of most pesticides, especially at low levels, whereas the other clarifying substances were ineffective. Wine and its byproducts (cake and lees) are used in the industry to produce alcohol and alcoholic beverages. Fenthion, quinalphos, and vinclozolin pass into the distillate from the lees only if present at very high concentrations, but with a very low transfer percantage (2, 1, and 0.1%, respectively). No residue passed from the cake into the distillate, whereas fenthion and vinclozolin pass from the wine, but only at low transfer percentages (13 and 5%, respectively).

Analysis by hplc of ryanodine and dehydroryanodine residues on fruits and in ryania powdery wood.

A simple and rapid HPLC method to evaluate residues of the major ryanoids (ryanodine and dehydroryanodine) on three fruits (olives, apples, and pears) has been developed. The pesticides were extracted from the fruits with hexane and acetone solution (1:1, v/v). Cleanup was carried out with aminopropyl-bonded silica cartridges. This method is characterized by recovery >75%, precision <11% RSD, and sensitivity of 0.020 mg/kg. The method can also be used to determine the level of active ingredients in ryania powdery wood.

Distribution of folpet on the grape surface after treatment.

Field trials were carried out to evaluate whether folpet sprayed on grapevines penetrated the epicuticular wax and cell walls of grapes. Folpet showed poor penetration into the epicuticular wax; it was found almost totally on the surface. Despite its low solubility in water, perhaps due to the presence of adjuvants, its residues showed such a high resistance to washing that the action of rain was negligible in decreasing residues.

Phenolic compounds in virgin olive oil. 2. Reappraisal of the extraction, HPLC separation, and quantification procedures.

The extraction procedures (solid/liquid SPE and liquid/liquid LLE) and HPLC separation and quantification methods of polyphenolic compounds have been checked in virgin olive oils in order to explain the differences in content reported in the literature. The work has been carried out on oils prepared from one cultivar and produced under the same protocol. The extraction methods are practically equivalent, but the SPE technique is more favorable because it is faster and simpler. It has been proved that the chromatographic features and the method of chemical expression of the concentrations may greatly affect the final values. Thus, under the same analytical method, the total concentration values of polyphenols of the same oil show variations from 18% to 80%, according to the formality of expression as gallic acid, caffeic acid, or tyrosol equivalents. The role of the nature and spectrophotometric features of the phenols and of the internal standard is also discussed, and it was found to be an important source of reported variation. A gradient separation with an eluent mixture acetonitrile-sulfuric acid (0.1 mol/L), detection at 225 nm, and quantitative calculation of polyphenolic compounds in oils (expressed as tyrosol equivalents, THY(eq)) is proposed.

Factors affecting imazalil and thiabendazole uptake and persistence in citrus fruits following dip treatments.

The effect of concentration, temperature, and length of treatment with imazalil (IMZ) and thiabendazole (TBZ) was studied with application to citrus fruit. The amount of residues retained by fruit after "home" washing was also monitored. IMZ uptake in citrus fruit was related to treatment duration, whereas TBZ residues was not. Residues of IMZ or TBZ fungicides were significantly correlated with dip temperature (r = 0.943 for IMZ; r = 0.911 for TBZ). Treatment at 50 degrees C produced a deposition approximately 8 and approximately 2.5 times higher than when treatments were carried out at 20 degrees C in IMZ and TBZ, respectively. No significant differences in terms of IMZ deposition were detected after treatments carried out alone or in combination. Uptake of the two fungicides was associated with their physicochemical characteristics as well as different formulation types. IMZ residues showed a great persistence during storage when applied separately, and >83% of active ingredient was present after 9 weeks of storage. IMZ residues increased with dip length, doubling when dip time increased from 0.5 to 3 min. In contrast, TBZ residues did not change with the different dip times. Following postharvest dip treatments of citrus fruit at 20 or 50 degrees C, home washing removed approximately 50% of the IMZ and approximately 90% of the TBZ.

Homogentisic acid: a phenolic acid as a marker of strawberry-tree (Arbutus unedo) honey.

