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Arcobacter butzleri is a zoonotic foodborne pathogen able to cause enteric and extraintestinal diseases. Its occurrence in foodstuff is well recognized worldwide but data on its presence in foods from Southern Italy are scarce. In this study the results on the occurrence and genotyping of Arcobacter spp. in bulk milk samples collected in Southern Italy are reported. Out of 484 samples, 64 (13.2%) resulted positive for the presence of Arcobacter spp. Using Real Time PCR but as few as 31.2% of these samples turned out as positive by using the cultural method, showing an overall prevalence of 4.1%. All isolates were identified as A. cryaerophilus using the biochemical identification whilst the sequencing of the atpA gene revealed that all the isolates were A. butzleri. Among the confirmed isolates, 16 different Sequence Types (ST) were identified using the Multi Locus Sequence Typing (MLST), 14 (87.5%) of which were previously unreported. Our survey reveals the presence of A. butzleri in bulk tank milk from Southern Italy and highlights the discrepancy between the two approaches used both for the detection (i.e., real time PCR vs cultural method) and the identification (i.e., biochemical test vs aptA sequencing) of Arcobacter spp In addition, a large genetic diversity among the isolates was detected and this makes the identification of source of the infections very challenging in outbreaks investigation.

The aim of the present study was to assess the occurrence of MRSA in buffalo dairy farms and in buffalo tank milk from Italy, and to provide information about the antimicrobial resistance profile and molecular characteristics of the isolates. We collected 75 bulk tank milk (BTM) samples from 75 farms and 24 nasal swabs from 24 farm operators. Three (4%) of the 75 BTM samples and 1 (4%) of the 24 human nasal swabs tested positive for MRSA. The milk isolates belonged to the genotypes ST1/t127/Va and ST72/t3092/V, while the human isolate was characterized as ST1/t127/IVa. All isolates were multidrug resistant but vancomycin susceptible; they carried the icaA gene but tested negative for the pvl and ses genes. ST72 is a CA-MRSA commonly found in South Korea, and this is the first report of its detection in Europe. Although we found a low prevalence of MRSA in the farms we surveyed, this study clearly demonstrates, for the first time in Europe, that MRSA can be found in dairy buffalo farms and in raw buffalo milk. Therefore, the risk of human colonization/infection with MRSA linked to the handling of raw milk or consumption of contaminated dairy products should not be ruled out.

Resumen del trabjo presentado a la 14th International Conference on Microbial Interactions and Microbial Ecology, celebrada en Viena (Austria) del 19 al 20 de agosto de 2019., [Statement of the Problem]: Intestinal mucus layer may provide a niche for many nosocomial pathogens, including S. aureus which can occasionally cause a Staphylococcal enterocolitis. Recent exciting researches support the notion that a healthy intestinal microbiota composition can promote resistance to invading pathogenic bacterial species., [Purpose]: The purpose of this study was to evaluate the survival of MRSA in simulated human ascendant colon conditions and its interaction with gut microbiota into the mucus layer., [Methodology and Theoretical Orientation]: The study was performed at ascendant colon environment: body-like temperature (37ºC), anaerobiosis (N2), pH 5.7, constant slow shaking (40 RPM). Mucin agar carriers stand for the intestinal mucus layer and a basic feed medium represented the intestinal lumen contents. A three-days long in vitro study was performed by using microbiota from pooled faeces of healthy individuals that were stabilized simulating ascendant colon conditions and a MRSA strain of animal origin (ST398-t011-SCCmecV; 107 UFC/mL). Each day we checked the viability of MRSA both into the mucin agar carriers and in the feed medium by using MRSA-SELECTplates (BioRad). The results were confirmed by quantitative PCR., [Findings]: MRSA population decreased as a function of time during the incubation with luminal colon microbiota where it was not viable after 24 h. Counts of 4 log cfu/g were still obtained in the mucin agar carriers after 72 h of incubation. On the other hand, counts of Bifidobacterium and Akkermansia increased in the mucin agar carriers as a function of time., [Conclusion and Significance]: The results support the hypothesis that a competitive microbiota may control MRSA intestinal colonization empathize the important role of specific groups which can inhibit the adhesion of/displace MRSA from the intestinal mucus layer.

