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1.
Res Vet Sci ; 165: 105052, 2023 Dec.
Article in English | MEDLINE | ID: mdl-37864907

ABSTRACT

Toxoplasma gondii is a major foodborne zoonotic pathogen that can be transmitted through the consumption of raw or undercooked meat of small ruminants, among others. Serology has been suggested as an epidemiological indicator and several tests are available nowadays. However, there is no comparative study with the most used ones. Therefore, the objective of this study was to develop and validate two in-house tests (Western blot -TgSALUVET WB- and ELISA -TgSALUVET ELISA 2.0-) and perform a comparative study including such tests and four commercial ELISA kits (IDScreen®, PrioCHECK®, Pigtype® and IDEXX). First, a specific pattern of recognition of immunodominant antigens by TgSALUVET WB was determined with serum panels of noninfected sheep and sheep infected with T. gondii or Neospora caninum. Next, TgSALUVET WB was used as a reference to preliminary validate TgSALUVET ELISA 2.0 using sera from sheep and goats naturally infected with T. gondii. Then, the abovementioned sheep serum panels were analyzed by all tests and subjected to TG-ROC analyses and agreement tests, and cross-reactivity with the anti-N. caninum IgGs was studied. All the techniques were accurate enough for the cutoff values initially suggested with all serum panels (Se and Sp ≥ 94%), except for PrioCHECK®, which showed 83% Sp. However, a cutoff readjustment improved their diagnostic performance. Additionally, cross-reactions between anti-N. caninum antibodies and T. gondii antigens were detected with all tests. Thus, a second cutoff readjustment was carried out and the use of both readjusted cutoff values is recommended to obtain comparable data and avoid false-positive results.


Subject(s)
Coccidiosis , Goat Diseases , Neospora , Toxoplasma , Toxoplasmosis, Animal , Sheep , Animals , Toxoplasmosis, Animal/diagnosis , Antibodies, Protozoan , Ruminants , Goats , Enzyme-Linked Immunosorbent Assay/veterinary , Serologic Tests/veterinary , Seroepidemiologic Studies , Coccidiosis/diagnosis , Coccidiosis/veterinary , Goat Diseases/diagnosis
2.
Appl Mater Today ; 32: 101828, 2023 Jun.
Article in English | MEDLINE | ID: mdl-37317691

ABSTRACT

Commercial polyurethane (PU) coating formulations have been modified with 1-(hydroxymethyl)-5,5-dimethylhydantoin (HMD) both in bulk (0.5 and 1% w/w) and onto the coatings surface as an N-halamine precursor, to obtain clear coatings with high virucidal activity. Upon immersion in diluted chlorine bleaching, the hydantoin structure on the grafted PU membranes was transformed into N-halamine groups, with a high surface chlorine concentration (40-43µg/cm2). Fourier transform infrared spectroscopy (FTIR) spectroscopy, thermogravimetric analysis (TGA), energy-dispersive X-ray (EDX), X-ray photoelectron spectroscopy (XPS) and iodometric titration were used to characterize the coatings and quantify the chlorine contents of the PU membranes after chlorination. Biological evaluation of their activity against Staphylococcus aureus (Gram-positive bacteria) and human coronaviruses HCoV-229E and SARS-CoV-2 was performed, and high inactivation of these pathogens was observed after short contact times. The inactivation of HCoV-229E was higher than 98% for all modified samples after just 30 minutes, whereas it was necessary 12 hours of contact time for complete inactivation of SARS-CoV-2. The coatings were fully rechargeable by immersion in diluted chlorine bleach (2% v/v) for at least 5 chlorination-dechlorination cycles. Moreover, the performance of the antivirus efficiency of the coatings is considered as long-lasting, because experiments of reinfection of the coatings with HCoV-229E coronavirus did not show any loss of the virucidal activity after three consecutive infection cycles without reactivation of the N-halamine groups.

