Genotypic and phenotypic profiling of 127 Legionella pneumophila strains: Insights into regional spread.

Given the recent global surge in Legionnaires' disease cases, the monitoring of Legionella pneumophila becomes increasingly crucial. Epidemiological cases often stem from local outbreaks rather than widespread dissemination, emphasizing the need to study the characteristics of this pathogen at a local level. This study focuses on isolates of L. pneumophila in the Italian region of Friuli Venezia Giulia to assess specific genotype and phenotype distribution over time and space. To this end, a total of 127 L. pneumophila strains isolated between 2005 and 2017 within national surveillance programs were analysed. Rep-PCR, RAPD, and Sau-PCR were used for genotypic characterization, while phenotypic characterization was conducted through fatty acids analysis. RAPD and Sau-PCR effectively assessed genetic characteristics, identifying different profiles for the isolates and excluding the presence of clones. Although Sau-PCR is rarely used to analyse this pathogen, it emerged as the most discriminatory technique. Phenotypically, hierarchical cluster analysis categorized strains into three groups based on varying membrane fatty acid percentages. However, both phenotypic and genotypic analyses revealed a ubiquitous profile distribution at a regional level. These results suggest an absence of correlations between strain profiles, geographical location, and isolation time, indicating instead high variability and strain dissemination within this region.

Draft genome sequences from 127 Legionella spp. strains isolated in water systems linked to legionellosis outbreaks.

Legionnaires' disease is a severe form of pneumonia caused by Legionella spp. bacteria. According to the European Centre for Disease Prevention and Control, problems related to this pathogen showed a significant surge in recent years, making its monitoring critical.

Self-Organizing Maps: An AI Tool for Identifying Unexpected Source Signatures in Non-Target Screening Analysis of Urban Wastewater by HPLC-HRMS.

(1) Background: Monitoring effluent in water treatment plants has a key role in identifying potential pollutants that might be released into the environment. A non-target analysis approach can be used for identifying unknown substances and source-specific multipollutant signatures. (2) Methods: Urban and industrial wastewater effluent were analyzed by HPLC-HRMS for non-target analysis. The anomalous infiltration of industrial wastewater into urban wastewater was investigated by analyzing the mass spectra data of "unknown common" compounds using principal component analysis (PCA) and the Self-Organizing Map (SOM) AI tool. The outcomes of the models were compared. (3) Results: The outlier detection was more straightforward in the SOM model than in the PCA one. The differences among the samples could not be completely perceived in the PCA model. Moreover, since PCA involves the calculation of new variables based on the original experimental ones, it is not possible to reconstruct a chromatogram that displays the recurring patterns in the urban WTP samples. This can be achieved using the SOM outcomes. (4) Conclusions: When comparing a large number of samples, the SOM AI tool is highly efficient in terms of calculation, visualization, and identifying outliers. Interpreting PCA visualization and outlier detection becomes challenging when dealing with a large sample size.

Summary recommendations on "Analytical methods for substances in the Watch List under the Water Framework Directive".

The Watch List (WL) is a monitoring program under the European Water Framework Directive (WFD) to obtain high-quality Union-wide monitoring data on potential water pollutants for which scarce monitoring data or data of insufficient quality are available. The main purpose of the WL data collection is to determine if the substances pose a risk to the aquatic environment at EU level and subsequently to decide whether a threshold, the Environmental Quality Standards (EQS) should be set for them and, potentially to be listed as priority substance in the WFD. The first WL was established in 2015 and contained 10 individual or groups of substances while the 4th WL was launched in 2022. The results of monitoring the substances of the first WL showed that some countries had difficulties to reach an analytical Limit of Quantification (LOQ) below or equal to the Predicted No-Effect Concentrations (PNEC) or EQS. The Joint Research Centre (JRC) of the European Commission (EC) organised a series of workshops to support the EU Member States (MS) and their activities under the WFD. Sharing the knowledge among the Member States on the analytical methods is important to deliver good data quality. The outcome and the discussion engaged with the experts are described in this paper, and in addition a literature review of the most important publications on the analysis of 17-alpha-ethinylestradiol (EE2), amoxicillin, ciprofloxacin, metaflumizone, fipronil, metformin, and guanylurea from the last years is presented.

