Multi-locus sequence typing of Mycoplasma bovis to assess its genetic diversity from 2009-2018 in Ningxia Hui Autonomous Region, China

doi:10.21203/rs.3.rs-23294/v1

Background: Mycoplasma bovis (M. bovis) is a highly contagious cattle pathogen spreading worldwide and especially in Ningxia province in China.

Methods: For investing epidemiology and population structure, we isolated 65 strains of M. bovis from infectious cattle with pneumonia or mastitis or arthritis. Multi-locus sequence typing (MLST) analysis were used to compare these 65 strains isolated from 2009 to 2018. The molecular epidemiology and population structure of M. bovis of these strains were further compared to PG45 strain and HB0801 reported for the first time in China and others strains isolated from different province of China through MLST analysis. Seven gene sequences of 65 strains in Ningxia and the strains of different STs in China and PG45 reference strains were concatenated to constructe a phylogenetic tree.

Results: Totally two types of ST, ST10 and ST134, were identified in Ningxia. 37 Ningxia strains were classified to ST10 and 28 Ningxia strains were classified to ST134. ST134 strain was a new ST and first found in 2009 and was only widely distributed in Ningxia at present. The M. bovis of ST10 was widely spread in many provinces in China and was widespread in Ningxia since 2010. It is speculated that the prevalence of M. bovis ST10 in Ningxia began in 2010.

Conclusion: This study is the first report on the genetic diversity of M. bovis from 2009 to 2018 in Ningxia Hui Autonomous Region and provides the epidemiological information, thus is meaningful for providing a scientific basis for the prevention and control of M.bovis in Ningxia, China.

Small Animal Medicine

Large Animal Medicine

Multi-locus sequence typing

Mycoplasma bovis

genetic diversity

Ningxia

Mycoplasma bovis (M. bovis) is an important pathogen causing severe pneumonia, mastitis and arthritis in the world. Especially, the pneumonia caused by M. bovis has high morbidity and mortality. It is becoming one of the most widely recognized pathogens in the world[1, 2]. The pathogen is highly contagious and can spread rapidly throughout the herd. M. bovis can not only cause pneumonia, mastitis, arthritis and otitis, but also induce postpartum infection of the uterus with a mortality rate of 80%[3]. Since there is no effective vaccines and drugs to prevent and cure the disease caused by the pathogen, the incidence of the disease is on the rise[4, 5].

With the completion of whole-genome sequencing of M. bovis,a variety of highly repeatable molecular typing methods have been developed for molecular epidemiology and population structure research, including arbitrarily primed PCR(AP-PCR), random amplified polymorphic DNA (RAPD)[6], amplified fragment length polymorphism (AFLP)[7], pulsed field gel electrophoresis (PFGE)[8], insertion sequence(IS)[9], variable number of tandem repeats(VNTR)[10, 11], multiple-locus variable number tandem repeat (MLVA)[10] and multi-locus sequence typing (MLST)[5]. Although AP-PCR, RAPD, AFLP, and PFGE methods can obtain a large amount of genetic information, they are subjective in the analysis of DNA fragments and require special equipment[5], so it is difficult to establish a standardized method.

MLST is a rapidly developing molecular biology analysis method with high resolution in recent years. It is suitable for both molecular epidemiological studies and molecular advancement studies. The MLST method compares the nucleic acid sequences of the core fragments of several housekeeping genes, and then compares the diversity of the alleles of the strains. Different strains correspond to different sequence types (ST)[4]. Through the STs of M. bovis pathogens to understand the genetic diversity, population structure and evolutionary trend, which will be beneficial to the control of M. bovis and the development of vaccines, as well as providing a theoretical basis for the prevention and control of M. bovis[4]. MLST is a typing technique based on the seven housekeeping genes of M. bovis to study the genetic diversity, population structure and evolutionary trend of M. bovis, including alcohol dehydrogenase-1 (adh-1), glutamate tRNA ligase (gltX), glycerol-3-phosphate dehydrogenase (gpsA), DNA gyrase subunit B (gyrB), phosphate acetyltrans-ferase-2 (pta-2), thymidine kinase (tdk) and transketolase(tkt)[12]. At present, MLST method is used in M. bovis and research data were uploaded to a public database (www.pubmlst.org/mbovis) for uploading and comparing the STs of M. bovis, and the new STs can also be uploaded to the website for review and registration[5]. MLST is a powerful, scalable and highly standardized method that makes it easy to clearly distinguish housekeeping genes among different strains[13].

