Epidemiological investigation and fiber gene genetic analysis of fowl adenovirus serotype 4 from 2017 to 2021 in Yunnan Province, China

doi:10.21203/rs.3.rs-3127901/v1

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Epidemiological investigation and fiber gene genetic analysis of fowl adenovirus serotype 4 from 2017 to 2021 in Yunnan Province, China

https://doi.org/10.21203/rs.3.rs-3127901/v1

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Background: In recent years, the prevalence of fowl adenovirus serotype 4 (FAdV-4) has caused huge economic losses to the Chinese poultry industry. The objective of this study was to determine the occurrence of FAdV-4 in Yunnan Province through an epidemiological investigation of poultry and wild birds from 2017 to 2021 and phylogenetic analysis of the isolated fiber genes.

Results: The results showed that the FAdV-4 positive rate in poultry and migratory birds was 11.34% and 0.3%, respectively. The infection rate of the poultry tissue samples was high in central and west-ern Yunnan Province, and the infected hosts were mainly local chickens. Via the detection of common poultry infectious diseases prone to co-infection, it was found that the birds were mainly co-infected with pathogens such as infectious laryngotracheitis, chicken infectious anemia virus, and avian influenza virus. Five poultry-source strains and one wild bird-source strain were isolated, and phylogenetic analysis of these strains using the fiber gene showed that they all had high homology with the early Chinese highly pathogenic strains and clustered with the same genetic and evolutionary clade as the Chinese reference strain. The fiber protein is characterized by loci that indicate highly pathogenic strains (G²¹⁹D, P³⁰⁷A, V³¹⁹I, and A³⁰⁸T).

Conclusions: The results of our study demonstrate that FAdV-4 continues to exhibit a high prevalence rate among poultry populations in Yunnan Province, China. Notably, we report the first detection of FAdV-4 in the feces of black-necked cranes. Our statistical analysis reveals that central Yunnan and western Yunnan experience a high incidence rate of FAdV-4, often accompanying co-infections with ILT, CIAV, and AIV.

FAdV-41

Epidemiology2

Infection rate3

fiber4

co-infection5

Fowl adenoviruses (FAdVs) are the main pathogens of viral infectious diseases associated with hydropericardium and hepatitis outbreaks in China in recent years, and they belong to the Aviadenovirus genus of the family Adenoviridae^[1]. Fowl adenoviruses are non-enveloped and non-segmented double-stranded DNA viruses with a globular genome of 26–43 kb^[2]. The virions of FAdVs possess 240 hexon proteins, 12 penton protein bases at the vertices, and fiber proteins projecting from each penton base. According to the specific antigen of FAdVs, they can be divided into Ⅰ, Ⅱ, and Ⅲ subgroups. Based on the profile of restriction enzyme digestion and serum cross-neutralization assay, FAdV-Ⅰ is clustered into 5 species (FAdV-A-E) with 12 serotypes (FAdV-1-8a, 8b-11)^[3–5]. The serotypes that have been reported to be prevalent in Asia are FAdV-4, FAdV-8a, FAdV-8b, and FAdV-11. Among them, FAdV-4 is the most prevalent and a highly pathogenic serotype^[6]. Studies have shown that the fiber proteins of FAdVs effectively induce virus-neutralizing antibodies and mediate viral attachment to the cellular receptor for entering the host cells and increase the rate of infection. Unlike the other FAdV serotypes, FAdV-4 has two fibers (fiber-1 and fiber-2). Fiber-1 is the key factor for directly triggering FAdV-4 infection, and fiber-2 is a key protein that controls the virulence of FAdV-4^[7–9].

The pathogen was first found in Angara Goth, Pakistan in 1987, so the infection is also called Angara disease. Based on the typical clinical symptoms of pericardial effusion, hyperemic, enlarged liver with petechial hemorrhages and punctiform areas in chickens, it is also called hepatitis-hydropericardium syndrome (HHS)^[10]. In China, FAdV-4 was first introduced in 2012, and outbreaks were reported in the Shandong, Liaoning provinces and other regions in 2015^[11–12]. FAdV-4 was first detected in Yunnan Province in 2016. By monitoring FAdV-4 infections in Yunnan Province, we found that there were FAdV-4 infections in poultry which caused huge economic losses to the poultry industry. However, the prevalence and genetic characteristics of FAdV-4 in Yunnan Province are not fully understood at present. Consequently, this study aimed to understand the epidemic situation of FAdV-4 in Yunnan Province and provide a reference for the prevention and control of the infection. Suspected samples of FAdV-4 were collected from poultry farms in Yunnan Province, and polymerase chain reaction (PCR) detection of FAdV-4 and related co-infection pathogens, pathogen isolation, and genetic evolution analysis of the fiber gene were carried out.

