Conjunctival polymerase chain reactions in COVID-19 patients and its correlation with clinical and paraclinical indexes

Background and Objectives: This study aimed to detect SARS-CoV-2 in conjunctival samples of COVID-19 patients to investigate the transmission route of COVID-19 and its correlation with laboratory indexes. Materials and Methods: In this cross-sectional study, 44 COVID-19 patients were tested for conjunctival PCR in Ayatollah Rouhani hospital of Babol, Iran, in January and February 2021. The conjunctival samples were collected using a conjunctival swab and suspended in a viral transport medium. After RNA extraction and cDNA synthesis, real-time PCR was performed to investigate the SARS-CoV-2 genome in samples. The ocular manifestations and laboratory indexes were evaluated for all patients. Results: Among 44 COVID-19 patients, 6 samples (13.63%) were positive in terms of conjunctival PCR. The mean ± SD age of conjunctival PCR-positive patients was 76.17 ± 16.61-year-old, while conjunctival PCR-negative COVID-19 pa- tients were aged 57.54 ± 13.61-year-old (p <0.05). D-dimer serum level is significantly higher in conjunctival PCR-positive COVID-19 patients (4001.00 ± 3043.36 µg/ml) compared to normal individuals (496.80 ± 805.92 µg/ml, p <0.01). Conclusion: Our study showed that the conjunctiva and tear contain the SARS-CoV-2 in COVID-19 patients as a possible transmission route.

Effect of domestic freezing on the viability of Toxoplasma gondii in raw and dry-cured ham from experimentally infected pigs

Toxoplasma gondii is the causative agent of the parasitic disease toxoplasmosis, which is an important food borne zoonosis. Eating undercooked meat of infected animals has been considered the major transmission route of T. gondii to humans. The present study evaluates the efficacy of domestic freezing on the inactivation of T. gondii bradyzoites in raw and dry-cured ham. Meat (raw and dry-cured ham) of a pig experimentally orally inoculated with 4,000 oocysts of T. gondii VEG strain was subjected to domestic freezing of -20 ºC at different days. The effect was evaluated by bioassay in mice followed by qPCR. In raw ham and dry-cured ham, temperature of -20 ºC for 7 and 14 days respectively did not inactivate T. gondii . More studies are needed to find the right temperature and time needed to render the bradyzoites non-infectious for human. Meanwhile, the recommendations of freezing to inactivate T. gondii in raw or dry-cured meats must be revisited considered that it does not reduce the risk of infection.

COVID-19: An Updated Insight of the Pandemic

Novel coronavirus (SARS-CoV-2) out-broke in the city of Wuhan in China and widely spread across the globe in a pandemic manner, causing societal and economic disruptions. Though the origin of the novel virus is still a debating topic, it is certain that SARS-CoV-2 acquired human to human transmission capacity. Regardless of aggressive containment and quarantine approaches, the number of confirmed cases continues to rise and being reported due to its highly infectious nature. As of the time, there is a little scope for the antiviral drugs or vaccines for the treatment of coronavirus infection; due to the vigorous mutation rate in the viral genome. However, existing anti-parasite drugs like ivermectin and chloroquine could effectively inhibit the virus has been reported. Few of the vaccines have come up with certain degree of efficacy and many are under the clinical trial phase. The research on novel coronavirus is still in the preliminary stage. In this chapter, we systematically summarize the origin, transmission route, molecular characterization, pathogenic mechanism, contagious nature, clinical symptoms, diagnosis, treatment, mutation and infection as well as prevention strategy of coronavirus disease based on the recently available literature. In addition to this, this chapter presents updated insights of the current state of knowledge pertaining to novel coronavirus and can be referred for potential future studies.