3.
ACS Appl Mater Interfaces ; 15(18): 22580-22589, 2023 May 10.
Article in English | MEDLINE | ID: mdl-37116104

ABSTRACT

The current global pandemic due to the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus has demonstrated the necessity to develop novel materials with antimicrobial and antiviral activities to prevent the infection. One significant route for the spread of diseases is by the transmission of the virus through contact with contaminated surfaces. Antiviral surface treatments can help to reduce or even avoid these hazards. In particular, the development of active-virucidal fabrics or paints represents a very important challenge with multiple applications in hospitals, public transports, or schools. Modern, cutting-edge methods for creating antiviral surface coatings use either materials with a metal base or sophisticated synthetic polymers. Even if these methods are effective, they will still face significant obstacles in terms of large-scale applicability. Here, we describe the preparation of fabrics and paints treated with a scaled-up novel nanostructured biohybrid material composed of very small crystalline phosphate copper(II) nanoparticles, synthesized based on a technology that employs the use of a small amount of biological agent for its formation at room temperature in aqueous media. We demonstrate the efficient inactivation of the human coronavirus 229E (HCoV-229E), the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus, and non-enveloped human rhinovirus 14 (HRV-14) (>99.9%) using an inexpensive, ecologically friendly coating agent. The reactive oxygen species produced during the oxidation of water or the more intensive reaction with hydrogen peroxide are believed to be the cause of the antiviral mechanism of the nanostructured material. In contrast to the release of a specific antiviral drug, this process does not consume the surface coating and does not need regeneration. A 12-month aging research that revealed no decline in antiviral activity is proof that the coating is durable in ambient circumstances. Also, the coated fabric can be reused after different washing cycles, even at moderate to high temperatures.


Subject(s)
COVID-19 , Coronavirus 229E, Human , Viruses , Humans , SARS-CoV-2 , Antiviral Agents/pharmacology , Antiviral Agents/chemistry , COVID-19/prevention & control
4.
Food Environ Virol ; 14(3): 304-313, 2022 09.
Article in English | MEDLINE | ID: mdl-35851946

ABSTRACT

The experience of COVID19 pandemic has demonstrated the real concern of biological agents dispersed in the air and surfaces environments. Therefore, the need of a fast and large-scale disinfection method has arisen for prevention of contagion. COUNTERFOG® is an innovative technology developed for large-scale decontamination of air and surfaces. The objective of this study is to assess experimentally the effectiveness of COUNTERFOG® in disinfecting viral-contaminated surfaces. We also aim to measure the necessary time to disinfect said surfaces. Stainless steel surfaces were contaminated with bacteriophage φ29 and disinfected using COUNTERFOG® SDR-F05A+, which uses a sodium hypochlorite solution at different concentrations and for different exposure times. A log reduction over 6 logs of virus titer is obtained in 1 min with 1.2% sodium hypochlorite when the application is direct; while at a radial distance of 5 cm from the point of application the disinfection reaches a reduction of 5.5 logs in 8 min. In the same way, a higher dilution of the sodium hypochlorite concentration (0.7% NaOCl) requires more exposure time (16 min) to obtain the same log reduction (> 6 logs). COUNTERFOG® creates, in a short time and at a distance of 2 m from the point of application, a thin layer of disinfectant that covers the surfaces. The selection of the concentration and exposure time is critical for the efficacy of disinfection. These tests demonstrate that a concentration between 0.7- 1.2% sodium hypochlorite is enough for a fast and efficient ɸ29 phage inactivation. The fact that ɸ29 phage is more resistant to disinfection than SARS-CoV-2 sustains this disinfection procedure.


