Study on Non-Limited Earth Pressures of Soilbag-Reinforced Retaining Structures with Surcharge Loads.

The earth pressure acting on soilbag-reinforced retaining structures subjected to surcharge loads under non-limited states is crucial for designing these structures. In this study, mode tests on soilbag-reinforced retaining walls were conducted to the earth pressure of the wall subjected to surcharge loads. The findings from these tests reveal a non-linear distribution of lateral earth pressure on the wall when subjected to surcharge loads in non-limited states, with an observed escalation in pressure corresponding to increased surcharge loads. Insights from the tests facilitated the development of a predictive method for estimating lateral pressure on soilbag-reinforced retaining walls under similar conditions, and its performance was fully validated by the model tests. Furthermore, the impact of the geometric dimensions and material properties of the soilbags on the earth pressure distribution was examined using the proposed method.

Detection and genetic evolution analysis of porcine parvovirus type 7 (PPV7) in Fujian Province.

Porcine parvovirus (PPV) is an important pathogen causing reproductive disorders in sows, with clinical symptoms including stillbirth, mummified fetuses, embryonic dysplasia and death, and sow infertility. Porcine parvovirus 7 (PPV7) is a recently discovered type of PPV and its widespread distribution and rapid evolution has caused huge economic losses in the pig industry. To investigate the molecular epidemiology of PPV7 in Fujian Province, China, we collected 491 blood samples and 72 tissue samples from diseased pigs in large-scale pig farms across selected areas of Fujian Province from 2019 to 2022. PPV7 infection was determined using real-time quantitative PCR, and positive samples underwent whole-genome amplification, sequencing, and subsequent homology, phylogenetic, and recombination analyses. The PPV7 positive detection rate was 25.73% (145/563) in Fujian Province, among which the positive rate of blood and tissue samples was 26.47% (130/491) and 20.83% (15/72), respectively. The nucleotide sequence homology among the 29 PPV7 whole-genome sequences obtained in this study was 90.0%-97.2%, whereas that with 128 reference strains from China and other countries was 88.9%-98.1%. Six strains had partial nucleotide deletions or insertions. Phylogenetic analysis based on the whole-genome sequences classified the 29 PPV7 strains and 128 reference strains into eight subtypes (PPV7a-PPV7h), and PPV7h was the predominant subtype in Fujian Province. Recombination analysis revealed evidence of inferred recombination events in the genomes of four strains. This study provides significant insights into the molecular characteristics of PPV7 in Fujian Province and serves as a crucial foundation for further advancements in PPV7 prevention and control strategies.

In Vitro, In Vivo, and In Silico Activities of Ginkgolic Acid C15:1 against Streptococcus agalactiae Clinical Isolates.

Streptococcus agalactiae is the major cause of invasive neonatal infections and is a recognized pathogen associated with various diseases in nonpregnant adults. The emergence and spread of antibiotic-resistant S. agalactiae necessitate the development of a novel antibacterial agent. Here, the potential antibacterial activities and mechanisms of ginkgolic acid C15:1 (GA (15:1)) from Ginkgo biloba against clinical S. agalactiae are characterized. The MIC50 and MIC90 values for GA (15:1) against 72 clinical S. agalactiae isolates were 6.25 and 12.5 µM, respectively. GA (15:1) showed a strong bactericidal effect against both planktonic bacteria and bacteria embedded in biofilms as well as significant effectiveness in suppressing the growth of S. agalactiae biofilms. Moreover, GA (15:1) possesses intracellular antibacterial activity and could significantly decrease the bacterial burden in the intraperitoneal infection model of S. agalactiae. Mechanistic studies showed that GA (15:1) triggers membrane damage of S. agalactiae through a unique dual-targeting mechanism of action (MoA). First, GA (15:1) targets phospholipids in the bacterial cytoplasmic membrane. Second, by using mass-spectrometry-based drug affinity responsive target stability (DARTS) and molecular docking, lipoprotein signaling peptidase II (lspA) was identified as a target protein of GA (15:1), whose role is crucial for maintaining bacterial membrane depolarization and permeabilization. Our findings suggest a potential therapeutic strategy for developing GA (15:1) to combat S. agalactiae infections.

Efficacy of a porcine reproductive and respiratory syndrome virus 1 (PRRSV-1) natural recombinant against a heterologous PRRSV-1 isolate both clustered within the subgroup of BJEU06-1-like isolates.

