Minimally invasive sampling to identify leprosy patients with a high bacterial burden in the Union of the Comoros

The World Health Organization (WHO) endorsed diagnosis of leprosy (also known as Hansen’s disease) entirely based on clinical cardinal signs, without microbiological confirmation, which may lead to late or misdiagnosis. The use of slit skin smears is variable, but lacks sensitivity. In 2017–2018 during the ComLep study, on the island of Anjouan (Union of the Comoros; High priority country according to WHO, 310 patients were diagnosed with leprosy (paucibacillary = 159; multibacillary = 151), of whom 263 were sampled for a skin biopsy and fingerstick blood, and 260 for a minimally-invasive nasal swab. In 74.5% of all skin biopsies and in 15.4% of all nasal swabs, M. leprae DNA was detected. In 63.1% of fingerstick blood samples, M. leprae specific antibodies were detected with the quantitative αPGL-I test. Results show a strong correlation of αPGL-I IgM levels in fingerstick blood and RLEP-qPCR positivity of nasal swabs, with the M. leprae bacterial load measured by RLEP-qPCR of skin biopsies. Patients with a high bacterial load (≥50,000 bacilli in a skin biopsy) can be identified with combination of counting lesions and the αPGL-I test. To our knowledge, this is the first study that compared αPGL-I IgM levels in fingerstick blood with the bacterial load determined by RLEP-qPCR in skin biopsies of leprosy patients. The demonstrated potential of minimally invasive sampling such as fingerstick blood samples to identify high bacterial load persons likely to be accountable for the ongoing transmission, merits further evaluation in follow-up studies.

Diagnosis of Helicobacter pylori Infection in a Routine Testing Workflow: Effect of Bacterial Load and Virulence Factors

Reliable diagnostic methods are mandatory for effective management of Helicobacter pylori infection. Histology and culture are the most common invasive methods in current practice, even if molecular methods are gaining in importance. The performance of these conventional methods varies significantly. We conducted a retrospective study of 1540 adults and 504 children with gastric biopsies taken during endoscopy to assess the impact of bacterial load and the cagA virulence factor on the performance of H. pylori infection testing. The association between virulence and histology findings was also investigated. With 23S rRNA qPCR confirmed by glmM amplification as the gold standard, culture and histology had lower sensitivity, 74.4% and 73.3%, respectively. However, their sensitivity was enhanced (>90%) in biopsies with high bacterial load (qPCR Ct < 30). Positive cagA status of the strain was associated with high bacterial load (94.9%), thus resulting in more frequent positive culture (94.3%) and H. pylori histology detection (91.7%) and more severe lesions on histology (p < 0.001). Conversely, the cagA status of the strains was negative in 110/119 (92.4%) of biopsies with low bacterial load (qPCR Ct < 30), 82/90 (91.1%) with negative H. pylori histology detection and 119/131 (90%) with negative culture findings (p < 0.001). This study highlights the low sensitivity of conventional culture and histology that may lead to false negative diagnosis if used alone. H. pylori quantification associated with cagA genotyping in routine workflow are essential for a sensitive and reliable diagnosis, to identify patients at high risk and to manage eradication therapies.

Synthesis and characterisation of super-paramagnetic iron oxide nanoparticles (SPIONs) for minimising Aeromonas hydrophila load from freshwater

Background: The current study was conducted to prepare an efficient super-paramagnetic iron oxide nanoparticle (SPIONs) to remove Aeromonas hydrophila from water. Methods: The nanoparticles were synthesized by the co-precipitation method and characterized by DLS, UV-Vis spectrophotometer, FT-IR, XRD, FEG-TEM, and VSM analysis. Results and Discussion: The results showed that the synthesized SPIONs were having a size range of 8-12nm with magnetic property. Bacteria removal efficiency and antibacterial activity of SPIONs were assessed in sterile distilled water by adding different concentrations of SPIONs viz. 0, 6.25, 12.5, 25, 50, 100, 200, 500, and 1000µM with different initial bacterial loads viz. 1×103, 1×104, 1×105, 1×106, and 1×107 CFU mL−1 at different time intervals 15, 30, 45, and 60 min. At low bacterial load (1×103 to 1×105 CFU mL−1), 95 to 99.99% of bacteria were removed by low SPIONs concentration (6.25-100µM) by 15min which was increased up to 100% by 30min. However, at high bacterial load (1×106 to 1×107 CFU mL−1), more than 87 to 95% of bacteria were removed by the highest SPIONs concentration (1000µM) by 15min, which was increased above 93 to 99.99% by increasing the exposure time to 60min. At low bacterial load (1×103 to1×105 CFU mL−1), the effective concentration was 3.21 to 6.42µM at 15-60 min intervals. Meanwhile, the effective concentration at high bacterial load was 267.81 µM at 15min, which was decreased to 104.09 µM with increasing exposure time to 60min. Conclusion: Based on the results, it is concluded that the antibacterial effect against A. hydrophila depends on the concentration as well as the exposure time of SPIONs. A low concentration of SPIONs is sufficient to remove 100% of bacterial load in lower exposure time and increasing concentration of SPIONs increases the antibacterial effect. However, further research requires to find the safe concentration of SPIONs for using it as a novel antibacterial agent for the treatment of aeromonads disease in aquaculture.

