Newly emerged sublineage 1.5 (NADC34-like) porcine reproductive and respiratory syndrome virus (PRRSV) has posed a direct threat to the Chinese pig industry since 2018. However, the prevalence and impact of NADC34-like PRRSV on Chinese pig farms is unclear. In the present study, we continuously monitored pathogens, including PRRSV, African swine fever virus (AFSV), classical swine fever (CSFV), pseudorabies virus (PRV) and porcine circovirus 2 (PCV2), on seven fattening pig farms with strict biosecurity practices located in five provinces of China from 2020 to 2021. The results showed that multiple types of PRRSVs commonly coexisted on a single pig farm. NADC30-like PRRSV was the predominant strain on most pig farms. Importantly, NADC34-like PRRSV, detected during the period of peak mortality, was one of the predominant strains on one pig farm in northern China. Sequence alignment suggested that these strains shared the same 100-aa deletion in the Nsp2 protein as IA/2014/NADC34 isolated from the United States (U.S.) in 2014. Phylogenetic analysis based on open reading frame 5 (ORF5) showed that the genetic diversity of NADC34-like PRRSV on this farm was relatively singular, but it had a relatively high rate of evolution. Restriction fragment polymorphism pattern (RFLP) analysis showed that almost all ORF5 RFLPs were 1-7-4, with one 1-4-4. In addition, two complete genomes of NADC34-like PRRSVs were sequenced. Recombination analysis and sequence alignment demonstrated that both viruses, with 98.9% nucleotide similarity, were nonrecombinant viruses. This study reports the prevalence and characteristics of NADC34-like PRRSV on a large-scale breeding farm in northern China for the first time. These results will help reveal the impact of NADC34-like PRRSV on Chinese pig farms and provide a reference for detection and further prevention and control of NADC34-like PRRSV.

Porcine reproductive and respiratory syndrome virus 1 (PRRSV-1) is one of the main pathogens causing porcine reproductive and respiratory syndrome (PRRS). In recent years, the detection number of PRRSV-1 in China has gradually increased, and the PRRSV-1 strains reported in China belong to subtype Ⅰ (Global; Clade A-L). In the present study, a novel PRRSV-1 strain, TZJ2134, was found during epidemiological surveillance of PRRSV-1 in Shandong Province in China. We obtained two fragments of TZJ2134: TZJ2134-L12 (located at nt 1672-nt 2112 in the partial Nsp2 gene) and TZJ2134-(A+B) (located at nt 7463-nt 11272 in the partial Nsp9, complete Nsp10 and partial Nsp11 genes). Phylogenetic and recombination analyses based on the two sequences showed that TZJ2134 is a recombinant strain derived from two commercial PRRSV-1 modified live vaccine (MLV) strains (the Amervac vaccine and DV vaccine strains), forming a new recombinant subgroup of DV+Amervac-like isolates with other strains. However, PRRSV-1 MLV is not currently allowed for use in China. We speculated that TZJ2134 might have been introduced from Europe. This study is the first to provide information on two recombinant PRRSV-1 MLV strains in China and provides new data for the epidemiological study of PRRSV-1 in China.

Porcine reproductive and respiratory syndrome virus (PRRSV) is an important pathogen that endangers the swine industry worldwide. Recently, lineage 1 PRRSVs, especially NADC30-like PRRSV, have become the major endemic strains in many pig-breeding countries. Since 2016, NADC30-like PRRSV has become the predominant strain in China. Unfortunately, current commercial vaccines cannot provide sufficient protection against this strain. Here, an attenuated lineage 1 PRRSV strain, named SD-R, was obtained by passaging NADC30-like PRRSV strain SD in Marc-145 cells for 125 passages. Four-week-old PRRSV-free piglets were vaccinated intramuscularly with 10 5.0TCID 50 SD-R and then challenged intramuscularly (2 ml) and intranasally (2 ml) with homologous NADC30-like PRRSV SD (1×10 5.0TCID 50/ml) and heterologous NADC30-like PRRSV HLJWK108-1711 (1×10 5.0TCID 50/ml). The results showed that 5 of 5 immunized piglets had a perceptible humoral immune response to vaccination and did not develop fever or clinical diseases after NADC30-like PRRSV challenge. Additionally, compared to challenge control piglets, vaccinated piglets gained significantly more weight and showed much milder pathological lesions. Furthermore, the viral replication levels of the immunized group were significantly lower than those of the challenge control group. These results demonstrate, for the first time, that lineage 1 PRRSV SD-R is a good candidate for an efficacious vaccine against NADC30-like PRRSVs.

NADC34-like PRRSV strains were first detected in China in 2017, with epidemic potential. In this study, the phylogenetic, epidemic, and recombinant properties of NADC34-like PRRSV in China were evaluated comprehensively. From 2020 to October 2021, 82 NADC34-like PRRSV isolates were obtained from 433 PRRSV-positive clinical samples. These strains accounted for 11.5% and 28.6% of positives in 2020 and 2021, respectively, and have spread to eight provinces. We selected 15 samples for whole-genome sequencing, revealing genome lengths of 15,009 to 15,113 nt. Phylogenetic analysis revealed that Chinese NADC34-like strains cluster with American sublineage 1.5 strains and do not form an independent branch. Recombination analysis revealed that six of fifteen complete genome sequences derived from recombination between NADC34-like and NADC30-like or HP-PRRSV; they all recombined with local strains in China, exhibiting a complex recombination pattern. Partial Nsp2 sequence alignment showed that nine of fifteen isolates have a continuous 100-aa deletion (similar to IA/2014/NADC34); other isolates have a 131-aa discontinuity deletion (similar to NADC30). Five of them also have additional amino acid deletions, all of which are reported for the first time here. In the last two years, NADC34-like PRRSV has become one of the main epidemic strains in some areas of China; it has changed significantly, its homology has decreased significantly, and it has undergone complex recombination with local Chinese strains. These results are of great significance for understanding the current epidemic situation of PRRSV in China.