Porcine reproductive and respiratory syndrome virus 1 (PRRSV-1) has been prevalent in more than 20 provinces of China. However, no PRRSV-1-specific vaccine is commercially available in China. To evaluate the feasibility of using a low virulent PRRSV-1 isolate against potential outbreaks caused by virulent Chinese PRRSV-1 isolates, here we evaluated the efficacy of a low virulent PRRSV-1 HLJB1 strain isolated in 2014 as live vaccine against a virulent PRRSV-1 SD1291 strain isolated in 2022. Genome-based phylogenetic analysis showed that both HLJB1 and SD1291 were grouped within BJEU06-1-like isolates. However, they shared only 85.27% genomic similarity. Piglet inoculation and challenge study showed that HLJB1 inoculation could reduce viremia but did not significantly alleviate clinical signs and tissue lesions. Virus neutralization test indicated that HLJB1 inoculation could induce homologous neutralizing antibodies (NAbs) but no heterologous NAbs at 42 dpi. In addition, flow cytometric analyses showed that no memory T follicular helper (Tfh) cells against SD1291 and SD1291-specific IFN-γ secreting cells were induced by HLJB1 pre-inoculation. These results supported that HLJB1 inoculation only provides partial cross-protection against SD1291 infection even though they are clustered within the same PRRSV-1 subgroup, which is closely related to the failure in conferring cross-protective adaptive immune responses.

Virus diversity, wildlife-domestic animal circulation and potential zoonotic viruses of small mammals, pangolins and zoo animals.

Wildlife is reservoir of emerging viruses. Here we identified 27 families of mammalian viruses from 1981 wild animals and 194 zoo animals collected from south China between 2015 and 2022, isolated and characterized the pathogenicity of eight viruses. Bats harbor high diversity of coronaviruses, picornaviruses and astroviruses, and a potentially novel genus of Bornaviridae. In addition to the reported SARSr-CoV-2 and HKU4-CoV-like viruses, picornavirus and respiroviruses also likely circulate between bats and pangolins. Pikas harbor a new clade of Embecovirus and a new genus of arenaviruses. Further, the potential cross-species transmission of RNA viruses (paramyxovirus and astrovirus) and DNA viruses (pseudorabies virus, porcine circovirus 2, porcine circovirus 3 and parvovirus) between wildlife and domestic animals was identified, complicating wildlife protection and the prevention and control of these diseases in domestic animals. This study provides a nuanced view of the frequency of host-jumping events, as well as assessments of zoonotic risk.

Detection and molecular characterisation of intestinal parasites in the South China tiger Panthera tigris amoyensis (Hilzheimer).

Parasitic infections of the South China tigers in the Meihua Mountains have not been explored previously. Faeces of 22 South China tigers from the China Tiger Park in the Meihua Mountains were examined. Eggs of ascaridoid nematodes and oocysts of coccidia were detected by Mini-FLOTAC assay. Morphological observation and molecular characterisation of the oocysts were carried out. The prevalence of Toxascaris leonina (von Linstow, 1902) was 18% (4/22), and the highest egg per gram (EPG) count in the faeces was 27,150. The prevalence of Cystoisospora sp. was 45% (1 0/22) and the highest oocysts per gram (OPG) in the faeces was 6,000. In addition, we found one ascaridoid nematode in the South China tiger's faeces and was molecularly and morphologically identified as T. leonina. The oocysts in the faeces were sporulated in vitro and identified as Cystoisospora sp. Amplification of full-length internal transcribed spacers (ITS) resulted in sequences 1,622 bp long. Using the sequences, Cystoisospora sp. of the South China tiger was closest to Isospora belli (Wenyon, 1923) and Cystoisospora suis (Biester, 1934).

Identification and molecular analysis of Ixodid ticks (Acari: Ixodidae) infesting wild boars (Sus scrofa) and tick-borne pathogens at the Meihua mountain of southwestern Fujian, China.

