Compliance of clinical microbiology laboratories with recommendations for the diagnosis of bloodstream infections: Data from a nationwide survey in Italy.

In 2014, the Italian Working Group for Infections in Critically Ill Patient of the Italian Association of Clinical Microbiologists updated the recommendations for the diagnostic workflow for bloodstream infections (BSI). Two years after publication, a nationwide survey was conducted to assess the compliance with the updated recommendations by clinical microbiology laboratories. A total of 168 microbiologists from 168 laboratories, serving 204 acute care hospitals and postacute care facilities, were interviewed during the period January-October 2016 using a questionnaire consisting of nineteen questions which assessed the level of adherence to various recommendations. The most critical issues were as follows: (a) The number of sets of blood cultures (BC) per 1,000 hospitalization days was acceptable in only 11% of laboratories; (b) the minority of laboratories (42%) was able to monitor whether BCs were over or under-inoculated; (c) among the laboratories monitoring BC contamination (80%), the rate of contaminated samples was acceptable in only 12% of cases;(d) the Gram-staining results were reported within 1 hr since BC positivity in less than 50% of laboratories. By contrast, most laboratories received vials within 2-4 hr from withdrawal (65%) and incubated vials as soon as they were received in the laboratory (95%). The study revealed that compliance with the recommendations is still partial. Further surveys will be needed to monitor the situation in the future.

Outbreak of severe Hepatitis A in Eastern Piedmont, Italy.

Hepatitis A (HA) is caused by a hepatovirus from the family Picornaviridae (Hepatitis A Virus, HAV). Transmission occurs mainly by the orofaecal route through food or water contaminated by faeces. Sexual transmission has also been reported among men who have sex with men (MSM). From February to May 2017, 14 patients with HA were hospitalized at the University Hospital "Maggiore della Carità", Novara (Eastern Piedmont), Italy. One patient was two years old and was therefore admitted to the Paediatric Unit, the remaining 13 to the Infectious Disease Unit. Two of the adults were female and the rest (11) were male. The male patients were MSM, and contracted the infection sexually; three of them were known to be HIV positive, while two had a new diagnosis of syphilis infection. Women contracted the infection from contaminated food.

From soil to blood: first human case of Sphingobacterium hotanense bacteraemia.

This report describes a case of Sphingobacterium hotanense bacteraemia in a patient scratched by a rooster on the right arm. Diagnostic, clinical and therapeutic features are discussed. To the best of our knowledge, this is the first case of Sphingobacterium hotanense bacteremia reported in the medical literature.

Climate, environment and transmission of malaria.

Malaria, the most common parasitic disease in the world, is transmitted to the human host by mosquitoes of the genus Anopheles. The transmission of malaria requires the interaction between the host, the vector and the parasite.The four species of parasites responsible for human malaria are Plasmodium falciparum, Plasmodium ovale, Plasmodium malariae and Plasmodium vivax. Occasionally humans can be infected by several simian species, like Plasmodium knowlesi, recognised as a major cause of human malaria in South-East Asia since 2004. While P. falciparum is responsible for most malaria cases, about 8% of estimated cases globally are caused by P. vivax. The different Plasmodia are not uniformly distributed although there are areas of species overlap. The life cycle of all species of human malaria parasites is characterised by an exogenous sexual phase in which multiplication occurs in several species of Anopheles mosquitoes, and an endogenous asexual phase in the vertebrate host. The time span required for mature oocyst development in the salivary glands is quite variable (7-30 days), characteristic of each species and influenced by ambient temperature. The vector Anopheles includes 465 formally recognised species. Approximately 70 of these species have the capacity to transmit Plasmodium spp. to humans and 41 are considered as dominant vector capable of transmitting malaria. The intensity of transmission is dependent on the vectorial capacity and competence of local mosquitoes. An efficient system for malaria transmission needs strong interaction between humans, the ecosystem and infected vectors. Global warming induced by human activities has increased the risk of vector-borne diseases such as malaria. Recent decades have witnessed changes in the ecosystem and climate without precedent in human history although the emphasis in the role of temperature on the epidemiology of malaria has given way to predisposing conditions such as ecosystem changes, political instability and health policies that have reduced the funds for vector control, combined with the presence of migratory flows from endemic countries.

