Use of a comprehensive diagnostic algorithm for Anisakis allergy in a high seroprevalence Mediterranean setting.

Summary: Background.Diagnosis of anisakis allergy (AA) is based on the skin prick test (SPT) and specific IgE (sIgE) determination. Anyway, false positivity cases are due to cross reactivity with numerous allergens. The aim of the study was to evaluate the reliability of a comprehensive diagnostic algorithm for the AA. Methods.An observational study was conducted on a sample of consecutive subjects accessing the allergology outpatient ambulatories of two hospitals located in Western Sicily. All the recruited outpatients were tested by Skin Prick Test performed using anisakis extracts by ALK-Abellò (Madrid, Spain). Specific IgE dosage for anisakis extracts was then performed by using ImmunoCAP250 (Immunodiagnostics Uppsala, Sweden). Consequently, outpatients who tested positive to first line tests underwent sIgE testing for ascaris and tropomyosin. Lastly, outpatients positive to the first line were invited to be further tested by basophil activation test (BAT) by using Flow CAST kit and anisakis commercial extract (Bühlmann Laboratories AG, Schönenbuch, Switzerland), as confirmatory analysis. Results.One hundred and eleven outpatients with an anamnesis suggestive of sensitization to anisakis (AS) and 466 subjects with chronic urticaria (CU) were recruited in the study. Of these, 22 with AS and 41 with CU showed a sensitization to anisakis allergens. The diagnostic algorithm revealed that 8.8% of outpatients who tested positive to sIgE determination were affected by CU, while 82.5% of all the sIgE positivity was related to cross-reactivity. Overall, a genuine anisakis seroprevalence of 2.3% was documented. Within a sub-sample of 15 subjects with clinical symptoms related to AA, n. 8 showed a real positivity after BAT. A greater response to A. pegreffii allergens as compared to A. simplex was reported. Conclusions.Our preliminary findings support the high clinical specificity of BAT for AA diagnosis, suggesting implementing this method in a comprehensive diagnostic algorithm.

An unusual case of positive sIgE to Galactose-alpha-1,3-galactose from South Italy.

Summary: We report the case of a 38-year-old man who was bitten several times during his life by a tick. He didn't report any previous history of anaphylaxis after the ingestion of red meat. The serum specific IgE showed positivity to α-Gal. The proximity of the bits didn't increase the titer of IgE antibodies to alpha-gal. We could hypothesize that the frequency of the exposure to the tick Corresponding author bites and the amount of tick bites during his lifetime induced a sort of tolerance in this patient.

Red foxes (Vulpes vulpes) bioindicator of lead and copper pollution in Sicily (Italy).

The aim of this study was to evaluate Pb and Cu accumulation in muscle and skin samples of red fox (Vulpes vulpes) from Sicily, for monitoring of environmental metals pollution. Metals determination, carried out by atomic absorption spectrophotometer (AAS), showed the presence of Pb and Cu in all samples analyzed. Pb concentrations were similar in muscle (0.04±0.009 mg/kg) and skin (0.03±0.004 mg/kg) samples, while Cu levels resulted higher in muscle (1.842±0.178 mg/kg) than in skin (1.22±0.151 mg/kg). In addition, a comparative analysis of Pb and Cu concentrations was carried out among samples from different areas of Sicily and between female and male, young and old, immature and mature foxes. Metals content found in all muscle and skin samples demonstrates that V. vulpes could be a valid "sentinel" species of rural and suburban areas to study the environmental metals pollution and the habitat quality.

Spirocerca lupi isolated from gastric lesions in foxes (Vulpes vulpes) in Sicily (Italy).

Spirocerca lupi (Rudolphi 1809) is a cosmopolitan nematode of dogs and wild carnivores. In the past it has been reported in Italy, mainly in southern regions and in Sicily, where the parasite was observed in foxes in 2005. The parasite typically produces nodular masses in the oesophagus and thoracic aorta. During the 2003-2004 hunting season, the authors investigated a total of 55 foxes (Vulpes vulpes) hunted or killed by car accidents in the provinces of Palermo and Agrigento. All the foxes were subjected to necropsy and 6 (9.16%) had S. lupi nodules located exclusively in the gastric wall. The nature of the nodules was determined by opening them and detecting the nematodes inside, which were identified as S. lupi. Some of the nodules were characterized anatomopathologically and histopathologically. The formation of the parasitic nodule in the stomach only suggests a deviation from the route commonly followed by the nematode to reach the oesophagus, the elective anatomical site for completion of its lifecycle. This survey gives a contribution to the epidemiology of this parasite which is severely outdated in Italy and highlights some distinctive features of the life cycle and parasite migration.

