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Eighty-one tetracycline-resistant Aeromonas sp. strains were isolated from farm-raised catfish. Morphological and biochemical characteristics indicated that 23 of the 81 aeromonads were Aeromonas hydrophila, 7 isolates were Aeromonas trota, 6 isolates were Aeromonas caviae, 42 isolates were Aeromonas veronii, and 3 isolates were Aeromonas jandaei. However, the AluI and MboI restriction fragment length polymorphism (RFLP) patterns of the PCR-amplified 1.4-kb 16S rRNA gene from all 81 tetracycline-resistant aeromonads from catfish were identical to the RFLP banding patterns of A. veronii ATCC 35626, indicating that all 81 isolates were strains of A. veronii. A multiplex PCR assay successfully amplified the 5 tetracycline-resistant genes (tetA to E) from the genomic DNA of all 81 isolates. The assay determined that tetE was the dominant gene occurring in 73/81 (90.0%) of the aeromonads. Plasmids (2.0 to 20 kb) were isolated from 33 of the 81 isolates. Dendrogram analysis of the SpeI pulsed-field gel electrophoresis identified 15 distinct macrorestriction patterns among the isolates. Our results indicate the need for use of 16S rRNA in the identification of Aeromonas spp. and the prevalence of catfish as a reservoir of tet genes.The United States produces more than $500 million per annum of pond-raised channel catfish (Ictalurus punctatus), primarily in the southeastern part of the country (4). Infectious diseases reduce catfish production by nearly 10% every year. Motile aeromonad septicemia, caused by Aeromonas hydrophila, is one of the common diseases accounting for the reduced production (5). Antimicrobials agents, such as oxytetracycline and Romet 30 (sulfadimethoxine-ormetoprim), are used to prevent the outbreak of infectious diseases (5, 32). However, widespread use of drugs may result in the selection of tetracycline-and sulfonamide-resistant bacteria in the aquaculture environment and may play a role in the dissemination of antibiotic resistance genes to clinical aeromonad strains (20,22).Aeromonads have been implicated in the cause of numerous human infections, such as gastroenteritis, cellulitis, meningitis, bacteremia, soft-tissue infections, peritonitis, and bronchopulmonary infections (8,15,(18)(19). Direct contact with contaminated water and soil is the most frequent cause of gastrointestinal and wound infections in humans (15). Broad-spectrum antibiotics, such as tetracycline, are prescribed clinically for the treatment of such infections.The molecular epidemiology of tetracycline-resistant aeromonads, especially Aeromonas salmonicida, has been well documented (2,3,29). These studies indicate that tetracycline resistance is plasmid-encoded and that, among the different classes of tetracycline-resistant genes, tetA is predominant. Schmidt et al. (29) reported the isolation and characterization of oxytetracycline-sulfonamide/trimethoprim-resistant aeromonads from Danish rainbow trout farms. Their PCR data indicate tetE as the predominant determinant, followed by tetA and tetD. Earlier, DePaola et al. ...

Background The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the cause of the ongoing global pandemic known as COVID-19. Based on the potential antiviral role of quercetin, and on its described anti-blood clotting, anti-inflammatory and antioxidant properties, we hypothesize that subjects with mild COVID-19 treated with Quercetin Phytosome ® (QP), a novel bioavailable form of quercetin, may have a shorter time to virus clearance, a milder symptomatology, and higher probabilities of a benign earlier resolution of the disease. Methods In our 2-week, randomized, open-label, and controlled clinical study, we have enrolled 42 COVID-19 outpatients. Twenty-one have been treated with the standard of care (SC), and 21 with QP as add-on supplementation to the SC. Our main aims were to check virus clearance and symptoms. Results The interim results reveal that after 1 week of treatment, 16 patients of the QP group were tested negative for SARS-CoV-2 and 12 patients had all their symptoms diminished; in the SC group, 2 patients were tested SARS-CoV-2 negative and 4 patients had their symptoms partially improved. By 2 weeks, the remaining 5 patients of the QP group tested negative for SARS-CoV-2, whereas in the SC group out of 19 remaining patients, 17 tested negatives by week 2, one tested negative by week 3 and one patient, still positive, expired by day 20. Concerning blood parameters, the add on therapy with QP, reduced LDH (−35.5%), Ferritin (−40%), CRP (−54.8%) and D-dimer (−11.9%). Conclusion QP statistically shortens the timing of molecular test conversion from positive to negative, reducing at the same time symptoms severity and negative predictors of COVID-19.

