Amplicon-based metagenomic association analysis of gut microbiota in relation to egg-laying period and breeds of hens.

BACKGROUND: The gut microbiota plays an essential role in maintaining gut homeostasis and improving performance, with the composition of microbial communities visibly differing across different laying stages in hens and significantly correlating with egg production. To gain further insights into the association between microbial community characteristics and laying periods in Hy-Line variety brown and Isa brown laying hens, we conducted a 16S rRNA amplicon sequencing survey. RESULTS: Our result revealed the diversity of bacteria in the early laying period was commonly higher than peak, and in Hy-Line variety brown laying hens were generally higher than Isa brown. Principal coordinate analysis (PCoA) and permutational multivariate analysis of variance (PERMANOVA) revealed that the structure and composition of the gut microbiota of laying hens exhibited significant differences among different groups. Phylum Firmicutes, Bacteroidota, Proteobacteria, and Fusobacteriota were found that dominant in the host's feces. Therein, the abundance of Fusobacteriota was higher in the peak period than in the early period, while the abundance of Cyanobacteria in the early period was higher in two breeds of hens. Furthermore, random forest based on machine learning showed that there were several distinctly abundant genera, which can be used as potential biomarkers to differentiate the different groups of laying periods and breeds. In addition, the prediction of biological function indicated the existing discrepancy in microbial function among the microbiota of four groups. CONCLUSIONS: Our findings offer new insights into the bacterial diversity and intestinal flora composition of different strains of laying hens during various laying periods, contributing significantly to the improvement of production performance and the prevention of chicken diseases.

Prevalence of bovine coronavirus in cattle in China: A systematic review and meta-analysis.

Bovine coronavirus (BCoV) is one of the important pathogens that cause calf diarrhea (CD), winter dysentery (WD), and the bovine respiratory disease complex (BRDC), and spreads worldwide. An infection of BCoV in cattle can lead to death of young animals, stunted growth, reduced milk production, and milk quality, thus bringing serious economic losses to the bovine industry. Therefore, it is necessary to prevent and control the spread of BCoV. Here, a systematic review and meta-analysis was conducted to assess the prevalence of BCoV in cattle in China before 2022. A total of 57 articles regarding the prevalence of BCoV in cattle in China were collected from five databases (PubMed, ScienceDirect, CNKI, VIP, and Wan Fang). Based on the inclusion criteria, a total of 15,838 samples were included, and 6,136 were positive cases. The overall prevalence of BCoV was 30.8%, with the highest prevalence rate (60.5%) identified in South China and the lowest prevalence (15.6%) identified in Central China. We also analyzed other subgroup information, included sampling years, sample sources, detection methods, breeding methods, age, type of cattle, presence of diarrhea, and geographic and climatic factors. The results indicated that BCoV was widely prevalent in China. Among all subgroups, the sample sources, detection methods, breeding methods, and presence or absence of diarrheal might be potential risk factors responsible for BCoV prevalence. It is recommended to strengthen the detection of BCoV in cattle, in order to effectively control the spread of BCoV.

Global Prevalence of Echinococcosis in Goats: A Systematic Review and Meta-Analysis.

Echinococcosis is a foodborne parasitic zoonosis caused by the larvae of Echinococcus. This disease can affect goats and other mammals. In this study, a systematic review and meta-analysis for echinococcosis in global goats were performed based on the following five databases (China National Knowledge Infrastructure [CNKI], VIP Chinese Journal Database, Wanfang Data, PubMed, and ScienceDirect). In total, 108,197 samples were collected. The global prevalence of echinococcosis in goats was identified to be 10.85% (3217/108,197). The prevalence of echinococcosis in goats was 6.16% (1369/22,208) and 13.27% (874/5932) in South America and Africa, respectively. The prevalence of echinococcosis in goats before 2010 (9.76%; 112/713) was significantly higher than that from 2010 to 2014 (1.44%; 45/32,145) or after 2014 (2.95%; 154/3889). The prevalence of echinococcosis in goats aged <12 months (4.48%; 70/2911) was higher than that in goats aged ≥12 months (2.88%; 36/819). We also investigated the effects of geographical factors and climates on the prevalence of echinococcosis in goats. The results showed that the prevalence of echinococcosis was higher in the areas with high altitude and cold climate. This meta-analysis indicated that echinococcosis was ubiquitous in goats. Thus, we should improve the feeding conditions for goats, and strengthen the control measures of echinococcosis epidemic in goats, with the aims of reducing the economic losses of animal husbandry and providing protection for humans in the aspects of food security and health.