[The mosquito-borne viruses in Europe]. / Virus trasmessi da zanzare in Europa.

Epidemiologic changes of vector-borne diseases in recent years have multiple causes, including climate change. There are about 3500 species of mosquitoes worldwide, three-quarters of which live in tropical and subtropical wetlands. Main viruses transmitted by mosquitoes in Europe belong to the genus Flavivirus; some of them have been recently reported in Italy (Usutu and Japanese encephalitis virus), while others have been circulating for years and autochthonous transmission has been documented (West Nile virus). Mosquito-borne viruses can be classified according to the vector (Aedes or Culex), which, in turn, is associated with different vertebrate host and pathology. The Flavivirus transmitted by Culex have birds as a reservoir and can cause meningoencephalitis, while viruses transmitted by Aedes have primates as reservoir, do not have neurotropism and mainly cause hemorrhagic diseases. Other arbovirus, potentially responsible of epidemics, are the Chikungunya virus (Alphavirus family), introduced for the first time in Europe in 2007, and the virus of Rift Valley fever (Phlebovirus family). The spread in non-endemic areas of vector-born diseases have highlighted the importance of surveillance systems and vector control strategies.

Multiple withdrawals from single-use vials: a study on sterility.

BACKGROUND: Reutilization of single-use vials containing medical drugs is still under discussion. This practice has been adopted as a standard to avoid drug wastage, particularly in developing countries and in the aftermath of disasters. Some studies have assessed sterility of medications stored in single-use vials after utilization as multiple doses; however, most of these were limited to one single drug, included a low number of samples and did not consider an intermediate transfer step from the vial to a disposable syringe. The purpose of this study was to assess microbial contamination of samples withdrawn over three days from disposable syringes prepared from single-use vials. METHODS: A prospective sterility study was conducted. A total of 600 initial samples were prepared from six-hundred 10 mL single-use vials of physiological solution into six-hundred 20 mL disposable syringes. Samples were prepared in three different standard operating rooms, on six different days and by the same operator, using basic sterile technique. All syringes were capped, placed together in a non-sterile steel container, covered with a clean drape and stored in the refrigerator at 4°C under non-sterile conditions. Using basic sterile technique, four samples were withdrawn daily and cultured from each syringe over the next 3 days. Microbial growth was examined on Sabouraud agar and chocolate agar culture media. RESULTS: A total of 7200 samples were collected and 14,400 cultures were performed. No evidence of microbial growth in any of the culture media plates was found. CONCLUSION: This study demonstrated that contents initially stored in single-use vials and subsequently transferred into disposable syringes in an operating room using sterile technique, maintain sterility after 4 withdrawals per day for a total of 3 days.

[Vector transmitted diseases and climate changes in Europe]. / Malattie trasmesse da vettorie cambiamenti climatici in Europa.

The increase in temperatures recorded since the mid-nineteenth century is unprecedented in the history of mankind. The consequences of climate changes are numerous and can affect human health through direct (extreme events, natural disasters) or indirect (alteration of the ecosystem) mechanisms. Climate changes have repercussions on ecosystems, agriculture, social conditions, migration, conflicts and the transmission mode of infectious diseases. Vector-borne diseases are infections transmitted by the bite of infected arthropods such as mosquitoes, ticks, triatomines, sand flies and flies. Epidemiological cornerstones of vector-borne diseases are: the ecology and behaviour of the host, the ecology and behaviour of the vector, and the population's degree of immunity. Mosquito vectors related to human diseases mainly belong to the genus Culex, Aedes and Mansonia. Climate changes in Europe have increased the spread of new vectors, such as Aedes albopictus, and in some situations have made it possible to sustain the autochthonous transmission of some diseases (outbreak of Chukungunya virus in northern Italy in 2007, cases of dengue in the South of France and in Croatia). Despite the eradication of malaria from Europe, anopheline carriers are still present, and they may allow the transmission of the disease if the climatic conditions favour the development of the vectors and their contacts with plasmodium carriers. The tick Ixodes ricinus is a vector whose expansion has been documented both in latitude and in altitude in relation to the temperature increase; at the same time the related main viral and bacterial infections have increased. In northern Italy and Germany, the appearance of Leishmaniasis has been associated to climatic conditions that favour the development of the vector Phlebotomus papatasi and the maturation of the parasite within the vector, although the increase of cases of visceral leishmaniasis is also related to host immune factors, particularly immunodepression caused by the human immunodeficiency virus (HIV). Despite the importance of global warming in facilitating the transmission of certain infectious diseases, due consideration must be taken of the role played by other variables, such as the increase in international travel, migration and trade, with the risk of importing parasites and vectors with the goods. In addition, the control of certain infections was possible in the past through improvements in socio-economic conditions of affected populations. However, the reduction in resources allocated to health care has recently led to the re-emergence of diseases that were considered eradicated.

