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Objective and importance: To verify the presence of beta-thalassemia in subjects showing hematologic phenotype of alpha-thalassemia, conduct normal molecular sequence analysis of the alpha-globin genes, and detect the absence of the most frequent alpha-thalassemia deletions. Clinical presentation: A patient from Apulia (Southern Italy) was referred to our institution for the occasional founding of hypochromic polyglobulia and microcytic red blood cells associated with normal levels of Hb A2 and Hb F and normal iron parameters. Intervention and technique: The patient has been investigated using Sanger sequencing, multiplex ligationdependent probe amplification (MLPA), quantitative real-time PCR, restriction analysis, and gap-PCR. A novel deletion, the Italian (epsilongammadeltabeta)0-thalassemia, has been identified. The 5' breakpoint was within a LINE element of 80 kb 3' of the epsilon-globin gene, and the 3' breakpoint was within a 160-bp palindrome of about 30 kb 5' of the beta-globin gene. The breakpoint region was characterized by the presence of a microhomology (5'-TCT-3') and of an insertion of 43 bp owing to the duplication of the 160-bp palindrome. Comparison of the Hb and Hb A2 values of (epsilongammadeltabeta)0-thalassemia from the literature with those of (molecularly known) thalassemia carriers indicated a higher level of Hb A2 with respect to alpha-thalassemia and a lower level of Hb with respect to beta0-thalassemia carriers. Conclusion: In this study, we report the first (epsilongammadeltabeta)0-thalassemia case identified in Italy. To avoid misdiagnosis of beta-thalassemia, we suggest verifying the presence of large deletions of the beta-globin gene cluster in subjects showing a higher border line level of Hb A2 and a lower level of Hb.

We report a novel alpha2-globin gene allele with the mutation cod 117 TTC>TCC or alpha 117(GH5)Phe>Ser detected in three carriers with alpha-thalassemia phenotype. The mutated mRNA was present in the reticulocytes in the same amount as the normal one, but no chain or hemoglobin variant were detected. Most likely the amino acid substitution impairs the interaction of the alpha-chain variant with the AHSP and prevents its stabilizing effect, thus leading to the alpha-chain pool reduction.

Extensive use of antimicrobial agents in finfish farming and the consequent selective pressure lead to the acquisition of antibiotic resistance in aquaculture environment bacteria. Vibrio genus represents one of the main pathogens affecting gilthead sea bream. The development of antibiotic resistance by Vibrio represents a potential threat to human health by exchange of resistant genes to human pathogens through food chain. The objective of the present study was to conduct a multisite survey on the antibiotic resistance of Vibrio spp. isolated from gilthead sea bream reared in Italian mariculture. Vibrio spp. strains were isolated from skin, gills, muscles and intestinal content of 240 gilthead sea bream. A random selection of 150 strains was sequenced for species identification. Resistance against 15 antimicrobial agents was tested by the broth microdilution method. Vibrio harveyi and Vibrio alginolyticus accounted for 36.7% and 33.3% of the isolates respectively. 96% of the strains showed multiple resistance to the tested drugs, with two strains, Vibrio aestuarianus and Vibrio harveyi resistant to 10 and 9 antibiotics, respectively. Ampicillin, amoxicillin, erythromycin and sulfadiazine showed low efficacy against Vibrio spp. Rational use of antimicrobial agents and surveillance on antibiotic administration may reduce the acquisition of resistance by microorganisms of aquatic ecosystems.

The study of the alleles of the delta-globin gene is relevant to the prevention of beta-thalassemia homozygosis; in fact, the increase of the HbA2 is an invaluable hematological marker of the beta-thalassemia heterozygosis and the double heterozygosis for alleles of delta- and beta-globin genes can cause the decrease of the HbA2 up to normal or borderline values. We carried out the characterization of alleles of the delta- and beta-globin genes, restriction fragment length polymorphism (RFLP) haplotype background, and hematologic phenotype in 23 double heterozygotes belonging to 18 unrelated families. A wide heterogeneity of the delta-globin alleles was detected; seven known alleles in trans to the beta-globin gene defects were revealed in 17 out of 18 families, while a new allele in cis to a beta-thalassemia allele was detected in one family. Moreover, the relative frequency of the delta-mutants was quite different from that found among heterozygotes. The new allele delta-cod 5 CCT>ACT, in cis to the allele beta+ thal IVS-I-110 G>A, was found in five carriers of a Sicilian family. The new variant delta5(A2)Pro>Thr, named HbA2-Partinico upon the origin of the family, was detected with high-performance liquid chromatography; it overlapped the HbA2 peak which was partially split. The double in cis heterozygotes had increased percentage of normal and variant HbA2 of comparable size. The variant originated most likely from a new mutational event because it was associated with RFLP haplotype I, commonly found with the beta+ thal IVS-I-110 G>A, even if crossing over or gene conversion cannot be excluded.