Prevalence of hepatitis E virus among swine in China from 2010 to 2019: A systematic review and meta-analysis.

Hepatitis E virus (HEV) is a zoonotic pathogen that has spread worldwide. The HEV reservoir associated with livestock hepatitis E poses a huge threat to public health. Awareness of the prevalence and spatial distribution of livestock hepatitis E is valuable to prevent and control diseases caused by HEV, especially human hepatitis E infection. Currently, swine, including pigs (Sus scrofa), are recognized as the major reservoir of HEV. Therefore, we conducted a systematic review and meta-analysis to evaluate the pooled prevalence of HEV among swine in China. A total of 71 published papers on HEV infection in swine in China (including data from 49,523 animals) from January 1, 2010 to December 31, 2019 met the standard after searching five databases including the Technology Periodical Database, the Wan Fang Database, the China National Knowledge Infrastructure, PubMed, and ScienceDirect. A random effects model was used to calculate the pooled prevalence of HEV in swine. The results showed that the seroprevalence was 48.0% (95% confidence interval (CI) 39.6-56.9) and the prevalence of HEV RNA was 14.4% (95% CI 10.7-18.5). The estimated overall prevalence was 34.1% (95% CI 27.2-41.4). Central China (68.0%, 95% CI 42.2-89.1) had a significantly higher prevalence than other regions. In the publication year subgroup, the prevalence in 2016 or later (27.2%, 95% CI 19.3-36.0) was significantly lower than that in 2011 or earlier (49.0%, 95% CI 36.2-61.8). The prevalence of IgG (42.9%, 95% CI 31.7-54.6) was significantly higher than that of IgM (4.9%, 95% CI 1.6-9.7). Suckling piglets (15.6%, 95% CI 6.6-27.1) had a lower prevalence compared with that in other age groups. In all sample types, body fluids showed the highest prevalence (50.5%, 95% CI 41.7-59.3). Moreover, the pooled prevalence of HEV in boars was higher than that in sows (35.4% > 17.3%). The analysis suggested that HEV infection is common among swine in China. Further strengthening HEV testing in boars, controlling environmental pollution, and reducing the mixed feeding of different stages could contribute to reducing HEV infection in pigs in China and the risk of porcine HEV infection in humans.

Intranasal delivery of plasmids expressing bovine herpesvirus 1 gB/gC/gD proteins by polyethyleneimine magnetic beads activates long-term immune responses in mice.

