A2 Strain Research Articles

Hop-like alfalfa as an object of introduction into culture

It was found that the most promising strains of nodule bacteria XM1 and 412b are the most promising for inoculation of the Mira variety, the former increased the collection of dry matter by 96%, the latter by 81%, seeds by 115 and 73%. Strain XM1 was isolated from the nodules of the same wild population of alfalfa in the hop-like Moscow region from which the variety Mira was formed by the method of chemical mutagenesis. Strains 412b and XM 1 can be recommended for pre-sowing inoculation of the Mira variety when cultivated for feed and seeds, and the XM6 strain - when cultivated for seeds. The latter increases the seed yield by 84%, and the green mass by only 31%.Strains XM2, XM5 and XM6 shift the metabolism of the alfalfa-rhizobial system towards an increase in the number of seeds. The proportion of seeds increases to 24-31% of the mass of the entire plant, in other symbiotic systems this figure is 19-22%.A promising breeding number of hop alfalfa VIC 26 has been created, which is 58% more productive than the Mira variety when grown in the traditional way and by 65% when inoculated with the A2 strain.

Nuclear-localized human respiratory syncytial virus NS1 protein modulates host gene transcription.

SUMMARYHuman respiratory syncytial virus (RSV) is a common cause of lower respiratory tract infections in the pediatric, elderly, and immunocompromised individuals. RSV non-structural protein NS1 is a known cytosolic immune antagonist, but how NS1 modulates host responses remains poorly defined. Here, we observe NS1 partitioning into the nucleus of RSV-infected cells, including the human airway epithelium. Nuclear NS1 coimmunoprecipitates with Mediator complex and is chromatin associated. Chromatin-immunoprecipitation demonstrates enrichment of NS1 that overlaps Mediator and transcription factor binding within the promoters and enhancers of differentially expressed genes during RSV infection. Mutation of the NS1 C-terminal helix reduces NS1 impact on host gene expression. These data suggest that nuclear NS1 alters host responses to RSV infection by binding at regulatory elements of immune response genes and modulating host gene transcription. Our study identifies another layer of regulation by virally encoded proteins that shapes host response and impacts immunity to RSV.

Immunogenicity and Protective Ability of Genotype I-Based Recombinant Japanese Encephalitis Virus (JEV) with Attenuation Mutations in E Protein against Genotype V JEV.

Genotype V (GV) Japanese encephalitis virus (JEV) has emerged in Korea and China since 2009. Recent findings suggest that current Japanese encephalitis (JE) vaccines may reduce the ability to induce neutralizing antibodies against GV JEV compared to other genotypes. This study sought to produce a novel live attenuated JE vaccine with a high efficacy against GV JEV. Genotype I (GI)-GV intertypic recombinant strain rJEV-EXZ0934-M41 (EXZ0934), in which the E region of the GI Mie/41/2002 strain was replaced with that of GV strain XZ0934, was introduced with the same 10 attenuation substitutions in the E region found in the live attenuated JE vaccine strain SA 14-14-2 to produce a novel mutant virus rJEV-EXZ/SA14142m-M41 (EXZ/SA14142m). In addition, another mutant rJEV-EM41/SA14142m-M41 (EM41/SA14142m), which has the same substitutions in the Mie/41/2002, was also produced. The neuroinvasiveness and neurovirulence of the two mutant viruses were significantly reduced in mice. The mutant viruses induced neutralizing antibodies against GV JEV in mice. The growth of EXZ/SA14142m was lower than that of EM41/SA14142m. In mouse challenge tests, a single inoculation with a high dose of the mutants blocked lethal GV JEV infections; however, the protective efficacy of EXZ/SA14142m was weaker than that of EM41/SA14142m in low-dose inoculations. The lower protection potency of EXZ/SA14142m may be ascribed to the reduced growth ability caused by the attenuation mutations.