Ningxia Hui Autonomous Region is one of the most important raising regions for cow and beef cattle in China[14]. The feeding level of bovine is currently ranked second in China. In 2009, our team isolated M. bovis for the first time from the lung of cows in Ningxia. To 2018, a total of 65 strains of M. bovis were obtained from samples of lung, synovial fluid, nasal swab and milk sample in Ningxia. However, there is no related study on the molecular epidemiology and population structure of M. bovis in Ningxia. In this study, MLST method was used to classify 65 isolates from different regions of Ningxia from 2009 to 2018, aiming to explore the molecular prevalence and to investigate the population structure of M. bovis[2, 5] and lay a foundation for further prevention and control of M. bovis in Ningxia and even in China.

Strains

The detail information of sixty-seven strains of M. bovis were listed in Table 1, including the host, isolation site, geographical location and number of strains. Sixty-five strains were isolated from nasal swabs, or milk, or joint fluid, or lungs of M.bovis-infected cattle in Ningxia from 2009 to 2018. HB0801 strain was isolated from the lungs of beef cattle in Hubei Province in 2008, when was first reported in China[15]. PG45 strain (ATCC 25523) was donated by Professor Aizhen Guo in Huazhong Agricultural University of China.

Table 1

M. bovis strains used in this study and their STs
Sample ID	Year of isolation	Origin	Host	Sample type	Clinical status	adh1	gltX	gpsA	gyrB	pta2	tdk	tkt	ST	Source
PG45	1961	Amercian	Cow	Milk	Clinical mastitis	3	2	4	2	1	3	2	17	This study
HB0801	2008	HuBei of China	Cattle	Lung	Pneumonia	4	3	2	3	5	3	4	10	This study
NX001	2009	Wuzhong	Cow	Milk	Clinical mastitis	4	3	2	3	17	3	4	134	This study
NX002	2010	Wuzhong	Cow	Lung	Pneumonia	4	3	2	3	5	3	4	10	This study
NX003	2010	Wuzhong	Dairy calf	Joint fluid	arthritis	4	3	2	3	5	3	4	10	This study
NX004	2010	Wuzhong	Dairy calf	Joint fluid	arthritis	4	3	2	3	17	3	4	134	This study
NX005	2010	Wuzhong	Cow	Nose Swab	Pneumonia	4	3	2	3	5	3	4	10	This study
NX006	2010	Shizuishan	Cow	Nose Swab	Pneumonia	4	3	2	3	5	3	4	10	This study
NX007	2010	Wuzhong	Dairy calf	Joint fluid	arthritis	4	3	2	3	5	3	4	10	This study
NX008	2011	Guyuan	Cattle	Lung	Pneumonia	4	3	2	3	17	3	4	134	This study
NX009	2011	Yinchuan	Cow	Nose Swab	Pneumonia	4	3	2	3	5	3	4	10	This study
