Matrix-Assisted Laser Desorption/Ionization Time Of Flight Mass Spectrometry (MALDI-TOF MS) technology is currently increasingly used in diagnostic laboratories as a cost effective, rapid and reliable routine technique for the identification and typing of microorganisms. In this study, we used MALDI-TOF MS to analyze a collection of 160 strains belonging to the Bacillus cereus group (57 B. anthracis, 49 B. cereus, 1 B. mycoides, 18 B. wiedmannii, 27 B. thuringiensis, 7 B. toyonensis and 1 B. weihenstephanensis) and to detect specific biomarkers which would allow an unequivocal identification. The Main Spectra Profiles (MSPs) were added to an in-house reference library, expanding the current commercial library which does not include B. toyonensis and B. wiedmannii mass spectra. The obtained mass spectra were statistically compared by Principal Component Analysis (PCA) that revealed seven different clusters. Moreover, for the identification purpose, were generated dedicate algorithms for a rapid and automatic detection of characteristic ion peaks after the mass spectra acquisition. The presence of specific biomarkers can be used to differentiate strains within the B. cereus group and to make a reliable identification of Bacillus anthracis, etiologic agent of anthrax, which is the most pathogenic and feared bacterium of the group. This could offer a critical time advantage for the diagnosis and for the clinical management of human anthrax even in case of bioterror attacks.

The coronavirus disease 2019 (Covid-19) pandemic is caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and presents a global health emergency that needs urgent intervention. Viruses constantly change through mutation, and new variants of a virus are expected to occur over time. In the United Kingdom (UK), a new variant called B.1.1.7 has emerged with an unusually large number of mutations. The aim of this study is to evaluate the level of protection of sera from 12 patients infected and later healed in Apulia Region (Italy) with Covid-19 between March and November 2020, when the English variant was not circulating in this territory yet, against the new VOC 202012/01 variant by seroneutralization assay. The sera of patients had already been tested before, using a virus belonging to the lineage B.1 and showed an antibody neutralizing titer ranging between 1:160 and 1:320. All the 12 sera donors confirmed the same titers of neutralizing antibodies obtained with a strain belonging to the lineage B.1.1.7 (VOC 202012/01). These data indicate that antibodies produced in subjects infected with variants of Sars-CoV-2 strain before the appearance of the English one, seem to have a neutralizing power also against this variant.

Salmonellosis is one of the most frequent foodborne zoonoses, and the consumption of contaminated poultry meat is considered the main source of Salmonella infections in humans. From 2021 to 2023, 384 chicken samples were collected from retailers in Apulia and Basilicata regions and analysed at the Istituto Zooprofilattico Sperimentale della Puglia e della Basilicata (IZSPB) laboratories. The Salmonella isolates were investigated to evaluate their phenotypic characteristics of antimicrobial resistance. A total of 125 (32.55%) samples tested positive for Salmonella spp. Three samples were simultaneously infected with strains of two different serotypes. Strains were classified into nine serotypes. Out of 128 strains, 112 (87.5%) isolates showed multidrug-resistant (MDR) profiles. Moreover, 16 isolates (12.5%) were ESBL producers with MDR profiles. Our data confirm that chicken products are a common source of Salmonella and highlight how S. Infantis was the most prevalent serotype (85.93%). Furthermore, Salmonella isolates showed antimicrobial resistance, which is very worrisome for animals and consumers. The high rate of antibiotic resistance along with the appearance of numerous MDR isolates is considered an actual concern and requires a robust surveillance network in a One Health concept.