Genotyping of Brucella strains isolated from humans and cattle of different geographical regions of Pakistan using MLVA‐15

BackgroundPrevalence of brucellosis and MLVA genotyping in animals and humans, isolated from different regions of Pakistan was performed. Animals having history of brucellosis from the field and local farms of Bannu, Mardan, Peshawar, Swat, Lahore and Islamabad were selected for blood collection. Humans that work with them were also selected for sampling in this study. Total of 600 samples were taken from cattle and humans and subjected to Rose Bengal plate Test (RBPT) for the initial screening of positive samples. Designed primers of B.abortus for cattle and B.melitensis for humans were utilised to perform PCR. Culturing and isolation was carried to further to perform MLVA genotyping assay through the selection of two panels of primer markers.ResultsRBPT showed more number of cases of brucellosis in animals and humans compared to the PCR findings. Genotyping findings based upon MLVA‐15 set of markers demonstrated that the isolated strains of B.abortus fall in the same clade with strain A1, P8 and A2 from Pakistan and also similar with BCCN#02‐45 strain from India. On the other hand, B.melitensis isolated from different districts of Pakistan shared the same clade with BwIM‐AFG 63, BwIM‐AFG 44 strains from Afghanistan and BwIM IRN 37 strain from Iran. Selected VNTR alleles were sequenced for calibration purposes.ConclusionIt is concluded that B rucella is prevalent in animals and humans in studied districts of Pakistan. Moreover, A1, P8, BwIM‐AFG 63, BwIM‐AFG 44 and A2 were found the common genotypes in Pakistan.

A safe and effective mucosal RSV vaccine in mice consisting of RSV phosphoprotein and flagellin variant.

Respiratory syncytial virus (RSV) is a major cause of serious acute lower respiratory tract infection in infants and the elderly. The lack of a licensed RSV vaccine calls for the development of vaccines with other targets and vaccination strategies. Here, we construct a recombinant protein, designated P-KFD1, comprising RSV phosphoprotein (P) and the E.-coli-K12-strain-derived flagellin variant KFD1. Intranasal immunization with P-KFD1 inhibits RSV replication in the upper and lower respiratory tract and protects mice against lung disease without vaccine-enhanced disease (VED). The P-specific CD4+ Tcells provoked by P-KFD1 intranasal (i.n.) immunization either reside in or migrate to the respiratory tract and mediate protection against RSV infection. Single-cell RNA sequencing (scRNA-seq) and carboxyfluorescein succinimidyl ester (CFSE)-labeled cell transfer further characterize the Th1 and Th17 responses induced by P-KFD1. Finally, we find that anti-viral protection depends on either interferon-γ (IFN-γ) or interleukin-17A (IL-17A). Collectively, P-KFD1 is a promising safe and effective mucosal vaccine candidate for the prevention of RSV infection.

HALOTOLERANT PHOSPHATE SOLUBILIZING IAA–SYNTHESIZING REPRESENTATIVES OF PANTOEA GENUS PROMOTE PROLINE SYNTHESIS IN WHEAT SEEDLINGS UNDER SALT STRESS

Two halotolerant bacterial strains A1 and A15 of Pantoea genus were isolated from the plants growing in saline soils near the waste banks of Solikamsk technogenic biotope (Perm krai, Russia). Microorganisms were able to produce indole-3-acetic acid (IAA) from L-tryptophan and solubilize insoluble phosphates. Maximum accumulation of IAA occurred at the stationary growth phase. In the experiment with wheat seedlings inoculated with isolated strains under salt stress condition, proline concentrations in fresh weight on day 3 were 3 and 2.4 times higher than the control ones reaching 616 and 485 µg/g for A1 and A15 with 1% NaCl respectively. However the higher values of protein content in the variants with salt were achieved only within 7 days, accounting 113% and 138% of the respective total protein amount in the control with 1% of salt for A1 and A15 respectively.

Enhanced inulin production by hairy root cultures of Cichoriumintybus in response to Pi and Fe starvation.

