Isolation and identification of the broad-spectrum high-efficiency phage vB_SalP_LDW16 and its therapeutic application in chickens.

BACKGROUND: Salmonella infection in livestock and poultry causes salmonellosis, and is mainly treated using antibiotics. However, the misuse use of antibiotics often triggers the emergence of multi-drug-resistant Salmonella strains. Currently, Salmonella phages is safe and effective against Salmonella, serving as the best drug of choice. This study involved 16 Salmonella bacteriophages separated and purified from the sewage and the feces of the broiler farm. A phage, vB_SalP_LDW16, was selected based on the phage host range test. The phage vB_SalP_LDW16 was characterized by the double-layer plate method and transmission electron microscopy. Furthermore, the clinical therapeutic effect of phage vB_SalP_LDW16 was verified by using the pathogenic Salmonella Enteritidis in the SPF chicken model. RESULTS: The phage vB_SalP_LDW16 with a wide host range was identified to the family Siphoviridae and the order Caudoviridae, possess a double-stranded DNA and can lyse 88% (22/25) of Salmonella strains stored in the laboratory. Analysis of the biological characteristics, in addition, revealed the optimal multiplicity of infection (MOI) of vB_SalP_LDW16 to be 0.01 and the phage titer to be up to 3 × 1014 PFU/mL. Meanwhile, the phage vB_SalP_LDW16 was found to have some temperature tolerance, while the titer decreases rapidly above 60 ℃, and a wide pH (i.e., 5-12) range as well as relative stability in pH tolerance. The latent period of phage was 10 min, the burst period was 60 min, and the burst size was 110 PFU/cell. Furthermore, gastric juice was also found to highly influence the activity of the phage. The clinical treatment experiments showed that phage vB_SalP_LDW16 was able to significantly reduce the bacterial load in the blood through phage treatment, thereby improving the pathological changes in the intestinal, liver, and heart damage, and promoting the growth and development of the chicken. CONCLUSIONS: The phage vB_SalP_LDW16 is a highly lytic phage with a wide host range, which can be potentially used for preventing and treating chicken salmonellosis, as an alternative or complementary antibiotic treatment in livestock farming.

Mouse adaptation of the H9N2 avian influenza virus causes the downregulation of genes related to innate immune responses and ubiquitin-mediated proteolysis in mice.

H9N2 avian influenza viruses sporadically infect humans worldwide. These viruses have also contributed internal genes to H5N1, H5N6, H7N9, and H10N8 viruses, which have been isolated from humans with infections and are a substantial public health threat. To investigate the potential pathogenic mechanism of the H9N2 virus, we performed serial lung-to-lung passage of an avirulent H9N2 avian influenza virus (A/Chicken/Shandong/416/2016 [SD/416]) in mice to increase the pathogenicity of this virus. We generated a mouse-adapted (MA) virus that exhibited increased viral titers in the lungs, caused severe lung damage in mice, and induced body weight loss in mice; however, the avirulent parental virus did not cause any clinical symptoms in infected mice. Global gene expression analysis was performed and indicated that the transcriptional responses of these viruses were distinct. The lungs of mice infected with the MA virus exhibited the downregulation of genes related to innate immunity and ubiquitin-mediated proteolysis, which was not seen in infections with the avirulent parental virus. These data indicated that the MA virus might evade immune surveillance and changed its replication capacity to increase the viral replication level and pathogenicity. Our study demonstrates that host factors play an important role in the adaptive evolution of influenza virus in new hosts.

Phages in sludge from the A/O wastewater treatment process play an important role in the transmission of ARGs.

Phages can influence the horizontal gene transfer (HGT) of antibiotic resistance genes (ARGs) through transduction, but their profiles and effects on the transmission of ARGs are unclear, especially in complex swine sludge. In this study, we investigated the characterization of phage and ARG profiles in sludge generated from anoxic/oxic (A/O) wastewater treatment processes on swine farms using metagenomes and viromes. The results demonstrated that 205-221 subtypes of ARGs could be identified in swine sludge, among which sul1, tet(M), and floR were the dominant ARGs, indicating that sludge is an important reservoir of ARGs, especially in sludge (S) tanks. The greater abundance of mobile genetic elements (MGEs) in the S tank could significantly contribute to the greater abundance of ARGs there compared to the anoxic (A) and oxic (O) tanks (P < 0.05). However, when we compared the abundances of ARGs and MGEs in the A and O tanks, we observed opposite significant differences (P < 0.05), suggesting that MGEs are not the only factor influencing the abundance of ARGs. The high proportion of lysogenic phages in sludge from the S tank can also have a major impact on the ARG profile. Siphoviridae, Myoviridae, and Podoviridae were the dominant phage families in sludge, and a network diagram of bacteria-ARG-phages revealed that dominant phages and bacteria acted simultaneously as potential hosts for ARGs, which may have led to phage-mediated HGT of ARGs. Therefore, the risk of phage-mediated HGT of ARGs cannot be overlooked.

Complete genome sequence and pathogenicity analysis of a highly pathogenic FAdV-4 strain.

Fowl adenovirus serotype 4 (FAdV-4) is a double-stranded DNA virus that mainly infects broiler chickens and has caused huge economic losses to the poultry industry. Recently, an FAdV-4 strain, SDLC202009, the causative pathogen of hydropericardium-hepatitis syndrome (HHS) in Liaocheng, Shandong, was isolated from commercial laying hens and propagated in specific pathogen free SPF chicken embryos. Pathogenicity studies showed that SDLC202009 could infect SPF chicken embryos and chickens, with a mortality rate of 100%. The complete genome was sequenced, and phylogenetic analysis showed that SDLC202009 belonged to the FAdV-4 cluster, with a genome length of 43, 077 bp. The SDLC202009 had 99.9% identity with the JSJ13 and SD1601, which were recently isolated in China. Compared to the recently isolated strain in China, SDLC202009 had deleted open reading frame 19 (ORF19), ORF27, ORF48, and ORF0. SDLC202009 harbored amino acid site mutations in the main structural proteins hexon, fiber1, and fiber2 similar with those in highly pathogenic strains. Furthermore, SDLC202009 showed unique mutations in hexon A571P, fiber1 E216K, and fiber2 N98K. In summary, our findings provide theoretical support for prevention and control of the HHS.

Enhanced in-plane thermoelectric figure of merit in p-type SiGe thin films by nanograin boundaries.

P-Type polycrystalline silicon-germanium (SiGe) thin films are grown by low-pressure chemical vapor deposition (LPCVD) and their thermoelectric properties are characterized from 120 K to 300 K for potential application in integrated microscale cooling. The naturally formed grain boundaries are found to play a crucial role in determining both the charge and thermal transport properties of the films. Particularly, the grain boundaries create energy barriers for charge transport which lead to different dependences of charge mobility on doping concentration and temperature from the bulk counterparts. Meanwhile, the unique columnar grain structures result in remarkable thermal conductivity anisotropy with the in-plane thermal conductivities of SiGe films about 50% lower than the cross-plane values. By optimizing the growth conditions and doping level, a high in-plane figure of merit (ZT) of 0.2 for SiGe films is achieved at 300 K, which is about 100% higher than the previous record for p-type SiGe alloys, mainly due to the significant reduction in the in-plane thermal conductivity caused by nanograin boundaries. The low cost and excellent scalability of LPCVD render these high-performance SiGe films ideal candidates for thin-film thermoelectric applications.