Subject(s)
Bacteriophages , COVID-19 , Disinfectants , Decontamination/methods , Disinfectants/pharmacology , Disinfection/methods , Humans , SARS-CoV-2 , Sodium Hypochlorite/pharmacology
5.
Article in English | MEDLINE | ID: mdl-36612897

ABSTRACT

The COVID-19 pandemic highlighted the dangers of airborne pathogen transmission. SARS-CoV-2 is known to be transmitted through aerosols; however, little is known about the dynamics of these aerosols in real environments, the conditions, and the minimum viral load required for infection. Efficiently measuring and capturing pathogens present in the air would help to understand the infection process. Air samplers usually take several hours to obtain an air sample. In this work a fast (1-2 min) method for capturing bioaerosols into a liquid medium has been tested in hospital rooms with COVID-19 patients. This fast sampling allows detecting transient levels of aerosols in the air. SARS-CoV-2 RNA is detected in aerosols from several hospital rooms at different levels. Interestingly, there are sudden boosts of the SARS-CoV-2 load in the air, suggesting that SARS-CoV-2 could be released abundantly at certain moments. These results show that the distribution of SARS-CoV-2-containing aerosols is not homogeneous in the hospital room. This technology is a fast and effective tool for capturing airborne matter in a very short time, which allows for fast decision-making any kind of hazard in the air is detected. It is also useful for a better understanding of aerosols dynamics.


Subject(s)
COVID-19 , SARS-CoV-2 , Humans , Pandemics , RNA, Viral , Respiratory Aerosols and Droplets , Hospitals
6.
Anim Sci J ; 92(1): e13667, 2021.
Article in English | MEDLINE | ID: mdl-34877736

ABSTRACT

The objectives of this study were to evaluate and compare the effect of working vacuum levels (35 and 44 kPa) and liners dimensions (mouthpiece lip diameter and overall length, 20-185 and 22-170 mm) on the main milking characteristics of goats. The results highlight that both the working vacuum level and the liner dimension have influenced the milk flow curve parameters. The maximum variations were found for peak flow rate, which increased significantly with liner dimensions of 20-185 mm at a working vacuum level of 44 kPa as well as average milk flow rate and for plateau phase duration. The incorrect adoption of operative parameters and unsuitable milking machine components, might affect the performance of the mechanical milking and negatively affecting animal productions and welfare.


Subject(s)
Goats , Milk , Animals , Dairying , Female , Lactation , Mammary Glands, Animal , Vacuum
7.
Sci Rep ; 11(1): 23494, 2021 12 06.
Article in English | MEDLINE | ID: mdl-34873184

ABSTRACT

Foot-and-mouth disease virus (FMDV) is a picornavirus that exhibits an extremely acid sensitive capsid. This acid lability is directly related to its mechanism of uncoating triggered by acidification inside cellular endosomes. Using a collection of FMDV mutants we have systematically analyzed the relationship between acid stability and the requirement for acidic endosomes using ammonium chloride (NH4Cl), an inhibitor of endosome acidification. A FMDV mutant carrying two substitutions with opposite effects on acid-stability (VP3 A116V that reduces acid stability, and VP1 N17D that increases acid stability) displayed a rapid shift towards acid lability that resulted in increased resistance to NH4Cl as well as to concanamicyn A, a different lysosomotropic agent. This resistance could be explained by a higher ability of the mutant populations to produce NH4Cl-resistant variants, as supported by their tendency to accumulate mutations related to NH4Cl-resistance that was higher than that of the WT populations. Competition experiments also indicated that the combination of both amino acid substitutions promoted an increase of viral fitness that likely contributed to NH4Cl resistance. This study provides novel evidences supporting that the combination of mutations in a viral capsid can result in compensatory effects that lead to fitness gain, and facilitate space to an inhibitor of acid-dependent uncoating. Thus, although drug-resistant variants usually exhibit a reduction in viral fitness, our results indicate that compensatory mutations that restore this reduction in fitness can promote emergence of resistance mutants.