Microscale tracking of coral-vibrio interactions

To improve our understanding of coral infection and disease, it is important to study host-pathogen interactions at relevant spatio-temporal scales. Here, we provide a dynamic microscopic view of the interaction between a coral pathogen, Vibrio coralliilyticus and its coral host Pocillopora damicornis. This was achieved using a microfluidics-based system facilitating control over flow, light and temperature conditions. Combined with time-resolved biochemical and microbial analyses of the system exudates, this approach provides novel insights into the early phases of a coral infection at unprecedented spatio-temporal resolution. We provide evidence that infection may occur through ingestion of the pathogen by the coral polyps, or following pathogen colonization of small tissue lesions on the coral surface. Pathogen ingestion invariably induced the release of pathogen-laden mucus from the gastrovascular cavity. Despite the high bacterial load used in our experiments, approximately one-third of coral fragments tested did not develop further symptoms. In the remaining two-thirds, mucus spewing was followed by the severing of calicoblastic connective tissues (coenosarc) and subsequently necrosis of most polyps. Despite extensive damage to symptomatic colonies, we frequently observed survival of individual polyps, often accompanied by polyp bail-out. Biochemical and microbial analyses of exudates over the course of symptomatic infections revealed that severing of the coenosarc was followed by an increase in matrix metaloprotease activity, and subsequent increase in both pathogen and total bacterial counts. Combined, these observations provide a detailed description of a coral infection, bringing us a step closer to elucidating the complex interactions underlying coral disease.

Feline comorbidities: What do we really know about feline triaditis?

Practical relevance: Feline triaditis describes concurrent pancreatitis, cholangitis and inflammatory bowel disease (IBD). The reported prevalence is 17–39% in ill referral patients. While the aetiology is poorly understood, it is known to include infectious, autoimmune and physical components. What is not known is whether different organs are affected by different diseases, or the same process; indeed, triaditis may be part of a multiorgan inflammatory disease. Feline gastrointestinal tract anatomy plays its role too. Specifically, the short small intestine, high bacterial load and anatomic feature whereby the pancreatic duct joins the common bile duct before entering the duodenal papilla all increase the risk of bacterial reflux and parenchymal inflammation. Inflammation may also be a sequela of bowel bacterial translocation and systemic bacteraemia. Diagnostic challenges: Cholangitis, pancreatitis and IBD manifest with overlapping, vague and non-specific clinical signs. Cholangitis may be accompanied by increased serum liver enzymes, total bilirubin and bile acid concentrations, and variable ultrasonographic changes. A presumptive diagnosis of pancreatitis is based on increased serum pancreatic lipase immunoreactivity or feline pancreas-specific lipase, and/or abnormal pancreatic changes on ultrasonography, though these tests have low sensitivity. Diagnosis of IBD is challenging without histopathology; ultrasound findings vary from normal to mucosal thickening or loss of layering. Triaditis may cause decreased serum folate or cobalamin (B12) concentrations due to intestinal disease and/or pancreatitis. Triaditis can only be confirmed with histopathology; hence, it remains a presumptive diagnosis in most cases. Evidence base: The literature on feline triaditis, pancreatitis, cholangitis and IBD is reviewed, focusing on histopathology, clinical significance and diagnostic challenges. Current management recommendations are provided. Further studies are needed to understand the complex pathophysiology, and in turn improve diagnosis and treatment.

Lung Microbiome Differentially Impacts Survival of Patients with Non-Small Cell Lung Cancer Depending on Tumor Stroma Phenotype

The link between a lung tumor and the lung microbiome is a largely unexplored issue. To investigate the relationship between a lung microbiome and the phenotype of an inflammatory stromal infiltrate, we studied a cohort of 89 patients with non-small cell lung cancer. The microbiome was analyzed in tumor and adjacent normal tissue by 16S rRNA amplicon sequencing. Characterization of the tumor stroma was done using immunohistochemistry. We demonstrated that the bacterial load was higher in adjacent normal tissue than in a tumor (p = 0.0325) with similar patterns of taxonomic structure and alpha diversity. Lung adenocarcinomas did not differ in their alpha diversity from squamous cell carcinomas, although the content of Gram-positive bacteria increased significantly in the adenocarcinoma group (p = 0.0419). An analysis of an inflammatory infiltrate of tumor stroma showed a correlation of CD68, iNOS and FOXP3 with a histological type of tumor. For the first time we showed that high bacterial load in the tumor combined with increased iNOS expression is a favorable prognostic factor (HR = 0.1824; p = 0.0123), while high bacterial load combined with the increased number of FOXP3+ cells is a marker of poor prognosis (HR = 4.651; p = 0.0116). Thus, we established that bacterial load of the tumor has an opposite prognostic value depending on the status of local antitumor immunity.

