Chlorine inactivation of adenovirus type 40 and feline calicivirus

Ct values, the concentration of free chlorine multiplied by time of contact with virus, were determined for free-chlorine inactivation experiments carried out with chloroform-extracted (dispersed) and non-chloroform-extracted (aggregated) feline calicivirus (FCV), adenovirus type 40 (AD40), and polio virus type 1 (PV-1). Experiments were carried out with high and low pH and temperature conditions. Ct values were calculated directly from bench-scale free-chlorine inactivation experiments and from application of the efficiency factor Hom model. For each experimental condition, Ct values were higher at pH 8 than at pH 6, higher at 5 degrees C than at 15 degrees C, and higher for dispersed AD40 (dAD40) than for dispersed FCV (dFCV). dFCV and dAD40 were more sensitive to free chlorine than dispersed PV-1 (dPV-1). Cts for 2 log inactivation of aggregated FCV (aFCV) and aggregated PV-1 (aPV-1) were 31.0 and 2.8 orders of magnitude higher than those calculated from experiments carried out with dispersed virus. Cts for 2 log inactivation of dFCV and dAD40 in treated groundwater at 15 degrees C were 1.2 and 13.7 times greater than in buffered-demand-free (BDF) water experiments at 5 degrees C. Ct values listed in the U.S. Environmental Protection Agency (EPA) Guidance Manual were close to, or lower than, Ct values generated for experiments conducted with dispersed and aggregated viruses suspended in BDF water and for dispersed viruses suspended in treated groundwater. Since the state of viruses in water is most likely to be aggregated and associated with organic or inorganic matter, reevaluation of the EPA Guidance Manual Ct values is necessary, since they would not be useful for ensuring inactivation of viruses in these states. Under the tested conditions, dAD40, dFCV, aFCV, dPV-1, and aPV-1 particles would be inactivated by commonly used free chlorine concentrations (1 mg/liter) and contact times (60 to 237 min) applied for drinking water treatment in the United States.     

Identification, expression, and purification of a pyrethroid-hydrolyzing carboxylesterase from mouse liver microsomes

Carboxylesterases are enzymes that catalyze the hydrolysis of a wide range of ester-containing endogenous and xenobiotic compounds. Although the use of pyrethroids is increasing, the specific enzymes involved in the hydrolysis of these insecticides have yet to be identified. A pyrethroid-hydrolyzing enzyme was partially purified from mouse liver microsomes using a fluorescent reporter similar in structure to cypermethrin (Shan, G., and Hammock, B. D. (2001) Anal. Biochem. 299, 54-62 and Wheelock, C. E., Wheelock, A. M., Zhang, R., Stok, J. E., Morisseau, C., Le Valley, S. E., Green, C. E., and Hammock, B. D. (2003) Anal. Biochem. 315, 208-222) and subsequently identified as a carboxylesterase (NCBI accession number BAC36707). The expressed sequence tag was then cloned, expressed in baculovirus, and purified to homogeneity. Kinetic constants for a large number of both type I and type II pyrethroid or pyrethroid-like substrates were determined. This esterase possesses similar kinetic constants for cypermethrin and its fluorescent-surrogate (k(cat) = 0.12 +/- 0.03 versus 0.11 +/- 0.01 s(-1)). Compared with their cis- counterparts, trans-permethrin and cypermethrin were hydrolyzed 22- and 4-fold faster, respectively. Of the four fenvalerate isomers the (2R)(alphaR)-isomer was hydrolyzed at least 1 order of magnitude faster than any other isomer. However, it is unlikely that this enzyme accounts for the total pyrethroid hydrolysis in the microsomes because both isoelectrofocusing and native PAGE indicate the presence of a second region of cypermethrin-metabolizing enzymes. A second carboxylesterase gene (NCBI accession number NM_133960), isolated during a cDNA mouse liver library screening, was also found to hydrolyze pyrethroids. Both these enzymes could be used as preliminary tools in establishing the relative toxicity of new pyrethroids.     

Tagging retrovirus vectors with a metal binding peptide and one-step purification by immobilized metal affinity chromatography

Retroviral vectors produced from packaging cells are invariably contaminated by protein, nucleic acid, and other substances introduced in the manufacturing process. Elimination of these contaminants from retroviral vector preparations is helpful to reduce unwanted side effects, and purified vector preparations are desirable to improve reproducibility of therapeutic effect. Here we report a novel approach to engineer a metal binding peptide (MBP)-tagged murine leukemia virus (MuLV), allowing for one-step purification of retroviral vectors by immobilized metal affinity chromatography (IMAC). We inserted a His6 peptide into an ecotropic envelope protein (Env) by replacing part of its hypervariable region sequence with a sequence encoding the His6 peptide. Display of the His6 tag on the surface of Env endowed the vectors with a high affinity for immobilized metal ions, such as nickel. We demonstrated that the His6-tagged MuLV could be produced to high titers and could be highly purified by one-step IMAC. The protein and DNA contaminants in the purified vector supernatants were below 7 microg/ml and 25 pg/ml, respectively, indicating a 1,229-fold reduction in protein contaminant level and a 6,800-fold reduction in DNA contaminant level. About 56% of the viral vectors were recovered in the IMAC purification. The purified vectors retained their functionality and infectivity. These results establish that an MBP can be functionally displayed on the surface of ecotropic retroviruses without interfering with their integrity, and MBP-tagged retroviral vectors can be highly purified by one-step IMAC.     

