Monitoring AMR in Campylobacter jejuni from Italy in the last 10 years (2011-2021): Microbiological and WGS data risk assessment.

Campylobacter jejuni is considered as the main pathogen in human food-borne outbreaks worldwide. Over the past years, several studies have reported antimicrobial resistance (AMR) in C. jejuni strains. In Europe, the official monitoring of AMR comprises the testing of Campylobacter spp. from food-producing animals because this microorganism is responsible for human infections and usually predominant in poultry. Food-producing animals are considered to be a major source of campylobacteriosis through contamination of food products. Concerns are growing due to the current classification of C. jejuni by the WHO as a 'high priority pathogen' due to the emergence of resistance to multiple drugs such as those belonging to the fluoroquinolones, macrolides and other classes, which limits the treatment alternatives. Knowledge about the contributions of different food sources to gastrointestinal disease is fundamental to prioritise food safety interventions and to establish proper control strategies. Assessing the genetic diversity among Campylobacter species is essential to the understanding of their epidemiology and population structure. Using a population genetic approach and grouping the isolates into sequence types within different clonal complexes, it is possible to investigate the source of the human cases. The work programme was aimed for the fellow to assess the AMR of C. jejuni isolated from humans, poultry and birds from wild and urban Italian habitats. Given the public health concern represented by resistant pathogens in food-producing animals and the paucity of data about this topic in Italy, the aim was to identify correlations between phenotypic and genotypic AMR and comparing the origin of the isolates. The work programme allowed the fellow to acquire knowledge, skills and competencies on the web-based tools used by IZSAM to process the NGS data and perform bioinformatics analyses for the identification of epidemiological clusters, the study of AMR patterns in C. jejuni isolates, and the assessment of the human exposure to such AMR pathogens. Furthermore, the fellow became able to transfer the acquired knowledge through innovative web-based didactical tools applied to WGS and clustering of specific food-borne pathogens, with particular reference to C. jejuni. To achieve this objective, 2,734 C. jejuni strains isolated from domestic and wild animals and humans, during the period 2011-2021 were analysed. The resistance phenotypes of the isolates were determined using the microdilution method with EUCAST breakpoints, for the following antibiotics: nalidixic acid, ciprofloxacin, chloramphenicol, erythromycin, gentamicin, streptomycin, tetracycline. The data were complemented by WGS data for each strain, uploaded in the Italian information system for the collection and analysis of complete genome sequence of pathogens isolated from animal, food and environment (GENPAT) developed and maintained at IZSAM; information like clonal complex and sequence type to understand the phylogenetical distance between strains according to their origins were also considered. This work underlines that a better knowledge of the resistance levels of C. jejuni is necessary, and mandatory monitoring of Campylobacter species in the different animal productions is strongly suggested.

Development of food safety risk assessment tools based on molecular typing and WGS of Campylobacter jejuni genome.

The 'learning-by-doing' EU-FORA fellowship programme in the development of risk assessment tools based on molecular typing and WGS of Campylobacter jejuni genome was structured into two main activities: the primary one focused on training on risk assessment methodology and the secondary one in starting and enhancing the cooperation between the hosting and home organisations, or other joint activities. The primary activities had three subsequent work packages (WPs): WP1 data organisation, WP2 cluster and association analyses, and WP3 development of risk assessment models. The secondary activities have branched into one workshop and the initiation of a cooperation programme between the hosting and home organisations. In the last quarter, the fellow had contributed to the characterisation of some pathogens in possible response to a changing climate, part of the CLEFSA project. The fellow attended various forms of training: online and on-site courses, and also participated at several conferences and meetings for improving his knowledge and skills, contributing to performing the Campylobacter risk assessment and source attribution.

Brucella suis biovar 2 multi locus sequence type ST16 in wild boars (Sus scrofa) from Abruzzi region, Italy. Introduction from Central-Eastern Europe?