Epidemiology of FAdV-4 in Yunnan Province during 2017–2021

In this study, a total of 2001 poultry tissue samples and 310 fecal samples of migratory birds were collected from Yunnan Province. Detailed information about the samples collected in this study is shown in Supplementary Table 3. Overall, 227 (11.34%) liver samples and one (0.3%) fecal sample tested positive for FAdV-4 nucleic acid, statistical analysis showed that the proportion of poultry infections was significantly higher than that for wild birds (χ² = 36.67; P < 0.001). Analysis of the poultry tissue samples showed that the annual infection positive rate was 13.14%, 9%, 9.5%, 5.56%, and 19.2% in 2017, 2018, 2019, 2020, and 2021, respectively (Table 1). When the Yunnan Province was divided into four regions (central, southern, western, and northern) and regional infection analysis was performed, it was found that the infection rate in central and western Yunnan Province was significantly higher (χ² = 55.26; P < 0.001) (Fig. 1). The host animals of the positive samples were mainly domestic chickens, with some broilers and layers (Table 1). The infection rate was the lowest in eastern Yunnan Province (χ² = 55.26; P < 0.001). Furthermore, the seasonal infection analysis showed that co-infections were more frequent in spring and single infections were more frequent in winter (χ² = 43.09; P < 0.001). In terms of overall infections, the proportion of co-infections was significantly greater than that of single infections (χ² = 54.41; P < 0.001). Seven pathogens susceptible to co-infection were detected, and 162 samples out of 227 samples were infected with 1–3 pathogens which were primarily ILT, CIAV, and AIV with co-infection rates of 37.8% (86/227), 19.8% (45/227), and 10.6% (24/227), respectively. The proportion of co-infections with pathogens such as NDV, IBV, MS, and MG was low (Table 2). The positive fecal samples of wild birds came from the black-necked crane population in Huize National Nature Reserve. There was no co-infection in this sample. Overall, our results revealed that the co-infection of FAdV-4 with immunosuppressive pathogens was widespread, especially with ILT and CIAV.

Table 1

Number of positive poultry samples tested and prevalence in Yunnan Province.
Year	Total sample	FAdV-4 positive sample	FAdV-4 positive rate	Domestic chickens positive sample	Broiler positive sample	Layers positive sample	Viral isolation
2017	411	54	13.14%	36	16	2	1
2018	389	35	9%	24	8	3	1
2019	429	41	9.56%	30	6	5	1
2020	378	21	5.56%	10	4	6	1
2021	394	76	19.29%	58	14	4	2

Table 2

Number of positive samples of pathogens co-infected with FAdV-4.
	2017	2018	2019	2020	2021	Total	co-infection rates
ILT	30	10	3	5	38	86	37.8%
CIAV	6	0	25	4	10	45	19.8%
AIV	4	10	3	11	0	28	12.3%
MG	0	0	6	0	0	6	2.6%
MS	4	0	0	0	0	4	1.7%
ND	0	0	0	4	0	4	1.7%
IBV	0	0	3	0	0	3	1.3%

Phylogenetic analysis

In this study, six Yunnan strains of FAdV-4 were isolated. The five strains were source poultry, named YN1605, YN1802, YN1904, YN2108 and YN2109 respectively and one strain was source wild bird black-necked crane, named YN2010. The six Yunnan strains had the typical genome organization of FAdV-4, with two adjacent fiber genes (fiber-1 and fiber-2). The fiber gene sequence has been successfully uploaded to GenBank, and the sequence number is shown in the supplementary Table 4. Nucleotide sequence analysis was conducted on the fiber genes of the six FAdV-4 isolates. The fiber-1 and fiber-2 gene identities were 96.2–99.5% and 95.8–100%. However, they showed a low sequence identity (< 43.6% and < 50.7%) with the fiber-1 and fiber-2 genes of other aviadenovirus species. Sequence identities for the Chinese highly pathogenic strain JSJ13, AH712, SD1511, HLJFAd15, and HB1510 were 99.1–100%. All six strains in Yunnan province belonged to FAdV-C and the same cluster as the FAdV-4 strains in China. Moreover, the FAdV strains of each subgroup of group I were less related to each other and were clustered in different clades (Fig. 2).