Subject(s)
Amino Acid Substitution/genetics , Capsid Proteins/genetics , Foot-and-Mouth Disease Virus/genetics , Foot-and-Mouth Disease/virology , Animals , Cell Line , Cricetinae , Endosomes/genetics , Mutation/genetics
8.
Pathogens ; 10(11)2021 Nov 20.
Article in English | MEDLINE | ID: mdl-34832673

ABSTRACT

Neospora caninum is an apicomplexan parasite that can cause abortions and perinatal mortality in sheep. Although ovine neosporosis has been described worldwide, there is a lack of information about the relationship between N. caninum serostatus and the reproductive performance. In this study, we described the infection dynamics in a dairy sheep flock with an abortion rate up to 25% and a N. caninum seroprevalence of 32%. Abortions were recorded in 36% and 9% of seropositive and seronegative sheep, respectively. Seropositive sheep were more likely to abort twice (OR = 4.44) or three or more times (OR = 10.13) than seronegative sheep. Endogenous transplacental transmission was the main route of transmission since 86% of seropositive sheep had seropositive offspring. Within dams that had any abortion, seropositive sheep were more likely than seronegative ones to have female descendants that aborted (OR = 8.12). The slight increase in seropositivity with the age, the low percentage of animals with postnatal seroconversion or with low avidity antibodies, and the seropositivity of one flock dog, indicated that horizontal transmission might have some relevance in this flock. A control programme based on selective culling of seropositive sheep and replacement with seronegative animals was effective in reducing the abortion rate to 7.2%.

9.
Vet Parasitol ; 293: 109432, 2021 May.
Article in English | MEDLINE | ID: mdl-33915322

ABSTRACT

Toxoplasma gondii is a worldwide distributed parasite causing abortions and fetal malformations in small ruminants. The aim of this study was to design and validate a new immunoassay based on the use of TgSAG1-GRA8 chimeric antigen for the detection of anti-T. gondii antibodies in serum of goats. First, a time-resolved fluorescence immunoassay (TgSAG1-GRA8-TRFIA) was developed. In addition, the diagnostic performance of TgSAG1-GRA8-TRFIA was compared with an optimized enzyme-linked immunosorbent assay (TgSALUVET-ELISA) and a Western Blot (WB), both based on whole T. gondii tachyzoite antigenic extract. The TgSAG1-GRA8-TRFIA has shown a high intra- and inter-assay precision, analytical sensitivity and accuracy. The ROC analysis of this assay showed an optimal cut-off of 217.4 Units of Fluorometry for T. gondii (UFT), with 92 % of sensitivity and 90.48 % of specificity. A positive and statistically significant Spearman's correlation with TgSALUVET-ELISA was detected, and kappa value was 0.83, presenting high agreement with both methods. However, TgSAG1-GRA8 protein showed cross-reactivity with specific anti-Neospora caninum antibodies. Thus, TgSAG-1-GRA8 chimeric antigen seems not to be an ideal option for the serodiagnosis of T. gondii infection in goats unless combined with the serodiagnosis of N. caninum infection in parallel. In the light of the results obtained, a comprehensive study on the existence of cross-reactivities between T. gondii antigens used in serological tests employed in animal health and specific antibodies directed against Toxoplasmatinae parasites should be performed.


Subject(s)
Antibodies, Protozoan , Enzyme-Linked Immunosorbent Assay , Goat Diseases , Toxoplasma , Toxoplasmosis, Animal , Animals , Antibodies, Protozoan/blood , Antigens, Protozoan , Coccidiosis/diagnosis , Coccidiosis/veterinary , Cross Reactions , Enzyme-Linked Immunosorbent Assay/standards , Enzyme-Linked Immunosorbent Assay/veterinary , Goat Diseases/diagnosis , Goats , Neospora/immunology , Seroepidemiologic Studies , Toxoplasma/immunology , Toxoplasmosis, Animal/diagnosis
10.
Animals (Basel) ; 10(5)2020 May 04.
Article in English | MEDLINE | ID: mdl-32375423

ABSTRACT

A dairy farmer chooses the number of milking groups in function of the herd size, stall type and milking system also in small cow dairies (number of animals lower than 100-120). In these dairies, there are different milking systems (bucket, trolley, pipeline, little autotandem, herringbone or parallel parlors) and each of them has a different work routine. The knowledge of the routine is the starting point for assessing the milking installation, because it determines the number of milked cows per hour. Different milking systems have common tasks (as pre-dipping, inspecting foremilk, udder preparation, attaching teat cups, post-dipping), but in the meantime there are different operations that characterize each specific routine (e.g., animal entry and exit if there is a parlor, bucket, trolley or milking group positioning if tie-stall). For this reason, we surveyed twenty small dairy farms located in the Piedmont Region (Italy) with different milking systems to correctly acquire the specific milking routines. Different models were therefore studied using the observed routines in in the examined farms. These models were then used to calculate the number of milked cows per hour and the number of milking groups. The main findings were simple equations, specific for each milking system, easily accessible by the farmer to correctly size his milking system.