Porcine brucellosis occurs in many countries where pigs are farmed, often representing an underrated problem. B. suis biovar 2 is the most common isolate in Europe, with high prevalence reported in wild boars in which it is generally isolated in the absence of gross lesions. In the last five years, we tested for Brucella spp. 389 lymph nodes of wild boars collected during hunting seasons or during necropsy procedures. In this paper, we describe the first case of isolation of B. suis biovar 2 from a wild boar aborted foetus, and we analyse the genomic relationships with B.suis biovar 2 strains isolated in the past five years in Abruzzi Region, Central Italy. The genetic fingerprint revealed that the isolates under study belong to the MLST ST16 and to the MLVA11 Gt 57, similar to the Central-Eastern European strains. Massive restocking (for hunting purpose) of wild boars from Eastern Europe have been done since 1950 in Italy contributing to the increasing of population size and distribution, as well as to the interbreeding between these foreign breeds and the local population. The contamination of pastures with infected material such as aborted wild boars foetuses can increase the risk of transmission of Brucella among wild and domestic animals. The contact of B. suis with domestic ruminants may also cause serological reactions to brucellosis serological testing, and even unapparent infection, thus hampering the efforts made in the brucellosis eradication campaign.

Bangladesh anthrax outbreaks are probably caused by contaminated livestock feed.

In Bangladesh from 1 July to 30 September 2010 there were 104 animal cases of anthrax and 607 associated human cases. This investigation was conducted in Sirajganj district in December 2010, on eight farms where animal cases had occurred. Bacillus anthracis was recovered from soil samples and turbinate bones on six farms. Canonical single nucleotide polymorphism (SNP) analysis showed that all the isolates belonged to the major lineage A, sublineage A.Br.001/002 of China and South East Asia while a multilocus variable-number tandem-repeat (VNTR) analysis (MLVA) with 15 VNTRs demonstrated three unique genotypes. The single nucleotide repeat (SNR) analyses showed two SNR types in 97 out of 99 isolates; nevertheless, due to its higher discriminatory power the presence of two isolates with different SNR-type polymorphisms were detected within two MLVA genotypes. The epidemic occurred during the monsoon season, a time of extensive flooding, suggesting that the source was contaminated feed, not grazing, which is supported by the genetic variance.

Development of a real time PCR Taqman assay based on the TPI gene for simultaneous identification of Clostridium chauvoei and Clostridium septicum.

In the present study, a Taqman allelic discrimination assay based on three SNPs of the TPI gene is described. It was used as a differential diagnostic tool to detect blackleg and malignant edema. Sudden deaths of grazing ruminants, such as cattle, sheep and goats, which show clinical signs related to hyperacute infective processes, encouraged the development of a rapid and precise diagnostic molecular method. Specific primers and probes for Clostridium septicum and Clostridium chauvoei were designed on the basis of the TPI gene sequence. The multiplex PCR was tested on the DNA of a total of 57 strains, including 24 Clostridium chauvoei, 20 Clostridium septicum, 1 Bacillus anthracis and 12 other Clostridium spp. The DNA samples from Clostridium chauvoei and Clostridium septicum strains were amplified. Amplification of other DNA samples was not observed, with the exception of Clostridium tertium, which showed a weak positive signal. To avoid misdiagnosis, a confirmatory assay based on a Sybr green real time PCR was proposed. The authors confirmed the efficacy and the specificity of the test used in this study, which proved to be a useful tool for the diagnosis of clostridiosis that are often diagnosed using only traditional tools.

Protective activity and immunogenicity of two recombinant anthrax vaccines for veterinary use.

In this study, the efficacy of two experimental vaccines against Bacillus anthracis toxinaemia was evaluated in the rabbit model. A recombinant Protective Antigen (rPA) mutant and a trivalent vaccine (TV) composed by the rPA, a inactive mutant of Lethal Factor (mLF-Y728A; E735A) and a inactive mutant of Edema Factor (mEF-K346R), both emulsified with mineral oils, were evaluated for their immunogenicity and protective activity in New Zealand white rabbits. Rabbits vaccinated subcutaneously with rPA and TV rapidly produced high level of anti-PA, anti-LF and anti-EF antibodies, which were still present 6 months later. In the efficacy test, these vaccines protected 100% of rabbits challenged with B. anthracis virulent strain 0843 one week after the vaccination. Moreover, all animals vaccinated twice with rPA and TV, resisted B. anthracis infection 6 months later. Our data indicate that rPA and TV could be good vaccine candidates for inducing protection against B. anthracis infection in target animal host. They could successfully be used in an emergency with simultaneous long-acting antibiotics to halt incubating infections or during an anthrax epidemic.