NX010	2011	Wuzhong	Cow	Lung	Pneumonia	4	3	2	3	5	3	4	10	This study
NX011	2011	Yinchuan	Cow	Nose Swab	Pneumonia	4	3	2	3	5	3	4	10	This study
NX012	2012	Wuzhong	Cow	Nose Swab	Pneumonia	4	3	2	3	5	3	4	10	This study
NX013	2012	Wuzhong	Cow	Nose Swab	Pneumonia	4	3	2	3	5	3	4	10	This study
NX014	2012	Guyuan	Cattle	Lung	Pneumonia	4	3	2	3	5	3	4	10	This study
NX015	2013	Wuzhong	Cattle	Lung	Pneumonia	4	3	2	3	5	3	4	10	This study
NX016	2013	Yinchuan	Cow	Milk	Clinical mastitis	4	3	2	3	17	3	4	134	This study
NX017	2013	Shizuishan	Cow	Milk	Clinical mastitis	4	3	2	3	17	3	4	134	This study
NX018	2013	Yinchuan	Cow	Milk	Clinical mastitis	4	3	2	3	17	3	4	134	This study
NX019	2013	Yinchuan	Cow	Milk	Clinical mastitis	4	3	2	3	17	3	4	134	This study
NX020	2013	Yinchuan	Cow	Nose Swab	Pneumonia	4	3	2	3	5	3	4	10	This study
NX021	2013	Guyuan	Cattle	Lung	Pneumonia	4	3	2	3	5	3	4	10	This study
NX022	2013	Yinchuan	Dairy calf	Joint fluid	arthritis	4	3	2	3	17	3	4	134	This study
NX023	2014	Yinchuan	Cow	Nose Swab	Pneumonia	4	3	2	3	17	3	4	134	This study
NX024	2014	Yinchuan	Cow	Milk	Clinical mastitis	4	3	2	3	5	3	4	10	This study
NX025	2014	Yinchuan	Cow	Lung	Pneumonia	4	3	2	3	17	3	4	134	This study
NX026	2014	Yinchuan	Cow	Nose Swab	Pneumonia	4	3	2	3	17	3	4	134	This study
NX027	2014	Yinchuan	Cow	Lung	Pneumonia	4	3	2	3	17	3	4	134	This study
NX028	2014	Yinchuan	Cow	Lung	Pneumonia	4	3	2	3	17	3	4	134	This study
NX029	2014	Shizuishan	Cattle	Lung	Pneumonia	4	3	2	3	5	3	4	10	This study
NX030	2014	Yinchuan	Cow	Nose Swab	Pneumonia	4	3	2	3	5	3	4	10	This study
NX031	2014	Yinchuan	Cow	Milk	Clinical mastitis	4	3	2	3	17	3	4	134	This study
NX032	2014	Yinchuan	Cow	Milk	Clinical mastitis	4	3	2	3	17	3	4	134	This study
NX033	2015	Wuzhong	Dairy calf	Joint fluid	arthritis	4	3	2	3	17	3	4	134	This study
NX034	2015	Wuzhong	Dairy calf	Joint fluid	arthritis	4	3	2	3	17	3	4	134	This study
NX035	2015	Wuzhong	Dairy calf	Joint fluid	arthritis	4	3	2	3	17	3	4	134	This study
NX036	2015	Wuzhong	Dairy calf	Joint fluid	arthritis	4	3	2	3	17	3	4	134	This study
NX037	2015	Wuzhong	Dairy calf	Joint fluid	arthritis	4	3	2	3	17	3	4	134	This study
NX038	2015	Shizuishan	Cow	Milk	Clinical mastitis	4	3	2	3	17	3	4	134	This study
NX039	2016	Guyuan	Cattle	Lung	Pneumonia	4	3	2	3	5	3	4	10	This study
NX040	2016	Guyuan	Cattle	Lung	Pneumonia	4	3	2	3	5	3	4	10	This study
NX041	2016	Guyuan	Cattle	Lung	Pneumonia	4	3	2	3	5	3	4	10	This study