BACKGROUND: DNA vaccine is one of the research hotspots in veterinary vaccine development. Several advantages, such as cost-effectiveness, ease of design and production, good biocompatibility of plasmid DNA, attractive biosafety, and DNA stability, are found in DNA vaccines. METHODS: In this study, the plasmids expressing bovine herpesvirus 1 (BoHV-1) gB, gC, and gD proteins were mixed at the same mass ratio and adsorbed polyethyleneimine (PEI) magnetic beads with a diameter of 50 nm. Further, the plasmid and PEI magnetic bead polymers were packaged into double carboxyl polyethylene glycol (PEG) 600 to use as a DNA vaccine. The prepared DNA vaccine was employed to vaccinate mice via the intranasal route. The immune responses were evaluated in mice after vaccination. RESULTS: The expression of viral proteins could be largely detected in the lung and rarely in the spleen of mice subjected to a vaccination. The examination of biochemical indicators, anal temperature, and histology indicated that the DNA vaccine was safe in vivo. However, short-time toxicity was observed. The total antibody detected with ELISA in vaccinated mice showed a higher level than PBS, DNA, PEI + DNA, and PBS groups. The antibody level was significantly elevated at the 15th week and started to decrease since the 17th week. The neutralizing antibody titer was significantly higher in DNA vaccine than naked DNA vaccinated animals. The total IgA level was much greater in the DNA vaccine group compared to other component vaccinated groups. The examination of cellular cytokines and the percentage of CD4/CD8 indicated that the prepared DNA vaccine induced a strong cellular immunity. CONCLUSION: The mixed application of plasmids expressing BoHV-1 gB/gC/gD proteins by nano-carrier through intranasal route could effectively activate long-term humoral, cellular, and mucosal immune responses at high levels in mice. These data indicate PEI magnetic beads combining with PEG600 are an efficient vector for plasmid DNA to deliver intranasally as a DNA vaccine candidate.

Molecular detection of Blastocystis in black bears and sika deer in northern China.

Protists of the Blastocystis genus are distributed worldwide and can infect a range of hosts. However, data concerning Blastocystis infection are limited for sika deer and are not available for black bears. Therefore, in the present study, a total of 312 black bears (Ursus thibetanus) from Heilongjiang Province and 760 sika deer (Cervus nippon) from four different northern Chinese provinces were investigated. Blastocystis infection in these animals was detected via PCR amplification of the small subunit rRNA gene in fecal samples. The prevalence of Blastocystis infection in black bears and sika deer was 14.4% (45/312 positive samples) and 0.8% (6/760 positive samples), respectively. Young black bears (18.3%) had a significantly higher Blastocystis prevalence than adult bears (9.1%). The prevalence of Blastocystis was significantly higher in black bears raised outdoors (24.6%) than in bears raised indoors (12.2%). Blastocystis-positive sika deer were only found in Jilin Province (1.3%, 6/480). Female sika deer (0%, 0/61) had a significantly lower Blastocystis prevalence than males (0.9%, 6/699). Sanger sequencing was used to determine the small subunit rRNA gene sequences of the Blastocystis-positive PCR products. A neighbor-joining phylogenetic tree based on the small subunit rRNA gene sequences showed that only Blastocystis subtype (ST)1 was identified in black bears, whereas ST10 and ST14 were found in sika deer. This is the first report of Blastocystis ST1 infection in black bears. These findings also extend the distribution information of Blastocystis subtypes, which will provide a foundation for further study of Blastocystis in different hosts in China.

Mapping a highly conserved linear neutralizing epitope at the N-terminus of the gD glycoprotein of bovine herpesvirus type I using a monoclonal antibody.

Bovine herpesvirus type 1 (BoHV-1), a member of the Alphaherpesvirinae subfamily, causes significant economic losses to the cattle industry worldwide. Envelope glycoprotein D (gD) of BoHV-1 plays an essential role in the viral entry into permissive cells and possibly cooperates with other envelope glycoproteins. The herpesvirus gD induces a protective immune response against diseases in cattle or animal models. Mapping epitopes on gD will facilitate the understanding of the BoHV-1 pathogenesis and development of alternative vaccines against various diseases associated with the virus. In this study, a monoclonal antibody (MAb), designated as 3C1, was generated using naive BoHV-1 in vaccination of mice, demonstrating that 3C1 was specific to gD and represents a neutralizing activity against BoHV-1 infection in Madin-Darby bovine kidney cells. Panels of overlapping gD recombinant proteins with glutathione S-transferase tag were prepared to define the epitope recognized by 3C1. The data demonstrated that the N-terminus of gD 23APRVTVYVD31 was recognized by 3C1. Furthermore, the 26VTVYVD31 motif was the minimal amino acid sequence for the recognition. The epitope identified in this study is highly conserved among the typical strains of BoHV-1 and BoHV-5, suggesting that this epitope may be useful in the diagnosis of diseases. In addition, the defined region on gD of BoHV-1 might be essential in viral entry upon comparison with the prototype virus in herpes simplex virus (Alphaherpesvirinae). The data will elucidate the roles of gD of BoHV-1 in viral entry and pathogenesis and its potential application for the development of vaccine candidates and diagnostic techniques based on the conserved epitopes on gD or in combination with those of other herpesvirus glycoproteins.

