The diagnosis of Q fever is challenging due to nonspecific symptoms and negative standard blood culture results. Serological testing through immunofluorescence assay (IFA) is the most commonly used method for diagnosing this disease. Polymerase chain reaction (PCR) tests can also be used to detect bacterial DNA if taken at an appropriate time. Once the presence of bacteria is confirmed in a sample, an enrichment step is required before characterizing it through sequencing. Cultivating C. burnetii is challenging as it can only be isolated by inoculation into cell culture, embryonated eggs, or animals. In this article, we describe the isolation of C. burnetii from a valve specimen in Vero cells. We conducted genome sequencing and taxonomy profiling of this isolate and were able to determine its taxonomic affiliation. Furthermore, Multispacer sequence typing (MST) analysis suggests that the infection originated from a local strain of C. burnetii found around northern Israel and Lebanon. This novel strain belongs to a previously described genotype MST6, harboring the QpRS plasmid, never reported in Israel.

Introduction Human adenoviruses are common causes of many acute illnesses, and keratoconjunctivitis is one of them. Acute infections, if left untreated, can progress to severity, thus causing morbidities and mortalities. It belongs to the mastadenovirus family and is characterized by seven subgenus, i.e., A-G; among those, Adenovirus D8 is the most common type associated with keratoconjunctivitis. Methodology A hospital-based study was conducted, and the samples were collected from GB Pant Hospital, Port Blair, Dr Agarwals Eye Hospital, Port Blair, from August 2017 to December 2022. Clinical data and demographic details were followed by conjunctival swab sample collection from suspected keratoconjunctivitis patients. Samples were subjected to molecular screening, and Sanger sequencing was carried out for positive samples. Results Out of 506 conjunctival samples, a prevalence of 24.9% (n=126) was observed, and the commonest type circulating among the population of Andaman was Adenovirus D8. The major symptoms associated were eye redness (87.30%, n=110), followed by watering (81.75%, n=103), eye pain (72.22%, n=91), eye itching (61.11%, n=77), and discharge (50%, n=63). Conclusion In clinical research, ocular infections are one of the underrated fields. However, the study revealed the high prevalence of adenoviral infection among the suspected patients. Thus, there is a need for proper surveillance and timely diagnosis of such infections, as their severity may lead to loss of vision.

Craniopharyngioma is a rare, low-grade tumor located in the suprasellar region of the brain, near critical structures like the pituitary gland. Here, we concurrently investigate the status of clinical and genomic data in a retrospective craniopharyngioma cohort and survey-based data to better understand patient-relevant outcomes associated with existing therapies and provide a foundation to inform new treatment strategies.
Clinical, genomic, and outcome data for a retrospective cohort of patients with craniopharyngioma were collected and reviewed through the Children\'s Brain Tumor Network (CBTN) database. An anonymous survey was distributed to patients and families with a diagnosis of craniopharyngioma to understand their experiences throughout diagnosis and treatment.
The CBTN repository revealed a large proportion of patients (40 - 70%) with specimens that are available for sequencing but lacked relevant quality of life (QoL) and functional outcomes. Frequencies of reported patient comorbidities ranged from 20-35%, which is significantly lower than historically reported. Survey results from 159 patients/families identified differences in treatment considerations at time of diagnosis versus time of recurrence. In retrospective review, patients and families identified preference for therapy that would improve QoL, rather than decrease risk of recurrence (mean 3.9 vs. 4.4 of 5) and identified endocrine issues as having the greatest impact on patients\' lives.
This work highlights the importance of prospective collection of QoL and functional metrics alongside robust clinical and molecular correlates in individuals with craniopharyngioma. Such comprehensive measures will facilitate biologically relevant therapeutic strategies that also prioritize patient needs.