11.
Vet Parasitol ; 276: 108994, 2019 Dec.
Article in English | MEDLINE | ID: mdl-31778941

ABSTRACT

Neospora caninum is a protozoan parasite (Phylum Apicomplexa) that has been recently suggested as a relevant cause of reproductive disorders in small ruminants. The aim of the present study is to develop and validate a new serological test based on time resolved fluorescency using N. caninum GRA7 recombinant antigen (GRA7-TRFIA) for the detection of N. caninum antibodies in sheep. A total of 346 serum samples (208 from experimentally infected sheep, 117 from a dairy farm with a previous history of Neospora-associated abortion, and 21 negative sera) were used. The validation of the new assay was performed by the evaluation of assay precision, analytical sensitivity (Se), accuracy and cross reactivity. In the experimentally infected sheep, antibody kinetics was compared between GRA7-TRFIA and an in house N. caninum tachyzoite soluble extract-based ELISA (NcSALUVET ELISA) by Wilcoxon matched-pairs signed rank test. The cut-off and diagnostic Se and specificity (Sp) of GRA7-TRFIA was estimated by ROC analysis with field samples. In addition, concordance and correlation between GRA7-TRFIA and a commercial ELISA and NcSALUVET ELISA were assessed by kappa value and Spearman correlation coefficient, respectively. Overall, GRA7-TRFIA showed an adequate precision, analytical Se and accuracy to detect anti-N. caninum antibodies in ovine serum, and no cross reactivity with the closely related protozoan Toxoplasma gondii. In naturally infected sheep, 100% Se and 95.35% Sp were obtained for a cut-off point of 62.68 Units of Fluorometry for N. caninum (UFN). Moreover, GRA7-TRFIA allowed earlier detection of N. caninum infection than NcSALUVET ELISA in experimentally infected sheep.


Subject(s)
Antibodies, Protozoan/blood , Antigens, Protozoan/immunology , Coccidiosis/veterinary , Neospora/immunology , Sheep Diseases/immunology , Animals , Coccidiosis/immunology , Cross Reactions , Enzyme-Linked Immunosorbent Assay/veterinary , Female , Fluorescence Polarization Immunoassay , Fluorometry/veterinary , Immunoglobulin G/blood , Sensitivity and Specificity , Sheep , Sheep Diseases/parasitology
12.
J Clin Med ; 8(9)2019 Sep 11.
Article in English | MEDLINE | ID: mdl-31514402

ABSTRACT

The objective of this study is to evaluate the risk of clinical infections by herpesviruses in patients exposed to valproic acid (VPA). We performed a case-control study nested in a primary cohort selected from the Spanish primary care population-based research database BIFAP (Base de datos para la Investigación Farmacoepidemiológica en Atención Primaria) over the period 2001-2015. The events of interest were those diseases caused by any herpesviruses known to infect humans. For each case, up to 10 controls per case matched by age, gender, and calendar date were randomly selected. A conditional logistic regression was used to compute adjusted odds ratios (OR) and their 95% confidence intervals (95% CI). Current use of VPA was associated with a trend towards a reduced risk of clinical infections by herpesviruses as compared with non-users (OR 0.84; CI 95% 0.7-1.0; p = 0.057). Among current users, a trend to a decreased risk with treatment durations longer than 90 days was also observed. The results show a trend to a reduced risk of clinical infection by herpesviruses in patients exposed to VPA. These results are consistent with those in vitro studies showing that, in cultured cells, VPA can inhibit the production of the infectious progeny of herpesviruses. This study also shows the efficient use of electronic healthcare records for clinical exploratory research studies.