Wildlife is essential to the biodiversity of the Meihua mountain, southwestern Fujian province, China. However, there have been few surveys of the distribution of ixodid ticks (Acari: Ixodidae) and tick-borne pathogens affecting wild animals at these locations. In this study, 1197 adult ixodid ticks infesting wild boars were collected from 10 sampling sites during 2019. Ticks were identified to species based on morphology, and the identification was confirmed based on mitochondrial 16S, ITS1 and ITS2 rRNA sequences. Eight tick species belonging to 2 genera were identified, including H. longicornis (n = 373, 31.1%), H. flava (n = 265, 22.1%), D. auratus (n = 153, 12.8%), H. hystricis (n = 119, 9.9%), D. silvarum (n = 116, 9.7%), H. bispinosa (n = 114, 9.5%), D. atrosignatus (n = 33, 2.8%), and D. taiwanensis (n = 24, 2.0%). DNA sequences of Rickettsia spp. (spotted fever group) and Babesia spp. were detected in these ticks. Phylogenetic analyses revealed the possible existence of Candidatus Rickettsia laoensis and Rickettsia raoultii. This study illustrates the potential threat to wild animals and humans from tick-borne pathogens.

Codon usage bias in the H gene of canine distemper virus.

Canine distemper virus (CDV), a non-segmented single negative-stranded RNA (ssRNA), is the etiological agent of canine distemper. Canine distemper is a highly contagious and lethal viral disease in domestic dogs and wild carnivores. Study of the evolution of CDV presents an essential key to improve the vaccine efficacy. In this study, a total of 328 full-length CDV hemagglutinin (H) gene sequences were subjected to phylogenetic, amino acid mutations, and codon usage analysis. In accordance with previous study, CDV genotypes consisted of fifteen lineages. The unique amino acid substitution sites in each CDV lineages have been identified for the first time, including America-1 (Q330H), America-2 (I585S), Asia-1 (A359V), Asia-2 (H61R), Asia-3 (P108Q), Asia-4 (K213T), India-1/Asia-5(S497P), Arctic (S20L), Africa-1(N489S), Colombian (V41I), EWL (I44V), Europe (D560E), Europe-1/South America-1(K161Q), South America-2 (R580Q), and East African (S214A). Codon usage analysis indicated that H gene exhibited low codon usage bias and further neutrality plot analysis demonstrated that natural selection played a dominated role in driving CPV evolution. The effective number of codons (ENC) plots show that all the different sequences are below the standard curve, indicating that mutational pressure is not the only factor affecting CUB but other forces, including natural selection. The neutrality analysis showed that the slope of the regression line was 0.1501, indicating natural selection dominates directional mutation pressure in driving the codon usage pattern. In addition, nucleotide composition, relative synonymous codon usage value, dinucleotide content, and geographical distribution have been proven to influence the codon usage bias of the CDV H gene. The novel findings enhanced the understanding of CDV evolution.

Identification of a multiple drug-resistance gene island in the Haemophilus parasuis chromosome.

OBJECTIVES: The aim of this study was to determine the mechanisms and molecular characterisation of one strain (HPS412) of Haemophilus parasuis, which exhibited high MICs of antimicrobial susceptibility. METHODS: A total of 113 H. parasuis strains isolated from pigs suffering from polyserositis, pneumonia or meningitis in China and screened them for antimicrobial susceptibility. Susceptibility testing of the minimal inhibitory concentrations (MIC) was determined in fastidious medium consisting of tryptone soya broth (TSB) containing 5% bovine serum and 10µg/mL NAD in 96-well microtiter plates. The genomic DNA was completely sequenced by combining PacBio RS II and Illumina HiSeq 4000 platforms. Gene prediction was performed using Glimmer v.3.02 with Hidden Markov models. RESULTS: One strain (HPS412) exhibited high MICs of sulfamethoxazole (256µg/mL), tetracycline (128µg/mL), streptomycin (128µg/mL), gentamicin (128µg/mL), amoxicillin (128µg/mL), chloramphenicol (64µg/mL), penicillin (64µg/mL) and cefaclor (64µg/mL). Sequence analysis showed that numerous drug-resistance genes including tet(B), blaROB-1, sul2, catIII, aph(3″)-Ib, aph(6)-Id and aph(3')-Ia were present in a chromosomal gene island as adjacent duplicate copies and the rep-orf3-blaROB-1 structure most likely had a direct plasmid origin. The tet(B) and blaROB-1 were flanked on one or both by ISApl1 elements. CONCLUSIONS: The acquisition of blaROB-1 and the other antibiotic resistance genes was related to the presence of ISApl1. ISApl1 plays important roles in the horizontal transmission of antibiotic resistance genes.

