Significant inactivation of SARS-CoV-2 by a green tea catechin, a catechin-derivative and galloylated theaflavins in vitro

Potential effects of teas and their constituents on SARS-CoV-2 infection were studied in vitro. Infectivity of SARS-CoV-2 was significantly reduced by a treatment with green tea, roasted green tea or oolong tea. Most remarkably, exposure to black tea for 1 min decreased virus titer to an undetectable level (less than 1/1,000 of untreated control). An addition of (-) epigallocatechin gallate (EGCG) significantly inactivated SARS-CoV-2, while theasinensin A (TSA) and galloylated theaflavins including theaflavin 3, 3’-di-gallate (TFDG) had more remarkable anti-viral activities. Virus treated with TSA at 500 μM or TFDG at 100 μM showed less than 1/10,000 infectivity compared with untreated virus. TSA and TFDG significantly inhibited interaction between recombinant ACE2 and RGD of S protein. These results strongly suggest that EGCG, and more remarkably TSA and galloylated theaflavins, inactivate the novel coronavirus.

Evidence for gastrointestinal infection of SARS-CoV-2

The new coronavirus (SARS-CoV-2) outbreak originating from Wuhan, China, poses a threat to global health. While it’s evident that the virus invades respiratory tract and transmits from human to human through airway, other viral tropisms and transmission routes remain unknown. We tested viral RNA in stool from 73 SARS-CoV-2-infected hospitalized patients using rRT-PCR. 53.42% of the patients tested positive in stool. 23.29% of the patients remained positive in feces even after the viral RNA decreased to undetectable level in respiratory tract. The viral RNA was also detected in gastrointestinal tissues. Furthermore, gastric, duodenal and rectal epithelia showed positive immunofluorescent staining of viral host receptor ACE2 and viral nucleocapsid protein in a case of SARS-CoV-2 infection. Our results provide evidence for gastrointestinal infection of SARS-CoV-2, highlighting its potential fecal-oral transmission route.

P437 Anti-TNF α antibodies in patients with inflammatory bowel disease naive to anti-TNF therapy treated with biosimilar infliximab (CT-P13): a prospective single-centre real-life study

Background There are several tests for detecting antibodies to infliximab. The tests differ in sensitivity and therefore the detection of antibodies varies significantly between clinical trials, and less data is available for induction compared with the maintenance therapy. The aim of the study was to determine the incidence of anti-drug antibodies (ADAs) in patients during induction and maintenance therapy using the Enzyme-Linked Immunosorbent Assay (ELISA) and to analyse the risk factors for their occurrence. Methods It was a prospective single-centre study involving 84 patients with IBD during treatment with biosimilar infliximab (CT-P13) at the Tertiary IBD Center in South-eastern Poland between 2016 and 2019. All participants were naive to previous anti-TNF α therapy (80 patients). ADAs serum testing was only performed when IFX levels below 3 µg/ml were found during induction or maintenance therapy. The incidence of ADAs was analysed, and the relationship between the presence of ADAs and selected variables was assessed: gender, type of disease, immunosuppressive therapy, and undetectable drug level at week 6. The relationship between response to induction treatment and high//low ADAs titers was also compared using Fisher’s exact test. Results The incidence of ADAs in the study population was 10.7%, which is 20.4% of patients with non-therapeutic and 50% of patients with undetectable levels of IFX. The percentage of people with ADAs in the induction treatment group was 11.3% (n = 6) and in the maintenance group 3.9% (n = 3). There was no statistically significant difference in the frequency of finding ADAs in both groups (p = 0.158). In contrast, a statistically significant relationship between the undetectable level of IFX at week 6 and the presence of ADAs was confirmed, 88.9% of patients with ADAs vs. 16.0% patients without ADAs (p <0.001. Comparison of the relationship between response to induction treatment and high//low ADAs titers did not show a statistically significant relationship between these variables (p = 0.381). Conclusion The overall incidence of ADAs in patients treated with biosimilar infliximab (CT-P13) naive to previous anti_TNF therapy was relatively low, but in the case of undetectable levels of the drug, the phenomenon of immunogenicity was detected in half of the patients. Undetectable level of infliximab at week 6 was a risk factor for the presence of ADAs. There was no significant association between response to induction therapy and ADAs titers.