13.
Article in English | MEDLINE | ID: mdl-31234466

ABSTRACT

European Directives 2002/44/EC and 2003/10/EC establish the exposure limit values for preventing operators' risks to vibration and noise transmitted by machines. Few studies studied noise and vibration of agricultural backpack powered machines (as mist blowers and blowers), but nobody critically studied them. This work analyzed the field back vibration, hand-arm vibration (HAV), and noise transmitted to ten operators by eight blowers and mist blowers. Unweighted and weighted vibration were analyzed, using the standards ISO 2631-1 (back), and ISO 5349-1 and ISO/TR 18570 (hand-arm system). The noise was evaluated by recording the acoustic pressure level at the operators' ears using the ISO 9612. With the ISO 2631-1, the vibration to the operators' back was low (0.38 ms-2), but the unweighted vibration measured along y and z-axes (not used by the ISO 2631-1) were high (>11 ms-2). HAV were also low when using the ISO 5349-1 (the highest value was 2.51 ms-2 in mist blowers), but high with the ISO/TR 18570 for the onset of vibration white finger (1446 ms-1.5 in blowers). Noise levels were always high: more than 100 dB(A), excluding the blower with the exhaust inside the blower hose. This last machine had noise levels lower than 86 dB(A), but its specific feature could increase environmental pollution.


Subject(s)
Equipment Design/standards , Noise , Occupational Exposure/standards , Vibration , Agriculture/instrumentation , European Union , Occupational Exposure/analysis
14.
J Sep Sci ; 41(21): 4047-4054, 2018 Nov.
Article in English | MEDLINE | ID: mdl-30168667

ABSTRACT

In this study, the viability of applying cork and montmorillonite clay modified with ionic liquid as biosorbents in the rotating-disk sorptive extraction technique was investigated. Specifically, this was aimed at the determination of methyl paraben, ethyl paraben, propyl paraben, and isobutyl paraben, with separation/determination by high-performance liquid chromatography coupled with mass spectrometry. The optimization of the method for both biosorbents was performed using multivariate procedures. The extraction efficiencies for the target compounds in aqueous matrices were compared to those obtained using the commercial sorbent Octadecil Silano-C18. The optimum extraction conditions for both natural sorbents were the use of an ammonia solution (pH 10) as desorption solvent and an extraction time of 30 min. The proposed methods show limits of quantification of 0.8 µg/L for cork, 3.0 µg/L for montmorillonite clay and 6.0 µg/L for Octadecil Silano-C18. The relative recoveries from river water and tap water samples ranged from 80.3 to 118.7% and 80.0 to 119.9% for cork and montmorillonite clay modified with ionic liquid, respectively. Relative standard deviations were obtained for intra- and interday precisions of <15% and <19% for cork and <19% and <17% for montmorillonite clay modified with ionic liquid.

15.
Viruses ; 10(2)2018 02 23.
Article in English | MEDLINE | ID: mdl-29473907

ABSTRACT

Flaviviruses are relevant animal and human pathogens that include West Nile virus (WNV), Japanese encephalitis virus, dengue virus, or Zika virus, among others. Currently, no licensed therapy is available to fight flaviviral infections. Protein kinases C (PKCs) constitute a family of multifunctional lipid-dependent isoenzymes that regulate a wide variety of cellular processes (apoptosis, differentiation, proliferation, cellular transformation, motility, adhesion, etc.) being currently considered at the front line of drug development for the treatment of diverse human disorders. PKCs have also been implicated in different steps during viral replication; however, nowadays, results regarding their role in flavivirus replication are controversial. Here we demonstrate that calphostin C and chelerythrine, two broad-PKC inhibitors that target conventional, novel and atypical PKCs, significantly inhibit WNV multiplication in cell culture without affecting cell viability. A reduction of viral yields was observed in treated cells when compared with mock-treated cells. Likewise, immunofluorescence detection of viral enveloped E protein was reduced in treated cells, as was the amount of viral RNA released to the supernatant, mainly in those treated with chelerythrine. On the other hand, two PKC inhibitors specific for conventional and novel isoforms (staurosporine and enzastaurine) did not show any significant effect in WNV multiplication. These results suggested that PKCs, more probably atypical PKCs, are likely involved in WNV multiplication, although both broad-spectrum tested drugs seem to act through different mechanisms, and point to them as potential antiviral candidates for WNV, as well as for other related flaviviruses.