Phylogenetic analysis

The fiber-1 protein for all six Yunnan FAdV-4 isolates had 431 amino acids. Amino acid sequence alignment with the reference strains showed a high similarity to the China highly pathogenic strains (99.1%) and a low similarity to with MX-SHP strain, ON1 strain and the KR5 strain (94.9–96.8%). When compared to the Chinese highly pathogenic strain JSJ13, the six Yunnan isolates had 21–24 amino acid mutations and an N/H⁴³⁰ amino acid deletion (Table 3A). Nuclear localization sequences (NLS) such as KKRP (17–20) and RRKP (37–40) were present in the isolates fiber-1 amino acid sequence. In addition, there was a conserved tail region VYPF (56–60) sequence that interacts with the penton protein. We found the same poly-G structure in the tail region of the fiber-1 amino acid sequence of the isolates as in the reference strain. It is known that the poly-G knob structure interacts with the penton base proteins to participate in the process of viral attachment. The fiber-2 protein contained 479 amino acids and showed high amino acid similarity (100%) with the Chinese highly pathogenic strains and low similarity with MX-SHP strain, ON1 strain and the KR5 strain (94.1–95.4%). In terms of the amino acid mutations, the site variants indicated highly pathogenic strains (G²¹⁹D, P³⁰⁷A, V³¹⁹I, and A³⁰⁸T), and there were 16 variant sites (Table 3B). Nevertheless, the strength of the virulence needs to be further studied. Fiber-2 had NLS that were different from fiber-1, such as KRAKR (28–32), ENGKP (11–15), and a VYPF sequence (21–24).

Table 3

Amino-acid differences in the *fiber* genes from highly pathogenic and low pathogenic FAdV-4 strains.
Amino acid position	14	28	39	44	46	89	102	119	126	154	186	196	204	251	255	262	263	310	329	374	376	383	401	430
HB1510	A	S	K	R	A	P	K	N	A	R	D	V	G	L	L	H	D	H	H	S	G	I	N	-
YN1605	.	.	.	.	.	.	.	.	.	.	.	.	.	.	F	.	.	.	.	.	E	L	Y	-
YN1802	.	.	.	P	.	.	.	.	.	.	.	.	.	.	F	.	.	.	.	P	.	L	.	-
YN1904	.	.	.	.	.	T	Q	.	.	.	.	.	.	.	F	.	.	.	.	P	.	L	Y	-
YN2011	.	.	.	.	T	.	.	.	.	.	.	.	.	.	.	.	.	.	.	P	.	L	Y	-
YN2109	.	.	E	P	.	.	.	.	.	.	.	.	.	.	F	.	.	.	.	P	E	L	Y	-
YN2110	.	.	.	.	T	.	.	.	.	.	.	.	.	.	.	.	.	.	.	P	.	L	Y	-
MX-SHP	V	I	K	P	A	P	K	D	V	H	N	.	A	I	L	Q	E	R	Q	P	G	L	Y	N
JSJ13	.	.	K	R	A	P	K	.	.	.	.	.	.	.	L	.	.	.	.	.	G	.	.	H
ON1	V	I	K	P	A	P	K	D	V	H	N	T	.	I	L	Q	E	R	Q	P	G	L	Y	H
KR5	.	I	K	P	A	P	K	.	.	H	.	.	.	I	L	Q	E	R	Q	P	G	L	Y	-

(A)

Amino acid position	22	25	109	139	219	227	319	324	329	334	338	343	378	380	393	400	427	435	453
HB1510	S	A	D	S	D	Q	I	V	Y	A	N	L	T	T	P	G	I	S	A
YN1605	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.
YN1802	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.
YN1904	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.
YN2011	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.
YN2109	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.
YN2110	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.
MX-SHP	Y	P	A	A	.	.	.	F	.	T	T	N		.	S	A	V	.	.
JSJ13	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.	.
ON1	.	P	.	.	G	E	V	F	V	T	T	N	A	A	.	A	.	T	S
KR5	.	.	.	.	G	E	V	F	V	T	T	N	.	A	.	A	.	T	S
(B)