Molecular detection of Cryptosporidium spp. and Enterocytozoon bieneusi in Longjiang Wagyu cattle in Northeastern China.

Cryptosporidium spp. and Enterocytozoon bieneusi are two important zoonotic pathogens that can cause diarrhea and other gastrointestinal illnesses in humans and animals. However, the prevalence and genotype of the parasites in Longjiang Wagyu cattle in Heilongjiang Province, Northeast China have not been reported. In the present study, a total of 423 fecal samples of Longjiang Wagyu cattle collected from different farms in Heilongjiang Province, Northeast China, were examined for Cryptosporidium spp. and E. bieneusi using nested PCR. The overall infection rates for Cryptosporidium spp. and E. bieneusi were 6.38% (n = 27) and 7.09% (n = 30), respectively. The prevalence in different age groups ranged from 3.80% (95% confidence interval (CI) 1.01-6.59) to 8.36% (95% CI 4.83-11.90) for Cryptosporidium spp. and 5.97% (95% CI 2.52-9.43) to 7.94% (95% CI 4.49-11.40) for E. bieneusi. By analyzing the DNA sequences of the small subunit (SSU) rRNA gene, two Cryptosporidium species were detected in this study, namely C. parvum (n = 25) and C. ryanae (n = 2). The IIdA20G1 subtype was further identified by using the 60-kDa glycoprotein (gp60) gene of C. parvum. E. bieneusi was identified using three known sequences through the analysis of internal transcribed spacer (ITS) sequences: J (n = 23), I (n = 5), and BEB4 (n = 2), and all belonged to group 2. The results indicated that some of the Cryptosporidium species and E. bieneusi genotypes identified in Longjiang Wagyu cattle in the study areas might have zoonotic potential.

Protective immunity following vaccination with a recombinant multiple-epitope protein of bovine herpesvirus type I in a rabbit model.

Bovine herpesvirus type 1 (BoHV-1) causes considerable economic losses to the cow industry. Vaccination remains an effective strategy to control the diseases associated with BoHV-1. However, live vaccines present safety concerns, especially in pregnant cows; thus, nonreplicating vaccines have been developed to control the disease. The envelope glycoproteins of BoHV-1 induce a protective immune response. In this work, selected epitopes on glycoproteins gD, gC, and gB were constructed in triplicate with linker peptides. Vaccination of rabbits demonstrated that P2-gD/gC/gB with AAYAAY induced higher specific antibodies than that with GGGGS linker. P2-gD/gC/gB with AAYAAY linker was fused with bovine interleukin-6 (BoIL-6) or rabbit IL-6 (RaIL-6) and bacterially expressed. Rabbits were intramuscularly immunized with 100 µg of P2-gD/gC/gB-BoIL-6, P2-gD/gC/gB-RaIL-6, P2-gD/gC/gB, P2-gD/gC/gB plus BoIL-6, P2-(gD-a)3-BoIL-6, or P2-(gD-a)3 emulsified with ISA 206 adjuvant thrice at 3-week intervals. P2-gD/gC/gB-BoIL-6 generated a higher titer of BoHV-1-specific antibodies, neutralizing antibodies, interferon (IFN)-γ, and IL-4 compared with P2-gD/gC/gB plus BoIL-6, P2-gD/gC/gB-RaIL-6, or other formulation. P2-gD/gC/gB-BoIL-6 triggered similar levels of antibodies and significantly higher titer of IFN-γ and IL-4 compared with inactivated bovine viral diarrhea (BVD)-infectious bovine rhinotracheitis (IBR) vaccine. Rabbits vaccinated with P2-gD/gC/gB-BoIL-6 dramatically reduced viral shedding and tissue lesions in lungs and trachea after viral challenge and reactivation compared with those with P2-gD/gC/gB plus BoIL-6 or P2-gD/gC/gB-RaIL-6. P2-gD/gC/gB-BoIL-6 provided protective effects against viral shedding and tissue pathogenesis similar to those of the inactivated vaccine. The data confirmed the safety and immunogenicity of multiple-epitope recombinant protein and a potential vaccine candidate to control the disease, especially for pregnant cattle.