Cichorium intybus is rich in inulin and has several pharmacological applications. Hairy roots culture is a valuable biotechnological tool used to produce plant secondary metabolites. Agrobacterium rhizogenes-mediated genetic transformation of chicory to hairy roots was investigated using Agrobacterium Strains A4, A13, A7, and ATCC15834. Several hairy roots were tested, from which 17 lines were selected based on their fast-growing characteristics. Results of PCR analysis revealed foreign DNA integration into the selected transgenic hairy root lines. Finally, four Adventitious roots that contained the highest ratio of total sugar to total weight (µg/gr DW), were selected. This study investigated the effects of various levels of minerals and sucrose on the production of inulin in Cichorium hairy root culture. Different levels of sucrose, phosphate (Pi) and Iron (Fe) were evaluated, separately. It was found that an increase in sucrose levels from 3 to 5% could decrease the root growth; however, 60 g/l sucrose remarkably enhanced the inulin production rate in all the examined lines. The highest biomass was achieved by applying 3.75 mM Pi but it ended in the decreasing the inulin content per unit weight. In contrast, the highest inulin accumulation and the lowest amount of biomass were observed in 0.5 mM Pi. Fe starvation caused the biomass decrease and a significant increase in inulin accumulation. Results of this study suggest a successfully optimized culture medium to initiate the induction of Cichorium intybus hairy root cells to produce inulin as a valuable medicinal secondary metabolite.

Development of a respiratory syncytial virus vaccine using human hepatitis B core-based virus-like particles to induce mucosal immunity

BackgroundRespiratory syncytial virus (RSV) is an important viral pathogen responsible for severe infection of the lower respiratory tract in children under the age of 5 years. No vaccines against RSV are currently in clinical use. Vaccine-associated enhanced respiratory disease (ERD) caused by excess Th2 type responses was observed in a clinical trial of formalin-inactivated RSV (FI-RSV) in antigen-naïve infants. Thus, inducing a balanced immune response is a crucial issue in the development of an RSV vaccine. MethodsIn this study, we constructed, expressed, and purified a recombinant RSV vaccine candidate (i.e., HRØ24) containing the two heptad repeat regions and the antigenic sites Ø, II, and IV of the RSV F protein. The RSV vaccine candidate was intranasally administrated to BALB/c and C57BL/6 mice in combination with virus-like particles (VLPs) derived from the core protein of the hepatitis B virus (HBc). Mucosal immunity to HRØ24 was then assessed. ResultsIntranasal administration of HBc VLPs in combination with HRØ24 induced serum IgGs against HRØ24 as well as lung HRØ24-specific sIgAs in both C57BL/6 and BALB/c mouse models. The secretion of IFN-γ from splenocyte re-stimulation and an elevated ratio of serum IgG2a to IgG1 indicated that the immune response induced by the HBc VLPs/HRØ24 mixture was Th1-biased. Weight loss of <5% and no to low eosinophil infiltration was observed in histological analysis of the lung following a challenge with the RSV A2 strain. These results suggest that the HBc VLPs/HRØ24 combination conferred substantial partial protection against RSV-induced illness in mice. ConclusionsLong-term immunity to RSV-induced illness was achieved via intranasal vaccination using a mixture of HBc VLPs and HRØ24 in mouse models.

Genetic Variation in the RSV fusion protein controls strain-dependent inflammasome activation and the development of a pathogenic Th17 response

Abstract Respiratory syncytial virus (RSV) is the leading cause of lower respiratory tract infections in young children worldwide. RSV-induced disease ranges from mild to severe, with severe disease correlating with increased levels of pro-inflammatory cytokines, including the Th17-associated cytokine, IL-17. To understand the viral factors that influence the induction of a pathogenic Th17 response, we compared the inflammatory responses elicited by different RSV strains. In contrast to the commonly used RSV A2 strain, RSV 2–20 infection resulted in increased protein levels of IL-6 and IL-1β, which were required to induce a pathogenic Th17 response. RSV 2–20 infection increased mucus production and airway hyperreactivity indicating strain-specific disease induction. RSV 2–20 and A2 induced similar expression of caspase-1, however, 2–20 infection resulted in increased levels of pro-IL-1β suggesting differential activation of inflammasome signal 1. Increased production of IL-1β following RSV 2–20 infection was dependent on TLR4 expression and signaling, a canonical inflammasome signal 1 activator. A recombinant A2 virus expressing the 2–20 fusion (F) protein induced IL-1β to similar levels as wild-type 2–20 indicating the 2–20 F protein is sufficient to increase pro-inflammatory cytokine production. Reversion of three-point mutations in a region likely to interact with host cells in the RSV A2/2–20 F recombinant virus to the A2 sequence significantly reduced IL-6 and IL-1β protein production. Our results demonstrate that genetic variation in the RSV F protein plays a crucial role in the induction of pro-inflammatory cytokines and modulating disease severity in the animal model and may contribute to differential RSV pathogenesis.

