FIRST RESULTS ON THE PRESENCE AND THE MOLECULAR CHARACTERIZATION OF ANISAKID NEMATODES IN MARINE FISH CAUGHT OFF NORTHERN SARDINIA

Anisakidosis is a parasitic zoonosis caused by nematodes of the family Anisakidae, belonging to the genera Anisakis, Contracaecum and Pseudoterranova. Molecular studies have shown that Anisakis larvae comprise a number of sibling species, which have different genetic structures, hosts and geographical distribution. A great variety of fish species can harbour infectious third stage larvae of this nematode. The preliminary results of a study carried out to evaluate the occurrence of this parasite in commercial fish caught off northern Sardinia are herein reported. From October 2008 to November 2009, 599 specimens of 8 commercial fish species were examined for anisakid larvae through visual inspection of body cavity and peptic digestion of the muscle. Isolated Anisakis sp. larvae were observed at light microscope and identified as Type I or Type II (sensu Berland, 1961). Out of 599 fish examined, 239 (40%) were infected by 1187 anisakid larvae, belonging to the genera Anisakis (1169 Type I and 18 Type II) and Hysterothylacium (692). The molecular identification of Anisakis spp. was carried out on a subsample of 30% of Type I larvae and all Type II larvae. Specimens were firstly examined using a species-specific PCR, with primers designed for Anisakis pegreffii (APEF) and Anisakis physeteris (APHF), and ITS-2 of nuclear rDNA. The results were confirmed by the analysis of the ITS region of nuclear rDNA (ITS-1, 5.8S and ITS-2) using the restriction enzymes HinfI and HhaI in PCR-RFLP. Type I larvae examined were all identified as A. pegreffii, and Type II were all A. physeteris. This is the first contribution to the epidemiology and molecular characterization of Anisakis spp. in commercial fish caught off Sardinia

Relationship of Late Lactation Milk Somatic Cell Count and Cathelicidin with Intramammary Infection in Small Ruminants

Late lactation is a critical moment for making mastitis management decisions, but in small ruminants the reliability of diagnostic tests is typically lower at this stage. We evaluated somatic cell counts (SCC) and cathelicidins (CATH) in late lactation sheep and goat milk for their relationship with intramammary infections (IMI), as diagnosed by bacteriological culture (BC). A total of 315 sheep and 223 goat half-udder milk samples collected in the last month of lactation were included in the study. IMI prevalence was 10.79% and 15.25%, respectively, and non-aureus staphylococci were the most common finding. Taking BC as a reference, the diagnostic performance of SCC and CATH was quite different in the two species. In sheep, receiver operating characteristic (ROC) analysis produced a higher area under the curve (AUC) value for CATH than SCC (0.9041 versus 0.8829, respectively). Accordingly, CATH demonstrated a higher specificity than SCC (82.92% versus 73.67%, respectively) at comparable sensitivity (91.18%). Therefore, CATH showed a markedly superior diagnostic performance than SCC in late lactation sheep milk. In goats, AUC was <0.67 for both parameters, and CATH was less specific than SCC (61.90% versus 65.08%) at comparable sensitivity (64.71%). Therefore, both CATH and SCC performed poorly in late lactation goats. In conclusion, sheep can be screened for mastitis at the end of lactation, while goats should preferably be tested at peak lactation. In late lactation sheep, CATH should be preferred over SCC for its higher specificity, but careful cost/benefit evaluations will have to be made

Milk cathelicidin and somatic cell counts in dairy goats along the course of lactation

This research communication reports the evaluation of cathelicidin in dairy goat milk for its relationship with the somatic cell count (SCC) and microbial culture results. Considering the limited performances of SCC for mastitis monitoring in goats, there is interest in evaluating alternative diagnostic tools. Cathelicidin is an antimicrobial protein involved in innate immunity of the mammary gland. In this work, half-udder milk was sampled bimonthly from a herd of 37 Alpine goats along an entire lactation and tested with the cathelicidin ELISA together with SCC and bacterial culture. Cathelicidin and SCC showed a strong correlation (r = 0.72; n = 360 milk samples). This was highest in mid-lactation (r = 0.83) and lowest in late lactation (r = 0.61), and was higher in primiparous (0.80, n = 130) than in multiparous goats (0.71, n = 230). Both markers increased with stage of lactation, but cathelicidin increased significantly less than SCC. Inaddition, peak level in late lactation was lower for cathelicidin (5.05-fold increase) than for SCC (7.64-fold increase). Twenty-one (5.8%) samples were positive to bacteriological culture, 20 for coagulase-negative staphylococci and one for Streptococcus spp.; 18 of them were positive to the cathelicidin ELISA (85.71% sensitivity). Sensitivity of SCC >500 000 and of SCC >1 000 000 cells/ml was lower (71.43 and 23.81%, respectively). Therefore, the high correlation of cathelicidin with SCC during the entire lactation, along with its lower increase in late lactation and good sensitivity indetecting intramammary infection (IMI), indicate a potential for monitoring subclinical mastitis in dairy goats. However, based on this preliminary assessment, specificity should be improved (40.41% for cathelicidin vs. 54.57 and 67.85% for SCC >500 000 and >1 000 000 cells/ml, respectively). Therefore, the application of cathelicidin for detecting goat IMI will require further investigation and optimization, especially concerning the definition of diagnostic thresholds

MICROBIOLOGICAL AND CHEMICAL TESTING OF FOOD SNAILS MARKETEDS IN SARDINIA

A total of 29 samples of snails belonging to the species Helix aspersa, Helix aperta and Helix vermiculata were analysed for the presence of Salmonella spp. and Clostridium perfringens; Cadmium and Lead concentrations were determined by ICP-MS. One sample contained Salmonella sp.; Clostridium perfringens were not detected. The means of toxic metals (expressed as mg/Kg wet weight) were the following: Cd 0,995, Pb 0,274. Key word

MOLECULAR CHARACTERIZATION OF ANISAKID NEMATODES IN FISHES OF NORTHERN SARDINIAN SEA

The authors report results of analysis carried out during 2008-2010 for identification and molecular characterization of larval Anisakis nematodes isolated from fishes of the northern Sardinian sea

Preliminary efficacy and safety data of nivolumab in never smoker patients with advanced squamous NSCLC: Experience from Italian sites participating in the Expanded Access Programme (EAP)

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