Molecular detection of Cryptosporidium and Enterocytozoon bieneusi in dairy calves and sika deer in four provinces in Northern China.

The protistan pathogens Cryptosporidium and Enterocytozoon bieneusi can cause significant intestinal diseases in animals and humans. However, limited information is available regarding prevalence and molecular characterization of Cryptosporidium and E. bieneusi in ruminants in Northern China. In this study, the overall prevalence of Cryptosporidium and E. bieneusi was 19.3% (62/321) and 28.97% (93/321) in dairy calves and 1.10% (9/818) and 13.57% (111/818) in sika deer (Cervus nippon) in four provinces in Northern China, respectively. The prevalence of Cryptosporidium and E. bieneusi in different factor groups was various. Five Cryptosporidium species/genotypes were identified, of which C. parvum, C. ryanae, C. bovis, and C. andersoni were only found in dairy calves, and only Cryptosporidium deer genotype was found in sika deer. Moreover, J, I, and BEB4 ITS genotypes of E. bieneusi were found in dairy calves, and six known genotypes (JLD-III, JLD-IX, JLD-VII, EbpC, BEB6, and I) and ten novel genotypes (namely LND-I and JLD-XV to JLD-XXIII) were found in sika deer in this study. Cryptosporidium parvum and E. bieneusi genotype J were identified as the predominant species/genotypes in dairy calves, whereas the predominance of Cryptosporidium spp. and E. bieneusi in sika deer was Cryptosporidium deer genotype and BEB6, respectively. The present study reported the prevalence and genotypes of Cryptosporidium and E. bieneusi in dairy calves and sika deer in four provinces in northern China. The present findings also suggest that investigated dairy calves and sika deer may play an important role in the transmission of E. bieneusi and Cryptosporidium to humans and other animals, and also in an effort to better understand the epidemiology of these enteric pathogens in China.

Preparation of porcine enterotoxigenic Escherichia coli (ETEC) ghosts and immunogenic analysis in a mouse model.

Enterotoxignenic Escherichia coli (ETEC)-associated colibacillosis causes high levels of morbidity and mortality in neonatal piglets. Vaccination is among effective strategy to fight against ETEC-related diseases. Bacterial ghosts (BGs) are empty bacterial envelopes, which substain subtle antigenic comformation in bacterial outer membrane. In this study, a BG vaccine was generated using porcine ETEC isolated strain DQ061 and evaluated its safety and immunogenicity in a mouse model. The recombinant bacteria were constructed by transformation of lysis plasmid pHH43 and generation of BGs was conducted in a lysis rate of 99.93% by incubation of the recombinant bacteria at 42 °C for 2 h. Mice were immunized subcutaneously twice in 2-week intervals with BGs, BGs emulsified with ISA 206 adjuvant, or formalin-inactivated ETEC vaccine after safety test. Mice with either of two BG vaccines developed higher titer of antibodies, secreted higher titer of interleukin 4, gamma interferon and alpha tumor necrosis factor after 2 doses than those with formalin-inactivated ETEC vaccine or those with adjuvant placebo (P < 0.01). The quantity of CD4+ and CD8+ T lymphocyte in spleen was higher in both BG groups than that in the inactivated vaccine group or adjuvant group 2 weeks post boost immunization (P < 0.05). The vaccinated mice were challenged intraperitoneally with 10 × LD50 dose of DQ061. Mice with the BGs plus adjuvant were completely protected against challenge, compared to 60% protection of mice with the inactivated vaccine. Mice exhibited decreased tissue lesion and reduced bacterial loads in the BGs groups by comparison with those with the inactivated vaccine or adjuvant only. Our results validated that the ETEC BGs bear high safety and immunogenicity in a mouse model, suggesting a potential of further evaluation in a pig model.