NX042	2016	Guyuan	Cattle	Lung	Pneumonia	4	3	2	3	5	3	4	10	This study
NX043	2017	Yinchuan	Dairy calf	Joint fluid	arthritis	4	3	2	3	5	3	4	10	This study
NX044	2017	Yinchuan	Dairy calf	Joint fluid	arthritis	4	3	2	3	5	3	4	10	This study
NX045	2017	Yinchuan	Dairy calf	Joint fluid	arthritis	4	3	2	3	5	3	4	10	This study
NX046	2018	Guyuan	Beef cattle	Lung	Pneumonia	4	3	2	3	5	3	4	10	This study
NX047	2018	Yinchuan	Dairy calf	Lung	Pneumonia	4	3	2	3	17	3	4	134	This study
NX048	2018	Yinchuan	Dairy calf	Lung	Pneumonia	4	3	2	3	17	3	4	134	This study
NX049	2018	Wuzhong	Dairy calf	Joint fluid	arthritis	4	3	2	3	5	3	4	10	This study
NX050	2018	Wuzhong	Dairy calf	Joint fluid	arthritis	4	3	2	3	5	3	4	10	This study
NX051	2018	Yinchuan	Dairy calf	Joint fluid	arthritis	4	3	2	3	5	3	4	10	This study
NX052	2018	Yinchuan	Dairy calf	Joint fluid	arthritis	4	3	2	3	5	3	4	10	This study
NX053	2018	Yinchuan	Dairy calf	Joint fluid	arthritis	4	3	2	3	5	3	4	10	This study
NX054	2018	Yinchuan	Dairy calf	Joint fluid	arthritis	4	3	2	3	5	3	4	10	This study
NX055	2018	Yinchuan	Dairy calf	Joint fluid	arthritis	4	3	2	3	5	3	4	10	This study
NX056	2018	Yinchuan	Dairy calf	Joint fluid	arthritis	4	3	2	3	5	3	4	10	This study
NX057	2018	Yinchuan	Dairy calf	Joint fluid	arthritis	4	3	2	3	5	3	4	10	This study
NX058	2018	Yinchuan	Dairy calf	Joint fluid	arthritis	4	3	2	3	5	3	4	10	This study
NX059	2018	Yinchuan	Dairy calf	Lung	Pneumonia	4	3	2	3	5	3	4	10	This study
NX060	2018	Wuzhong	Dairy calf	Lung	Pneumonia	4	3	2	3	5	3	4	10	This study
NX061	2018	Yinchuan	Dairy calf	Nose Swab	Pneumonia	4	3	2	3	17	3	4	134	This study
NX062	2018	Yinchuan	Dairy calf	Nose Swab	Pneumonia	4	3	2	3	17	3	4	134	This study
NX063	2018	Yinchuan	Dairy calf	Nose Swab	Pneumonia	4	3	2	3	17	3	4	134	This study
NX064	2018	Yinchuan	Dairy calf	Nose Swab	Pneumonia	4	3	2	3	17	3	4	134	This study
NX065	2018	Yinchuan	Dairy calf	Nose Swab	Pneumonia	4	3	2	3	17	3	4	134	This study
Hubei-1	2008	Hubei	Cattle	lung	pneumonia	4	3	2	3	5	4	4	26	MLST web
EZ-8-NHD0962	2008	Hubei	bovine	lung	pneumonia	4	5	2	3	5	3	4	32	MLST web
EZ-2	2008	Hubei	bovine	lung	pneumonia	4	3	2	12	5	3	4	43	MLST web
NHD0986	2008	Hunan	bovine	lung	pneumonia	4	3	2	12	5	3	4	43	MLST web
NMH7	2018	Inner Mongolia	bovine	milk	mastitis	10	3	6	13	21	6	10	173	MLST web
HBHS01	2018	Hubei	bovine	milk	mastitis	4	3	2	3	5	7	4	172	MLST web
Shaanxi04	2018	Shaanxi	bovine	milk	mastitis	4	3	2	3	5	7	4	172	MLST web