Early diagnosis of invasive pulmonary aspergillosis in a young immunocompetent patient.

A 22-year-old insulin-dependent diabetic male was admitted for diabetic ketoacidosis. He developed hospital-acquired pneumonia (HAP) for which empirical antibiotic and antifungal therapy was started on the ward. On day 6, clinical and laboratory findings worsened, and bronchoalveolar lavage (BAL) was performed. Serum real time-polymerase chain reaction (RT-PCR) indicated invasive pulmonary aspergillosis (IPA) and led to antifungal therapy being initiated 48 hours before the results of the BAL culture were available. Despite early appropriate antifungal therapy, however, the patient died on day 22 while being supported by venovenous extracorporeal membrane oxygenation.

Direct identification of bacteria in blood culture by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry: a new methodological approach.

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has recently been demonstrated to be a powerful tool for the rapid identification of bacteria from growing colonies. In order to speed up the identification of bacteria, several authors have evaluated the usefulness of this MALDI-TOF MS technology for the direct and quick identification bacteria from positive blood cultures. The results obtained so far have been encouraging but have also shown some limitations, mainly related to the bacterial growth and to the presence of interference substances belonging to the blood cultures. In this paper, we present a new methodological approach that we have developed to overcome these limitations, based mainly on an enrichment of the sample into a growing medium before the extraction process, prior to mass spectrometric analysis. The proposed method shows important advantages for the identification of bacterial strains, yielding an increased identification score, which gives higher confidence in the results.

CMY-16, a novel acquired AmpC-type beta-lactamase of the CMY/LAT lineage in multifocal monophyletic isolates of Proteus mirabilis from northern Italy.

We report multifocal detection (four different cities in northern Italy) of Proteus mirabilis isolates resistant to both oxyimino- and 7-alpha-methoxy-cephalosporins and producing a novel acquired AmpC-like beta-lactamase. The enzyme, named CMY-16, is a variant of the CMY/LAT lineage, which differs from the closest homologues, CMY-4 and CMY-12, by a single amino acid substitution (A171S or N363S, respectively) and from CMY-2 by two substitutions (A171S and W221R). Expression of the cloned bla(CMY-16) gene in Escherichia coli decreased susceptibility to penicillins, cephalosporins, and aztreonam. Tazobactam was more effective than clavulanate at antagonizing the enzyme activity. Genotyping, by random amplification of polymorphic DNA and pulsed-field gel electrophoresis of genomic DNA digested with SfiI, showed that isolates were clonally related to each other, although not identical. The bla(CMY-16) gene was not transferable to E. coli by conjugation or transformation. In all isolates, it was chromosomally located and inserted in a conserved genetic environment. PCR mapping experiments revealed that the bla(CMY-16) was flanked by ISEcp1 and the blc gene, similar to other genes of this lineage from plasmids of Salmonella enterica, Klebsiella spp., and E. coli. Overall, these results revealed multifocal spreading of a CMY-16-producing P. mirabilis clone in northern Italy. This finding represents the first report of an acquired AmpC-like beta-lactamase in Proteus mirabilis from Italy and underscores the emergence of similar resistance determinants in the European setting.