Generation of porcine Pasteurella multocida ghost vaccine and examination of its immunogenicity against virulent challenge in mice.

Pasteurella multocida (PM) causes a varity of clinical manifestation in domestic animals, even acute death. Vaccination is among effective strategy to prevent and control PM-related diseases. Bacterial ghosts (BGs) are empty bacterial envelopes, which sustain subtle antigenic comformation in bacterial outer-membrane and exhibit higher efficacy compared to inactivated vaccines. Here, a BG vaccine generated from the porcine PM reference strain CVCC446 (serotype B:2) was prepared upon lysis by E protein of bacteriophage PhiX174, and the safety and immunogenicity were evaluated its in a mouse model. Lysis rate was in 99.99% and the BG vaccine was completely inactivated by addition of freeze-dry procedure. Mice were immunized subcutaneously twice in 2-week intervals with BGs, or BGs plus adjuvant, or formalin-inactivated PM or an adjuvant control. Mice inoculated twice with BGs vaccines generated higher titer of antibodies, interleukin 4 and gamma interferon than those in the inactivated vaccine group or adjuvant placebo group (P < 0.05). CD4+ and CD8+ T lymphocyte levels in spleen were higher in both BG groups than inactivated vaccine group or adjuvant group. Mice administered with the BGs plus adjuvant were completely protected against intraperitoneal challenge with 10 × LD50 dose of virulent isolate and exhibited decreased tissue lesion and lower bacterial loads, which was superior to the inactivated vaccine. The results demonstrated safety of the BG vaccine and primary immunogenicity in a mouse model, suggesting a potential of further evaluation in a pig model and vaccine candidate.

Epidemiology of porcine epidemic diarrhea virus among Chinese pig populations: A meta-analysis.

BACKGROUND: Porcine epidemic diarrhea results from infection with porcine epidemic diarrhea virus (PEDV). It is an acute and highly contagious enteric disease in swine characterized by watery diarrhea and vomiting. Here, we performed a systematic review and meta-analysis in order to assess the prevalence of PEDV infection in pig populations from mainland China. METHODS: We conducted a literature search on the prevalence of PEDV infection in pigs between Jan 1, 1988 and Aug 20, 2018 in English and Chinese databases, including PubMed, Google scholar, Cochrane library, Clinical Trials, VIP, CNKI and WanFang database. Selections were made based on the title and the abstract of paper, and duplicated literature was excluded along with other host studies, and data incomplete literature according to the exclusion criteria we formulated. Finally, we extracted the number of swine with PEDV infection from the obtained studies and provided information that permitted us to estimate the prevalence of PEDV infection in pigs in mainland China. RESULTS: A total of 45 studies (including data from 15,990 pigs) met our evaluation criteria. In China, the overall estimated prevalence of PEDV infection in pigs was 44% (7113/15,990), while the estimated prevalence of PEDV infection in pigs from northern China was 37% (793/2136), lower than those in other regions of China. The prevalence of PEDV infection was associated with sampling season, category of pigs and clinical signs (diarrhea) in pigs. However, the prevalence of PEDV among pigs in China was not significantly associated with the effect of detected target genes, nor was it associated with date of study publication. CONCLUSION: Our findings suggest that PEDV infection is common among pigs in China. It is therefore necessary to carry out further research and monitor the prevalence of PEDV infection. Furthermore, powerful and effective regulatory measures should be taken in order to prevent the transmission and spread of PEDV among pig populations.