Sporadic outbreaks of HHS in China have not attracted much attention in the past. However, by 2015, the FAdV-4 widespread outbreak in China had caused huge economic losses to China's poultry industry^{[6, 13, 14]}. Although most poultry farms have been immunized with the FAdV-4 vaccine, the disease still occurs in poultry. Therefore, to prevent and control the occurrence and epidemic of FAdV-4, it was necessary to conduct an epidemiological investigation and sequence variation analysis of the fiber gene. It was also necessary to further investigate the prevalence of FAdV-4 and the molecular characteristics of the fiber genes. The results of this study showed that the average positive rate of FAdV-4 in the 2001 samples from 2017 to 2021 was 11.34%. However, there was a large difference between this value and the FAdV-4 positive detection rate which was reported by Tian et al. (23.4%), which may be related to their increased viral load in the LMH cell culture^[15]. In addition, we found high levels of FAdV-4 infection in the central and western regions of Yunnan Province, which may be related to a large number of poultry farms in the region and the relatively complex introduction channels but this still needs further analysis of the related factors. The pathogen FAdV-4 can infect a wide range of host animals, including broilers, laying hens, and breeding hens^[16]. The positive samples that were identified in this study were mainly domestic chickens, with some broilers, layers, and wild birds. In addition to poultry infections, FAdV-4 infection has also been reported in wild bird samples including Antarctic skuas, crows, and Wild Black Kites^[17–19]. It shows that FAdV-4 has the possibility of cross-population transmission. At the same time, wild birds may act as intermediate hosts in the process of virus transmission. Therefore, we increased the collection of fecal samples from migratory birds from 2020 to 2021, and one sample was found to be FAdV-4 nucleic acid positive out of the 301 fecal samples. Then, by isolating the strain using LMH cells, the FAdV-4 black-necked crane strain was identified. This is the first reported isolation of a FAdV-4 strain from black-necked crane feces in China. The results showed that the black-necked crane had the ability to infect and carry FAdV-4. The phylogenetic analysis showed that the poultry-derived FAdV-4 strain and black-necked crane-derived FAdV-4 strain had a high similarity but the transmission route of FAdV-4 between the black-necked crane source and poultry source needs further investigation. Currently, co-infection of FAdV-4 and other pathogens is very common, especially before infection with FAdV-4 or for simultaneous infection with an immune suppression disease. With co-infections, pathological changes are typical, and in 3–5 weeks they can lead to the broilers or layers dying suddenly with pericardial effusion, hepatitis clinical symptoms, and inhibited humoral immunity, thus, affecting the immune efficacy of NDV and AIV-H9 vaccines and increasing the incidence of respiratory diseases^[2]. Studies have shown that FAdV co-infection with immunosuppressive pathogens may lead to viral re-combination and the emergence of a more pathogenic virus such as IBDV, CIAV, or AIV, which can increase mortality and cause severe clinical symptoms^{[20, 21]}. In this study, we detected 162 cases of co-infections, mainly with a mix of pathogens such as ILT, CIAV, and AIV. ILT is a respiratory virus with high morbidity in Yunnan Province in recent years, and it frequently occurs in the cold seasons of winter and spring, especially in farms with poor feeding environment, high feeding density and irregular management. When combined with the statistical conclusion of a high morbidity associated with co-infection in spring in this study, it can be inferred that the probability of FAdV-4 and ILT co-infection in Yunnan Province is greater. This suggests that to prevent and control FAdV-4 infection, comprehensive prevention and control measures such as vaccine immunization should be strengthened against immunosuppressive diseases such as ILT, CIAV and AIV.