Cultivation and identification ofM. bovis

The M. bovis strains were cultured in PPLO broth(BD DifcoTM, US California)[4, 16]. M.bovis genomic DNA was extracted using a bacterial DNA extraction kit (Tiangen,Beijing,China)[17]. 16S rRNA gene[18] were amplificated using two pairs of primers (Forward:5'-GAATTCCGAGAGTTTGATCCTGGCT-3'; Reverse:5'-AAGCTTGAGGTAATC CATCCCCACGTTC-3') and uvrC gene[19] primers (Forward:5'-GAATTCAATGTGTCTAC TAGTCCTGG − 3'; Reverse:5'-AAGCTTAGCGTCATAGATTTTTGCATA-3'). The PCR reaction mixture was 50µL (5 µL 10 × PCR Buffer, 4 µL dNTP Mixture, 0.25 µL rTaq, 2µL primers, 1µL DNA, plus ddH2O to 50µL). The reaction mixture was incubated at 95℃ for 2 min, 35 cycles of 95℃ for 30 s, 50℃ for 20 s and 72℃ for 2 min, then a final incubation at 72℃ for 8 mins[18, 19].

Multi-locus Sequence Typing

The gene of 67 strains of M. bovis isolates were amplified by PCR using MLST scheme (adh-1, gltX, gpsA, gyrB, pta-2, tdk and tkt)[4, 5] (Table 2). The PCR reaction mixture was 50µL (5 µL 10 × PCR Buffer, 4 µL dNTP Mixture, 0.25 µL rTaq, 2µL primers, 1µL DNA, plus ddH2O to 50µL). The reaction mixture was incubated at 95 ℃ for 2 min, 35 cycles of 95 ℃ for 30 s, 55 ℃ for 30 s and 72 ℃ for 1 min, then a final incubation at 72 ℃ for 5 mins[5]. The PCR amplification products were sent to Shanghai Bioengineering Co., Ltd for sequencing. The assembly sequences were aligned by the http://pubmlst.org/mbovis/ database to obtain the allele number and STs.

Table 2

Primers used for amplification of MLST loci.
Name	Sequence	Amplicon size (bp)
adh-1-1	5’- GGA GTA ACT AGT TAC AAA GCA CTT A -3’	546
adh-1-2	5’- TGC TAG TTG TTC AAA CAC GT -3’	546
gltx-3	5’- TGG TGA GTA TTC AAT AAG GT-3’	530
gltX-4	5’- GTT TTG AGA ATC ATT GCA − 3’	530
gpsA-3	5’- AAA ATG TGA GGA ATT GAT CA -3’	521
gpsA-2	5’- CCA ATT CCA ATT GCT AAA AC -3’	521
gyrB-1	5’- AGC TTG CTA ATT GCA CCA − 3’	678
gyrB-2	5’- TAT TTT GAA CAA ATT TTG CAT − 3’	678
pta-2-1	5’- AAT TCG TAA TGG CAA AGA AG -3’	490
pta-2-2	5’- CTT AGC TTT TCT TAC ATT TAG GT -3’	490
tdk-3	5’ –ATG TAT TTA AAA AGT GGA TTA GG -3’	572
tdk-4	5’- TAT CTC ATA GCT TTT TTA GC -3’	572
tkt-1	5’- CCA ACT TAT ATT ATG GTG CA -3’	533
tkt-2	5’- CCA CCA TAT AAA TTA ATG CC -3’	533

Phylogenetic Analysis

The phylogenetic tree constructed from the concatenated sequences of the seven target genes revealed two distinct lineages (Fig. 1). The PG45 (ST17) and NMH7 (ST173) were in the same lineage. The others STs were in the other lineage including ST26, ST32, ST43, ST 172 and the Ningxia isolates of ST173.

Strains identification

67 strains were identified by PCR amplification using 16S rRNA primers and uvrC primers and sequencing of amplified products, which confirmed that 67 strains were all M. bovis.

MLST analysis ofM. bovis isolates

A total of 3 STs were identified among the 67 strains of M. bovis. There were 2 STs among 65 isolates from Ningxia, which are ST10 (n = 37) and ST134 (n = 28). HB0801 strain was ST10 and PG45 strain was ST17 (Table 1).

According to Table 1, ST10 and ST134 strains could be isolated from samples of clinical mastitis (n = 9), arthritis (n = 22), and pneumonia (n = 34). The main clinical symptoms of calves were arthritis and pneumonia. The clinical symptoms of dairy cattle were mainly mastitis and pneumonia, while beef cattle were mainly pneumonia and arthritis.