Addition of Olive Leaf Extract (OLE) for Producing Fortified Fresh Pasteurized Milk with An Extended Shelf Life

An olive leaf extract (OLE) has been tested in vitro for its antibacterial activity and ability to inhibit α-glucosidase enzyme. OLE was also evaluated for its potential, when added to pasteurized milk, to preserve nutritional parameters and to limit microbial growth, thus prolonging shelf life. In vitro assays demonstrated a strong antibacterial efficacy of OLE mainly against Bacillus cereus and the capacity to inhibit α-glucosidase enzyme (IC50) when used at 0.2 mg oleuropein/mL. The milk fortification with OLE at 3.6 mg of oleuropein/mL of milk reduced total mesophilic bacteria at undetectable level after 6 d (expiration date) and by 1 log CFU/mL after 10 d. Moreover, OLE addition at 1.44 and 3.6 mg of oleuropein/mL of milk significantly reduced fat and lactose losses up to 10 d. The results motivate the use of the OLE to make a new functional milk with an extended shelf life.

Heterologous Replicase from Cucumoviruses can Replicate Viral RNAs, but is Defective in Transcribing Subgenomic RNA4A or Facilitating Viral Movement

Interspecific exchange of RNA1 or RNA2 between the cucumoviruses cucumber mosaic virus (CMV) and tomato aspermy virus (TAV) was reported to be non-viable in plants previously. Here we investigated viability of the reassortants between CMV and TAV in Nicotiana benthamiana plants by Agrobacterium-mediated viral inoculation. The reassortants were composed of CMV RNA1 and TAV RNA2 plus RNA3 replicated in the inoculated leaves, while they were defective in viral systemic movement at the early stage of infection. Interestingly, the reassortant containing TAV RNA1 and CMV RNA2 and RNA3 infected plants systemically, but produced RNA4A (the RNA2 subgenome) at an undetectable level. The defect in production of RNA4A was due to the 1a protein encoded by TAV RNA1, and partially restored by replacing the C-terminus (helicase domain) in TAV 1a with that of CMV 1a. Collectively, exchange of the replicase components between CMV and TAV was acceptable for viral replication, but was defective in either directing transcription of subgenomic RNA4A or facilitating viral long-distance movement. Our finding may shed some light on evolution of subgenomic RNA4A in the family Bromoviridae.

Biodiesel Byproduct and Biofuel Synthesizing Clostridium acetobutylicum

In the manufacture of biodiesel, glycerol is a byproduct. Acetone-butanol-ethanol (ABE) fermentation by anaerobe Clostridium acetobutylicum ia also a source of biofuel. Growth of C. acetobutylicum in the presence of waste biodiesel glycerol byproduct was examined in this investigation. In cultures containing added glucose or a mixture of glucose and byproduct the optical density (OD) was increased and a positive fermentation reaction was exhibited while results in cultures containing byproduct or pure glycerol were similar to those in the base medium. The pH of cultures containing glucose markedly decreased. The concentration of glucose dropped to an undetectable level. All cultures were determined to contain alcohol. The potential applications for biodiesel glycerol byproduct need to e explored.

Congenital primary hypothyroidism in a cat

A five and a half-month-old, male domestic shorthair of 1.4 kg was presented with severe constipation. Physical examination showed a dull, small cat with a poor hair coat and excessive scaling, hypothermia and a large amount of feces in the abdomen. Body proportions showed disproportional dwarfism with a large head and a short neck and limbs. Radiographs revealed marked epiphyseal dysgenesis with delayed maturation and ossification. Megacolon was present. Based on an undetectable level of TT4 and an elevated TSH level in serum, congenital primary hypothyroidism was diagnosed. On scintigraphic examination, the diagnosis was confirmed. After several months of levothyroxine therapy, the cat was bright and alert, showed no signs of constipation and developed normally.

Longitudinal follow-up of Zika virus RNA in semen of a traveller returning from Barbados to the Netherlands with Zika virus disease, March 2016

We report the longitudinal follow-up of Zika virus (ZIKV) RNA in semen of a traveller who developed ZIKV disease after return to the Netherlands from Barbados, March 2016. Persistence of ZIKV RNA in blood, urine, saliva and semen was followed until the loads reached undetectable levels. RNA levels were higher in semen than in other sample types and declined to undetectable level at day 62 post onset of symptoms.