Bats, monkeys and plants in the time of Covid-19 pandemic at Angkor monuments.

Knowledge of biodeterioration and protection of cultural heritage depends on the scientific understanding of the substratum materials, the ambient environment, the fauna and flora including the microorganisms so an overall picture can be constructed to serve as a basis for protection and management. Over the past more than 20 years of survey and research, an accumulated dataset is available on the mechanisms on the (bio)deterioration of stone monuments in Cambodia, involving interactions among water cycling and salt dynamics with the presence of a rich surface microbiome, the biofilms. However, during the Covid-19 period (2020-2022), because of a drastic drop on tourist population, the number of bats and monkeys are on the rising, which have an impact on the on-going protection efforts. At the same time, large trees around and on the cultural heritage sites are being managed by trimming and removal to decrease the potential risk and negative impacts from them. The new management scheme needs scientific results for the long-term successful protection of these cultural heritage. A close examination of these issues is also meaningful and important to the research new initiatives and policy to be implemented not only in Cambodia but also elsewhere.

The Flow Cytometry Study of Cellular Immunity in Rhesus Monkeys after Experimental Infection with SARS-CoV-2 Virus

Cellular immunity plays an important role in the pathogenesis and formation of protective immune defense against the SARS-CoV-2 virus. The aim of the work was to study the cellular immunity of rhesus monkeys applying flow cytometry after experimental infection with the SARS-CoV-2 virus. Materials and methods. Male rhesus monkeys were intranasally inoculated with the SARS-CoV-2 virus, Isolate B strain and hCoV-19/Russia/SP48-1226/2020 strain (abbreviated name U-2), at a dose of 5.0 lg PFU. Using flow cytometry, the levels of 21 populations/subpopulations of mononuclear cells in the peripheral blood of animals were determined before experimental infection with the pathogen and on day 14 after infection. SARS-CoV-2 coronavirus RNA was assessed using real-time polymerase chain reaction. Determination of the titer of virus-neutralizing antibodies to the SARS-CoV-2 virus in the blood sera of animals was conducted through neutralization test evaluating the ability to suppress negative colonies. Results and discussion. Infection with Isolate B strain culture has led to an increase in the relative content of total T-lymphocytes (p<0.2), cytotoxic T-lymphocytes (p<0.1), as well as monocytes expressing the early activation marker CD25 (p<0.2). The decrease in levels has been observed for total B-lymphocytes (p<0.2) and T-helper cells (p<0.1). Infection with the U-2 strain culture revealed an increase in the relative content of monocytes expressing the early activation marker CD25 (p<0.2). Thus, for the first time in the Russian Federation, flow cytometry was used to study the cellular immunity of rhesus monkeys before and after experimental infection with the SARS-CoV-2 virus. The obtained information can be used for studying the pathogenesis of SARS-CoV-2 infection, course, and outcome of the disease, and developing strategies for vaccination and treatment. © 2022 Russian Research Anti-Plague Institute. All rights reserved.

No toxic reaction observed after 2-week repeated administration of recombinant whole human anti-SARS-CoV-2 monoclonal antibody 2811 injection in rhesus monkeys

OBJECTIVE To find out whether 2811 will cause toxic reactions in animals, and determine the safe dose of without any toxic reactions and the relationship between the dose, time of administration and results of the toxicity test. METHODS Thirty healthy rhesus monkeys were selected and randomly divided into 3 groups, 1 O in each group (half males and half females), namely, the solvent control group, 2811 100 and 400 mg* kg-1 groups, respectively. The solvent control group was iv given 0.9% sodium chloride injection, while the experimental groups were iv given 2811 100 and 400 mg* kg-1 , respectively, once every 6 days, 3 times in 2 weeks, and allowed to recover for 9 weeks after drug withdrawal. During the test, the body weight, food intakes, body temperatures, ophthalmology, blood pressure, electrocardiograms, blood routine, anticoagulation, blood biochemistry, electrolytes, urine, systemic anatomy, organ weight and coefficient were observed, while histopathology and immunology tests were performed. At the same time, the anti-drug antibody (ADA) and plasma concentrations were determined, and toxikinetic parameters were analyzed. RESULTS During the experiment, the observation of general symptoms, body mass, food intakes, body temperatures, ophthalmology, blood pressure, electrocardiograms, blood routine, anticoagulation, blood biochemistry, electrolytes, urine, organ weight and coefficient, histopathology and immunology of animals in each dose group showed no significant changes related to the tested animals. ADA was not detected in any of the groups. Plasma drug concentrations in 2B11 100 and 400 mg* kg-1 groups were basically consistent and proportional to the dose, so was the ratio of peak concentration and exposure. 2B11 injection showed linear kinetics in vivo. CONCLUSION Under the conditions set in this test, the 2-week repeated administration toxicity test of 2B11 in rhesus monkeys is safe, and no clinical adverse reactions are observed at the dose level of 400 mg*kg-1, which provides reference for the follow-up clinical study of 2B11. Copyright © 2022 Chinese Journal of Pharmacology and Toxicology. All rights reserved.