Impacts of vitamin A deficiency on mucosal immunity to respiratory syncytial virus infection

Abstract Human respiratory syncytial virus (RSV) is a leading cause of severe acute lower respiratory tract disease in infants and young children worldwide. Vitamin A (VA) deficiency (VAD) is one of the most prevalent nutrition-related health problems globally. Acute childhood infections have been linked to an increased risk of VAD, while conversely, VAD leads to increased susceptibility to severe respiratory infections. To date, however, the impact of VAD on RSV infection specifically, or on adaptive immune responses in the respiratory tract, remains unexplored. Here, we used the BALB/c mouse model to determine the impact of VAD on RSV infection and lung virus-specific CD8 T cell immunity. Groups of VA sufficient (VAS) and VAD mice were infected with RSV strain A2. On day 8 after infection, antigen-specific CD8 T cell responses were evaluated using tetramer and intracellular cytokine staining and ELISA assays for IFNγ secretion in cell supernatants. Although no changes in morbidity and mortality were observed between groups, CD8 T cells from VAD animals showed significantly impaired production of IFNγ, suggesting changes in the Th1 balance of the lungs in the context of VAD. Future experiments will focus on changes in the innate and adaptive immune systems in the lungs during VAD and downstream effects on the response to RSV infection.

First Draft Genome Sequence Resource of Colletotrichum liriopes Causing Leaf Anthracnose on Ophiopogon japonicus.

Leaf anthracnose of Ophiopogon japonicus is an important disease that can significantly reduce the quality and economic value of this traditional Chinese medicinal plant. The disease is caused by Colletotrichum liriopes, a necrotrophic fungus that belongs to the Glomerellaceae family of the Sordariomycetes class. Here, we present the draft whole-genome sequence of the C. liriopes strain A2 that caused leaf anthracnose on O. japonicus. The assembly consists of 407 contigs with an estimated genome size of 53.1 Mb. Furthermore, we identified 670 carbohydrate-active enzymes, 1,377 secreted proteins, and 60 secondary metabolite gene clusters, which may be associated with the pathogenicity of this pathogen. This genome resource will provide a valuable resource for future research on the pathogenesis of C. liriopes and comparative genome analyses within genus Colletotrichum.

Biological Treatment of Real Textile Effluent Using Aspergillus flavus and Fusarium oxysporium and Their Consortium along with the Evaluation of Their Phytotoxicity.

Twenty-one fungal strains were isolated from dye-contaminated soil; out of them, two fungal strains A2 and G2-1 showed the highest decolorization capacity for real textile effluent and were, hence, identified as Aspergillus flavus and Fusarium oxysporium based on morphological and molecular methods. The highest decolorization percentage of 78.12 ± 2.1% was attained in the biotreatment with fungal consortium followed by A. flavus and F. oxysporium separately with removal percentages of 54.68 ± 1.2% and 52.41 ± 1.0%, respectively. Additionally, ultraviolet-visible spectroscopy of the treated effluent showed that a maximum peak (λmax) of 415 nm was reduced as compared with the control. The indicators of wastewater treatment efficacy, namely total dissolved solids, total suspended solids, conductivity, biological oxygen demand, and chemical oxygen demand with removal percentages of 78.2, 78.4, 58.2, 78.1, and 77.6%, respectively, demonstrated a considerable decrease in values due to fungal consortium treatment. The reduction in peak and mass area along with the appearance of new peaks in GC-MS confirms a successful biodegradation process. The toxicity of treated textile effluents on the seed germination of Vicia faba was decreased as compared with the control. The shoot length after irrigation with effluents treated by the fungal consortium was 15.12 ± 1.01 cm as compared with that treated by tap-water, which was 17.8 ± 0.7 cm. Finally, we recommended the decrease of excessive uses of synthetic dyes and utilized biological approaches for the treatment of real textile effluents to reuse in irrigation of uneaten plants especially with water scarcity worldwide.