Infection of wild-type mice by SARS-CoV-2 B.1.351 variant indicates a possible novel cross-species transmission route

COVID-19 is identified as a zoonotic disease caused by SARS-CoV-2, which also can cross-transmit to many animals but not mice. Genetic modifications of SARS-CoV-2 or mice enable the mice susceptible to viral infection. Although neither is the natural situation, they are currently utilized to establish mouse infection models. Here we report a direct contact transmission of SARS-CoV-2 variant B.1.351 in wild-type mice. The SARS-CoV-2 (B.1.351) replicated efficiently and induced significant pathological changes in lungs and tracheas, accompanied by elevated proinflammatory cytokines in the lungs and sera. Mechanistically, the receptor-binding domain (RBD) of SARS-CoV-2 (B.1.351) spike protein turned to a high binding affinity to mouse angiotensin-converting enzyme 2 (mACE2), allowing the mice highly susceptible to SARS-CoV-2 (B.1.351) infection. Our work suggests that SARS-CoV-2 (B.1.351) expands the host range and therefore increases its transmission route without adapted mutation. As the wild house mice live with human populations quite closely, this possible transmission route could be potentially risky. In addition, because SARS-CoV-2 (B.1.351) is one of the major epidemic strains and the mACE2 in laboratory-used mice is naturally expressed and regulated, the SARS-CoV-2 (B.1.351)/mice could be a much convenient animal model system to study COVID-19 pathogenesis and evaluate antiviral inhibitors and vaccines.

Risk Analysis of the Transmission Route for the African Swine Fever Virus in Mainland China

African swine fever first broke out in mainland China in August 2018 and has caused a substantial loss to China’s pig industry. Numerous investigations have confirmed that trades and movements of infected pigs and pork products, feeding pigs with contaminative swills, employees, and vehicles carrying the virus are the main transmission routes of the African swine fever virus (ASFV) in mainland China. However, which transmission route is more risky and what is the specific transmission map are still not clear enough. In this study, we crawl the data related to pig farms and slaughterhouses from Baidu Map by writing the Python language and then construct the pig transport network. Following this, we establish an ASFV transmission model over the network based on probabilistic discrete-time Markov chains. Furthermore, we propose spatiotemporal backward detection and forward transmission algorithms in semi-directed weighted networks. Through the simulation and calculation, the risk of transmission routes is analyzed, and the results reveal that the infection risk for employees and vehicles with the virus is the highest, followed by contaminative swills, and the transportation of pigs and pork products is the lowest; the most likely transmission map is deduced, and it is found that ASFV spreads from northeast China to southwest China and then to west; in addition, the infection risk in each province at different times is assessed, which can provide effective suggestions for the prevention and control of ASFV.

A need of COVID19 vaccination for children aged <12 years: Comparative evidence from the clinical characteristics in patients during a recent Delta surge (B.1.617.2)

Objective: To evaluate the necessity of Covid-19 vaccination in children aged < 12 y by comparing the clinical characteristics in unvaccinated children aged < 12 y with vaccinated patients aged > 12y during the Delta surge (B.1.617.2) in Putian, Fujian, China. Methods: A total of 226 patients with SARS-Cov-2 Delta variant (B.1.167.2; confirmed by Realtime PCR positive and sequencing) were enrolled from Sep 10th to Oct 20th, 2021, including 77 unvaccinated children (aged < 12y) and 149 people aged > 12y, mostly vaccinated. The transmission route was explored and the clinical data of two groups were compared;the effect factors for the time of the nucleic acid negativization (NAN) were examined by R statistical analysis. Results: The Delta surge in Putian spread from children in schools to factories, mostly through family contact. Compared with those aged; 12y, patients aged < 12y accounted for 34.07% of the total and showed milder fever, less cough and fatigue; they reported higher peripheral blood lymphocyte counts [1.84(1.32,2.71)× 10^9/L vs. 1.31(0.94,1.85)× 10^9/L; p<0.05), higher normal CRP rate (92.21% vs. 57.72%), lower IL-6 levels [5.28(3.31,8.13) vs. 9.10(4.37,15.14); p<0.05]. Upon admission, their COVID19 antibodies (IgM and IgG) and IgG in convalescence were lower [0.13(0.00,0.09) vs. 0.12(0.03,0.41), p<0.05; 0.02(0.00,0.14) vs. 1.94(0.54,6.40), p <0.05; 5.46(2.41,9.26) vs. 73.63 (54.63,86.55), p<0.05, respectively], but longer NAN time (18 days vs. 16 days, p=0.13). Conclusion: Children aged < 12y may be critical hidden spreaders, which indicates an urgent need of vaccination for this particular population.