Poultry is the main reservoir of Campylobacter spp. and most human cases result from consuming undercooked poultry or handling raw meat. In 2022, a total of 55 samples, including neck skin, cecal contents, and processing waters, were collected at two poultry slaughterhouses in Italy and analysed according to ISO 10272-2:2017 at the Istituto Zooprofilattico Sperimentale della Puglia e della Basilicata laboratories. Overall, 51/55 (92.72%) samples tested positive for Campylobacter. Among the isolates, 64.71% were identified as C. coli, and 35.29% as C. jejuni. Phenotypic and genotypic analysis were performed to assess antimicrobial resistance and virulence characteristics. All C. jejuni isolates and 72.72% of C. coli showed resistance to fluoroquinolones. Resistances to tetracycline and carbapenem were observed in 60.78% and 45.09% of isolates, respectively. Genomic analysis confirmed the presence of the tet(O) gene, conferring tetracycline resistance. In addition, OXA-450 and OXA-466 genes, conferring beta-lactam resistance, were detected in 78.43% and 3.92% of isolates. Virulence-associated genes were detected. Specifically, the ciaB gene was found in 50/51 (98.04%) of isolates, whereas jlpA, cdtA, cdtB, and ctdC genes were exclusively identified in C. jejuni strains. The high prevalence of pathogenic and antimicrobial-resistant Campylobacter strains highlights the need for strengthened control measures along the poultry production chain.

Purpose. During the 2024-2025 hunting season, sampling was carried out to obtain a preliminary estimate of the circulation of Salmonella spp. in wild boars hunted in Puglia and Basilicata, with the aim of collecting useful data to better define the infectious risk associated with the handling and consumption of the meat derived from them. Methods. The survey was conducted on 148 wild boar specimens shot during the 2024-2025 hunting season. Specifically, 106 samples were collected from carcasses and 42 rectal swabs were collected using a non-destructive method (sponges). Carcass sampling was carried out at a Game Processing Centre (CLS), while rectal swabs were taken in the field. The isolation of Salmonella spp. was carried out following the protocol set out in ISO 6579-1:2017. An isolate attributable to Salmonella spp. per positive sample was confirmed by MALDI-TOF MS (Matrix-Assisted Laser Desorption/Ionisation Time-of-Flight Mass Spectrometry). Each strain identified as Salmonella spp. was subjected to serotyping according to ISO/TR 6579-3:2014 and to minimum inhibitory concentration (MIC) determination using commercial Sensititre™ EUVSEC 3 plates (Thermo Scientific). The MIC results were interpreted using the reference breakpoints reported in EUCAST 2025 version 15. Results. Overall, Salmonella spp. was detected in 5/148 samples (3.37%). Specifically, in 3 out of 78 carcasses (3.84%) from Basilicata (PZ), it was possible to isolate 2 strains of S. Kasenyi (38:e,h:1.5) and 1 strain of S. Kottbus (6.8:e,h:1.5). Two rectal swabs out of 42 (4.76%) tested positive for S. ser. II (41:z:1.5) (Puglia - FG) and for S. Kasenyi (Basilicata - PZ). All isolates were sensitive to the antimicrobials tested. Conclusions. The results of this preliminary investigation highlighted the presence of Salmonella spp. in 3.37% of the samples examined, confirming that wild boar is a species capable of spreading this microorganism in the environment and, potentially, transferring it to consumers. With regard to the identification of isolates, the detection of S. Kasenyi and S. Kottbus in carcasses is of interest. While there are no reports of human cases for S. Kasenyi, already isolated from wild boars from Campania, there are several reports of outbreaks caused by S. Kottbus in the United States, the Canary Islands and Germany. In addition, Denmark has reported a case of S. Kottbus carrying the blaNDM-1 gene, which confers resistance to carbapenems. In light of the current evidence, there are no data available on the presence of Salmonella ser. II (41:z:1,5) in wild boars hunted in Italy, nor are there any reports on this in European scientific literature. Finally, with regard to antimicrobial resistance potential, in our investigation all isolates showed a high sensitivity profile to the molecules tested. In conclusion, the identification of S. Kasenyi and S. Kottbus in wild boar is of interest for the ecological surveillance of salmonellosis and highlights the potential role of this species as a reservoir of minor serotypes, with epidemiological implications yet to be clarified.