Supply delays hit planned Ebola vaccine stockpile

Speed read Gavi and partners announce global stockpile of Ebola vaccine However only 6,890 doses available because of “supply bottlenecks” Target could take up to three years to reach because of supply issues Supply chain delays must be addressed to speed up creation of a stockpile of vaccines against deadly Ebola disease, researchers say. “By creating a stockpile of 500,000 doses of the Ebola vaccine, available to all countries, we can help prevent loss of life and swiftly end Ebola outbreaks in the future,” Gavi CEO Seth Berkley The new Ebola vaccine was used to tackle a two-year long outbreak in the eastern Democratic Republic of Congo, which was declared over last June after 300,000 people were immunised. The first deliveries of doses into the stockpile are being funded through a US$20 million contribution from the United States Agency for International Development (USAID). Since Ebola outbreaks are relatively rare and unpredictable, there is no natural market for the vaccine, the WHO explained.

2 weeks of repeated administration of recombinant fully human anti-new coronavirus Viral monoclonal antibody 2B11 injection did not cause toxicity in rhesus monkeys

Objective To observe the toxic reaction of recombinant fully human monoclonal antibody against novel coronavirus (2B11) injection to rhesus monkeys after repeated administration for 2 weeks, and to determine the non-toxic reaction The relationship between the safe dose and administration dose, administration time and toxicity can provide reference for clinical medication. Methods Thirty healthy rhesus monkeys were randomly divided into 3 groups, 10 in each group, half male and half male. The vehicle control group was given 0.9% sodium chloride injection, and the experimental group was given 2B11 100 and 400 mg·kg, respectively. -1 . Administer once every 6 d iv, 3 times in 2 weeks, and recover for 9 weeks after drug withdrawal. During the experiment, general symptom observation, body weight, food intake, body temperature, ophthalmological examination, blood pressure, electrocardiogram, blood routine, hemostasis, blood biochemistry and electrolytes, urine, system anatomy, organ coefficient, histopathology and immunology were tested. At the same time, anti-drug antibody (ADA) and blood drug concentrations were detected, and toxicokinetic parameters were analyzed. Results The general symptoms, body weight, food intake, body temperature, ophthalmological examination, blood pressure, electrocardiogram, blood routine, hemostasis, blood biochemistry and electrolytes, urine, organ coefficient, histopathology and No obvious changes related to the test substance were found in the indicators such as immunological detection. 2B11 No ADA was detected in the two dose groups. The changes of plasma drug concentrations were basically the same and proportional to the administered dose. The ratio of peak concentration and exposure was also proportional to the administered dose. Linear dynamic characteristics. Conclusion Under the conditions of this experiment, it is safe for rhesus monkeys to be given 2B11 3 times by iv for 2 weeks, and no clinical adverse reactions were observed at 400 mg·kg -1.

Immunogenicity and protective efficacy of SARS-CoV-2 recombinant S-protein vaccine S-268019-b in cynomolgus monkeys.

The vaccine S-268019-b is a severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike (S)-protein vaccine consisting of full-length recombinant SARS-CoV-2 S-protein (S-910823) as antigen, mixed with the squalene-based adjuvant A-910823. The current study evaluated the immunogenicity of S-268019-b using various doses of S-910823 and its vaccine efficacy against SARS-CoV-2 challenge in cynomolgus monkeys. The different doses of S-910823 combined with A-910823 were intramuscularly administered twice at a 3-week interval. Two weeks after the second dosing, dose-dependent humoral immune responses were observed with neutralizing antibody titers being comparable to that of human convalescent plasma. Pseudoviruses harboring S proteins from Beta and Gamma SARS-CoV-2 variants displayed approximately 3- to 4-fold reduced sensitivity to neutralizing antibodies induced after two vaccine doses compared with that against ancestral viruses, whereas neutralizing antibody titers were reduced >14-fold against the Omicron variant. Cellular immunity was also induced with a relative Th1 polarized response. No adverse clinical signs or weight loss associated with the vaccine were observed, suggesting safety of the vaccine in cynomolgus monkeys. Immunization with 10 µg of S-910823 with A-910823 demonstrated protective efficacy against SARS-CoV-2 challenge according to genomic and subgenomic viral RNA transcript levels in nasopharyngeal, throat, and rectal swab specimens. Pathological analysis revealed no detectable vaccine-dependent enhancement of disease in the lungs of challenged vaccinated monkeys. The current findings provide fundamental information regarding vaccine doses for human trials and support the development of S-268019-b as a safe and effective vaccine for controlling the current pandemic, as well as general protection against SARS-CoV-2 moving forward.

