Whole-genome analysis reveals possible sources of ALV-J infection in an anyi tile-like gray chicken flock.

Avian leukosis virus (ALV) induces multiple tumors in chicken and is still prevalent in a lot of local flocks in China. In this study, we analyzed the ALV infection status in an Anyi tile-like gray chicken flock by DF1-cells isolation, virus identification, and genome sequencing. Results showed a 29% (29/100) ALV positive rate in this flock. Homology analysis based on env genes illustrated that all these stains belong to subgroup J (92-100% identities) and can be further divided into 5 batches, suggesting a higher diversity of ALV-J within the same flock. The whole-genome analysis of representative stains from each batch confirmed the close relationship between these isolated strains with previously reported strains from different regions (Guangxi, Shandong, and Heilongjiang), revealing the enrichment of different strains in Anyi tile-like grey chickens. This study provides the epidemiological data of ALV-J in a special chicken flock and a reference for the further eradication of ALV in China.

Differential metabolism-associated gene expression of duck pancreatic cells in response to two strains of duck hepatitis A virus type 1.

Several outbreaks of duck hepatitis A virus type 1 (DHAV-1), which were characterized by yellow coloration and hemorrhage in pancreatic tissues, have occurred in China. The causative agent is called pancreatitis-associated DHAV-1. The mechanisms involved in pancreatitis-associated DHAV-1 infection are still unclear. Transcriptome analysis of duck pancreas infected with classical-type DHAV-1 and pancreatitis-associated DHAV-1 was carried out. Deep sequencing with Illumina-Solexa resulted in a total of 53.9 Gb of clean data from the cDNA library of the pancreas, and a total of 29,597 unigenes with an average length of 993.43 bp were generated by de novo sequence assembly. The expression levels of D-3-phosphoglycerate dehydrogenase, phosphoserine aminotransferase, and phosphoserine phosphatase, which are involved in glycine, serine, and threonine metabolism pathways, were significantly downregulated in ducks infected with pancreatitis-associated DHAV-1 compared with those infected with classical-type DHAV-1. These findings provide information regarding differences in expression levels of metabolism-associated genes between ducks infected with pancreatitis-associated DHAV-1 and those infected with classical-type DHAV-1, indicating that intensive metabolism disorders may contribute to the different phenotypes of DHAV-1-infection.

A novel group of avian Avastrovirus in domestic geese, China.

Using an ORF1b-based astrovirus-specfic reverse transcription (RT)-PCR assay, a novel astrovirus-like was detected from domestic geese in China. Pairwise comparisons and phylogenetic analyzes suggested that a novel group of goose astrovirus, different with previously known astroviruses in the genus Avastrovirus, was found circulating in geese. This study has expanded our understanding about the role of domestic waterfowls as reservoirs for diverse astroviruses.

Effect of dexmedetomidine-induced anesthesia on the postoperative cognitive function of elder patients after laparoscopic ovarian cystectomy.

