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Additional file 1. Appendix A. Supplementary data.

An autonomous adaptive MPC architecture is presented for control of heating, ventilation and air condition (HVAC) systems to maintain indoor temperature while reducing energy use. Although equipment use and occupant changes with time, existing MPC methods are not capable of automatically relearning models and computing control decisions reliably for extended periods without intervention from a human expert. We seek to address this weakness. Two major features are embedded in the proposed architecture to enable autonomy: (i) a system identification algorithm from our prior work that periodically re-learns building dynamics and unmeasured internal heat loads from data without requiring re-tuning by experts. The estimated model is guaranteed to be stable and has desirable physical properties irrespective of the data; (ii) an MPC planner with a convex approximation of the original nonconvex problem. The planner uses a descent and convergent method, with the underlying optimization problem being feasible and convex. A year long simulation with a realistic plant shows that both of the features of the proposed architecture - periodic model and disturbance update and convexification of the planning problem - are essential to get the performance improvement over a commonly used baseline controller. Without these features, though MPC can outperform the baseline controller in certain situations, the benefits may not be substantial enough to warrant the investment in MPC., Comment: 12 pages, 7 figures

Our present study aimed to identify host cell proteins that may interact with avian reovirus (ARV) σA protein and their potential effect on ARV replication. The ARV structural protein σA has been demonstrated to suppress interferon production and confirmed to activate the PI3K/Akt pathway. However, host cell factors interacting with σA to affect ARV replication remain unknown. In current study, a cDNA library of chicken embryo fibroblasts (CEFs) was constructed, and host cell proteins interacting with σA were screened by a yeast two-hybrid system. We identified four candidate cellular proteins that interact with ARV σA protein. Among them, Gallus NME/NM23 nucleoside diphosphate kinase 2 (NME2) was further validated as a σA-binding protein through co-immunoprecipitation. The key interaction domain was identified at amino acids (aa) 121-416 in NME2 and at aa 71-139 in σA, respectively. We demonstrated that overexpression of NME2 substantially inhibited ARV replication. In addition silencing NME2 by small interfering RNAs (siRNAs) resulted in marked enhancement of ARV replication. Our work has demonstrated that NME2 is a σA-binding protein that may affect ARV replication in CEF cells.

An electrochemical immunoassay for the ultrasensitive detection of Newcastle disease virus (NDV) was developed using graphene and chitosan-conjugated Cu(I)/Cu(II) (Cu(I)/Cu(II)-Chi-Gra) for signal amplification. Graphene (Gra) was used for both the conjugation of an anti-Newcastle disease virus monoclonal antibody (MAb/NDV) and the immobilization of anti-Newcastle disease virus polyclonal antibodies (PAb/NDV). Cu(I)/Cu(II) was selected as an electroactive probe, immobilized on a chitosan-graphene (Chi-Gra) hybrid material, and detected by differential pulse voltammetry (DPV) after a sandwich-type immune response. Because Gra had a large surface area, many antibodies were loaded onto the electrochemical immunosensor to effectively increase the electrical signal. Additionally, the introduction of Gra significantly increased the loading amount of electroactive probes (Cu(I)/Cu(II)), and the electrical signal was further amplified. Cu(I)/Cu(II) and Cu(I)/Cu(II)-Chi-Gra were compared in detail to characterize the signal amplification ability of this platform. The results showed that this immunosensor exhibited excellent analytical performance in the detection of NDV in the concentration range of 100.13 to 105.13 EID50/0.1 mL, and it had a detection limit of 100.68 EID50/0.1 mL, which was calculated based on a signal-to-noise (S/N) ratio of 3. The resulting immunosensor also exhibited high sensitivity, good reproducibility and acceptable stability.