Seroprevalence and risk factors of Toxoplasma gondii infection in domestic raccoon dogs in four provinces in northern China.

Toxoplasma gondii is an obligate intracellular apicomplexan parasite that can infect almost all homoiothermal animals, including domestic raccoon dogs (Nyctereutes procyonoides). However, related reports on T. gondii infection in domestic raccoon dogs were limited in China. Therefore, a serological investigation was undertaken to investigate the seroprevalence and risk factors for T. gondii infection in domestic raccoon dogs. A total of 962 serum samples were collected from Jilin, Liaoning, Heilongjiang and Hebei provinces, northern China between April 2016 and November 2017, and were detected by the indirect hemagglutination test (IHA). The seroprevalence of T. gondii infection was 7.28% in the overall surveyed raccoon dogs by IHA, which was different among the four provinces ranging from 6.54% to 7.57%. The seroprevalence of T. gondii infection in male and female raccoon dogs was 6.62% and 7.79%, respectively. Based on statistical analysis, age was regarded as an important risk factor for T. gondii infection in raccoon dogs in this study (P < 0.05). This study reported the seroprevalence and risk factors of T. gondii infection in domestic raccoon dogs in northern China, which provided essential data for prevention and control of T. gondii infection in raccoon dogs in Jilin province, Liaoning province, Heilongjiang province and Hebei province.

Correction to: Brucellosis seroprevalence in ovine and caprine flocks in China during 2000-2018: a systematic review and meta-analysis.

The original article [1] contained a minor error whereby Table 1 was typeset incorrectly.

Brucellosis seroprevalence in ovine and caprine flocks in China during 2000-2018: a systematic review and meta-analysis.

BACKGROUND: Brucellosis remains one of the most common zoonotic diseases globally, with more than half million human cases reported annually. Brucellosis is an emerging and re-emerging disease in China since the 1990s. An infectious reservoir constituted by domestic animals with brucellosis, especially ovine and caprine herds, poses a significant threat to public health. The seroprevalence of brucellosis in sheep and goat flocks in a national context is unavailable so far. Therefore, we conducted this systematic review and meta-analysis to assess the overall status of brucellosis in sheep and goats in China in almost two decades. RESULTS: The pooled prevalence of brucellosis in ovine and caprine flocks in China increased in 2000-2009 (1.00%; 95% CI, 0.70-1.30) to 2010-2018 (3.20%; 95% CI, 2.70-3.60). The seroprevalence of brucellosis in sheep and goat flocks was higher in Eastern China, with 7.00% of positive rate, than that in any other region, especially Shandong province (18.70%). Brucellosis is highly endemic in some local regions. The high prevalence of brucellosis in agricultural regions is suggestive of a shift of geographic distribution. The pooled prevalence of brucellosis is higher in goat flocks than in sheep flocks in China. CONCLUSIONS: The overall data in this meta-analysis demands comprehensive intervention measures and further surveillance to facilitate the control of brucellosis in livestock. Further studies aimed at evaluating the risk factors associated with spreads of brucellosis in domestic animals unaddressed so far, and sufficient epidemiological data is important to the exploration and understanding of the prevalent status of brucellosis throughout the country and to disease control.

Genomic Sequencing and Analysis of Enzootic Nasal Tumor Virus Type 2 Provides Evidence for Recombination within the Prevalent Chinese Strains.