OBJECTIVE: To investigate the effect of dexmedetomidine at maintenance dose on the postoperative function of elder patients after general anesthesia for laparoscopic ovarian cystectomy. METHODS: We enrolled a total of 96 elder patients who were admitted to this hospital for laparoscopic ovarian cystectomy under general anesthesia between March 2015 and March 2017, and divided them into two groups, Group A (n = 48) and B (n = 48). Patients in both groups received the same methods for anesthesia induction and maintenance drugs. At the beginning of operation, patients in Group A received the intravenous injection of dexmedetomidine (0.8 µg/kg) followed by maintenance dose [0.5 µg/(kg h)] to the end of operation, while those in Group B underwent intravenous injection of 0.9% normal saline at the same rate, during which blood pressure, heart rate, oxyhemoglobin saturation and dosage of anesthetics at T1 (5 min after being delivered into the operation room), T2 (immediately after anesthesia induction), T3 (immediately after intubation), T4 (immediately after operation), T5 (immediately after end of operation) and T6 (immediately after extubation). Then, the levels of NSE, IL-6, CRP and HMGB1 were compared between two groups at 24 h before the operation, at the end, 24 h, 3 d and 7 d after operation. Besides, we also compared the postoperative cognitive functions and incidence of adverse reactions at 1 d before, 1, 2, 3 and 7 d after operation through MOCA scales. RESULTS: At T3, T4 and T6, comparisons of the average arterial pressure and heart rate showed that the differences between the two groups had statistical significance (p < .05). At the end of operation, and at 24 h, 3 d and 7 d after operation, we found that the levels of IL-6 and CRP in patients of two groups were all significantly elevated when compared with those before operation; at the end of operation and at 24 h and 3 d after operation, the levels of IL-6 and CRP in the Group B were higher than those in the Group A, and the differences had statistical significance (p < .05). At the end of operation and at 24 h and 3 d after operation, the levels of NSE and HMGB1 in two group were higher than those before operation, and a more significant elevation was identified in Group B with statistically significant differences (p < .05); at 7 d after operation, a decreasing trend was found in the level of HMGB1, which, however, remained higher than the preoperative level, and the level in Group B was still higher than that in Group A with statistically significant differences (p < .05). At 2 d after operation, we found that the scores of MOCA in the Group B were remarkably decreased in comparison with the scores in Group A with a statistically significant difference (p < .05). Moreover, the incidence rate of postoperative cognitive dysfunction (POCD) in the Group A was significantly lower than that in the Group B, and the difference had statistical significance (p < .05). CONCLUSION: Dexmedetomidine can ameliorate the postoperative cognitive functions of elder patients who received the laparoscopic ovarian cystectomy under general anesthesia, and effectively decrease the incidence rate of POCD without any obvious or severe adverse reaction. Thus, it can serve as a kind of adjuvant drug for general anesthesia in clinical practice.

Development of a restriction length polymorphism combined with direct PCR technique to differentiate goose and Muscovy duck parvoviruses.

A restriction fragment length polymorphism combined with direct PCR technique to differentiate goose and Muscovy duck parvoviruses (GPV and MDPV) was developed based on comparison of the NS gene of GPV and MDPV. Both GPV and MDPV genomic DNA can be amplified with 641 bp using the specific PCR primers. The PCR fragments can be cut into 463 bp and 178 bp only in the case of MDPV-derived PCR products, whereas the GPV-derived PCR products cannot. The method established in this study can be used to differentiate GPV and MDPV with high specificity and precision, by using a direct PCR kit and QuickCut enzyme, as quickly as conventional PCR.

[Geographic distribution and gene sequencing of Paragonimus westermani in some areas of Guangdong Province].

OBJECTIVE: To investigate the current distribution of Paragonimus westermani in Guangdong Province. METHOD: Snails and crabs collected from mountain streams in regional survey sites were dissected to detect cercarial and metacercarial infections of P. westermani. Domestic cats and dogs artificially infected with the collected metacercariae were also dissected to detect adult worms of P. westermani. The COI and ITS2 gene sequences of those adult worms were compared with those of known Paragonimus specimen deposited in the GenBank. RESULTS: All of the first intermediate hosts in five survey sites of Liangkou, Nankun, Mountain, Dadong, Muxi, Guowu, were identified as Semisulcospira libertina, whose cercariae infection rates were 0.33%, 0.15%, 0.058%, 0.10%, and 0.05%, respectively; the second intermediate hosts in above five sites were all identified as Sinopotamon denticulatum, whose metacercariae infection rates were 100%, 100%, 38.09%, 55.36%, and 65.26%, respectively. The numbers of metacercariae in the five sites were 79.4, 105.66, 9.16, 16.18, and 15.6 per positive crab, respectively, and 11.12, 7.87, 0.58, 0.69, and 0.85 per gram of crab, respectively. All the metacercariae were identical to those of P. westermani. Adult worms and eggs of P. westermani were found in both reservoir hosts of domestic cats and dogs infected artificially. By comparing the COI genes of five representative samples from each survey site with that of Paragonimus #AF219379.21, AF540958.1 from GenBank, we found out the homology to be 99%, 99%, 99%, 98%, and 99%, respectively. In addition, a comparison of the ITS2 gene sequences between the above five samples and Paragonimus #DQ836243.1, DQ351845.1, AB354217.1 from GenBank revealed 98%, 99%, 98%, 98%, and 98% gene homology, respectively. CONCLUSION: Two ultra-high and three high endemic areas of P. westermani are discovered in Guangdong Province. No obvious differences were found among the types of P. westermani in the above five endemic areas.