The H3 subtype of avian influenza virus (AIV) is widespread in avian species and is frequently isolated in surveillance projects; thus, we have developed a more effective diagnostic approach of a monoclonal antibody (mAb)-based sandwich ELISA for the H3 AIV detection. First, we have produced the essential reagent of mAb against AIV H3 strains with the development of an mAb-Mouse immunization with a purified H3-subtype AIV strain and cell fusion to generate hybridoma cells. These cells were screened with hemagglutination inhibition (HI) tests, and optimal cells were subcloned. We chose a hybridoma cell line that steadily secreted a specific H3-subtype AIV mAb, designated 9F12, that belongs to the IgG1 subclass and has a K-type light chain. 9F12 was shown to bind specifically to the H3-subtype AIV antigen by both immunofluorescence assay and Western blot analysis. Finally, a 9F12-based sandwich ELISA was successfully developed and used to specifically test for this antigen. The sandwich ELISA conditions were optimized, and the specificity and sensitivity were validated. The results for clinical sample detection were consistent with viral isolation. Consequently, the 9F12-based sandwich ELISA is a specific, sensitive, robust, rapid and versatile diagnostic tool for H3-subtype AIV and provides a promising strategy for effective influenza virus prevention and control.

The aim of the current study was to determine the genomic sequence of parvovirus strain GX-Tu-PV-1, which was isolated from a turkey in Guangxi Province, South China. The analysis showed that the genome sequence of GX-Tu-PV-1 was 81.3% to ∼99.3% similar to those of other turkey parvoviruses (TuPVs) and 79.8% to ∼92.1% related to chicken parvovirus (ChPV). This study will help in understanding the epidemiology and molecular characteristics of parvovirus in turkeys.

Immunochromatography test strips that use metal particles constructed from Au, Fe3O4, and Au/Fe3O4 nanoparticles were developed for the rapid detection of avian influenza virus subtype H7 (AIV H7). The principle of this immunochromatography test strip was based on a sandwich immunoreaction in which AIV H7 antigens bind specifically to their corresponding antibodies on a nitrocellulose membrane. An antibody–metal (Au, Fe3O4 or Au/Fe3O4) nanoparticle conjugate was used as a label and coated onto a glass fiber membrane, which was used as a conjugate pad. To create a test and a control zone, an anti-H7 polyclonal antibody and an anti-IgG antibody were immobilized onto the nitrocellulose membrane, respectively. Positive samples displayed brown/red lines in the test and control zones of the nitrocellulose membrane, whereas negative samples resulted in a brown/red line only in the control zone. The limit of detection (LOD) of the Au/Fe3O4 nanoparticle-based immunochromatography test strips was found to be 103.5 EID50 (EID50: 50% Egg Infective Dose), which could be visually detected by the naked eye within 15 min. In addition, 200 clinical samples were tested using the Au/Fe3O4 nanoparticle-based immunochromatography test strip to estimate its performance, and seven were positive for AIV H7. In summary, the Au/Fe3O4 nanoparticle-based immunochromatography test strip offers a simple and cost-effective tool for the rapid detection of AIV H7.

Sequence analysis of duck plague virus (DPV) revealed that there was a 528bp (B fragment) deletion within the UL2 gene of DPV attenuated vaccine strain in comparison with field virulent strains. The finding of gene deletion provides a potential differentiation test between DPV virulent strain and attenuated strain based on their UL2 gene sizes. Thus we developed a polymerase chain reaction (PCR) assay targeting to the DPV UL2 gene for simultaneous detection of DPV virulent strain and attenuated strain, 827bp for virulent strain and 299bp for attenuated strain. This newly developed PCR for DPV was highly sensitive and specific. It detected as low as 100fg of DNA on both DPV virulent and attenuated strains, no same size bands were amplified from other duck viruses including duck paramyxovirus, duck tembusu virus, duck circovirus, Muscovy duck parvovirus, duck hepatitis virus type I, avian influenza virus and gosling plague virus. Therefore, this PCR assay can be used for the rapid, sensitive and specific detection of DPV virulent and attenuated strains affecting ducks.