Between April 2018 and August 2019, a total of 135 strains of Enterobacter cloacae complex (ECC) were randomly collected at the University Hospital Center of Guadeloupe to investigate the structure and diversity of the local bacterial population. These nosocomial isolates were initially identified genetically by the hsp60 typing method, which revealed the clinical relevance of E. xiangfangensis (n = 69). Overall, 57/94 of the third cephalosporin-resistant strains were characterized as extended-spectrum-β-lactamase (ESBL) producers, and their whole-genome was sequenced using Illumina technology to determine the clonal relatedness and diffusion of resistance genes. We found limited genetic diversity among sequence types (STs). ST114 (n = 13), ST1503 (n = 9), ST53 (n = 5) and ST113 (n = 4), which belong to three different Enterobacter species, were the most prevalent among the 57 ESBL producers. The blaCTXM-15 gene was the most prevalent ESBL determinant (56/57) and was in most cases associated with IncHI2/ST1 plasmid replicon carriage (36/57). To fully characterize this predominant blaCTXM-15/IncHI2/ST1 plasmid, four isolates from different lineages were also sequenced using Oxford Nanopore sequencing technology to generate long-reads. Hybrid sequence analyses confirmed the circulation of a well-conserved plasmid among ECC members. In addition, the novel ST1503 and its associated species (ECC taxon 4) were analyzed, in view of its high prevalence in nosocomial infections. These genetic observations confirmed the overall incidence of nosocomial ESBL Enterobacteriaceae infections acquired in this hospital during the study period, which was clearly higher in Guadeloupe (1.59/1000 hospitalization days) than in mainland France (0.52/1,000 hospitalization days). This project revealed issues and future challenges for the management and surveillance of nosocomial and multidrug-resistant Enterobacter in the Caribbean.

Coconut oil cake (COC), a byproduct of oil extraction, contains high levels of cellulose. The aim of this study was to isolate a cellulose-degrading yeast from rotten dahlia that can effectively use COC as the only carbon source for cellulase secretion. Based on screening, Meyerozyma guillermondii CBS 2030 (M. guillermondii) was identified as a potential candidate, with the highest cellulolytic activity among the yeast strains isolated, with the carboxymethyl cellulase (CMCase) activity reaching 102.96 U/mL on day 5. The cellulose in COC samples was evaluated before and after degradation by M. guillermondii. Analysis based on field emission scanning electron microscopy (FESEM) revealed that the COC structure was changed significantly during the treatment, indicating effective hydrolysis. Fourier transform infrared spectroscopy (FTIR) of the modified functional groups indicated successful depolymerization of coconut cake. X-ray diffraction (XRD) and analysis of color differences established effective degradation of COC by M. guillermondii. The results demonstrate that M. guillermondii effectively secretes CMCase and degrades cellulose, which has important practical significance in COC degradation.

Juvenile hormone epoxide hydrolase (JHEH) plays an important role in the metabolism of juvenile hormone III (JH III) in insects. To study the role that JHEH plays in female Aedes aegypti JHEH 1, 2, and 3 complementary DNA (cDNAs) were cloned and sequenced. Northern blot analyses show that the three transcripts are expressed in the head thorax, the gut, the ovaries, and the fat body of females. Molecular modeling shows that the enzyme is a homodimer that binds JH III acid (JH IIIA) at the catalytic groove better than JH III. The cDNA of JHEH 1 and 2 are very similar indicating close relationship. Knocking down of jheh 1, 2, and 3 in adult female and larval Ae. aegypti using double-stranded RNA (dsRNA) did not affect egg development or caused adult mortality. Larvae that were fed bacterial cells expressing dsRNA against jheh 1, 2, and 3 grew normally. Treating blood-fed female Ae. aegypti with [12-3 H](10R) JH III and analyzing the metabolites by C18 reversed phase chromatography showed that JHEH preferred substrate is not JH III but JH IIIA. Genomic analysis of jheh 1, 2, and 3 indicate that jheh 1 and 2 are transcribed from a 1.53 kb DNA whereas jheh 3 is transcribed from a 10.9 kb DNA. All three genes are found on chromosome two at distinct locations. JHEH 2 was expressed in bacterial cells and purified by Ni affinity chromatography. Sequencing of the recombinant protein by MS/MS identified JHEH 2 as the expressed recombinant protein.