Table S1. Group, stage, generation and genotype of ticks genetically identified in this study. (DOCX 18 kb)

The capability to produce biofilm is an important persistence and dissemination mechanism of some foodborne bacteria. This paper investigates the relationship between some molecular characteristics (SCCmec, ST, spa-type, agr-type, cna, sarA, icaA, icaD, clfA, fnbA, fnbB, hla, hlb) of 22 food-related methicillin-resistant Staphylococcus aureus (MRSA) strains and their ability to form biofilm on stainless steel and polystyrene. Five (22.7%, 5/22) strains were able to synthesize biofilm on polystyrene, and one of these (4.5%, 1/22) strains was also able to synthesize biofilm on stainless steel. The largest amount of biofilm was formed on polystyrene by 2 MRSA strains isolated from cows' milk, thus raising concern about the dairy industry. The majority of MRSA biofilm producers carried SCCmec type IVa, suggesting that the presence of SCCmecIVa and/or agr type III could be related to the ability to form biofilm. In conclusion, in order to achieve an acceptable level of food safety, Good Hygiene Practices should be strictly implemented along the food chain to reduce the risk of colonization and dissemination of MRSA biofilm-producing strains in the food industry.In this study, some assayed isolates of food-related MRSA demonstrated the capacity to form biofilm. Biofilm formation differed according to surface characteristics and MRSA strains. A relationship was observed between some molecular characteristics and the ability to form biofilms. Few studies have investigated the ability of MRSA to form biofilms, and the majority of these studies have investigated clinical aspects. This work was performed to investigate whether or not there is a difference between MRSA food isolates and MRSA clinical isolates in their ability to form biofilm. These initial findings could provide information that will contribute to a better understanding of these aspects.

Isolate and subtyping data of the 48 analyzed L. monocytogenes isolates collected from 2006 to 2014 in the Lombardy Region in Italy from clinical (n = 43), food (n = 4), and environmental (n = 1) sources. All clinical isolates were previously serotyped as 1/2a and typed as sequence type 38 (ST38), which belongs to clonal complex 101 (CC101). All isolates share the pulsotypes identified in Cluster 11 in Mammina et al. [7]. aBG – Bergamo, BS – Brescia, CO – Como, CR – Cremona, LC – Lecco, LO – Lodi, MB – Monza Brianza, MI – Milan, MN – Mantova, PV – Pavia, SO – Sondrio, VA – Varese. bCSF: cerebrospinal fluid. cVT: Virulence Type. (DOCX 18 kb)

Epitopes predicted by bioinformatics and those reported in the literature for other organisms mapped to Onchocerca lupi paramyosin. Residues underlined indicate T-cell epitopes whilst those in bold indicate B-cell epitopes. Of the 874 predicted antibody epitopes, when compared with those predicted for Acanthocheilonema viteae, Brugia malayi, Dirofilaria immitis, Loa loa, Onchocerca volvulus and Wuchereria bancrofti, 621 were unique to Onchocerca lupi. (DOCX 32Â kb)

Epitopes predicted by bioinformatics and those reported in the literature for other organisms mapped to Onchocerca lupi paramyosin. Residues underlined indicate T-cell epitopes whilst those in bold indicate B-cell epitopes. Of the 874 predicted antibody epitopes, when compared with those predicted for Acanthocheilonema viteae, Brugia malayi, Dirofilaria immitis, Loa loa, Onchocerca volvulus and Wuchereria bancrofti, 621 were unique to Onchocerca lupi. (DOCX 32Â kb)

Since the beginning in December 2019, the SARS-CoV-2 outbreak appeared to affect mostly the adult population, sparing the vast majority of children who only showed mild symptoms. The purpose of this investigation is to assess the status on the mechanisms that give children and infants this variation in epidemiology compared to the adult population and its impact on therapies and vaccines that are aimed towards them. A literature review, including in vitro studies, reviews, published guidelines and clinical trials was performed. Clinical trials concerned topics that allowed a descriptive synthesis to be produced. Four underlying mechanisms were found that may play a key role in providing COVID-19 protection in babies. No guidelines are available yet for therapy due to insufficient data; support therapy remains the most used. Only two vaccines are approved by the World Health Organization to be used in children from 12 years of age, and there are currently no efficacy or safety data for children below the age of 12 years. The COVID-19 clinical frame infection is milder in children and adolescents. This section of the population can act as vectors and reservoirs and play a key role in the transmission of the infection; therefore, vaccines are paramount. More evidence is required to guide safely the vaccination campaign.