Isolation and degradation characteristics of bacteria in black and odorous water

Abstract Bioremediation is one of the treatment technologies for the black-odorous water, and obtaining functional bacteria is the key step to its success. In this study, a number of highly efficient pollutant degrading strains were isolated from the sediment of black-odorous river, and were identified by phenotypic and phylogenetic analysis. The composite bacterial agent J1 was mixed by strains A1, A2, A5 and A7, with a volume ratio of 4:4:2:1. And the final compound bacteria injected into black-odorous water were composed of J1 and NS3, with the volume ratio of 1:1. Optimized degradation conditions of compound bacterium agent were as follows: pH 7.5, DO 2.5 mg/L, temperature 30 °C. Under optimized conditions, add 1% by volume to the black-odorous water for a 50-day experimental operation. Finally, the overlying water ammonia nitrogen, chemical oxygen demand (COD), and total phosphorus has been significantly degraded. The research is expected to contribute to the use of bioremediation methods to repair black-odorous water, and the application of isolates can be carried out in situ for water types similar to pollute waterways.

Evaluation of Antagonism Activity and Control of Vibrio alginolyticus in Artemia Culture Using Mixed Probiotic

Supplementation with mixed probiotic in aquaculture has been proven to benefit the hosts as disease resistance tool. In this study, a mixed probiotic which consisted of three isolated strains (Lysinibacillus fusiformis strain SPS11, A2, and Bacillus megaterium strain I24) was formulated for the in vitro assays against Vibrio alginolyticus and in vivo preliminary study towards Artemia nauplii. These strains showed antagonism activities against V. alginolyticus in in vitro assay. An increase in biofilm formation of this mixed probiotic was observed which indicated that the strains could work synergistically with each other to confer benefits to the hosts. Enrichment of Artemia nauplii with the formulated mixed probiotic was done to investigate its role in enhancing resistance against the V. alginolyticus. Artemia nauplii were cultured in two different concentrations of mixed probiotic (106 and 108 CFU mL-1) and challenged via immersion method. The mixed probiotic at both concentrations resulted in significantly higher survival of Artemia compared to the challenged group with no probiont added (106 CFU mL-1, 65.00 ± 0.00 % and 108 CFU mL-1, 77.50 ± 3.53 %). Significant reduction of Vibrio loads was observed in Artemia and its culture water supplemented with mixed probiotic at 108 CFU mL-1 whereas there was no reduction of Vibrio at 106 CFU mL-1. This study suggests that the usage of formulated mixed probiotic at high concentration (108 CFU mL-1) as opposed to single-strain probiotic can confer protection against V. alginolyticus infection towards Artemia.

Acetate Triggers Antiviral Response Mediated by RIG-I in Cells from Infants with Respiratory Syncytial Virus Bronchiolitis

Background: Gut microbiota-derived short-chain fatty-acid (SFCA) acetate protects mice againstRSV A2 strain infection by increasing interferon-β production and expression of interferonstimulated genes (ISGs). However, the role of SFCAs in RSV infection using strains isolated from patients is unknown.Methods: We first used RSV clinical strains isolated from infants hospitalized with RSV bronchiolitis to investigate the effects of in vitro acetate treatment in human pulmonary epithelial cells. We next examined whether acetate treatment is beneficial in a mouse model of RSV infection using clinical isolates. We sought to investigate the relationship of gut microbiota and fecal acetate with disease severity among infants hospitalized with RSV bronchiolitis, and whether treating their respiratory epithelial cells with acetate ex-vivo impacts upon viral load and ISG expression. We further treated epithelial cells from SARS-CoV-2 infected patients with acetate.Findings: In vitro pre-treatment of A549 cells with acetate reduced RSV load after infection with clinical isolates and increased the expression of RIG-I and ISG15. Animals treated with acetate intranasally recovered significantly faster, with reduction in the RSV clinical isolates viral load, and increased lung expression of IFNB1 and the RIG-I receptor. Experiments in RIG-I knockout A549 cells demonstrated that the protection relies on RIG-I presence. Gut microbial profile was associated with bronchiolitis severity and with acetate in stool. Increased acetate levels were associated with increasing oxygen saturation at admission, and shorter duration of fever. Ex-vivo treatment of patients’ respiratory cells with acetate reduced RSV load and increased expression of ISGs OAS1 and ISG15, and virus recognition receptors MAVS and RIG-I, but not IFNB1. These acetate effects were not found on cells from SARS-CoV-2 infected patients.Interpretation: Acetate reduces the severity of RSV infection and RSV viral load through modulation of RIG-I expression.Funding: This study was supported by Rio Grande do Sul Research Foundation FAPERGS (FAPERGS/MS/CNPq/SESRS no. 03/2017 - PPSUS 17/2551-0001380-8 and COVID-19 20/2551-0000258-6), CNPq 312504/2017-9 and by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Finance Code 001. P.B., C.V.B. and L.A.B. would like to acknowledge financial support given by CNPq/FAPERGS/CAPES/BNDES to the National Institute of Science and Technology on Tuberculosis (INCT-TB), Brazil [grant numbers: 421703- 2017-2/17-1265-8/14.2.0914.1).Declaration of Interest: The authors declare no competing interests.Ethical Approval: All animal procedures were performed in accordance with protocols approved by CEUA/UNICAMP (protocols 4022-1 and 4599-1).

