samples were related to two differe

samples were related to two different production systems,
with one production system presenting only one MLVA
type and the other production system presenting 11
different MLVA types found in 15 different farms. The
limited number of samples originated from Mexico could
have influenced the observation of these two clusters. In
Brazil samples were originated from a diagnostic laboratory
that receives clinical specimens from various states
and one frequent type was found in 8 different states.
Furthermore, it was possible to observe heterogeneity of
M. hyopneumoniae within state, in all states analyzed. In
Spain the samples were related to the main pig production
areas, located in Catalonia and Aragon, which can explain
the higher heterogeneity found in that country. Samples
that were obtained from diagnostic laboratories (Brazil,
Mexico, and USA) showed higher heterogeneity in the P97
gene when compared to the isolates originated from Spain.
Spanish samples had limited variation in the P97 locus,
with only five types. The P146 locus had demonstrated
higher variation in both set of samples, strains and clinical
specimens. In total, 17 different types were observed in the
P97 locus, and these types ranged from 2 to 18 repeats. On
the other hand, P146 presented 34 different types ranging
from 7 to 48 repeats. The types that presented more than
30 repeats in P146 were found only in Spain and Brazil.
This observation is in agreement with reports from De
Castro et al. (2006), in which Brazilian samples ranged
from 18 to 44 repeats in the same repeat region.
Nevertheless, no relationship has been demonstrated
between number of repeats and bacterial virulence. It is
important to note that various DNA extraction methods
were used for samples originating in different countries,
and this could have biased the comparison among
countries.
In the results presented by Vranckx et al. (2011) and
Charlebois et al. (2014) the MLVA types for each sample
were not disclosed. Therefore, it was not possible to
compare the number of repeats found in our study with
those previously published. The large number of MLVA
types found in this investigation suggests a large diversity
in M. hyopneumoniae circulating in the swine herds. Also,
evidence of multiple M. hyopneumoniae variants was
observed in single samples (13.8%; 49/355), which was
observed by the amplification of more than one peak in the
capillary electrophoresis analysis. These results suggest
infection with more than one strain in the same pig, as
previously suggested (Vranckx et al., 2011; Charlebois
et al., 2014). The high numbers of M. hyopneumoniae
variants identified in this investigation are in agreement
with other reports which identified clonal variants in the
single herds and in different herds (Nathues et al., 2011;
Vranckx et al., 2012; Charlebois et al., 2014). Clonal
variants present in the same herd showed a low variability
among them, and shared the same number of repeats in
one locus and variable number of repeats in the other
locus. In large production systems, a different pattern of
clonality was observed which is in agreement with the
results of Vranckx et al. (2012).
The M. hyopneumoniae reference strain used to standardize
this MLVA is the parental strain of US232, a
reference strain in the USA (Minion et al., 2004). Both
samples showed the same type (14–21) when evaluated
with this method. This type was only identified in the USA,
which could be explained by the origin of the isolate. There
was not a common MLVA type across all countries.
However, some types appeared to be shared among
countries.
5. Conclusion
The MLVA typing method developed in this study is an
improved assay for differentiation of M. hyopneumoniae
variants in clinical specimens. This assay revealed a high
discriminatory index, suggestive of high diversity of M.
hyopneumoniae in the investigated herds, indicating that
multiple M. hyopneumoniae variants are circulating in
swine herds in the USA, Brazil, Mexico and Spain. Further
analysis of samples collected longitudinally from diverse
geographic locations and clinical presentation is necessary
to investigate if a nonrandom distribution of genotypes is
present among strains.
Competing interest
The authors declare that they have no competing
interest.
Acknowledgments
The authors thank Dr. Anna Perez de Rozas from CReSA
for providing Spanish isolates, Jason Daniels for his help
with sample processing, the UMN-VDL for providing USA
and Mexican samples, and Microvet for its support in
providing samples from Brazil. This project was funded by
the Minnesota Pork Board and the Pork Checkoff, and the
Swine Disease Eradication Center. L.F. Dos Santos was
supported by Capes Foundation, Ministry of Education of
Brazil (proc. No: BEX17617/12-0) and Fundac¸a˜o de Amparo
a` Pesquisa do estado de Minas Gerais FAPEMIG. M.A.S.
Moreira was supported by CNPQ.