Detailed analysis of antibody responses to SARS-CoV-2 vaccination and infection in macaques

Macaques are a commonly used model for studying immunity to human viruses, including for studies of SARS-CoV-2 infection and vaccination. However, it is unknown whether macaque antibody responses resemble the response in humans. To answer this question, we employed a phage-based deep mutational scanning approach (Phage-DMS) to compare which linear epitopes are targeted on the SARS-CoV-2 Spike protein in convalescent humans, convalescent (re-infected) rhesus macaques, mRNA-vaccinated humans, and repRNA-vaccinated pigtail macaques. We also used Phage-DMS to determine antibody escape pathways within each epitope, enabling a granular comparison of antibody binding specificities at the locus level. Overall, we identified some common epitope targets in both macaques and humans, including in the fusion peptide (FP) and stem helix-heptad repeat 2 (SH-H) regions. Differences between groups included a response to epitopes in the N-terminal domain (NTD) and C-terminal domain (CTD) in vaccinated humans but not vaccinated macaques, as well as recognition of a CTD epitope and epitopes flanking the FP in convalescent macaques but not convalescent humans. There was also considerable variability in the escape pathways among individuals within each group. Sera from convalescent macaques showed the least variability in escape overall and converged on a common response with vaccinated humans in the SH-H epitope region, suggesting highly similar antibodies were elicited. Collectively, these findings suggest that the antibody response to SARS-CoV-2 in macaques shares many features with humans, but with substantial differences in the recognition of certain epitopes and considerable individual variability in antibody escape profiles, suggesting a diverse repertoire of antibodies that can respond to major epitopes in both humans and macaques. Differences in macaque species and exposure type may also contribute to these findings.

Susceptibility of cell lines from different animal species to Porcine Deltacoronavirus

[Objective] To explore whether porcine deltacoronavirus (PDCoV) can infect and proliferate in different animal species-derived cell lines. [Methods] The Sichuan isolate CHN-SC2015of PDCoV was inoculated in twelve cell lines derived from hamster,poultry,monkey, human and swine. After at least five blindly passages in each cell line, the virus was identified by RT-PCR,RT-q PCR, indirect immunofluorescence assay (IFA), and sequencing. [Results] PDCoV caused distinct cytopathic effect (CPE) in Vero,PAM,PK15,ST, and LLC-PK1 cells at the 1st passage (P1) and proliferated to various degrees in PAM,PK15,ST, and LLC-PK1 cells, while the CPE gradually disappeared during subsequent passages in Vero and PAM cells. Except that in the three susceptible cell lines (PK15,LLC-PK1, and ST), the viral copies of the infected cell lines gradually decreased with the increase in passages, and PDCoV could not be detected at P4 or P5 of DEF,Marc-145,HEK-293,ZYM-SIEC02, and PAM cells. PCR results showed that PDCoV could be detected only in CEF and Vero cells at P5. The IFA results showed that PDCoV could infect other cell lines except BHK-21 and ZYM-SIEC02, and specific immunofluorescence was observed in PK15,LLC-PK1, and ST cells at P1,P3, and P9. Therefore, only three cell lines (PK15,LLC-PK1, and ST) were suitable for serial passage, with the virus titers up to 107.11,107.00, and 107.37 TCID50/mL at P9,respectively. After passage in different cell lines,CHN-SC2015 accumulated 14 nucleotide mutations corresponding to 12 amino acid mutations. [Conclusion] This study indicates that PDCoV can infect a variety of cells in vitro, suggesting that it may have the potential of cross-species transmission.

Safety, Pharmacokinetics, and Activity of High Dose Ivermectin and Chloroquine against the Liver Stage of Plasmodium cynomolgi Infection in Rhesus Macaques