L'identificazione delle mutazioni del gene delta-globinico è importante sia per la prevenzione della beta thalassemia omozigote sia per la conoscenza della variabilità genetica in studi popolazionistici. Centoquarantasei famiglie sono state selezionate mediante dosaggio dell'HB A2 TAG) o per l'alterazione del consenso del sito di poliadenilazione (AATAAA>AATTAA). Negli altri 3 casi la mutazione porta ad una sostituzione aminocidica; tuttavia in nessun caso la nuova variante attesa è presente sul cromatogramma HPLC. Il confronto con il fenotipo delle mutazioni beta-globiniche suggerisce che nel caso delle mutazioni Hb A2-Catania ed Hb A2-Scarano l'assenza dell'Hb A2 variante potrebbe essere dovuta ad una comigrazione con la normale HbA; infatti, la variante beta con la stessa sostituzione da origine ad una "fast-mooving-band". Pertanto, i valori bassi di Hb A2 nei rispettivi portatori potrebbero essere dovuti soltanto a problemi tecnici di detection dell'Hb A2 variante. Nel caso della mutazione Hb A2-Foggia, invece, l'assenza della variante dovrebbe essere dovuta alla sua instabilità considerato che la variante beta corrispondente è associata ad un fenotipo di anemia emolitica (corpi di Heinz e riduzione della percentuale di variante circolante al 24%).

Listeriosis is a foodborne disease characterized by high hospitalization and fatality rates, especially in vulnerable groups including elderly subjects, pregnant women, etc. We report on the first case of Listeria monocytogenes ST-219 meningo-encephalitis in a woman aged 83 years. An epidemiological and molecular investigation was performed to detect the source of infection and the virulence factors associated with L. monocytogenes invasiveness in this patient. All environmental- and clinical-associated isolates were found to belong to serotype 4b and ST-219 as well as possessing actA, prfA, hlyA, and rrn virulence genes. Antibiotic susceptibility testing also detected resistance to cotrimoxazole, clindamycin, erythromycin, and oxacillin in these isolates. Conventional and molecular surveillance of listeriosis cases, based on the systematic assessment of spatio-temporal trends, virulence genes, and antimicrobial susceptibility testing patterns, are key to preventing and controlling the emergence and spread of L. monocytogenes strains, including hypervirulent clones.

This study aimed to evaluate Arcobacter species contamination of industrial sheep ricotta cheese purchased at retail and to establish if the dairy plant environment may represent a source of contamination. A total of 32 sheep ricotta cheeses (1.5 kg/pack) packed in a modified atmosphere were purchased at retail, and 30 samples were collected in two sampling sessions performed in the cheese factory from surfaces in contact with food and from surfaces not in contact with food. Seven out of 32 samples (21.9%) of ricotta cheese collected at retail tested positive for Arcobacter butzleri at cultural examination; all positive samples were collected during the same sampling and belonged to the same batch. Ten surface samples (33.3%) collected in the dairy plant were positive for A. butzleri . Cluster analysis identified 32 pulsed-field gel electrophoresis (PFGE) patterns. The same PFGE pattern was isolated from more than one ricotta cheese sample, indicating a common source of contamination, while more PFGE patterns could be isolated in single samples, indicating different sources of contamination. The results of the environmental sampling showed that A. butzleri may be commonly isolated from the dairy processing plant investigated and may survive over time, as confirmed by the isolation of the same PFGE pattern in different industrial plant surface samples. Floor contamination may represent a source of A. butzleri spread to different areas of the dairy plant, as demonstrated by isolation of the same PFGE pattern in different production areas. Isolation of the same PFGE pattern from surface samples in the dairy plant and from ricotta cheese purchased at retail showed that plant surfaces may represent a source of A. butzleri postprocessing contamination in cheeses produced in industrial dairy plants.