Rabbit meat has outstanding dietetic and nutritional properties. However, few data on microbiological hazards associated with rabbit productions are available. In this study, the presence of Listeria monocytogenes was determined in 430 rabbit carcasses, 256 rabbit meat cuts and products, and 599 environmental sponges collected from four Italian rabbit slaughterhouses over a period of 1 year. Prevalence of L. monocytogenes among the 1285 rabbit meat and environmental samples was 11%, with statistically significant differences between slaughterhouses. The highest prevalence (33.6%) was observed in rabbit meat cuts and products; the majority of positive environmental samples were collected from conveyor belts. Overall, 27.9% and 14.3% of rabbit cuts and carcasses, respectively, had L. monocytogenes counts higher than 1 colony-forming unit (CFU)/10 g. A selection of 123 isolates from positive samples was genotyped and serotyped to determine genetic profiles and diversity among L. monocytogenes isolates contaminating different slaughterhouses and classes of products investigated. Discriminatory power and concordance among the results obtained using multilocus variable-number tandem-repeat analysis (MLVA), multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE), automated EcoRI ribotyping, and serotyping were assessed. The isolates selected for typing were classified into serotypes 1/2a (52.8%), 1/2c (32.5%), and 1/2b (14.6%). The majority of the isolates were classified as ST14 (34.1%), ST9 (35.5%), ST121 (17.9%), and ST224 (14.6%). The greatest discriminatory power was observed with the MLVA typing, followed by MLST, PFGE, and ribotyping. The best bidirectional concordance was achieved between PFGE and MLST. There was 100% correlation between both MLST and MLVA with serotype. Moreover, a high unidirectional correspondence was observed between MLVA and both MLST and PFGE, as well as between PFGE and both MLST and serotyping. The results of this study show for the first time in Italy prevalence and genetic profiles of L. monocytogenes isolated in rabbit products and slaughterhouses.

In this study, the isolation of Acinetobacter baumannii in a dog with clinical bilateral otitis externa is described. Moreover, to investigate the zoonotic potential of the isolate, microbiological examinations on the family members were performed. An A. baumanniistrain was isolated from nasal swab in one of the dog owners. The identity of bacterial strains, either from dog and owner, was confirmed by phenotypic and molecular typing (wgMLST). Furthermore, to assess the pathogenic potential of the isolates a deep characterization of virulence and antibiotic resistance genes was done by Whole Genome Sequencing (WGS). Finally, the susceptibility towards a wide panel of antimicrobials was investigated. In our knowledge, this is the first recorded case of A. baumanniiisolation from canine auricular swabs in Italy. And interestingly, this study underlines the possible spread of this microorganism from human to animal.

Aims The present study aimed to determine, by multilocus sequence type (MLST), the heterogeneity level of Arcobacter butzleri isolates and to compare MLST and pulsed-field gel electrophoresis (PFGE) in terms of discriminatory power (DI) as well as unidirectional and bi-directional concordance. Methods and Results Arcobacter butzleri isolates (N = 133) from dairy products and environmental samples, collected from dairy plants, were characterized by MLST and PFGE with SacII and classified in 29 sequence types (STs), 47 PFGE and 62 type strains (TS). Among the 119 alleles, 19 were previously unreported and the same for all the STs but two. A significant linkage disequilibrium was detected when the complete ST data set was analysed The DIs of MLST, PFGE and their combination were 0·937, 0·953 and 0·965 respectively. The adjusted Wallace coefficients between MLST and PFGE as well as PFGE and MLST were 0·535 and 0·720 respectively; the adjusted Rand coefficient was 0·612. Conclusions The A. butzleri studied population showed recombination to some degree. PFGE showed a DI higher than MLST. Both methods presented good concordance. The TS analysis seems to show persistence of the same strain on time and possible cross-contaminations between food and environmental sites. Significance and Impact of the Study This study provides insights in the A. butzleri population found in raw milk, cheese, and dairy production plants. The data suggest that MLST and PFGE genotypes correlate reasonably well, although their combination results in optimal resolution.