The NX001 strain isolated from Ningxia for the first time in 2009 was ST134, and the NX002 strain isolated in 2010 was ST10. Since then, the ST10 and ST134 have been isolated from different lesions of bovine in different cities in Ningxia. In China, there were six STs that have reported including ST10, ST26, ST32, ST43, ST172 and ST173. Therefore, ST134 was a new type isolated in 2009.

Molecular epidemiological studies are of great significance to reveal the population structure, genetic diversity and prevalence of M. bovis[4, 20], which facilitates the formulation of effective prevention and control measures, including the development of vaccines and diagnostic methods[4, 21]. It has been reported that MLST studies of 44 strains isolated from 9 provinces of China from 2008 to 2014[4] including Hubei province (n = 25) ST10, ST32, ST43, Anhui province (n = 1) ST10, Fujian province (n = 2) ST10, Hunan province (n = 1) ST10, Henan province (n = 8) ST10, Inner Mongolia (n = 1) province ST10, Jiangxi province (n = 3) ST10, Guangzhou province (n = 2) ST10 and Shandong province (n = 1) ST10. However, there is no reports about the MLST study of M. bovis in Ningxia. Therefore, this study is the first report on the molecular epidemiology of M. bovis from 2009 to 2018 in Ningxia Hui Autonomous Region.

At present, there are seven STs in M. bovis in China including ST10, ST26, ST32, ST43, ST134, ST172 and ST173[4]. ST10 is widely prevalent in all provinces reported in China, including Ningxia (n = 37, 56.9%) where are no MLST reports. After the ST10 strain was first isolation in 2010 in Ningxia, the ST10 of M. bovis could be isolated and identified every year. And it has been widely diffuse in different cities in Ningxia. At present, the reason for the widespread prevalence of M. bovis in China is that the north of China is mainly cattle breeding area[4]. With the transportation of northern cattle, ST10 strains are widely distributed in different province of China. The reason for the widespread prevalence of ST10 strain in Ningxia may be that the Ningxia government strongly supports the construction of large-scale cattle breeding Park. Expansion of intensive cattle farms had to purchase cattle from different province of China and M. bovis was ignored, which led to the widespread presence of exogenous M. bovis in Ningxia. It is speculated that the prevalence of M. bovis ST10 in Ningxia began in 2010.

In 2009, our lab isolated M.bovis from the lung tissue of cows for the first time, which was ST134. From 2009 to 2018, ST134 (n = 28, 43.1%) can be isolated and identified from different cities of Ningxia. However, it has not been identified in other provinces in China. Therefore, it confirmed that ST134 strains were unique strains in Ningxia, China, and has been widely popular in Ningxia for many years.

In order to evaluate the evolutionary relationship of M. bovis between isolates from Ningxia and isolates from other provinces in China. Evolutionary analysis showed that all ST10 strains were in the same lineage and were the same lineage as ST26, ST32, ST43 and ST172, even include Ningxia strains of ST134. However, the strain with ST173 was single lineage and was the same as the PG45 strain with ST17. The study results indicate that the strains were the same source except the ST173 strain.

This study is the first report on the genetic diversity of M. bovis from 2009 to 2018 in Ningxia Hui Autonomous Region. There were only two types of M.bovis that were ST10 strains and ST134 strains in Ningxia from 2009 to 2018. ST134 strain was first found in 2009 in China and was only widely distributed in Ningxia Hui Autonomous Region (n = 28, 43.1%) since 2009. ST10 strain was widely prevalent in all provinces reported in China.

M.bovis: Mycoplasma bovis; MLST: Multi-locus sequence typing; ST: Sequence types; adh-1: Alcohol dehydrogenase-1; gltX: Glutamate tRNA ligase; gpsA: Glycerol-3-phosphate dehydrogenase; gyrB: DNA gyrase subunit B; pta-2: Phosphate acetyltrans-ferase-2; tdk: thymidine kinase; tkt: Transketolase.

Ethics approval and consent to participate

This study was conducted in accordance with the Law on Animal Protection and Welfare of Ningxia Hui Autonomous Region of China. Samples were recovered after acquiring the permission from study participants. All owners of farmers who participated in this provided their verbal consent to participate in this study. This study was submitted to and approved by the Ethics Committee of Animal Study in Ningxia University.