In recent years, the clinical cases of ENTV-2 infection have increased and become prevalent in several provinces of China. In this study, we reported the occurrence of ENTV-2 in one goat farm in Chongqing, southwest China. The complete genome of an emerged ENTV-2 isolate (designated as CQ2) was sequenced with 7468 bp in length. Phylogenetic analysis revealed that ENTV-2 consisted of two main lineages. Lineage 1 was composed of Chinese strains and could be subdivided into five sublineages. CQ2 and the other six recent isolates from China were clustered in sublineage 1.5; however, CQ2 was significantly different from the other six isolates. Furthermore, recombination analysis suggested that CQ2 might be a recombinant variant derived from sublineage 1.5 and sublineage 1.2 strains, with the recombination region in areas of pro and pol genes. In conclusion, we sequenced and analyzed the complete genome of a potential ENTV-2 recombinant, which may contribute to our understanding of the genetic variation and evolution of ENTV-2 in China.

Subtypes of Blastocystis in Tibetan Antelope (Pantholops hodgsonii).

Blastocystis is a protist that is distributed in the gut tract of humans and animals. However, the reports about Blastocystis infection in Tibetan antelope are scarce. We collected 173 Tibetan antelope feces samples from Xinjiang, Qinghai and Xizang, and amplified the SSU rRNA gene of 600 bp region of Blastocystis in our research. Fifty-one samples in total were positive for Blastocystis, with all subtypes being ST31. The lowest prevalence of Blastocystis was observed in Xizang (2/20, 9.1%), followed by Qinghai (18/92, 16.4%), Xinjiang (31/61, 33.7%). The highest prevalence of Blastocystis in Tibetan antelope was detected during the summer was (19/30, 38.8%). This is the first research work regarding the Blastocystis subtypes ST31 in Tibetan antelope. Our research provides information for future researches on the distribution of this Blastocystis subtype and the control of Blastocystis infection.

The prevalence and genotypes of Cryptosporidium spp. in bar-headed goose (Anser indicus) in China.

Cryptosporidium spp. is an important foodborne and waterborne pathogen in humans and animals, causing diarrhoea in humans and respiratory and gastrointestinal diseases in birds. However, reports of Cryptosporidium infection in bar-headed goose are limited. To determine the infection rate and species/genotypes of Cryptosporidium in bar-headed goose in China, a total of 358 fecal samples were collected from 3 regions. Nested PCR was used to amplify Cryptosporidium SSU rRNA regions from the fecal extracted-DNA samples. The total infection rate of Cryptosporidium in bar-headed in China was 3.9 % (14/358), with 4.2 % (5/120) in Aba (Ngawa) Tibetan and Qiang Autonomous Prefect, Sichuan province, 7.6 % (9/119) in Maqu county, Gansu province, and 0.0 % (0/119) in Caohai, Wei ning county, Guizhou province. The differences in prevalence rate by region were statistically significant. All positive samples were identified as Cryptosporidium goose genotype I (n = 14). This is the first systematic investigation of the epidemiological status and dominant species/genotypes of Cryptosporidium in bar-headed goose in China, thereby enhancing our understanding of the epidemiology of Cryptosporidium infection in wild migratory birds.

Global prevalence of Plasmodium infection in wild birds: A systematic review and meta-analysis.

Avian malaria is a vector-borne parasitic disease caused by Plasmodium infection transmitted to birds by mosquitoes. The aim of this systematic review was to analyze the global prevalence of malaria and risk factors associated with infection in wild birds. A systematic search of the databases CNKI, WanFang, VIP, PubMed, and ScienceDirect was performed from database inception to 24 February 2023. The search identified 3181 retrieved articles, of which 52 articles met predetermined inclusion criteria. Meta-analysis was performed using the random-effects model. The estimated pooled global prevalence of Plasmodium infection in wild birds was 16%. Sub-group analysis showed that the highest prevalence was associated with adult birds, migrant birds, North America, tropical rainforest climate, birds captured by mist nets, detection of infection by microscopy, medium quality studies, and studies published after 2016. Our study highlights the need for more understanding of Plasmodium prevalence in wild birds and identifying risk factors associated with infection to inform future infection control measures.