Effect of shearing and environmental conditions on physiological mechanisms in ewes.

The aim of this study was to establish the influence of shearing in dairy sheep during summer season. Several physiological and haematological parameters, which are closely related to thermoregulatory potential, have been investigated with the purpose to assess shearing influence on thermoregulation. Forty dairy sheep, clinically healthy and well-fed, were used. They were divided into two groups of 20 subjects each. Twenty sheep were let unshorn as a control group (Group A), and twenty sheep were shorn (Group B). On each subject of group A and Group B, rectal temperature, respiration and heart rates were recorded and blood samples were collected in order to asses the following parameters: red blood cells, white blood cells, platelets, haematocrit and haemoglobin. All measurements taken on day 0 (before shearing), were repeated after 1, 15, 30, 45 and 60 days after shearing. The statistical analysis, the ANOVA followed by the Bonferroni's test, showed statistical differences between two groups for rectal temperature (F(1.90) = 5.57, p < 0.0001), respiratory rate (F(1.90) = 18.40, p < 0. 0001) and white blood cells (F(1.190) = 7.61, p < 0.0001). These results suggest that the shearing induce adaptative responses in the organism.

Anisakis spp. larvae in three mesopelagic and bathypelagic fish species of the central Mediterranean Sea.

In this work 437 fish samples of species belonging to the families Myctophidae (Electrona risso and Diaphus metopoclampus) and Phosichthyidae (Vinciguerria attenuata) were examined for the presence of Anisakidae larvae. The study was performed with fishes in the central Mediterranean Sea, particularly in the Strait of Sicily and in the Strait of Messina. The visual inspection and chloro-peptic analysis revealed the presence of nematode parasites with prevalence values between 2.9% in Electrona risso samples and 5.4% in Vinciguerria attenuata samples. A positive correlation was found between standard length (SL) and prevalence of infestation in D. metopoclampus samples (p<0.05). The larvae examined were morphologically ascribed, at genus level, to Anisakis morphotypes I and II and molecularly identified as Anisakis pegreffii, Anisakis ziphidarum and Anisakis physeteris, in 67%, 9% and 24% of the fish samples examined. Overall, A. pegreffii and A. ziphidarum larvae were isolated in 14 and 2 specimens of D. metopoclampus respectively, A. physeteris larvae were found in 3 E. risso and 2 V. attenuata. A positive correlation was found between standard length and prevalence of infestation in D. metopoclampus samples (p<0.05). First information is provided on the presence of Anisakis spp. larvae of the myctophid fish species E. risso, D. metopoclampus and V. attenuata from the Central Mediterranean. It is also confirmed the role of lanternfishes (Myctophidae) as paratenic hosts for Anisakis spp.

Climatic conditions and gastrointestinal nematode egg production: observations in breeding sheep and goats.

Parasitic egg production was studied in sheep and goats affected by parasitic gastroenteritis. The herds studied were located at different altitudes and in different climatic conditions. Samples were taken every month and the number of eggs per grams of feces was calculated. Observation of preliminary data shows that the maximum peak of egg production was during the winter period, whereas in other countries winter is a period of hypobiosis. This study shows that understanding peak time of infection related to different climatic and environmental conditions will help improve anthelminthic treatments and animal health strategies.

Identification of a novel bluetongue virus vector species of Culicoides in Sicily.

The vectors of bluetongue virus are certain species of Culicoides biting midges, and in the Mediterranean area Culicoides imicola has long been considered to be the only field vector. In Sicily an entomological and serological surveillance programme has been in operation since the autumn of 2000, which has shown that the prevalence and abundance of C. imicola is lower than in many other Italian regions. Moreover, in 2002, there were outbreaks of bluetongue in the absence of C. imicola, and in these regions bluetongue viral RNA was detected by means of a nested reverse-transcriptase PCR in wild-caught, non-blood-engorged, parous Culicoides pulicaris. Furthermore, bluetongue virus serotype 2 was isolated on five occasions from extracts of non-blood-engorged parous C. pulicaris by using embryonated hens eggs and BHK-21 cells as assay systems. These findings suggest that in parts of Italy and possibly in other areas of Europe, where C. imicola is absent or rare, C. pulicaris may act as a fully competent vector of bluetongue virus.