Consent for publication

Not applicable.

Availability of data and materials

The datasets used and analyzed during the current study are available from the corresponding author on reasonable request. Some of the DNA sequences during the current study are available in the MLST database(www.pubmlst.org/mbovis).

Competing interests

The authors declare no co mpeting financial interests of this article.

Funding

This work was funded by Ningxia Department of Science and technology for supporting the project funding (2019BBF02005).

Authors' contributions

YNG, SHH and YL conceived and designed the experiments; YNG and HFL analyzed the data and drafted the manuscript; YNG, YYL and SQG performed experiments and acquired data. YNG, SHH and HFL revised the manuscript. All authors read and approved the final manuscript.

Acknowledgements

The authors thank the staff of Clinical Veterinary Laboratory, School of Agricultural of Ningxia University for their unremitting efforts in the isolation and identification of M. bovis since 2009. Thanks to Professor Aizhen Guo from Huazhong Agricultural University for donating PG45 and HB0801 strains. Thanks to the Ningxia Science and Technology Department for providing funds for this work.

Authors' information

YNG, HFL and SHH: Department of Veterinary Clinical Sciences, School of Agriculture, Ningxia University, Yinchuan 750021, Ningxia, China. SQG: Technology Center Department, Yinchuan Customs, Yinchuan 750000, Ningxia, China. YYL: Animal Disease Prevention and Control Center of Yan’an, Yan’an 716000, Shaanxi, China YL: Department of Life Science and Technology, Ningxia Polytechnic College, Yinchuan 750021, Ningxia, China