Analysis of organic acids in strawberry-tree (Arbutus unedo) honey showed the presence of an unknown acid as the main constituent. This compound was isolated and identified as homogentisic acid (2, 5-dihydroxyphenylacetic acid) by MS and NMR techniques. Its average content in honey was 378 +/- 92 mg/kg. Analysis of nectar confirmed the floral origin of the compound found in honey. Since this acid was not detected in any of the different monofloral honeys, it could be used as a marker of strawberry-tree (A. unedo) honey.

Fate of quinoxyfen residues in grapes, wine, and their processing products.

Quinoxyfen is a new fungicide that belongs to the family of the quinolines, recently introduced to control powdery mildew (Uncinula necator). In this paper the fate of quinoxyfen residues from vine to wine and in their processing products was studied. After the last of four applications at the recommended rate, 0.38 mg/kg of residue was found on the grapes, which is under the legal limit fixed in Italy (0.5 mg/kg). The degradation rate was according to a pseudo-first-order kinetics (r = 0.964) and the half-life was 7.24 days. Vinification was carried out with and without maceration. During the vinifications without maceration <50% of the residues passed from the grapes to the musts. Separation of the lees (8%) from the must by centrifugation caused no detectable residues in centrifuged must. At the end of fermentation with and without maceration no quinoxyfen residues were determinable in the wine. No effect on the alcoholic or malolactic fermentation was observed even in the presence of higher quinoxyfen concentrations than those found in the grapes at harvest time. During fermentation, the yeasts partially degraded the pesticides and completly adsorbed them. Bacteria, on the other hand, do not have any degradative effect on the pesticides. The raisins obtained by sun-drying did not contain any residues, whereas those obtained by oven-drying show the same amount of residues as in the fresh grapes. During the sun-drying process the fruit weight decreased by a factor of 4; the decrease in the oven-drying was equivalent. Samples of dregs and liquid lees, fortified with high levels of quinoxyfen. were double-distilled. The first dregs distillate, with an alcohol content of 32.1%, did not show any residues, whereas the first lees distillate, with an alcohol content of 34.5%, showed 7% of the initial residues. After the second lees distillation, the obtained product showed an alcoholic content of 81.2% and no residues of quinoxyfen (<0.01 mg/kg).

Pesticides in fermentative processes of wine.

The influence of six fungicides (azoxystrobin, cyprodinil, fludioxonil, mepanipyrim, pyrimethanil, and tetraconazole) on the fermentative activity of two yeasts (Saccharomyces cerevisiae and Kloeckeraapiculata) and two lactic bacteria (Leuconostoc oenos and Lactobacillus plantarum) was studied. The possibility of their being degraded by these yeasts and bacteria was also investigated. The presence of the pesticides did not affect alcoholic fermentation, not even with levels higher than those normally found in grapes in field experiments. On the contrary, their presence stimulated the yeast, especially K. apiculata, to produce more alcohol. The fermentative process did not affect the amount of pesticides either by degradation or by adsorption. During malolactic fermentation by Le. oenos, malic acid decreased slightly less (by approximately 15%) in the presence of all pesticides, except mepanipyrim. A lower effect ( approximately 5%) was found during the fermentative process with La. plantarum. The bacteria studied did not show a degradative effect on pesticides during malolactic fermentation.

Quality of life and overweight: the obesity related well-being (Orwell 97) questionnaire.

The development and validation of a self-reported measure of obesity-related quality of life, the Obesity Related Well-Being (ORWELL 97), were undertaken to examine the intensity and the subjective relevance of physical and psychosocial distress. The questionnaire was validated in a sample of 147 obese patients (99 females, 48 males). The Eating Disorder Examination 12.0D interview, a structured diagnostic interview for DSM-III-R (DSM-IV criteria for binge eating disorder), Beck Depression Inventory, Binge Eating Scale, and the State-Trait Anxiety Inventory 1 and 2 scales were also applied. Internal consistency and test-retest reliability were satisfactory. Factor analysis allowed the identification of two subscales: ORWELL 97-1 related to psychological status and social adjustment, and ORWELL 97-2 related to physical symptoms impairment. Obese female patients showed a lower quality of life, and the severity of obesity appeared to interfere with physical functioning rather than psychological status and social adjustment. The ORWELL 97 questionnaire appears to be a simple and reliable measure of obesity-related quality of life, which can be used in current clinical practice.