Effect of chirality of cysteine on the cell growth and photo-fermentative hydrogen production by using acetate as substrate

Cysteine can participate in the essential metabolism of photo-fermentation and plays an important role in biological hydrogen production. However, D-cysteine is usually ignored in current researches despite its potential amounts and unknown effects. This study firstly investigated the photo-fermentative cell growth and hydrogen production with two chiralities of cysteine (L-cysteine and D-cysteine). This and previous studies illustrated proper L-cysteine concentration could improve hydrogen production performance of Rhodopseudomonas sp. nov. strain A7. However, D-cysteine inhibited the cells proliferation and hydrogen production. Groups of initial D-cysteine concentration above 0.3 g/L postponed reactor start-up for several days and obtained little hydrogen production and bacterial biomass. Groups of initial D-cysteine concentration under 0.3 g/L could operate reactor normally in time and obtained about 1000 mL/L hydrogen and 2.0 g/L biomass. Addition time was another key factor, and groups of adding D-cysteine on day 1 influenced little on cells growth but decreased hydrogen production. Other groups with adding time on day 2–5 obtained normal biomass and hydrogen production. Besides, 0.3 g/L functioned as the limit concentration in different initial biomass concentrations for D-cysteine. This work concluded whether the cells could grow and produce hydrogen depended on the addition time and initial D-cysteine concentrations. D-cysteine determined the duration of adapt phase and was regarded as the quorum sensing factor. Furthermore, although biofilm reactors could improve hydrogen production compared to control reactor, biofilm showed little influence on D-cysteine limit concentration. This study would pave the way for the future photo-fermentative hydrogen production.

Effectsof growth-promoting rhizobacteria on maize growth and rhizosphere microbial community under conservation tillage in Northeast China.

Conservation tillage in conjunction with straw mulching is a sustainable agricultural approach. However, straw mulching reduces the soil temperature, inhibits early maize growth and reduces grain yield in cold regions. To address this problem, we investigated the effects of inoculation of plant growth-promoting rhizobacteria (PGPR) on maize growth and rhizosphere microbial communities under conservation tillage in Northeast China. The PGPR strains Sinorhizobium sp. A15, Bacillus sp. A28, Sphingomonas sp. A55 and Enterobacter sp. P24 were isolated from the maize rhizosphere in the same area and inoculated separately. Inoculation of these strains significantly enhanced maize growth, and the strains A15, A28 and A55 significantly increased grain yield by as much as 22%-29%. Real-time quantitative PCR and high-throughput sequencing showed that separate inoculation with the four strains increased the abundance and species richness of bacteria in the maize rhizosphere. Notably, the relative abundance of Acidobacteria_Subgroup_6, Chloroflexi_KD4-96, and Verrucomicrobiae at the class level and Mucilaginibacter at the genus level were positively correlated with maize biomass and yield. Inoculation with PGPR shows potential for improvement of maize production under conservation tillage in cold regions by regulating the rhizosphere bacterial community structure and by direct stimulation of plant growth.