Subject(s)
Antiviral Agents/pharmacology , Enzyme Inhibitors/pharmacology , Protein Kinase C/antagonists & inhibitors , Virus Replication/drug effects , West Nile Fever/metabolism , West Nile Fever/virology , West Nile virus/drug effects , West Nile virus/physiology , Animals , Chlorocebus aethiops , Humans , Vero Cells
16.
Emerg Microbes Infect ; 6(9): e81, 2017 Sep 20.
Article in English | MEDLINE | ID: mdl-28928416

ABSTRACT

Flaviviruses are RNA viruses that constitute a worrisome threat to global human and animal health. Zika virus (ZIKV), which was initially reported to cause a mild disease, recently spread in the Americas, infecting millions of people. During this recent epidemic, ZIKV infection has been linked to serious neurological diseases and birth defects, specifically Guillain-Barrè syndrome (GBS) and microcephaly. Because information about ZIKV immunity remains scarce, we assessed the humoral response of immunocompetent mice to infection with three viral strains of diverse geographical origin (Africa, Asia and America). No infected animals showed any sign of disease or died after infection. However, specific neutralizing antibodies were elicited in all infected mice. Considering the rapid expansion of ZIKV throughout the American continent and its co-circulation with other medically relevant flaviviruses, such as West Nile virus (WNV), the induction of protective immunity between ZIKV and WNV was analyzed. Remarkably, protection after challenge with WNV was observed in mice previously infected with ZIKV, as survival rates were significantly higher than in control mice. Moreover, previous ZIKV infection enhanced the humoral immune response against WNV. These findings may be relevant in geographical areas where both ZIKV and WNV co-circulate, as well as for the future development of broad-spectrum flavivirus vaccines.


Subject(s)
Antibodies, Neutralizing/immunology , Antibodies, Viral/immunology , West Nile Fever/immunology , West Nile virus/immunology , Zika Virus Infection/immunology , Zika Virus/immunology , Africa/epidemiology , Americas/epidemiology , Animals , Antibodies, Neutralizing/biosynthesis , Antibodies, Neutralizing/blood , Antibodies, Viral/biosynthesis , Antibodies, Viral/blood , Asia/epidemiology , Immunity, Humoral , Immunocompetence , Mice , West Nile Fever/virology , West Nile virus/isolation & purification , Zika Virus/genetics , Zika Virus Infection/epidemiology
17.
Front Microbiol ; 8: 1314, 2017.
Article in English | MEDLINE | ID: mdl-28744282

ABSTRACT

The Flavivirus genus contains important pathogens, such as West Nile virus (WNV), Zika virus (ZIKV), and Dengue virus (DENV), which are enveloped plus-strand RNA viruses transmitted by mosquitoes and constitute a worrisome threat to global human and animal health. Currently no licensed drugs against them are available, being, thus, still necessary the search for effective antiviral molecules. In this line, a novel antiviral approach (economical, simple to use, and environmental friendly) is the use of natural compounds. Consequently, we have tested the antiviral potential of different polyphenols present in plants and natural products, such as wine and tea, against WNV, ZIKV, and DENV. So that, we assayed the effect of a panel of structurally related polyphenols [delphinidin (D), cyanidin (Cy), catechin (C), epicatechin (EC), epigallocatechin (EGC), and epigallocatechin gallate (EGCG)] on WNV infection, and found that D and EGCG inhibited more effectively the virus production. Further analysis with both compounds indicated that they mainly affected the attachment and entry steps of the virus life cycle. Moreover, D and EGCG showed a direct effect on WNV particles exerting a virucidal effect. We showed a similar inhibition of viral production of these compounds on WNV variants that differed on acidic pH requirements for viral fusion, indicating that their antiviral activity against WNV is produced by a virucidal effect rather than by an inhibition of pH-dependent viral fusion. Both polyphenols also reduced the infectivity of ZIKV and DENV. Therefore, D and EGCG impair the infectivity in cell culture of these three medically relevant flaviviruses.