Correlation of Respiratory Aerosols and Metabolic Carbon Dioxide

Respiratory aerosols from breathing and talking are an important transmission route for viruses, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Previous studies have found that particles with diameters ranging from 10 nm to 145 μm are produced from different regions in the respiratory system and especially smaller particles can remain airborne for long periods while carrying viral RNA. We present the first study in which respiratory aerosols have been simultaneously measured with carbon dioxide (CO2) to establish the correlation between the two concentrations. CO2 concentrations are easily available through low-cost sensors and could be used to estimate viral exposure through this correlation, whereas source-specific aerosol measurements are complicated and not possible with low-cost sensors. The increase in both respiratory aerosols and CO2 was linear over ten minutes in a 2 m3 chamber for all participants, suggesting a strong correlation. On average, talking released more particles than breathing, with 14,600 ± 16,800 min−1 (one-σ standard deviation) and 6210 ± 5630 min−1 on average, respectively, while CO2 increased with 139 ± 33 ppm min−1 during talking and 143 ± 29 ppm min−1 during breathing. Assuming a typical viral load of 7×106 RNA copies per mL of oral fluid, ten minutes of talking and breathing are estimated to produce 1 and 16 suspended RNA copies, respectively, correlating to a CO2 concentration of around 1800 ppm in a 2 m3 chamber. However, viral loads can vary by several orders of magnitude depending on the stage of the disease and the individual. It was therefore concluded that, by measuring CO2 concentrations, only the number and volume concentrations of released particles can be estimated with reasonable certainty, while the number of suspended RNA copies cannot.

Inactivation of Polyomavirus SV40 as Surrogate for Human Papillomaviruses by Chemical Disinfectants

Human papillomaviruses (HPV) are non-enveloped DNA viruses infecting cutaneous and mucosal squamous epithelia. Sexually transmitted HPV-types that are carcinogenic to humans such as HPV16 can induce cervical and other anogenital cancers. Virus transmission through fomites such as inadequately disinfected gynecological equipment is a further potential transmission route. Since HPV cannot be easily grown in cell culture, polyomavirus SV40 has been used as a surrogate virus when testing the virucidal activity of chemical disinfectants. So far, studies that have compared the virucidal activity of different disinfectants against HPV and SV40 are lacking. Here, we evaluated the susceptibility of HPV16 pseudovirus and SV40 to seven active biocidal substances using quantitative suspension tests. Ethanol, glutaraldehyde (GTA), dodecyldipropylentriamin (DPTA), and ortho-phthalaldehydes (OPA) were able to reduce the infectivity of HPV16 pseudovirus >99.99% after 5 min. In contrast, isopropanol, peracetic acid (PAA), and quaternary ammonium compounds with alkylamines (QAC) only led to a slight or no reduction in infectivity. Concerning SV40, only GTA (60 min contact time), PAA, and OPA had virus-inactivating effects. In conclusion, the virucidal activity of three out of seven disinfectants tested was different for HPV16 pseudovirus and SV40. In this study, SV40 was shown to be a reliable surrogate virus for HPV when testing isopropanol-, GTA-, QAC-, and OPA-based disinfectants.

Is Fresh Produce in Tigray, Ethiopia a Potential Transmission Vehicle for Cryptosporidium and Giardia?

In rural Ethiopia, where people often share their homes with their livestock, infections of humans and animals with Cryptosporidium and Giardia are relatively common. One possible transmission route is consumption of contaminated fresh produce; this study investigated the occurrence of Cryptosporidium and Giardia in fresh produce in four districts of rural Tigray in Ethiopia. Fresh produce samples (n = 55) were analysed using standard laboratory procedures. Overall, 15% (8/55) of samples were found to be contaminated. Although contamination levels were mostly low, a few samples had high numbers of Giardia cysts (up to around 70 cysts per 30 g sample). Molecular analyses were largely unsuccessful, but Giardia Assemblage A was identified in one sample. Contamination with these parasites was identified in two of the four districts, but, although a similar pattern has already been described for water contamination, this may be at least partially explained by sampling bias. Nevertheless, we speculate that access to clean water sources may be an important factor for reducing the occurrence of these pathogens. Given the public health and veterinary burden associated with both parasites, the factors which are of importance for their circulation in the communities and environments deserve further investigation.