ผลลัพธ์ (อังกฤษ) 2:

Two different samples were related to Production Systems,
Production System with one presenting only one MLVA
Type Other Production System and the presenting 11
different types MLVA Found in 15 different Farms. The
Limited Number of samples originated from Mexico could
have influenced the Observation Two of these clusters. In
Brazil samples were originated from a Diagnostic Laboratory
that receives clinical specimens from Various States
and was one Frequent Type Found in 8 different States.
Furthermore, it was possible to Observe heterogeneity of
M. hyopneumoniae within state, in all states analyzed. In
Spain the samples were related to the Main Pig Production
areas, located in Catalonia and Aragon, which Can Explain
the Higher heterogeneity Found in that Country. Samples
that were obtained from Diagnostic Laboratories (Brazil,
Mexico, and USA) Showed Higher heterogeneity in the P97
Gene when compared to the isolates originated from Spain.
Spanish Limited variation in the samples had P97 Locus,
with only Five types. Locus had demonstrated the P146
Higher variation of SET in both samples, strains and clinical
specimens. In total, 17 different types were observed in the
P97 Locus, and these types ranged from 2 to 18 Repeats. On
the Other Hand, P146 Presented 34 different types ranging
from 7 to 48 Repeats. Presented the types that more than
30 Repeats in P146 were Found only in Spain and Brazil.
This is in Agreement with Observation reports from De
Castro et al. (Two thousand and six), in which Brazilian samples ranged
from 18 to 44 in the Repeats Same Repeat Region.
Nevertheless, no has been demonstrated Relationship
between Number of Repeats and bacterial virulence. It is
important to note that DNA extraction Various methods
were used for samples originating in different countries,
and this could have biased the comparison among
countries.
In the results Presented by Vranckx et al. (2,011th) and
Charlebois et al. (The 2,014th) the types MLVA for each sample
were not disclosed. Therefore, it was not possible to
compare the Number of Repeats Found in our Study with
those previously Published. The Large Number of MLVA
types Found in this Investigation suggests a Large Diversity
in M. hyopneumoniae circulating in the Swine herds. Also,
M. hyopneumoniae Evidence of multiple variants was
observed in single samples (13.8%; 49/355), which was
observed by the amplification of more than one in the Peak
capillary electrophoresis Analysis. These results suggest
infection with more than one strain in the Same Pig, as
previously SUGGESTED (Vranckx et al., 2011th; Charlebois
et al., 2014). The High Numbers of M. hyopneumoniae
variants identified in this Investigation are in Agreement
with Other reports which identified the clonal variants in
single herds and herds in different (Nathues et al., 2011th;
Vranckx et al., 2012; Charlebois et al.,. 2014). Clonal
variants present in the Same Herd Showed a low variability
among them, and the Shared Same Number of Repeats in
one and Locus Variable Number of Repeats in the Other
Locus. Production in Large Systems, a different Pattern of
clonality was observed which is in Agreement with the
results of Vranckx et al. (The 2,012th).
The M. hyopneumoniae strain used to Standardize Reference
MLVA this is the Parental strain of US232, a
Reference strain in the USA (Minion et al., 2,004th). Both
samples Showed the Same Type (14-21) when evaluated
with this method. Type this was only identified in the USA,
which could be the origin of the isolate by expLAineD. There
was not a common MLVA Type Across all countries.
However, Some types appeared to be Shared among
countries.
5. Conclusion
Typing MLVA The method developed in this Study is an
improved assay for differentiation of M. hyopneumoniae
variants in clinical specimens. This assay revealed a High
discriminatory index, Suggestive of High Diversity of M.
hyopneumoniae in the investigated herds, Indicating that
multiple variants are circulating in M. hyopneumoniae
Swine herds in the USA, Brazil, Mexico and Spain. Further
Analysis of Collected samples longitudinally from diverse
locations and clinical Geographic Presentation is necessary
to Investigate if a nonrandom Distribution of genotypes is
present among strains.
Competing interest
that they have no competing Declare The Authors
interest.
Acknowledgments
The Authors Thank Dr. Anna Perez de Rozas. from Cresa
for providing Spanish isolates, Jason Daniels for his Help
with sample Processing, UMN-VDL for providing the USA
and Mexican samples, and for Microvet ITS Support in
providing samples from Brazil. This Project was funded by
the Minnesota Pork Board and the Pork checkoff, and the
Swine Disease Eradication Center. Dos Santos LF was
supported by Capes Foundation, Ministry of Education of
Brazil (proc. No: BEX17617 / 12-0) and Fundac¸a~o de Amparo
Pesquisa do Estado de Minas Gerais A `FAPEMIG. MAS
Moreira was supported by CNPQ.