18.
Vet Microbiol ; 203: 275-279, 2017 May.
Article in English | MEDLINE | ID: mdl-28619156

ABSTRACT

Foot-and-mouth disease virus (FMDV) is the etiological agent of a highly contagious disease that affects important livestock species. Vaccines based on inactivated FMDV virions provide a useful tool for the control of this pathogen. However, long term storage at 4°C (the temperature for vaccine storage) or ruptures of the cold chain, provoke the dissociation of virions, reducing the immunogenicity of the vaccine. An FMDV mutant carrying amino acid replacements VP1 N17D and VP2 H145Y isolated previously rendered virions with increased resistance to dissociation at 4°C. We have evaluated the immunogenicity in swine (a natural FMDV host) of a chemically inactivated vaccine based on this mutant. The presence of these amino acid substitutions did not compromise the immunological potential, including its ability to elicit neutralizing antibodies. These results support the feasibility of this kind of mutants with increased capsid stability as suitable viruses for producing improved FMDV vaccines.


Subject(s)
Antibodies, Viral/immunology , Capsid Proteins/immunology , Foot-and-Mouth Disease Virus/immunology , Foot-and-Mouth Disease/prevention & control , Swine Diseases/prevention & control , Viral Vaccines/immunology , Amino Acid Substitution , Animals , Antibodies, Neutralizing , Capsid Proteins/genetics , Foot-and-Mouth Disease/virology , Foot-and-Mouth Disease Virus/genetics , Models, Molecular , Mutation , Swine , Swine Diseases/virology , Vaccines, Inactivated/immunology , Virion
19.
Prog Lipid Res ; 64: 123-137, 2016 10.
Article in English | MEDLINE | ID: mdl-27702593

ABSTRACT

Flaviviruses are emerging arthropod-borne pathogens that cause life-threatening diseases such as yellow fever, dengue, West Nile encephalitis, tick-borne encephalitis, Kyasanur Forest disease, tick-borne encephalitis, or Zika disease. This viral genus groups >50 viral species of small enveloped plus strand RNA virus that are phylogenetically closely related to hepatitis C virus. Importantly, the flavivirus life cycle is intimately associated to host cell lipids. Along this line, flaviviruses rearrange intracellular membranes from the endoplasmic-reticulum of the infected cells to develop adequate platforms for viral replication and particle biogenesis. Moreover, flaviviruses dramatically orchestrate a profound reorganization of the host cell lipid metabolism to create a favorable environment for viral multiplication. Consistently, recent work has shown the importance of specific lipid classes in flavivirus infections. For instances, fatty acid synthesis is linked to viral replication, phosphatidylserine and phosphatidylethanolamine are involved on the entry of flaviviruses, sphingolipids (ceramide and sphingomyelin) play a key role on virus assembly and pathogenesis, and cholesterol is essential for innate immunity evasion in flavivirus-infected cells. Here, we revise the current knowledge on the interactions of the flaviviruses with the cellular lipid metabolism to identify potential targets for future antiviral development aimed to combat these relevant health-threatening pathogens.


Subject(s)
Flavivirus/metabolism , Lipid Metabolism/physiology , Animals , Dengue Virus/metabolism , Dengue Virus/pathogenicity , Encephalitis/metabolism , Encephalitis/pathology , Encephalitis/virology , Flavivirus/pathogenicity , Flavivirus Infections/metabolism , Flavivirus Infections/pathology , Flavivirus Infections/virology , Humans , Phosphatidylethanolamines/metabolism , Phosphatidylserines/metabolism , Sphingolipids/metabolism , West Nile virus/metabolism , West Nile virus/pathogenicity , Zika Virus/metabolism , Zika Virus/pathogenicity
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