Complete genomic sequence of the virulent bacteriophage RAP44 of Riemerella anatipestifer.

A virulent Riemerella anatipestifer bacteriophage, RAP44, belonging to the Siphoviridae family of tailed phages, was previously isolated from feces of healthy Muscovy ducks in China. A complete genomic sequence analysis indicates that the phage's genome consists of a linear, double-stranded DNA molecule of 49,329 nucleotides. Eighty open reading frames (ORF) were identified. Putative functions could be assigned to 24 of the ORFs. The location of these genes was consistent with organization of the genome in a modular format which includes modules for host cell lysis, tail morphogenesis, head morphogenesis, and DNA replication and modification modules. Until now, no R. anatipestifer phage genome sequence has been reported in the literature. Therefore, this study represents the first complete genomic and molecular description of the R. anatipestifer phage.

[Sequence comparison of the hemagglutinin gene of the duck-origin H9N2 subtype avian influenza viruses].

To demonstrate the phylogenetic evolution, the molecular characteristics of the motif of HA protein cleavage site and the varieties at the receptor binding sites of the hemagglutinin gene of the duck-origin H9N2 subtype avian influenza viruses, sequence alignment and phylogenetic analysis were performed by MEGA 4.1 Neighbor-Joining method.. The results revealed that the duck-origin H9N2 AIV viruses originated from CK/BJ/1/94-like and North-Ame-like, all the duck-origin H9N2 AIV viruses from mainland China belonged to CK/BJ/1/94-like and formed multiple genotypes through complicated re-assortment, while other duck-origin H9N2 AIV, isolated from other countries in Aisa, American and European such as Korea, Japan, Alberta, Austria, Switzerland, Iran, belonged to the North-Ame-like phylogenetic lineage. The amino acids at positions 183, 190, and 226 of the receptor binding sites of North-Ame-like group isolates had highly conserved H, E and Q respectively. In contrast with duck-origin H9N2 AIV viruses isolates from mainland China, the amino acids had N at positions 183, A, T, or V at 190, L or Q at 226, which was the same as the chicken-origin H9N2 AIV from mainland China. Most newly isolated chicken-origin H9N2 AIV in Fujian Province in Southern China had L at position 226 emphasized the higher risk of cross-infection between the chicken-origin and duck-origin H9N2 AIV in China.

Epidemiological investigation and genome analysis of duck circovirus in Southern China.

Duck circovirus (DuCV), a potential immunosuppressive virus, was investigated in Southern China from March 2006 to December 2009 by using a polymerase chain reaction (PCR) based method. In this study, a total of 138 sick or dead duck samples from 18 different farms were examined with an average DuCV infection rate of ∼35%. It was found that ducks between the ages of 40∼60 days were more susceptible to DuCV. There was no evidence showing that the DuCV virus was capable of vertical transmission. Farms with positive PCR results exhibited no regularly apparent clinical abnormalities such as feathering disorders, growth retardation or lower-than-average weight. The complete genomes of 9 strains from Fujian Province and 1 from Zhejiang Province were sequenced and analyzed. The 10 DuCV genomes, compared with others genomes downloaded from GenBank, ranged in size from 1988 to 1996 base pairs, with sequence identities ranging from 83.2% to 99.8%. Phylogenetic analysis based on genome sequences demonstrated that DuCVs can be divided into two distinct genetic genotypes, Group I (the Euro-USA lineage) and Group II (the Taiwan lineage), with approximately 10.0% genetic difference between the two types. Molecular epidemiological data suggest there is no obvious difference among DuCV strains isolated from different geographic locations or different species, including Duck, Muscovy duck, Mule duck, Cheery duck, Mulard duck and Pekin duck.

[Genome cloning and sequence analysis of duck circovirus].

To reveal the molecular biological characteristics of genome of circovirus in infected ducks, two nucleotide fragments were amplified by overlapping PCRs using DNA extracted from various tissues of ducks. After they had been assembled together, the nucleotide components, the genome organization and the phylogenetic scale of the sequence were analyzed. The results showed that the obtained sequence is a circular DNA with a total length of 1995nt. It contains 6 open reading frames (ORFs), and shares a high identity of 97.4% with the MuDCV circovirus sequence presented in GenBank (AY228555). These results indicate that the amplified product stems from duck circovirus sequence.