Previously, ivermectin (1 to 10 mg/kg of body weight) was shown to inhibit the liver-stage development of Plasmodium berghei in orally dosed mice. Here, ivermectin showed inhibition of the in vitro development of Plasmodium cynomolgi schizonts (50% inhibitory concentration [IC50], 10.42 micro M) and hypnozoites (IC50, 29.24 micro M) in primary macaque hepatocytes when administered as a high dose prophylactically but not when administered in radical cure mode. The safety, pharmacokinetics, and efficacy of oral ivermectin (0.3, 0.6, and 1.2 mg/kg) with and without chloroquine (10 mg/kg) administered for 7 consecutive days were evaluated for prophylaxis or radical cure of P. cynomolgi liver stages in rhesus macaques. No inhibition or delay to blood-stage P. cynomolgi parasitemia was observed at any ivermectin dose (0.3, 0.6, and 1.2 mg/kg). Ivermectin (0.6 and 1.2 mg/kg) and chloroquine (10 mg/kg) in combination were well-tolerated with no adverse events and no significant pharmacokinetic drug-drug interactions observed. Repeated daily ivermectin administration for 7 days did not inhibit ivermectin bioavailability. It was recently demonstrated that both ivermectin and chloroquine inhibit replication of the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in vitro. Further ivermectin and chloroquine trials in humans are warranted to evaluate their role in Plasmodium vivax control and as adjunctive therapies against COVID-19 infections.

Crossing (Conceptual) Boundaries of Transnational Environmental Law

The case comment reflects on the 2019 decision of the Constitutional Court of Basel-Stadt, which ruled that citizens should be allowed to vote on whether to ‘expand the circle of rights holders beyond the anthropological barrier’,23 and the subsequent decision of the Swiss Federal Supreme Court to uphold the validity of the citizens’ initiative.24 Blattner and Fasel explain why including rights for non-human primates in a cantonal constitution could add value to their protection in comparison with the traditional animal welfare protection measures.25 While acknowledging that the change of law advocated by the initiative might have limited practical implications, they posit that the mere symbolism of the initiative is worthwhile.26 These two decisions form part of a recent judicial trend of challenging the absence of basic rights for non-human beings.27 However, it emerges from the case comment that these decisions are particularly original in three ways. [...]the courts addressed, possibly for the first time, the relationship between animal rights and federalism in order to evaluate whether the primate rights initiative would be inconsistent with federal law. The courts responded in the negative, finding that while the Swiss Civil Code precludes animals from having fundamental rights, the initiative sought to reform Swiss public law to alter the relationship between individuals and the state: as a result, cantons were free to extend rights to non-human animals.28 Secondly, the decision of the Federal Supreme Court departed from existing animal rights scholarship, which concentrates on the overlaps between human and animal rights. [...]it declared that the initiative ‘does not aim to extend existing human constitutional rights to animals, but instead seeks to create special fundamental rights for non-human primates’.29 Thirdly, the case resulted in an important opportunity for citizens to participate in lawmaking processes as it paved the way for ‘the first ever direct democratic vote on whether some non-human animals should be granted basic rights to life and to bodily and mental integrity’.30 While the two contributions adopt a different starting point – one grounded in a theoretical exercise, the other in the commentary of a judicial decision – they nevertheless converge in their claims that our legal systems need to be reconceptualized to better account for the non-human in our worlds. 3.

SARS-CoV-2 and wastewater: What does it mean for non-human primates?

In most of our lifetimes, we have not faced a global pandemic such as the novel coronavirus disease 2019. The world has changed as a result. However, it is not only humans who are affected by a pandemic of this scale. Our closest relatives, the non-human primates (NHPs) who encounter researchers, sanctuary/zoo employees, and tourists, are also potentially at risk of contracting the virus from humans due to similar genetic susceptibility. "Anthropozoonosis"-the transmission of diseases from humans to other species-has occurred historically, resulting in infection of NHPs with human pathogens that have led to disastrous outbreaks. Recent studies have assessed the susceptibility of NHPs and predict that catarrhine primates and some lemurs are potentially highly susceptible to infection by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus. There is accumulating evidence that a new factor to consider with the spread of the virus is fecal-oral transmission. The virus has been detected in the watersheds of countries with underdeveloped infrastructure where raw sewage enters the environment directly without processing. This may expose NHPs, and other animals, to SARS-CoV-2 through wastewater contact. Here, we address these concerns and discuss recent evidence. Overall, we suggest that the risk of transmission of SARS-CoV-2 via wastewater is low. Nonetheless, tracking of viral RNA in wastewater does provide a unique testing approach to help protect NHPs at zoos and wildlife sanctuaries. A One Health approach going forward is perhaps the best way to protect these animals from a novel virus, the same way that we would protect ourselves.

SARS-CoV-2 Aerosol Exhaled by Experimentally Infected Cynomolgus Monkeys.

We analyzed size of severe acute respiratory coronavirus 2 (SARS-CoV-2) aerosol particles shed by experimentally infected cynomolgus monkeys. Most exhaled particles were small, and virus was mainly released early during infection. By postinfection day 6, no virus was detected in breath, but air in the isolator contained large quantities of aerosolized virus.