To obtain superior performance in adhesion, polyvinyl chloride (PVC) substrates were modified by excimer laser direct writing with different operating wavelengths, scanning speeds, and laser fluences. The induced morphological and electrochemical changes were detailedly tested and analyzed. Microchannels were formed on the surfaces of the PVC substrates due to the laser ablation, where the melted-resolidified droplet-like structures were distributed uniformly and can significantly improve the mechanical interlock. Furthermore, according to the Fourier transform infrared spectroscopy and x-ray photoelectron spectroscopy analyses, Lewis bases such as hydroxyl and carbonyl were formed after laser treatment, which is beneficial to the adhesion strength. These mechanical and chemical modifications may play positive roles in enhancing the bonding strength of the PVC edge bandings.

Recent studies have demonstrated that at least eight subtypes of avian influenza virus (AIV) can infect humans, including H1, H2, H3, H5, H6, H7, H9 and H10. A GeXP analyser-based multiplex reverse transcription (RT)-PCR (GeXP-multiplex RT-PCR) assay was developed in our recent studies to simultaneously detect these eight AIV subtypes using the haemagglutinin (HA) gene. The assay consists of chimeric primer-based PCR amplification with fluorescent labelling and capillary electrophoresis separation. RNA was extracted from chick embryo allantoic fluid or liquid cultures of viral isolates. In addition, RNA synthesised via in vitro transcription was used to determine the specificity and sensitivity of the assay. After selecting the primer pairs, their concentrations and GeXP-multiplex RT-PCR conditions were optimised. The established GeXP-multiplex RT-PCR assay can detect as few as 100 copies of premixed RNA templates. In the present study, 120 clinical specimens collected from domestic poultry at live bird markets and from wild birds were used to evaluate the performance of the assay. The GeXP-multiplex RT-PCR assay specificity was the same as that of conventional RT-PCR. Thus, the GeXP-multiplex RT-PCR assay is a rapid and relatively high-throughput method for detecting and identifying eight AIV subtypes that may infect humans.

Mycoplasma bovis (MB) and bovine herpes virus 1 (BHV-1) are two important pathogens that cause bovine respiratory disease in the beef feedlot and dairy industries. The aim of this study was to develop and validate a duplex fluorescence-based loop-mediated isothermal amplification (DLAMP) assay for simultaneous detection of MB and BHV-1. Two sets of specific primers for each pathogen were designed to target the unique sequences of the MB uvrC gene and the BHV-1 gB gene. The inner primer for BHV-1 was synthesized with the fluorophore FAM at the 5′ end to detect the BHV-1 gB gene, and the inner primer for MB was synthesized with the fluorophore CY5 at the 5′ end to detect the MB uvrC gene. The DLAMP reaction conditions were optimized for rapid and specific detection of MB and BHV-1. The DLAMP assay developed here could specifically detect MB and BHV-1 without cross-reaction with other known non-target bovine pathogens. The sensitivity of this DLAMP assay was as low as 2 × 102 copies for recombinant plasmids containing the MB and BHV-1 target genes. In a detection test of 125 clinical samples, the positive rates for MB, BHV-1 and co-infection were 44.8%, 13.6% and 1.6%, respectively. Furthermore, the sensitivity and specificity of DLAMP were determined as 95%–96.6% and 100%, respectively, of those of field sample detection by the real-time polymerase chain reaction (PCR) assay recommended by the World Organisation for Animal Health. Overall, DLAMP provides a rapid, sensitive and specific assay for the identification of MB and BHV-1 in clinical specimens and for epidemiological surveillance.

Infections with low pathogenic avian influenza viruses (LPAIVs) can be mild or asymptomatic in poultry; however, in humans, LPAIVs can cause severe infections and death, as demonstrated by the H7N9 and H10N8 human infection outbreaks in 2013 in China. In this study, we conducted an epidemiological survey of LPAIVs at live poultry markets (LPMs) in Guangxi Province, Southern China, which is near several Southeast Asian countries. From January 2012 to December 2015, we collected 3,813 swab samples from poultry at LPMs in Guangxi. Viral isolation, hemagglutination inhibition assay and viral sequencing were utilized to identify LPAIVs in the collected samples. Among the samples, 622 (16.3%) were positive for LPAIVs. Six subtypes (H1, H3, H4, H6, H9 and H11) were individually isolated and identified. Of these subtypes, H3, H6 and H9 were predominant in ducks, geese and chickens, respectively. Among the 622 positive samples, 160 (25.7%) contained more than one subtype, and H8, H10, H12, H13, and H16 were identified among them, which highlights the continuous need for enhanced surveillance of AIVs. These results provide detailed information regarding the epidemic situation of LPAIVs in the area, which can aid efforts to prevent and control AIV transmission in humans and animals.