BACKGROUND: Information regarding the microbiome in sinusitis using genetic sequencing is lacking and more-in-depth understanding of the microbiome could improve antimicrobial selection and treatment outcomes for cases of primary sinusitis.
OBJECTIVE: To describe sinus microbiota in samples from horses with sinusitis and compare microbiota and the presence of antimicrobial resistance genes between primary, dental-related and other secondary causes of sinusitis.
METHODS: Retrospective case series.
METHODS: Records of equine sinusitis from 2017 to 2021 were reviewed and historical microbial amplicon sequence data were obtained from clinical diagnostic testing of sinus secretions. Following bioinformatic processing of bacterial and fungal sequence data, the sinus microbiota and importance of sinusitis aetiology among other factors were investigated from the perspectives of alpha diversity (e.g., number of operational taxonomic units [OTUs], Hill1 Diversity), beta diversity, and differentially abundant taxa. Quantitative PCR allowed for comparisons of estimated bacterial abundance and detection rate of common antibiotic resistance-associated genes. In a smaller subset, longitudinal analysis was performed to evaluate similarity in samples over time.
RESULTS: Of 81 samples analysed from 70 horses, the bacterial microbiome was characterised in 66, and fungal in five. Only sinusitis aetiology was shown to significantly influence microbiome diversity and composition (p < 0.05). Dental-related sinusitis (n = 44) was associated with a significantly higher proportion of obligate anaerobic bacteria, whereas primary sinusitis (n = 12) and other (n = 10) groups were associated with fewer bacteria and higher proportions of facultative anaerobic and aerobic genera. Antimicrobial resistance genes and fungal components were exclusively identified in dental-related sinusitis.
CONCLUSIONS: Retrospective nature, incomplete prior antimicrobial administration data.
CONCLUSIONS: Molecular characterisation in sinusitis identifies microbial species which may be difficult to isolate via culture, and microbiome profiling can differentiate sinusitis aetiology, which may inform further treatment, including antimicrobial therapy.
UNASSIGNED: Es fehlen Informationen über das Mikrobiom bei Sinusitis, die durch genetische Sequenzierung gewonnen wurden, und ein tieferes Verständnis des Mikrobioms könnte die Auswahl antimikrobieller Mittel und die Behandlungsergebnisse bei primärer Sinusitis verbessern.
UNASSIGNED: Beschreibung der Sinus-Mikrobiota in Proben von Pferden mit Sinusitis und Vergleich der Mikrobiota und des Vorhandenseins antimikrobieller Resistenzgene zwischen primären, zahnbedingten und anderen sekundären Ursachen der Sinusitis.
METHODS: Retrospektive Betrachtung.
METHODS: Aufzeichnungen über Sinusitis bei Pferden aus den Jahren 2017-2021 wurden überprüft und historische mikrobielle Amplikon-Sequenzdaten aus klinischen diagnostischen Tests von Sinus-Sekreten gewonnen. Nach der bioinformatischen Verarbeitung von Bakterien- und Pilzsequenzdaten wurden die Mikrobiota der Nasennebenhöhlen und die Bedeutung der Ätiologie der Sinusitis unter anderem unter dem Gesichtspunkt der Alpha-Diversität (z. B. Anzahl der operativen taxonomischen Einheiten [OTUs], Hill1-Diversität), der Beta-Diversität und der unterschiedlich häufigen Taxa untersucht. Die quantitative PCR ermöglichte einen Vergleich der geschätzten Bakterienhäufigkeit und der Nachweisrate von Genen, die mit der Antibiotikaresistenz in Verbindung stehen. Bei einer kleineren Untergruppe wurde eine Längsschnittanalyse durchgeführt, um die Ähnlichkeit der Proben im Laufe der Zeit zu bewerten.
UNASSIGNED: Von 81 analysierten Proben von 70 Pferden wurde das bakterielle Mikrobiom von 66 und das Pilzmikrobiom von fünf Pferden charakterisiert. Nur die Ätiologie der Sinusitis zeigte einen signifikanten Einfluss auf die Diversität und Zusammensetzung des Mikrobioms (p < 0,05). Zahnbedingte Sinusitis (n = 44) war mit einem signifikant höheren Anteil an obligat anaeroben Bakterien verbunden, während primäre Sinusitis (n = 12) und andere (n = 10) Gruppen mit weniger Bakterien und einem höheren Anteil an fakultativ anaeroben und aeroben Gattungen verbunden waren. Antimikrobielle Resistenzgene und Pilzkomponenten wurden ausschließlich bei zahnbedingter Sinusitis identifiziert. Serielle Proben ähnelten den Ausgangsproben tendenziell mehr als Proben von anderen Pferden.
UNASSIGNED: Retrospektiver Charakter, unvollständige Daten über die vorherige Verabreichung antimikrobieller Mittel.
UNASSIGNED: Durch die molekulare Charakterisierung der Sinusitis werden Mikrobenarten identifiziert, die sich möglicherweise nur schwer über eine Kultur isolieren lassen, und durch die Erstellung von Mikrobiomprofilen kann die Ätiologie der Sinusitis differenziert werden, was wiederum Aufschluss über die weitere Behandlung, einschließlich einer antimikrobiellen Therapie, geben kann.