Fiber proteins are important structural proteins involved in FAdV viral infection, with type-specific and subgenus-specific neutralizing and type-specific non-neutralizing epitopes^[22]. Furthermore, the adenovirus fiber is responsible for binding to the cell surface and is related to tissue tropism^{[23, 24]}. In FAdV-4, the fiber protein had two consecutive segments (fiber-1 and fiber-2), which is consistent with our previous sequencing and splicing results^[25]. It is known that the fiber-1 protein plays an important role in viral capsids and adhesion, and we found the same poly-G structure in the tail region of the fiber-1 amino acid sequence as in the reference strain. This structure interacts with penton base proteins to participate in the process of viral attachment^[26]. Additionally, the fiber-2 protein can affect viral virulence, and it was identified as one of the main virulence determinants of the highly pathogenic FAdV-4 strains in China^[26]. The phylogenetic analysis of the fiber genes of the isolates showed that the prevalent FAdV-4 strains in Yunnan Province had a high identity with the early pathogenic FAdV-4 strains in Chinese chickens, suggesting that the Yunnan isolates originated from the early Chinese pathogenic strains. Recently, the signature sites were reported with the non-pathogenic strain variants and pathogenic strains in the fiber protein amino acid site mutations at G219D, P307A, V319I, and A308T, indicating that the variants are highly pathogenic strains^{[27, 28]}. Alignment of the fiber-1 and fiber-2 amino acid sequences of the six Yunnan FAdV-4 isolates with the pathogenic and non-pathogenic strains from different geographical regions identified multiple amino acid substitutions, especially in the fiber-2 protein, which was consistent with other pathogenic strain indicating that the molecular characteristics of the virulent strains have been obtained. This was in agreement with our previous results on the molecular characteristics of hexon gene and pathogenicity of Yunnan poultry FAdV-4 isolates. This study provides an understanding of the prevalence of FAdV-4 in Yunnan Province, which could be used as a reference for the prevention and treatment of the disease.

In conclusion, this study comprehensively investigated the prevalence and genetic features of FAdV-4 in Yunnan Province from 2017 to 2021, showing that FAdV-4 remains highly prevalent among poultry populations in Yunnan Province, China, and FAdV-4 was detected from the feces of black-necked cranes for the first time. Statistical analysis shows that the incidence of FAdV-4 in central Yunnan and western Yunnan is high, and it is easy to be co-infection with ILT, CIAV and AIV. Phylogenetic analysis showed that all FAdV-4 Yunnan strains belonged to genotype C, and they clustered in the same branch as the new highly pathogenic strains in China. At the same time, the importance of continuing to monitor the molecular epidemiology of FAdV-4 strains in the future was emphasized, which provided scientific basis for comprehensive prevention and control of FAdV-4 infection in poultry.

Sample collection and treatment

In this study, a total of 2001 suspected FAdV-4 infected liver samples from poultry and 310 feces samples of wild birds from overwintering habitat for migratory birds in 10 regions (Kunming, Qujing, Yuxi, Zhaotong, Baoshan, Lincang, Dehong, Dali, Chuxiong, and Honghe) and the Yunnan Black-necked Crane National Nature Reserve (Huize, Zhaotong) in Yunnan Province were collected from January 2017 to December 2021.Where about 80% of the Yunnan poultry industry is located. Liver tissue sample were collected from poultry presenting with fluid accumulation in the pericardial sac and an enlarged discolored liver with foci of hemorrhage and necrosis. When sampling wild birds' feces, firstly, the species and approximate population of wild birds in the wetland beach of the plateau in the nature reserve were observed from a distance through a high-powered telescope. Collect fresh feces within the range of wild birds' activities for 1 hour, and immediately put them into a 5 mL frozen storage tube filled with sample preservation solution. According to the morphological characteristics of feces, determine the species and genus of wild birds from which the samples come. The samples’ host sources included broilers, layers, domestic chickens, black-necked cranes, common crane, Bar-headed Goose, Greylag Goose and Black-headed Gull. The specimens were collected using standard procedures and all the samples that were submitted for inspection were stored and transported at a low temperature. All samples were homogenized and diluted using a 1:10 dilution in phosphate-buffered saline, and they underwent three freeze-thaw cycles. The supernatants were collected and centrifuged at 8000 g for 10 min. Then, 200 µL of the supernatant was transferred into a new tube and kept at − 80 ℃ until RNA or DNA extraction. The extracts were used for the detection of FAdV-4 and other viral pathogens.

Detection of the viral pathogens in the samples

The supernatants were processed using a TaKaRa MiniBEST Viral RNA/DNA Extraction Kit Ver according to the manufacturer’s instructions (TaKaRa, Shanghai, China). The DNA or RNA was used to detect FAdV-4, Newcastle disease virus (NDV), avian influenza virus (AIV), chicken infectious anemia virus (CIAV), infectious bronchitis virus (IBV), infectious laryngotracheitis (ILT), Mycoplasma synoviae (MS), and Mycoplasma galliscepticum (MG) pathogens by PCR or reverse transcription-PCR. The primers were designed based on the conserved sequences (Supplementary Table 1), and references were designed from GenBank to test the primers of the FAdV-4, NDV, AIV, CIAV, IBV, ILT, MS, and MG strains. The amplified product was visualized using 1% agarose gel electrophoresis. The amplification products with positive results were verified by the first generation sequencing to ensure the accuracy of the detection results.