Unilateral Urogenital Disontogeny in a Dog.

The purpose of this report was to describe an uncommon congenital anomaly in a dog. An 8-year-old, mixed-breed, male dog, was referred because of progressive difficulties on defecation. A complete diagnostic work-up (hematological analysis, radiology, ultrasound, and computed tomography), followed by surgery and histopathology, allowed us to diagnose the condition as unilateral urogenital disontogeny. The disorder was characterized by unilateral anomalies of the urinary tract (ectopic and dilated hydroureter, hydronephrosis, and renal dysplasia) associated with ipsilateral anomalies of the genital system (partial permanence of the duct of Wolff evolved into an epididymal-like structure and testicular agenesis). En bloc surgical excision of the complex of urogenital anomalies was performed with no complications during or after surgery. Surgery was considered to be effective in this dog since he no longer showed clinical signs of illness.

Evaluation of cranial tibial translation in dogs: Diagnostic accuracy of radiographic method using a simple device.

A hand-made, radiolucent, custom-designed device having a mobile and a non-mobile platforms was used to objectively quantify the in vivo cranial tibial translation, in order to assess the functional status of cranial-cruciate-ligament (CrCL) in dogs. The hypothesis was that changes in CrCL integrity would result in detectable changes in tibial translation. To validate the diagnostic method, data from injured (PA, n = 32), contralateral (CO, n = 32) and healthy stifles (HE, n = 32) were compared. Normalized tibial translation (ΔN) of each stifle was measured in medio-lateral radiographic projection obtained before and during standard thrust force application, in PA (43.59±12.97%), CO (20.32±6.69%) and HE (12.22±3.77%). Comparing PA with HE and CO (ΔN cut-off value: 29.73%), diagnosis could be issued with very high probability. Comparing HE with CO (ΔN cut-off value: 14.80%), high performance was obtained. The translator device could be a useful tool to objectively quantify the in vivo tibial translation in dogs with CrCL rupture, before surgery and during post-operatory follow-up.

Environmental surveillance and spatio-temporal analysis of Legionella spp. in a region of northeastern Italy (2002-2017).

Legionella spp. are considered an important cause of potentially preventable morbidity and mortality, making environmental surveillance a crucial component of risk assessment plans. In this work, 20,319 water samples were collected in 3,983 environmental surveys during a 16-year period by ARPA, the Regional Agency for Environmental Protection, Friuli Venezia Giulia, and the results were studied to better understand the diffusion mechanisms of Legionella. The data showed a strong seasonal signal, a prevalence of L. pneumophila serogroup 2-15 in most environments (63% of positive samples), a prevalence of L. pneumophila serogroup 1 in swimming pool-associated environments (82% of positive samples), a persistent presence of Legionella in hospitals and a recurrent presence of Legionella in other facilities such as hotels, possibly years after interventions, highlighting the difficulty of eradicating the bacteria. Retrospective spatio-temporal analyses on geocoded historical data were carried out with SaTScan using an ordinal model with risk as a covariate to identify potential clusters with an excess of cases in the higher-risk categories. Although no outbreaks occurred during the period of study, such analyses identified spatially restricted zones with unusual contamination, which sometimes were also areas in which several surveys triggered by notifications of clinical cases were performed. Simulations of periodic prospective analyses permitted the assessment of the efficacy of the method in early detection of such clusters. The proposed method may be a useful tool in environmental surveillance, prevention and control of Legionella.

Platelet-rich Plasma and Other Hemocomponents in Veterinary Regenerative Medicine.

Platelet-rich plasma, platelet-rich fibrin, fibrin glue, and platelet lysate are the most widely used nontransfusional hemocomponents that, as biological and therapeutic aids, enhance the physiological reactions after an injury to facilitate the repair and regeneration processes. Recently, this type of therapy also has significantly expanded in veterinary medicine. Due to many similarities, the animal patient could be a good reference as a study model for humans, especially for chronic and difficult-to-heal injuries. This review discusses the current state of hemocomponent use for topical application in veterinary medicine, with a comparison with human medicine.