UNASSIGNED: In moving towards the elimination of hepatitis C virus (HCV) infection among people living with HIV, understanding HCV transmission patterns may provide insights to guide and evaluate interventions. In this study, we evaluated patterns of, and factors associated with HCV phylogenetic clustering among people living with HIV/HCV co-infection in Australia in the direct-acting antiviral era.
UNASSIGNED: HCV RNA was extracted from dried blood spot (DBS) samples collected between 2014 and 2018 in the CEASE cohort study. The HCV Core-E2 region was amplified by a polymerase chain reaction and Sanger sequenced. Maximum likelihood phylogenetic trees (1000 bootstrap replicates) were used to identify patterns of clustering (3% genetic distance threshold). Mixed-effects logistic regression was used to determine correlates of phylogenetic clustering. Factors assessed were sexual risk behavior, education, injecting drug use, housing, employment, HIV viral load, age, sex, and sexuality.
UNASSIGNED: Phylogenetic trees were reconstructed for HCV subtype 1a (n = 139) and 3a (n = 63) sequences, with 29% (58/202) in a pair or cluster. Overall (n = 202), phylogenetic clustering was positively associated with younger age (under 40; adjusted odds ratio [aOR] 2.52, 95% confidence interval [CI] 1.20-5.29), and among gay and bisexual men (n = 168), was positively associated with younger age (aOR 2.61, 95% CI 1.10-6.19), higher education (aOR 2.58, 95% CI 1.09-6.13), and reporting high-risk sexual behavior (aOR 3.94, 95% CI 1.31-11.84). During follow-up, five reinfections were observed, but none were in phylogenetic clusters.
UNASSIGNED: This study found a high proportion of phylogenetic relatedness, predominantly among younger people and gay and bisexual men reporting high-risk sexual behavior. Despite this, few reinfections were observed, and reinfections demonstrated little relationship with known clusters. These findings highlight the importance of rapid HCV treatment initiation, together with monitoring of the phylogeny.

OBJECTIVE: Invasive listeriosis is a severe foodborne infection caused by Listeria(L.)monocytogenes. The aim of this investigation was to verify and describe a molecular cluster of listeriosis patients and identify factors leading to this outbreak.
METHODS: Whole genome sequencing and core genome multilocus sequence typing were used for subtyping L. monocytogenes isolates from listeriosis cases and food samples in Germany. Patient interviews and investigational tracing of foodstuffs offered in health-care facilities (HCF), where some of the cases occurred, were conducted.
RESULTS: We identified a German-wide listeriosis outbreak with 39 genetically related cases occurring between 2014 and 2019. Three patients died as a result of listeriosis. After identification of HCF in different regions of Germany for at least 13 cases as places of exposure, investigational tracing of food supplies in six prioritized HCF revealed meat products from one company (X) as a commonality. Subsequently the outbreak strain was analysed in six isolates from ready-to-eat meat products and one isolate from the production environment of company X. No further Sigma1 cases were detected after recall of the meat products from the market and closure of company X (as of August 2020).
CONCLUSIONS: Interdisciplinary efforts including whole genome sequencing, epidemiological investigations in patients and investigational tracing of foods were essential to identify the source of infections, and thereby prevent further illnesses and deaths. This outbreak underlines the vulnerability of hospitalized patients for foodborne diseases, such as listeriosis. Food producers and HCF should minimize the risk of microbiological hazards when producing, selecting and preparing food for patients.

Here we report on the infection of captive crested geckos Correlophus ciliatus Guichenot (Reptilia: Diplodactylidae), with adults of the ascaridoid nematode, Hexametra angusticaecoides Chabaud & Brygoo, 1960 (Ascarididae). A population of captive crested geckoes became ill and died within a short period of time. Nematodes were recovered from the crested geckoes examined from within the coelomic cavity, penetrating various organs and migrating through subcutaneous tissues, as well as emerging through the geckos\' skin. One gecko was treated with levamisole following surgical excision of nematodes from under the skin; this gecko survived. The potential source of the nematode infection in the captive geckoes is discussed. It is most likely that wild-caught Madagascan mossy geckoes, Uroplatus sikorae Boettger (Reptilia: Gekkonidae), introduced the infection to the colony. Molecular sequences of the nematodes are the first produced for the members of this genus. A redescription of the species and its genetic characterization based on the internal transcribed spacer sequence data is provided, suggesting some of the morphological criteria that have been used in the past to distinguish between Hexametra spp. may have been intraspecific morphological variations.