Virus isolation

Two representative FAdV-4 positive samples were selected from each year, and they were filtered through 0.22 µm filters, inoculated on monolayer LMH cells, and adsorbed for 1h. Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12 was used a culture medium which was supplemented with 2% newborn bovine serum, 100 U/mL penicillin, and 100 µg/mL streptomycin. The cells were maintained at 37 ℃ in a humidified 5% CO2 incubator for 96 h for at least eight passages. The supernatants and cells with obvious cytopathic effects were harvested for molecular identification by PCR assays and the presence of FAdV-4 viruses was confirmed by sequence analysis to obtain stable strains.

Phylogenetic analysis of the fiber gene

Using the supernatants that were identified to be FAdV-4 positive as the DNA template, the full length of the fiber gene was amplified by PCR using the primers fiber1F/1R, fiber2F/2R, and fiber3F/3R. The PCR products were analyzed on a 1% agarose gel followed by DNA purification and sequencing (Sangon, Shanghai, China). The DNAstar 7.0 software was used to assemble the fiber gene sequences of the FAdV-4 isolates. The analysis was performed with reference to the fiber gene sequences of 72 FAdV-4 strains that were obtained from GenBank and detailed information on the FAdV-4 reference strains is shown in Supplementary Table 2. The Clustal W model in MEGA 6.0 was used for the alignment of the homologous sequences. The phylogenetic tree of the fiber genes of the FAdV-4 isolates and the reference strains were constructed using the neighbor-joining method with 1000 bootstrap replicates.

Data analyses

The differences in the FAdV-4 positive rates between the different groups were analyzed using a Chi-square test in SPSS 21.0 software. A difference with a significance value of P < 0.05 was considered statistically significant.

Full name	Abbreviation
Fowl adenovirus serotype 4	FAdV-4
Fowl adenoviruses	FAdVs
Hepatitis-hydropericardium syndrome	HHS
Polymerase chain reaction	PCR
Newcastle disease virus	NDV
Avian influenza virus	AIV
Chicken infectious anemia virus	CIAV
Infectious bronchitis virus	IBV
Infectious laryngotracheitis	ILT
Mycoplasma synoviae	MS
Mycoplasma galliscepticum	MG

Ethics approval and consent to participate

Based on the Animal Ethics Laws and Regulations in P.R. China, the Animal Ethics Committee of Yunnan Animal Science and Veterinary Institute has approved the feces sampling from Yunnan Black-necked Crane National Nature Reserve (Huize, Zhaotong) Yunnan Province China, on 3 Dec 2020.

Consent for publication

not applicable

Availability of data and materials

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Competing interests

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Funding

This study was financially supported by the epitope localization of avian adenovirus type 4 and molecular mechanism of AS-ONs inhibiting infection, grant number 202201AS070022; The key project of Yunnan Provincial Applied Basic Re-search Plan, grant number 2016FA013; The Yunnan Center for Animal Disease Control and Prevention, grant number 1963037.

Authors' contributions

Resources, J.S. and S.L.; methodology, Z.Z. and W.W.; software, Q.Y.; writing—original draft preparation, X.X.; Q.Y. and Z.Z; writing—review and editing, J.S and X.X.; supervision, J.S.; project administration, Z.Z.; funding acquisition, J.S. and S.L. All authors have read and agreed to the published version of the manuscript.

Acknowledgements

We gratefully acknowledge all data contributors, the Authors and their Originating laboratories responsible for obtaining the sample, and their Submitting laboratories for generating the genetic sequence and metadata.

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Epidemiological investigation and fiber gene genetic analysis of fowl adenovirus serotype 4 from 2017 to 2021 in Yunnan Province, China

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Abstract

Figures

Background

Results

Epidemiology of FAdV-4 in Yunnan Province during 2017–2021

Phylogenetic analysis

Phylogenetic analysis

Discussion

Conclusion

Methods

Sample collection and treatment

Detection of the viral pathogens in the samples

Virus isolation

Data analyses

Abbreviations

Declarations

References

Additional Declarations

Supplementary Files

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