An H4N6 subtype avian influenza virus was isolated from a pockmark duck in southern China in November 2013 and named A/duck/Guangxi/149D24/2013 (H4N6). All eight gene segments of the strain were sequenced. Sequence analysis indicated that this H4N6 virus was a natural reassortant virus. This H4N6 virus has two basic amino acids in the cleavage site of hemagglutinin 1 (HA1) and HA2, and the amino acid motif of cleavage site was PEKASRGLF, which is the typical characteristic of the low-pathogenic avian influenza virus. This study will help understand the epidemiology and molecular characteristics of avian influenza virus in pockmark ducks.

The complete mitochondrial genome sequence of the Luchuan pig was measured by PCR-based methods, primer-walking sequencing, and fragment cloning. The entire mitochondrial genome of the Luchuan pig was identified as a circular molecule consisting of 16,730 bp (GenBank accession number: KP126954). The contents of A, T, C, and G in the mitochondrial genome were found to be 34.65%, 25.80%, 26.19%, and 13.36%, respectively. The complete mitochondrial genome of the Luchuan pig contains a typical structure, including 13 protein-coding genes, two rRNA genes, 22 tRNA genes, and one control region (D-loop region). This complete mitochondrial genome sequence provides essential information in understanding phylogenetic relationships among Sus scrofa domestic mitochondrial genomes.

A previously unidentified chicken parvovirus (ChPV) strain, associated with runting-stunting syndrome (RSS), is now endemic among chickens in China. To explore the genetic diversity of ChPV strains, we determined the first complete genome sequence of a novel ChPV isolate (GX-CH-PV-7) identified in chickens in Guang Xi, China, and showed moderate genome sequence similarity to reference strains. Analysis showed that the viral genome sequence is 86.4 %–93.9 % identical to those of other ChPVs. Genetic and phylogenetic analyses showed that this newly emergent GX-CH-PV-7 is closely related to Gallus gallus enteric parvovirus isolate ChPV 798 from the USA, indicating that they may share a common ancestor. The complete DNA sequence is 4612 bp long with an A+T content of 56.66 %. We determined the first complete genome sequence of a previously unidentified ChPV strain to elucidate its origin and evolutionary status.

The present study was conducted to identify avian reovirus (ARV) proteins that can activate the phosphatidylinositol 3-kinase (PI3K)-dependent Akt pathway. Based on ARV protein amino acid sequence analysis, σA, σNS, μA, μB and μNS were identified as putative proteins capable of mediating PI3K/Akt pathway activation. The recombinant plasmids σA-pcAGEN, σNS-pcAGEN, μA-pcAGEN, μB-pcAGEN and μNS-pcAGEN were constructed and used to transfect Vero cells, and the expression levels of the corresponding genes were quantified by immunofluorescence and Western blot analysis. Phosphorylated Akt (P-Akt) levels in the transfected cells were measured by flow cytometry and Western blot analysis. The results showed that the σA, σNS, μA, μB and μNS genes were expressed in Vero cells. σA-expressing and σNS-expressing cells had higher P-Akt levels than negative control cells, pcAGEN-expressing cells and cells designed to express other proteins (i.e., μA, μB and μNS). Pre-treatment with the PI3K inhibitor LY294002 inhibited Akt phosphorylation in σA- and σNS-expressing cells. These results indicate that the σA and σNS proteins can activate the PI3K/Akt pathway.

Here, we report the complete genome sequence of an H6N6 avian influenza virus (AIV) isolated from a pigeon in Guangxi, southern China, in 2014. The eight RNA segment genes shared a high nucleotide identity (97 to 99%) with H6N6 subtypes of AIV isolated from ducks in the regions around Guangxi Province. The finding of this study will help us understand the ecology and molecular characteristics of H6 avian influenza virus in wild birds in southern China.