Three-decade failure to eradication of refractory Helicobacter pylori infec-tion and recent efforts to eradicate the infection

Background: Helicobacter pylori causes dangerous and deadly diseases such as gastric cancer and duodenal ulcers. Eradication and treatment of this bacterium are very important due to the deadly diseases caused by H. pylori and the high cost of treatment for countries. So, we present a complete list of the most important causes of failure in the treatment and eradication of H. pylori, and addresses new therapeutic methods that may be effective in controlling this bacterium in the future. Results: Many efforts have been made to control and eradicate this bacterium over the years, but no success has been achieved since its eradication is a complex process affected by the bacterial properties and host factors. Previous studies have shown that various factors are involved in failure to eradicate H. pylori, such as new genotypes of the bacterium with higher pathogenicity, inappropriate patient cooperation, mutations, biofilm formation and dormant forms that cause antibiotic resistance, acidic stomach pH, high bacterial load, smoking, immunosuppressive features and intracellular occurrence of H. pylori. On the other hand, recent studies reported that the use of probiotics, nanoparticles, antimicrobial peptides, natural product and vaccine can be helpful in the treatment and eradication of H. pylori infections. Conclusion: Eradication of H. pylori is crucial for the treatment of important diseases such as gastric cancer. Therefore, it seems that identifying the failure causes of treating this bacterium can be helpful in controlling the infections. Besides, further studies on new therapeutic strategies may help eradicate H. pylori in the future.

A case of purple urine bag syndrome in an elderly patient

Purple urine bag syndrome is a rare condition that occurs predominantly in the elderly, immobilized and bearer of chronic bladder catheter. It is a phenomenon usually associated with urinary tract infection, particularly in the presence of high bacterial load. The presence of alkaline urine, constipation, high-protein diet and dehydration are predisposing factors. In most cases it is an asymptomatic condition. We described the case of an elderly patient admitted to the geriatric Department in which purple urine was found in the urine collection bag. The culture of urine showed the presence of infection with Klebsiella pneumoniae. Targeted antibiotic treatment and proper hydration determined the resolution of the phenomenon and the resolution of the infection. Although it is an alarming phenomenon due to the particularity of the color of the urine, it is a treatable and solvable condition with an adequate specific antibiotic treatment. Prevention measures are equally effective and consist in eliminating the risk factors for this condition.

Organized Framework of Main Possible Applications of Sheep Wool Fibers in Building Components

Greasy sheep wool is currently considered a special waste for its high bacterial load, with expensive disposal costs for sheep breeders. For this reason, wool is often burned or buried, with serious consequences for the environment. On the other hand, sheep wool is well regarded as one of the most performative insulating natural fibers due to its thermo-hygrometric and acoustic properties. In the building sector, sheep wool meets the requirements of green building components because it is an eco-friendly material, there is a surplus of it, it is annually renewable, and totally recyclable. If used instead of common insulation materials (e.g., fiberglass, rock wool, polyurethane foam, polystyrene), sheep wool offers significant benefits for sustainability such as a reduction in the production costs for new insulating materials and in environmental pollution. Mechanical and physical properties of sheep wool investigated in previous studies were assessed and discussed with the aim of providing an organized framework of possible applications of wool fibers in building components. This paper highlights in detail aspects that have not yet been investigated enough to detect new potential uses of sheep wool fibers in rural buildings and the reuse of traditional ones.

Swine conjunctivitis outbreaks associated with Mycoplasma hyorhinis

Conjunctivitis is an uncommon finding in commercial swine herds, and the etiology of the disease is rarely studied. We investigated cases of conjunctivitis in 3 wean-to-finish swine farms. Eye swabs and tissues were obtained from clinically affected pigs (8–22 wk of age), from unaffected pigs in contact with affected pen-mates, and from age-matched pigs from an unaffected herd. Real-time PCR (rtPCR) testing for Mycoplasma hyorhinis demonstrated consistent detection and high bacterial load in samples from affected herds (clinically affected animals and non-clinical pen-mates). Ct values in affected pigs were 18.9–25.3; values were 36.4–38.6 in unaffected pigs from unaffected herds. Additionally, M. hyorhinis was identified within inflamed palpebral conjunctivae by in situ hybridization. The association of rtPCR and in situ detection of M. hyorhinis, along with the lack of detection of other potential pathogens and noninfectious causes, suggests the involvement of M. hyorhinis in the etiology and pathogenesis of the reported swine conjunctivitis.