Gille L, Pilo P, Valgaeren BR, Van Driessche L, Van Loo H, Bodmer M, Burki S, Boyen F, Haesebrouck F, Deprez P et al: A new predilection site of Mycoplasma bovis: Postsurgical seromas in beef cattle. Vet Microbiol 2016, 186:67-70.
Register KB, Woodbury MR, Davies JL, Trujillo JD, Perez-Casal J, Burrage PH, Clark EG, Windeyer MC: Systemic mycoplasmosis with dystocia and abortion in a North American bison (Bison bison) herd. J Vet Diagn Invest 2013, 25(4):541-545.
Burki S, Spergser J, Bodmer M, Pilo P: A dominant lineage of Mycoplasma bovis is associated with an increased number of severe mastitis cases in cattle. Vet Microbiol 2016, 196:63-66.
Menghwar H, He C, Zhang H, Zhao G, Zhu X, Khan FA, Faisal M, Rasheed MA, Zubair M, Memon AM et al: Genotype distribution of Chinese Mycoplasma bovis isolates and their evolutionary relationship to strains from other countries. Microb Pathog 2017, 111:108-117.
Register KB, Thole L, Rosenbush RF, Minion FC: Multilocus sequence typing of Mycoplasma bovis reveals host-specific genotypes in cattle versus bison. Vet Microbiol 2015, 175(1):92-98.
Butler JA, Pinnow CC, Thomson JU, Levisohn S, Rosenbusch RF: Use of arbitrarily primed polymerase chain reaction to investigate Mycoplasma bovis outbreaks. Vet Microbiol 2001, 78(2):175-181.
Soehnlen MK, Aydin A, Murthy KS, Lengerich EJ, Hattel AL, Houser BA, Fenton GD, Lysczek HR, Burns CM, Townsend AM et al: Epidemiology of Mycoplasma bovis in Pennsylvania veal calves. J Dairy Sci 2012, 95(1):247-254.
Arcangioli MA, Aslan H, Tardy F, Poumarat F, Le Grand D: The use of pulsed-field gel electrophoresis to investigate the epidemiology of Mycoplasma bovis in French calf feedlots. Vet J 2012, 192(1):96-100.
Miles K, McAuliffe L, Persson A, Ayling RD, Nicholas RA: Insertion sequence profiling of UK Mycoplasma bovis field isolates. Vet Microbiol 2005, 107(3-4):301-306.
Pinho L, Thompson G, Rosenbusch R, Carvalheira J: Genotyping of Mycoplasma bovis isolates using multiple-locus variable-number tandem-repeat analysis. J Microbiol Methods 2012, 88(3):377-385.
Nava Vargas A, Milian Suazo F, Canto Alarcon GJ, Rubio Venegas Y, Guerrero Solorio R, Rodriguez Hernandez E, Pizano Martinez O: Genetic diversity based on MIRU-VNTR profile of isolates of Mycobacterium bovis from Mexican cattle. Prev Vet Med 2016, 131:75-78.
Manso-Silvan L, Dupuy V, Lysnyansky I, Ozdemir U, Thiaucourt F: Phylogeny and molecular typing of Mycoplasma agalactiae and Mycoplasma bovis by multilocus sequencing. Vet Microbiol 2012, 161(1-2):104-112.
Yeo M, Mauricio IL, Messenger LA, Lewis MD, Llewellyn MS, Acosta N, Bhattacharyya T, Diosque P, Carrasco HJ, Miles MA: Multilocus sequence typing (MLST) for lineage assignment and high resolution diversity studies in Trypanosoma cruzi. PLoS Negl Trop Dis 2011, 5(6):e1049.
Tan C, Yang J, Li M: Temporal-Spatial Variation of Drought Indicated by SPI and SPEI in Ningxia Hui Autonomous Region, China. Atmosphere 2015, 6(10):1399-1421.
Qi J, Guo A, Cui P, Chen Y, Mustafa R, Ba X, Hu C, Bai Z, Chen X, Shi L et al: Comparative geno-plasticity analysis of Mycoplasma bovis HB0801 (Chinese isolate). PLoS One 2012, 7(5):e38239.
Rasheed MA, Qi J, Zhu X, Chenfei H, Menghwar H, Khan FA, Zhao G, Zubair M, Hu C, Chen Y et al: Comparative Genomics of Mycoplasma bovis Strains Reveals That Decreased Virulence with Increasing Passages Might Correlate with Potential Virulence-Related Factors. Front Cell Infect Microbiol 2017, 7:177.
Li J, Zuo X, Lv X, Kong F, Xu W, Yang S: Association of DNA repair gene polymorphisms with response to chemotherapy and prognosis of gastric cancer in a Chinese population. Tumour Biol 2014, 35(8):7569-7574.
Konigsson MH, Bolske G, Johansson KE: Intraspecific variation in the 16S rRNA gene sequences of Mycoplasma agalactiae and Mycoplasma bovis strains. Vet Microbiol 2002, 85(3):209-220.
Subramaniam S, Bergonier D, Poumarat F, Capaul S, Schlatter Y, Nicolet J, Frey J: Species identification of Mycoplasma bovis and Mycoplasma agalactiae based on the uvrC genes by PCR. Mol Cell Probes 1998, 12(3):161-169.
Olaogun OM, Kanci A, Barber SR, Tivendale KA, Markham PF, Marenda MS, Browning GF: Genetic diversity of Mycoplasma arginini isolates based on multilocus sequence typing. Vet Microbiol 2015, 180(1-2):123-128.
Dos Santos LF, Sreevatsan S, Torremorell M, Moreira MA, Sibila M, Pieters M: Genotype distribution of Mycoplasma hyopneumoniae in swine herds from different geographical regions. Vet Microbiol 2015, 175(2-4):374-381.

Multi-locus sequence typing of Mycoplasma bovis to assess its genetic diversity from 2009-2018 in Ningxia Hui Autonomous Region, China

Status:

Journal Publication

Version 1

Abstract

Figures

Background

Methods

Strains

Cultivation and identification ofM. bovis

Multi-locus Sequence Typing

Phylogenetic Analysis

Results

Strains identification

MLST analysis ofM. bovis isolates

Discussion

Conclusions

List of Abbreviations

Declarations

References

Status:

Journal Publication

Version 1