Detection of PCV2e strains in Southeast China.

Porcine circovirus 2 (PCV2) has been prevalent in swine herds in China since 2002, causing severe economic loss to the pig industry. The number of live pigs in southeast China is > 20 million. Since information on the genetic variation of PCV2 in the Fujian province is limited, the objective of the present work was to investigate the epidemiological and evolutionary characteristics of PCV2 in southeast China from 2013 to 2017. Of the 685 samples collected from 90 different swine herds from 2013 to 2017, 356 samples from 84 different swine herds were positive for PCV2. PCV2a, PCV2b, PCV2d, and PCV2e co-existed in the Fujian province, with PCV2d being the predominant circulating strain in swineherds and PCV2e being reported for the first time in China. Strikingly, PCV2-FJ-water DNA comes from contaminated river water and not infected animals. Sequence comparison among all isolates indicated that 95 isolates shared approximately 78.7%-100% nucleotide identity and 74.5%-100% amino acid identity for open reading frame 2 (ORF2). Amino acid alignment showed that the Cap protein of PCV2e differed markedly from those of PCV2a, PCV2b, PCV2c, and PCV2d. These results indicated that various PCV2 genotypes exist in China, and that PCV2 is continuously evolving, leading to rapid emergence of new variant stains.

Complete genomic characterization of two European-genotype porcine reproductive and respiratory syndrome virus isolates in Fujian province of China.

Porcine reproductive and respiratory syndrome (PRRS) is considered one of the most devastating swine diseases worldwide, resulting in immense economic losses. PRRS virus (PRRSV) is divided into two major genotypes, European (type 1) and the North American (type 2). Type 1 PRRSV have recently emerged in Fujian province (South China), and this might have a significant impact on the Chinese pig industry. From 2013 to 2014, two type 1 PRRSV strains, named FJEU13 and FJQEU14, were isolated from piglets and sows with respiratory problems and reproductive disorders in Fujian province. The full genome length of the two isolates was 14,869-15,062 nucleotides (nt), excluding the poly(A) tail. These isolates shared 86.0-89.9% sequence identity with the prototypic strains Lelystad virus (LV) and 82.8-92% with Chinese type 1 PRRSV strains, but only 59.9-60.1% with the North American reference strain VR-2332. However, they were 82.9% identical to each other. Nonstructural protein 2 (Nsp2) and ORF3-ORF5 were the most variable regions when compared to other type 1 PRRSV strains. Nsp2 and ORF3 contained multiple discontinuous deletions and a 204-bp deletion in NSP2 in isolate FJQEU14, which has never been described in other Chinese type 1 PRRSV strains. All of these results might be useful for understanding the epidemic status of type 1 PRRSV in China.

Role of Itk signalling in the interaction between influenza A virus and T-cells.

Although the T-cell-mediated immune response to influenza virus has been studied extensively, little information is available on the direct interaction between influenza virus and T-cells that pertains to severe diseases in humans and animals. To address these issues, we utilized the BALB/c mouse model combined with primary T-cells infected with A/WSN/33 influenza virus to investigate whether influenza virus has an affinity for T-cells in vivo. We observed that small proportions of CD4(+) T-cells and CD8(+) T-cells in spleen and thymus expressed viral proteins in infected mice. A significant proportion of mouse primary T-cells displayed expression of α-2,6 sialic acid-linked influenza virus receptor and were infected directly by influenza A virus. These experiments reveal that there exists a population of T-cells that is susceptible to influenza A virus infection. Furthermore, we employed human Jurkat T-cells to investigate the virus-T-cell interaction, with particular emphasis on understanding whether Itk (interleukin-2-inducible T-cell kinase), a Tec family tyrosine kinase that regulates T-cell activation, is involved in virus infection of T-cells. Interestingly, influenza virus infection resulted in an increased recruitment of Itk to the plasma membrane and an increased level of phospholipase C-γ1 (PLC-γ1) phosphorylation, suggesting that Itk/PLC-γ1 signalling is activated by the virus infection. We demonstrated that depletion of Itk inhibited the replication of influenza A virus, whereas overexpression of Itk increased virus replication. These results indicate that Itk is required for efficient replication of influenza virus in infected T-cells.