In the domain of Earth Explorer satellites nanoengineered nonlinear crystals can optimize UV tunable solid-state laser converters. Lightweight sources can be based on Lithium Niobate (LN) domain engineering by electric field poling and guided wave interactions. In this Communication we report the preliminary experimental results and the very first demonstration of UltraViolet second-harmonic generation by first-order quasi-phase-matching in a surface-periodically-poled proton-exchanged LN waveguide. The pump source was a Ti-Sapphire laser with a tunability range of 700- 980 nm and a 40 GHz linewidth. We have measured UV continuous-wave light at 390 nm by means of a lock-in amplifier and of a photodiode with enhanced response in the UV. Measured conversion efficiency was about 1%W-1cm-2. QPM experiments show good agreement with theory and pave the way for a future implementation of the technique in materials less prone to photorefractive damage and wider transparency in the UV, such as Lithium Tantalate.

Antonella Accardo1,2*, Giuseppina Salsano3*, Anna Morisco4, Michela Aurilio4, Antonio Parisi5, Francesco Maione5, Carla Cicala5, Diego Tesauro1,2, Luigi Aloj4, Giuseppe De Rosa3, Giancarlo Morelli1,21CIRPeB, Department of Biological Sciences and IBB CNR, University of Naples “Federico II”, 2Invectors srl, 3Department of Pharmaceutical Chemistry, University of Naples “Federico II”, 4Department of Nuclear Medicine, Istituto Nazionale per lo Studio e la Cura dei Tumori, Fondazione “G. Pascale”, 5Department of Experimental Pharmacology, University of Naples “Federico II”, Napoli, Italy*These authors contributed equally to this workObjectives: Drug delivery systems consisting of liposomes displaying a cell surface receptor-targeting peptide are being developed to specifically deliver chemotherapeutic drugs to tumors overexpressing a target receptor. This study addresses novel liposome composition approaches to specifically target tissues overexpressing bombesin (BN) receptors.Methods: A new amphiphilic peptide derivative (MonY-BN) containing the BN(7–14) peptide, the DTPA (diethylenetriaminepentaacetate) chelating agent, a hydrophobic moiety with two C18 alkyl chains, and polyethylene glycol spacers, has been synthesized by solid-phase methods. Liposomes have been generated by co-aggregation of MonY-BN with 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC). The structural and biological properties of these new target-selective drug-delivery systems have been characterized.Results: Liposomes with a DSPC/MonY-BN (97/3 molar ratio) composition showed a diameter of 145.5 ± 31.5 nm and a polydispersity index of 0.20 ± 0.05. High doxorubicin (Dox) loading was obtained with the remote pH gradient method using citrate as the inner buffer. Specific binding to PC-3 cells of DSPC/MonY-BN liposomes was obtained (2.7% ± 0.3%, at 37°C), compared with peptide-free DSPC liposomes (1.4% ± 0.2% at 37°C). Incubation of cells with DSPC/MonY-BN/Dox showed significantly lower cell survival compared with DSPC/Dox-treated cells, in the presence of 100 ng/mL and 300 ng/mL drug amounts, in cytotoxicity experiments. Intravenous treatment of PC-3 xenograft-bearing mice with DSPC/MonY-BN/Dox at 10 mg/kg Dox dose produced higher tumour growth inhibition (60%) compared with nonspecific DSPC/Dox liposomes (36%) relative to control animals.Conclusion: The structural and loading properties of DSPC/MonY-BN liposomes along with the observed in-vitro and in-vivo activity are encouraging for further development of this approach for target-specific cancer chemotherapy.Keywords: bombesin peptide, doxorubicin delivery, gastrin-releasing peptide receptors, PC-3 cells, theranostic applications