Behavioral audiogram of two Arctic foxes (Vulpes lagopus)

With increased polar anthropogenic activity, such as from the oil and gas industry, there are growing concerns about how Arctic species will be affected. Knowledge of species’ sensory abilities, such as auditory sensitivities, can be used to mitigate the effects of such activities. Herein, behavioral audiograms of two captive adult Arctic foxes (Vulpes lagopus) were measured using a yes/no paradigm and descending staircase method of signal presentation. Both foxes displayed a typical mammalian U-shaped audiometric curve, with a functional hearing range of 125 Hz–16 kHz (sensitivity ≤ 60 dB re: 20 μPa) and average peak sensitivity of 24 dB re: 20 μPa at 4 kHz. The foxes had a lower frequency range and sensitivity than would be expected when compared to previous audiograms of domestic dogs (Canis familiaris) and other carnivores. These differences indicate Arctic foxes (V. lagopus) may have a lower frequency range than previously expected, which was similar to the only other fox species tested to date, kit foxes (Vulpes macrotis). Alternatively, differences may be due to testing constraints, such as masking of test signals by ambient noise and/or an unintentionally trained conservative response bias, which most likely resulted in underestimated hearing curves. While results of this study should be interpreted with caution due to its limitations, findings indicate that foxes have a narrower frequency range than formerly presumed. Anthropogenic activities near fox habitats can mitigate their impacts by reducing noise at frequencies within the functional hearing range and peak sensitivities of this species.     

Blue photobiomodulation LED therapy impacts SARS-CoV-2 by limiting its replication in Vero cells

The study of any intervention able to counteract SARS-CoV-2 pandemic is considerably envisaged. It was previously shown, in in vitro models of infections, that the LED blue light is able to decrease the viral load of HSV-1 and ZIKV. In our study, LED photobiomodulation therapy (PBMT) at blue wavelengths (450, 454 and 470 nm) was tested in an in vitro model of SARS-CoV-2 infection, employing three experimental settings: SARS-CoV-2 was irradiated and then transferred to cells; already infected cells were irradiated; cells were irradiated prior to infection. A decrement of the viral load was observed when previously infected cells were irradiated with all three tested wavelengths and relevant effects were registered especially at 48 hours post-infection, possibly suggesting that the blue light could interfere with the intracellular viral replication machinery. Our in vitro findings could represent the starting point for translational applications of PBMT as a supportive approach to fight SARS-CoV-2.     

Estimating temperature effects on Drosophila suzukii life cycle parameters

1. Drosophila suzukii is an invasive, polyphagous pest of soft-skinned fruits, having huge impact on fruit production in Asia, North and South America and Europe including Germany.
2. To investigate the effect of temperature on oviposition, egg-to-adult development success and duration, as well as immature heat survival and adult cold survival for a German D. suzukii population several experiments were conducted under different constant temperatures in the laboratory. The resulting life cycle data were described mathematically as functions of temperature and compared with experimental results of other researchers in a summary table.
3. Curve fittings used herein revealed that minimum, optimum and maximum temperatures are: 13.2, 26.7 and 33.6 °C for oviposition, 14.1, 22.6 and 30.0 °C for egg-to-adult development success, and 9.6, 27.3 and 35.7 °C for egg-to-adult development duration. Eggs and larvae of D. suzukii showed a reduced heat survival within the tested temperature range of 29 to 41 °C and exposure durations from 1 to 8 h. A cold survival rate of 50% was measured at e.g. −6 °C for 4 h in summer morph adults and at e.g. −6 °C for 45 h in winter morph adults confirming that the latter are more cold tolerant.
4. Results obtained in this study for a German population of D. suzukii are similar to those obtained for populations of other origins such as Canada, Japan, Spain and USA. Thus, presumably, present data based on a German D. suzukii population can be used for a new or fine-tune of already existing population dynamics models of D. suzukii in order to support an effective pest management strategy.

     

Effect of near-infrared and blue laser light on vero E6 cells SARS-CoV-2 infection model

Photobiomodulation therapy (PBMT) employing laser light has been emerging as a safe strategy to challenge viruses. In this study the effect of blue and near-infrared (NIR) laser light was assessed in an in vitro model of SARS-CoV-2 infection. PBMT at blue wavelength inhibited viral amplification when the virus was directly irradiated and then transferred to cell culture and when cells already infected were treated. The NIR wavelength resulted less efficacious showing a minor effect on the reduction of the viral load. The cells receiving the irradiated virus or directly irradiated rescued their viability to level comparable to not treated cells. Virion integrity and antigenicity were preserved after blue and NIR irradiation, suggesting that the PBMT antiviral effect was not correlated to viral lipidic envelope disruption. Our results suggested that PBMT can be considered a valid strategy to counteract SARS-CoV-2 infection, at least in vitro.     

A review on temperature and humidity effects on Drosophila suzukii population dynamics

1. Drosophila suzukii is an invasive polyphagous pest of wild and cultivated soft‐skinned fruits, which can cause widespread economic damage in orchards and vineyards.
2. The simulation and prediction of D. suzukii's population dynamics would be helpful for guiding pest management. Therefore, we reviewed and summarized the current knowledge on effects of air temperature and relative humidity on different life cycle parameters of D. suzukii.
3. The literature summary presented shows that high oviposition rates can occur between 18 and 30 °C. Temperatures between 16 and 25 °C resulted in fast and high egg‐to‐adult development success of more than 80%. Oviposition and adult life span were positively affected by high relative humidity; however, the factor humidity is so far rarely investigated.
4. We assume that this is one reason why relative humidity usually is not considered in modelling approaches, which are summarized herein. The high number of recently published research articles on D. suzukii's life cycle suggests that there is already a lot of knowledge available on its biology. However, there are still considerable research gaps mentioned in the literature, which are also summarized herein.
5. Nevertheless, we conclude that sufficient temperature data in the literature are suitable to understand and predict population dynamics of D. suzukii, in order to assist pest management in the field.