ผลลัพธ์ (อังกฤษ) 3:

Samples were related to two different, production systemsWith one production system presenting only one MLVA.Type and the other production system presenting 11.Different MLVA types found in 15 different farms. The.Limited number of samples originated from Mexico could.Have influenced the observation of these two clusters. In.Brazil samples were originated from a diagnostic laboratory.That receives clinical specimens from various states.And one frequent type was found in 8 different states.Furthermore it was, possible to observe heterogeneity of.M. Hyopneumoniae within state in all, states analyzed. In.Spain the samples were related to the main pig production.Areas located in, Catalonia, and Aragon which can explain.The higher heterogeneity found in that country. Samples.That were obtained from diagnostic, laboratories (BrazilMexico and USA), showed higher heterogeneity in the P97.Gene when compared to the isolates originated from Spain.Spanish samples had limited variation in the, P97 locusWith only five types. The P146 locus had demonstrated.Higher variation in both set of samples strains and, clinical.Specimens. In total 17 different, types were observed in the.P97 locus and these, types ranged from 2 to 18 repeats. On.The, other hand P146 presented 34 different types ranging.From 7 to 48 repeats. The types that presented more than.30 repeats in P146 were found only in Spain and Brazil.This observation is in agreement with reports from De.Castro et al. (2006), in which Brazilian samples ranged.From 18 to 44 repeats in the same repeat region.Nevertheless no relationship, has been demonstrated.Between number of repeats and bacterial virulence. It is.Important to note that various DNA extraction methods.Were used for samples originating in, different countriesAnd this could have biased the comparison among.Countries.In the results presented by Vranckx et al. (2011 and.)Charlebois et al. (2014) the MLVA types for each sample.Were not disclosed. Therefore it was, not possible to.Compare the number of repeats found in our study with.Those previously published. The large number of MLVA.Types found in this investigation suggests a large diversity.In M. Hyopneumoniae circulating in the swine, Also herds.Evidence of multiple M. Hyopneumoniae variants was.Observed in single samples (13.8%; 49 / 355), which was.Observed by the amplification of more than one peak in the.Capillary electrophoresis analysis. These results suggest.Infection with more than one strain in the, same pig as.Previously suggested (Vranckx et al, 2011; Charlebois.Et al, 2014). The high numbers of M. Hyopneumoniae.Variants identified in this investigation are in agreement.With other reports which identified clonal variants in the.Single herds and in different herds (Nathues et al, 2011;Vranckx et al, 2012; Charlebois et al, 2014). Clonal.Variants present in the same herd showed a low variability.Among them and shared, the same number of repeats in.One locus and variable number of repeats in the other.Locus. In large, production systems a different pattern of.Clonality was observed which is in agreement with the.Results of Vranckx et al. (2012).The M. Hyopneumoniae reference strain used to standardize.This MLVA is the parental strain, of US232 a.Reference strain in the USA (Minion et al, 2004). Both.Samples showed the same type (14 - 21) when evaluated.With this method. This type was only identified in, the USAWhich could be explained by the origin of the isolate. There.Was not a common MLVA type across all countries.However some types, appeared to be shared among.Countries.5. Conclusion.The MLVA typing method developed in this study is an.Improved assay for differentiation of M. Hyopneumoniae.Variants in clinical specimens. This assay revealed a high.Discriminatory index suggestive of, high diversity of M.Hyopneumoniae in the investigated herds indicating that,,Multiple M. Hyopneumoniae variants are circulating in.Swine herds in, the USA Brazil Mexico and, Spain. Further.Analysis of samples collected longitudinally from diverse.Geographic locations and clinical presentation is necessary.To investigate if a nonrandom distribution of genotypes is.Present among strains.Competing interest.The authors declare that they have no competing.Interest.Acknowledgments.The authors thank Dr. Anna Perez de Rozas from CReSA.For providing Spanish isolates Jason Daniels, for his help.With sample processing the UMN-VDL, for providing USA.And, Mexican samples and Microvet for its support in.Providing samples from Brazil. This project was funded by.The Minnesota Pork Board and the Pork Checkoff and the,,Swine Disease Eradication Center. L.F. Dos Santos was.Supported by Capes Foundation Ministry of, Education of.Brazil (proc. No: BEX17617 / 12-0) and Fundac a ˜ o de Amparo. "A "Pesquisa do estado de Minas Gerais FAPEMIG. M.A.S.Moreira was supported by CNPQ.