Objectives In order to develop a multiplex RT‐PCR assay using the GeXP analyser for the simultaneous detection of four different NA serotypes of H5‐subtype AIVs, effective to control and reduce H5 subtype of avian influenza outbreak. Design Six pairs of primers were designed using conserved and specific sequences of the AIV subtypes H5, N1, N2, N6 and N8 in GenBank. Each gene‐specific primer was fused at the 5′ end to a universal sequence to generate six pairs of chimeric primers, and one pair of universal primers was used for RT‐PCR, and PCR product separation and detection were performed by capillary electrophoresis using the GenomeLab GeXP genetic analysis system. Setting Single and mixed avian pathogen cDNA/DNA templates were employed to evaluate the specificity of a multiplex assay with a GeXP analyser. Corresponding specific DNA products were amplified for each gene, revealing amplification peaks for M, H5, N1, N2, N6 and N8 genes from four different NA subtypes of influenza A H5 virus. Sample A total of 180 cloacal swabs were collected from poultry at live bird markets. Main outcome measures The multiplex PCR assay demonstrated excellent specificity, with each pair of specific primers generating only products corresponding to the target genes and without cross‐amplification with other NA‐subtype influenza viruses or other avian pathogens. Using various premixed ssRNAs containing known AIV target genes, the detection limit for the multiplex assay was determined to be 102 copies/μl. The GeXP assay was further evaluated using 180 clinical specimens and compared with RRT‐PCR (real‐time reverse transcriptase PCR) and virus isolation. Conclusions This GeXP analyser‐based multiplex assay for four different NA subtypes of H5 HPAI viruses is both highly specific and sensitive and can be used as a rapid and direct diagnostic assay for testing clinical samples.

A field strain of H3N6 avian influenza virus (AIV), A/duck/Guangxi/175D12/2014(H3N6), was isolated from a native duck in Guangxi Province, southern China, in 2014. All of the eight AIV gene segments were sequenced, and sequence results revealed that there were 11 amino acid deletions at the NA stalk region. The NA, PB2, and NP genes showed highest homology to H5N6 AIV, and the PA gene showed highest homology to H7N2 AIV. Phylogenetic analysis indicated that the eight AIV gene segments belonged to the Eurasian lineage. These findings provide scientific evidence of possible or potential mutations of H3N6 AIV circulating in waterfowl in southern China.

The objective of this study was to obtain the complete mitochondrial DNA sequence of chukar partridge, and to provide reference data for protection and utilization of these resources of chukar partridge. The complete mitochondrial genome sequence of the China chukar partridge was measured by PCR-based methods and analysed in detail. Our research findings reveal that the entire mitochondrial genome of the chukar partridge is a circular molecule consisting of 16,688 bp (GenBank accession number: KY829450). The contents of A, T, C, and G in the mitochondrial genome were found to be 30.44%, 24.43%, 31.57%, and 13.56%, respectively. The complete mitochondrial genome of the chukar partridge has a typical structure, including 13 protein-coding genes, two rRNA genes, 22 tRNA genes, and one control region (D-loop region). This complete mitochondrial genome sequence provides essential information in understanding phylogenetic relationships among Galliformes mitochondrial genomes.

The H3 subtype of avian influenza virus can provide genes for human influenza virus through gene reassortment, which has raised great concerns about its potential threat to human health. An H3N6 subtype of avian influenza virus was isolated from Guangxi Province, China, in 2009. All eight gene segments of the strain were sequenced. The sequence analysis indicated that this H3N6 virus was a nature reassortant virus. The genome sequences now can be used to understand the epidemiological and molecular characteristics of the H3N6 influenza virus in southern China.

The complete mitochondrial genome sequence of the Cenxi classical three-buff chicken was measured by PCR-based methods and analyzed in detail. Our research findings reveal that the entire mitochondrial genome of the Cenxi classical three-buff chicken is a circular molecule consisting of 16,786 bp (GenBank accession number: KM433666). The contents of A, T, C, and G in the mitochondrial genome were found to be 30.27%, 23.75%, 32.49% and 13.49%, respectively. The complete mitochondrial genome of the Cenxi classical three-buff chicken contains a typical structure, including 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes and 1 control region (D-loop region). This complete mitochondrial genome sequence provides essential information in understanding phylogenetic relationships among Gallus gallus domesticus mitochondrial genomes.