Evaluation of mercury levels in pangasius and cod fillets traded in Sicily (Italy).

Predator fishes at the top of the aquatic food chain can accumulate large concentrations of metals and their consumption, consequently, makes a significant contribution, in particular, to mercury intake. The aim of this study was to determine mercury levels in fillets of two predatory species: pangasius (Pangasius hypophthalmus) from the Vietnam region of Megong and Chao Pharayai and cod (Gadus morhua) from the Baltic and North Sea, both being commercially important in the Italian market. A comparative analysis of these two imported fish species was carried out as a risk assessment for consumer safety. The results showed the presence of higher mercury levels in pangasius (0.41 ± 0.08 mg kg(-1)) than in cod (0.11 ± 0.004 mg kg(-1)) fillets. These data underline the importance of monitoring on imported fish before marketing, to evaluate better the risk of mercury exposure through fish and seafood consumption, and of selecting safer fishes for consumption by those groups more sensitive to the toxic effects of this metal.

Salmonella infection in illegally imported spur-thighed tortoises (Testudo graeca).

The prevalence of Salmonella infection was determined in a group of spur-thighed tortoises (Testudo graeca) seized during two smuggling attempts and in a population of captive Hermann's tortoises (Testudo hermanni) sheltered in a wildlife rescue centre. Salmonella spp. was isolated in 81 of 220 (36.8%) and in 17 of 67 (25.4%) cloacal swabs collected from the T. graeca and T. hermanni tortoises respectively. Overall, a total of 21 different Salmonella serotypes were found. Some of these serotypes are common to terrestrial chelonians while others have never been reported. All cultured serotypes were non-typhoidal but nonetheless many of these have been previously reported as source of human outbreaks of reptile-related salmonellosis. Eighty-two per cent and 5.3% of the isolates were resistant to two and three anti-microbial agents respectively. However, the isolates were highly susceptible to the anti-microbials of choice for the treatment of salmonellosis such as cephalosporins and fluoroquinolones. Our findings confirm that tortoises can be considered a reservoir for Salmonella and that care should be employed when handling and breeding these animals. Tight surveillance should be enforced to avoid illegal importation and prevent the trading of live tortoises, carriers of zoonotic pathogens.

Analysis of serum proteins in clinically healthy goats (Capra hircus) using agarose gel electrophoresis.

BACKGROUND: Electrophoretic patterns of serum proteins provide useful information on pathological conditions in ruminants. Their reference values, however, are dissimilar to those of other species. Reference values for goats using agarose gel as the supporting matrix have not been reported. OBJECTIVE: The aim of this study was to evaluate serum concentrations of total protein and protein fractions (albumin and globulins) by means of agarose gel electrophoresis (AGE) in goats in order to establish electrophoretic reference intervals and to evaluate potential changes associated with aging. METHODS: Blood was collected from 105 clinically healthy Girgentana goats by means of jugular venipuncture. Serum protein concentrations were assessed by AGE. Three age groups were compared: 1-1.5 years, 2-4 years, and 5-12 years. RESULTS: Values (mean ± SD) were determined for concentrations of total protein (72.26 ± 6.40 g/L), albumin (31.80 ± 4.00 g/L), α-globulins (6.40 ± 1.23 g/L), ß(1) -globulins (10.50 ± 2.58 g/L), ß(2) -globulins (5.18 ± 1.60 g/L), and γ-globulins (18.65 ± 5.90 g/L) and for albumin/globulin (A/G) ratio (0.82 ± 0.20). One-way ANOVA showed statistically significant age-related differences for total protein and α-globulin concentrations and A/G ratios. Age influenced protein concentrations with the 5-12-year-old group having higher total protein and α-globulin concentrations and lower albumin concentration and A/G ratios than the 2-4-year-old group. CONCLUSIONS: This study provides reference values for total protein concentrations and protein fractions obtained by AGE in goats. Some values vary with age. Age-specific reference intervals are reported in order to provide clinicians with an additional diagnostic aid.