English

In vitro antiviral activity of three ellagitannins - castalagin, vescalagin and grandinin was investigated against replication of three of the most common herpes viruses infecting animals, namely bovine herpesvirus-1 (BoHV-1), suid alphaherpesvirus-1 (SuHV-1), and caprine herpesvirus-1 (CapHV-1). The cytopathic effect inhibition test via the neutral red uptake assay in Madin-Darby bovine kidney (MDBK) cell culture was applied. The highest activity was demonstrated by castalagin followed by vescalagin against replication of SuHV-1, strain A2, with selectivity index values of 336.8 and 309, respectively, which are close to that of acyclovir (SI = 540). The activity of grandinin against the replication of SuHV-1 A2 strain (SI = 40.8) as well as the activity of all three substances against the replication of BoHV-1 strain K22 was lower but still significant (castalagin SI = 45; vescalagin SI = 42.5; grandinin SI = 32.3). Against the E/CH strain of CapHV-1, the three ellagitannins showed a moderate to weak activity: Castalagin SI = 19.3, vescalagin SI = 18.8, grandinin SI = 11.8. The results obtained characterize compounds tested as perspective antivirals. Key words: bovine herpesvirus-1, suid alphaherpesvirus-1, caprine herpesvirus-1, ellagitannins, alpha acyclovir, antiviral activity.

Phylogenetic characterization and screening of halophilic bacteria from Algerian salt lake for the production of biosurfactant and enzymes

Environments containing significant concentration of NaCl such as salt lakes harbor extremophiles microorganisms which have a great biotechnology interest. To explore the diversity of Bacteria in Chott Tinsilt (Algeria), an isolation program was performed. Water samples were collected from the saltern during the pre-salt harvesting phase. This Chott is high in salt (22.47% (w/v). Seven halophiles Bacteria were selected for further characterization. The isolated strains were able to grow optimally in media with 10–25% (w/v) total salts. Molecular identification of the isolates was performed by sequencing the 16S rRNA gene. It showed that these cultured isolates included members belonging to the Halomonas, Staphylococcus, Salinivibrio, Planococcus and Halobacillus genera with less than 98% of similarity with their closest phylogenetic relative. The halophilic bacterial isolates were also characterized for the production of biosurfactant and industrially important enzymes. Most isolates produced hydrolases and biosurfactants at high salt concentration. In fact, this is the first report on bacterial strains (A4 and B4) which were a good biosurfactant and coagulase producer at 20% and 25% ((w/v)) NaCl. In addition, the biosurfactant produced by the strain B4 at high salinity (25%) was also stable at high temperature (30-100°C) and high alkalinity (pH 11).

First molecular detection and complete sequence analysis of porcine circovirus type 3 (PCV3) in Peninsular Malaysia.

Porcine circovirus type 3 (PCV3) is a newly emerging virus in the swine industry, first reported recently in 2016. PCV3 assembles into a 2000 bp circular genome; slightly larger than PCV1 (1758–1760 bp), PCV2 (1766–1769 bp) and PCV4 (1770 bp). Apart from being associated with porcine dermatitis and nephropathy syndrome (PDNS), PCV3 has been isolated from pigs with clinical signs of reproductive failures, myocarditis, porcine respiratory disease complex (PRDC) and neurologic disease. Given that PCV3 is increasingly reported in countries including Thailand and U.S. with whom Malaysia shares trade and geographical relationship; and that PCV3 is associated with several clinical presentations that affect productivity, there is a need to study the presence and molecular characteristics of PCV3 in Malaysian swine farms. Twenty-four commercial swine farms, three abattoirs and retail shops in Peninsular Malaysia were sampled using convenience sampling method. A total of 281 samples from 141 pigs, including 49 lung archive samples were tested for PCV3 by conventional PCR. Twenty-eight lung samples from wild boar population in Peninsular Malaysia were also included. Nucleotide sequences were analyzed for maximum likelihood phylogeny relationship and pairwise distances. Results revealed that PCV3 is present in Peninsular Malaysia at a molecular prevalence of 17.02%, with inguinal lymph nodes and lungs showing the highest molecular detection rates of 81.82% and 71.43% respectively. Despite wide reports of PCV3 in healthy animals and wild boars, no positive samples were detected in clinically healthy finishers and wild boar population of this study. PCV3 strain A1 and A2 were present in Malaysia, and Malaysian PCV3 strains were found to be phylogenetically related to Spanish, U.S. and Mexico strains.