We report the complete genomic sequence of the full Muscovy duck parvovirus (MDPV) strain, designated GX2011-5, isolated from a Muscovy duck in Guangxi Province, China. The complete genomic sequence was 5,132 bp in length and contained two major open reading frames encoding a 1,844-nucleotide (nt) nonstructural protein and a 2,199-nt capsid protein. Comparison of the complete sequence of GX2011-5 with other published sequences of Muscovy duck parvovirus revealed that this strain exhibited 90.4% to 95.1% sequence homology. This report will advance our understanding of the epidemiology and molecular characteristics of MDPV in the Muscovy duck population in Guangxi, China.

We report here two complete genome sequences of porcine circovirus 2. The complete genomes of BH5 and BH6 were amplified and analyzed. Sequence analysis demonstrated that strains BH5 and BH6 belonged to PCV2d and PCV2b, respectively. Knowledge regarding the complete genome sequences of strains BH5 and BH6 will be useful for epidemiological surveillance.

We report here the complete genome sequence of a novel reassortant H4N2 avian influenza virus strain, A/duck/Guangxi/125D17/2012(H4N2) (GX125D17), isolated from a duck in Guangxi Province, China in 2012. We obtained the complete genome sequence of the GX125D17 virus isolation by PCR, cloning, and sequencing. Sequence analysis revealed that this H4N2 virus strain was a novel reassortant avian influenza virus (AIV). Information about the complete genome sequence of the GX125D17 virus strain will be useful for epidemiological studies.

The complete mitochondrial genome sequence of the Rongshui Xiang duck was measured by PCR-based methods. Our research findings reveal that the entire mitochondrial genome of the Rongshui Xiang duck is a circular molecule consisting of 16,605 bp (GenBank accession number: KJ833587). The contents of A, T, C, and G in the mitochondrial genome were found to be 29.20%, 22.20%, 32.82% and 15.79%, respectively, similar to the majority of avian species. The complete mitochondrial genome of the Rongshui Xiang duck contains 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes, and 1 control region. The characteristics of the mitochondrial genome were analyzed in detail. Our complete mitochondrial genome sequence should provide essential information for understanding phylogenetic relationships among duck mitochondrial genomes.

The complete mitochondrial genome sequence of the Xilin small partridge duck was measured by PCR-based methods. Our research findings revealed that the entire mitochondrial genome of the Xilin small partridge duck was 16,604 bp (GenBank accession number: KJ833586). The contents of A, T, C, and G in the mitochondrial genome were 29.20%, 22.19%, 32.82% and 15.79%, respectively, which were similar to the majority of most avian species. The complete mitochondrial genome of the Xilin small partridge duck contains 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes, and 1 control region. Components of the Xilin small partridge duck’s mitochondrial genome were similar to those of other Anatidae in gene arrangement and composition. The complete mitochondrial genome of the Xilin small partridge duck should provide essential information for understanding phylogenetic relationships of duck mitochondrial genome.

We report here the complete genome sequence of a duck Tembusu virus (DTMUV) strain, GX2013H, isolated from a duck from Cheery Valley in the Guangxi Province of southern China in 2013. We obtained the strain GX2013H from a Cheery Valley duck with severely decreased egg production and neurological signs. The genome of GX2013H is 10,990 nucleotides (nt) in length and contains a single open reading frame encoding a putative polyprotein of 3,425 amino acids (aa). A comparison of the complete sequence and the deduced amino acid sequence of GX2013H with published sequences of 15 other chicken anemia viruses from China showed that the homologies of the nucleotides are approximately 96.5% to 97.5% and the homologies of the deduced amino acid sequences are approximately 98.9% to 99.3%. This report will help to understand the epidemiology and molecular characteristics of TMUV in Guangxi.