How to effectively use consultants.

Apart from a few pioneering medical centers, most medical organizations lack the internal knowledge and technical expertise to develop a comprehensive digital imaging business plan, create medical imaging architectures, select vendor(s), and implement picture archiving and communication systems (PACS) and other information systems. Many of these processes are only done once (eg, vendor selection and initial implementation) and developing such one-time expertise in-house is wasteful and inefficient. It is highly unlikely that an inexperienced hospital-based group undertaking their first and only PACS project will be able to match the experience and depth of a well-organized consulting group with prior experience. Organizations therefore are looking to consulting groups to fill their internal resource gaps. This report presents the benefits of engaging consultants, guidelines for selecting a consulting group, and insight on effectively using the consultants. The presentation is based on actual experiences using consultants for planning, selecting, and implementing a PACS at Children's Hospital Medical Center, Cincinnati.

Demystifying the hospital information system/radiology information system integration process.

Most organizations planning to implement picture archiving and communications systems (PACS) are aware of the need to integrate the hospital information system (HIS) and radiology information system (RIS) with the PACS, yet few are acutely aware of the challenges associated with this requirement. This report highlights the results of collaborative efforts between Children's Hospital Medical Center-Cincinnati (CHMC) applications specialists with expertise in the HIS and CHMC information system, radiology staff familiar with the enterprise and radiology workflow and data flow requirements; and General Electric integration engineers familiar with the SMS HIS and RIS, and GE PACS. CHMC received Board approval, including full funding of the entire PACS project, in October 1998. An aggressive time frame for installation was established, as CHMC's PACS leadership committed to the selection, design, and implementation of PACS and computed radiography (CR) within 18 to 20 months. CHMC selected GE (Milwaukee, WI) as its PACS vendor in July 1999, and began its implementation in November 1999. We will present the four-stage integration process undertaken at CHMC: (1) planning the integration effort, (2) designing the Interface, (3) building the interface, and (4) testing the Interface.

[Pulmonary hamartoma: a case report]. / L'amartoma polmonare: contributo casistico.

The authors describe a case of lung hamartoma recently observed. Diagnosis occurred by chance through routine chest X-rays. A stationary radiological aspect of the hamartoma, regularly followed up, suggested to defer surgery up to the moment of a visible radiologic enlargement of the lesion.

[Critical considerations on a case of nephroblastoma in a horseshoe kidney]. / Considerazioni critiche su un caso di nefroblastoma in un rene a ferro di cavallo.

In a review of the literature, the association nephroblastoma-horseshoe kidneys has been demonstrated in 34 patients. Radiologic signs of horseshoe kidney may be difficult to evaluate when a large mass is present. Treatment of Wilms' tumor in horseshoe kidneys does not differ, on principle, from that of Wilms' tumor occurring in the normally separated kidneys.

[Primary infarction of the greater omentum: our experience in 3 cases]. / Infarto primitivo del grande omento: nostra esperienza su tre casi.

The infarction of the greater omentum is an unusual cause of acute abdominal pain. The Authors report their experience in the management of three cases. It is stressed that clinical manifestations are those of an appendicitis or cholecystitis; therefore, the diagnosis is usually intraoperative.

[A-mode echography in surgery. Study of nodular lesions of the breast]. / L'ecografia A-mode in chirurgia. Studio delle lesioni nodulari della mammella.

In the context of researches for studying the possibilities of ultrasonic A-mode research for a diagnosis of thyroid, mamma and abdominal organs affections, in this first part the authors make known the results obtained studying nodulary lesions. After a short notice about ultra-sounds, echoscopic apparati and methods for echoscopic research, they outline the semeiologic characters for echoscopic A-mode diagnostic and they stop on echoscopic obtained pictures: liquid crops, nodulary images with homogeneus echostructure scarcely ecogena and nodulary images with not homogeneus echostructure highly ecogena. Then they ended with some considerations about the principles and the indications of this method, in the light of personal experience.