Background: Quercetin, a well-known naturally occurring polyphenol, has recently been shown by molecular docking, in vitro and in vivo studies to be a possible anti-COVID-19 candidate. Quercetin has strong antioxidant, anti-inflammatory, immunomodulatory, and antiviral properties, and it is characterized by a very high safety profile, exerted in animals and in humans. Like most other polyphenols, quercetin shows a very low rate of oral absorption and its clinical use is considered by most of modest utility. Quercetin in a delivery-food grade system with sunflower phospholipids (Quercetin Phytosome ® , QP) increases its oral absorption up to 20-fold. Methods: In the present prospective, randomized, controlled, and open-label study, a daily dose of 1000 mg of QP was investigated for 30 days in 152 COVID-19 outpatients to disclose its adjuvant effect in treating the early symptoms and in preventing the severe outcomes of the disease. Results:The results revealed a reduction in frequency and length of hospitalization, in need of non-invasive oxygen therapy, in progression to intensive care units and in number of deaths. The results also confirmed the very high safety profile of quercetin and suggested possible anti-fatigue and pro-appetite properties. Conclusion: QP is a safe agent and in combination with standard care, when used in early stage of viral infection, could aid in improving the early symptoms and help in preventing the severity of COVID-19 disease. It is suggested that a double-blind, placebo-controlled study should be urgently carried out to confirm the results of our study.

S evere fever with thrombocytopenia syndrome (SFTS) is an emerging tickborne disease caused by the SFTS virus (SFTSV; genus Banyangvirus, family Phenuiviridae, order Bunyavirales). The disease is prevalent in East Asia countries. It was first detected in China in 2009 and later in Japan and South Korea (1) and is suspected to be widely spread across other parts of the world (2). The recent identification of SFTSV in Xinjiang, China (3), expanded our awareness of epidemic areas of SFTS and suggested the possibility of SFTSV spreading to bordering countries like Pakistan. However, the presence of SFTSV in Pakistan has been unclear. We investigated the seroprevalence of SFTSV in humans in Pakistan. The Study For this study, we randomly collected human serum samples (n = 1,657) from 4 provinces in Pakistan during 2016-2017 (Figure). All participants were farmers of livestock (sheep, goats, cattle, buffaloes, and camels). We recorded and summarized testing results by sex, age, and geographic location (Table). The collection of human serum samples and subsequent tests were reviewed and approved by the Ethics Committees of

A fast, reliable, and inexpensive Triton X-100 boiling procedure for RNA isolation from both the Gram-positive and Gram-negative bacteria was developed. The yield of RNA was 0.2-2 mg per 10 ml bacterial culture. The method was tested on Gram-positive and Gram-negative bacteria of eight genera and nine species and yielded reproducible results. In parallel experiments, the Qiagen and hot phenol extraction methods both yielded RNA that contained contaminating 16S and 23S rRNA. The Triton X-100 boiling method reported here yielded RNA that was free from 16S and 23S rRNA, contained full-length transcripts and did not require additional purification. The presence of specific mRNA in one of the RNA samples obtained by this procedure was demonstrated by partial amplification of a 732 bp vancomycin resistance gene, vanA, by reverse transcription-polymerase chain reaction (RT-PCR). The presence of a full-length transcript (1031 bases) of the vanA gene was verified by Northern hybridization and probing with a digoxigenin (DIG)-labeled vanA PCR partial product. The method provides a rapid, reliable, and simple tool for the isolation of good quality RNA suitable for various molecular biology experiments.

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The purpose of this study was to simultaneously screen for Extended-spectrum β-lactamases (ESBL) and AmpC β-lactamases in gram negative clinical isolates from four tertiary care hospitals and further to compare two detection methods three-dimensional extraction method and AmpC disk test for AmpC β-lactamases. A total of 272 isolates were screened for ESBL and AmpC ß-lactamase by modified double disk approximation method (MDDM). Synergy observed between disks of ceftazidime/cefotaxime and clavulanate were considered as ESBL producer. Isolates showing reduced susceptibility to either of the test drugs (ceftazidime or cefotaxime) and cefoxitin were considered as presumptive AmpC producers and further confirmed by three-dimensional extraction method and AmpC disk test. A total of 173 (64%) of the isolates were found to be ESBL positive and 61 (23%) showed resistant to cefoxitin. ESBL was detected in 80 (62%) isolates of E. coli and 71 (73%) of Klebsiella spp. The occurrence of AmpC β-lactamases was found to be 8% (22) of the total isolates and the two detection methods for AmpC β -lactamase showed concordant results. Screening for ESBL and AmpC can be simultaneously done by MDDM method and confirmation for AmpC ß-lactamase should be carried out routinely in tertiary care hospitals by AmpC disk test, as it is a simple and rapid procedure.