Innovative, intra-articular, prosthetic technique for cranial cruciate ligament reconstruction in dogs: a cadaveric study.

The purpose of this study was to describe and assess the feasibility of a new intra-articular approach in the treatment of cranial cruciate ligament deficiency in dogs using an artificial ligament and a new bone-anchor system. Twelve canine cadavers weighting 26 to 45 kg were used in this ex-vivo study. Special tibial and femoral screws, two helicoils, and a high resistance artificial fiber compose the implant. Surgery was performed using the cranio-lateral approach to the stifle joint. Helicoil and tibial screw, connected to the fiber, were inserted in the center of the tibial insertion area of the cranial cruciate ligament. The fiber was passed over-the-top, tensioned, and fixed to the femoral screw, previously inserted with the helicoil in the distal part of the femur. Surgery was completed in all the cases. Occasional problems found during the insertion of the helicoils and screws were resolved with simple procedures. Post-operative clinical assessment showed negative cranial drawer test, negative cranial tibial thrust, and normal range of motion. Radiographic evaluation showed an appropriate positioning of both tibial and femoral implants in all the cases. The results of the first surgical appraisal of this new technique are encouraging, although further studies are necessary to demonstrate the in vivo efficacy of this procedure.

Platelet-rich plasma to treat experimentally-induced skin wounds in animals: A systematic review and meta-analysis.

The objective of the study was to review current literature to determine whether the topical application of platelet-rich plasma (PRP) promotes healing in experimentally-induced full-thickness skin wounds in animals. The hypothesis was that the adjunct of PRP has a positive effect on wound healing. An electronic search was carried out on the following databases: Web of Science, Cochrane Library, PubMed, Research Gate, Cochrane Wounds Group, Veterinary Information Network. No publication date nor language restrictions were applied. Randomised and not randomised controlled clinical trials comparing PRP with placebo or with other treatments were included. The reduction of open wound area in PRP-treated (test) wounds compared to control wounds was the primary outcome. Secondary outcomes were healing time and number of healed cases in test group compared to control. The following effect sizes were calculated: the Hedges' g for continuous variables; the odds ratio for binary data. Eighteen controlled clinical trials were included in the qualitative and quantitative synthesis, with a total of 661 wounds. All studies were published in the period 2007-2016. Eight studies were carried out on rodent/lagomorph mammals and 10 on non-rodent/lagomorph mammals. In all included studies, control wounds underwent placebo or were left untreated. The PRP group showed a better healing performance than the control group in each outcome. The effect size was statistically significant considering the primary outcome and the overall aggregation of the three outcomes. The effect size, although in favour of the treatment with PRP, was not significant considering the healing time and the number of healings. The overall heterogeneity was mild or moderate. Five studies reported a high risk of selection bias. The publication bias was always mild or absent. The results support the hypothesis of the positive effects of the PRP when compared to control groups in the treatment of experimentally-induced full-thickness skin wounds in animals. PRP can therefore be considered an effective adjunctive therapy in stimulating second intention healing of acute wounds in healthy animals.

Bilateral cervical ribs in a mixed breed dog.

A 4-year-old intact female, mixed breed dog was presented with a complaint of dyspnea. Clinical examination revealed symptoms related to disease of the upper airways. Radiographic findings were consistent with tracheal collapse associated with anomalies involving the seventh cervical vertebra and the first ribs bilaterally. Radiographs were highly suggestive of cervical ribs; computed tomography and ultrasound examination allowed complete characterization and better localization of the anomalies with relationship to the adjacent muscle and vasculature. Cervical ribs are malformations widely described in human medicine, but only sporadically in dogs. Herein, we discuss etiological, clinical, diagnostic and therapeutic aspects of cervical ribs and possible correlations between the cervical ribs and other anatomical anomalies noted in this dog.