Limits...
Prevalence and molecular heterogeneity of Bartonella bovis in cattle and Haemaphysalis bispinosa ticks in Peninsular Malaysia.

Kho KL, Koh FX, Jaafar T, Nizam QN, Tay ST - BMC Vet. Res. (2015)

Bottom Line: Isolation of B. bovis was successful from all PCR-positive cattle blood samples, except one.At least six genotypes of B. bovis were found circulating in the cattle understudied.New MLST sequence types were identified in Malaysian B. bovis isolates.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Microbiology, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia. khokailing@yahoo.com.

ABSTRACT

Background: Bartonellosis is an emerging zoonotic infection responsible for a variety of clinical syndromes in humans and animals. Members of the genus Bartonella exhibit high degrees of genetic diversity and ecologic plasticity. The infection is usually transmitted to animals and humans through blood-feeding arthropod vectors such as fleas, lice, ticks and sandflies. This study was conducted to investigate the prevalence of Bartonella species in 184 beef cattle, 40 dairy cattle, 40 sheep and 40 goats in eight animal farms across Peninsular Malaysia. Bartonella-specific PCR assays and sequence analysis of partial fragments of the citrate synthase gene were used for detection and identification of B. bovis. Isolation of B. bovis was attempted from PCR-positive blood samples. Molecular heterogeneity of the isolates was investigated based on sequence analysis of gltA, ITS, rpoB genes, ERIC-PCR, as well as using an established multilocus sequence typing (MLST) method. The carriage rate of B. bovis in ticks was also determined in this study.

Results: B. bovis was detected using Bartonella gltA-PCR assays from ten (4.5 %) of 224 cattle blood samples, of which three (1.3 %) were from beef cattle and seven (3.1 %) were from dairy cattle. None of the blood samples from the sheep and goats understudied were positive for B. bovis. Haemaphysalis bispinosa and Rhipicephalus (Boophilus) microplus were the predominant tick species identified in this study. B. bovis was detected from eight of 200 H. bispinosa ticks and none from the R. microplus ticks. Isolation of B. bovis was successful from all PCR-positive cattle blood samples, except one. Strain differentiation of B. bovis isolates was attempted based on sequence analysis of gltA, ITS, rpoB, and ERIC-PCR assay. B. bovis isolates were differentiated into six genotypes using the approach. The genetic heterogeneity of the isolates was confirmed using MLST method. Of the six MLST sequence types identified, five were designated new sequence types (ST23-27), while one (ST18) had been reported previously from Thailand isolates. All except one isolates were segregated into lineage II. A new lineage (IIa) is proposed for a single isolate obtained from a dairy cow.

Conclusions: The current study reported the first detection of B. bovis infection in the cattle and H. bispinosa ticks in Peninsular Malaysia. At least six genotypes of B. bovis were found circulating in the cattle understudied. New MLST sequence types were identified in Malaysian B. bovis isolates. Further study is necessary to explore the zoonotic potential of B. bovis and the vector compatibility of H. bispinosa ticks.

No MeSH data available.


Related in: MedlinePlus

Phylogenetic placement of Malaysian B. bovis isolates based on the concatenated sequences of ftsZ, gltA, groEL, nuoG, ribC, rpoB, ssrA, and ITS genes. Reference sequences were retrieved from Bai et al. [14] for comparison. Bootstrap analysis was performed with 1000 replications. Numbers in brackets are GenBank accession numbers. Scale bar indicates the nucleotide substitutions per sites
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4502507&req=5

Fig2: Phylogenetic placement of Malaysian B. bovis isolates based on the concatenated sequences of ftsZ, gltA, groEL, nuoG, ribC, rpoB, ssrA, and ITS genes. Reference sequences were retrieved from Bai et al. [14] for comparison. Bootstrap analysis was performed with 1000 replications. Numbers in brackets are GenBank accession numbers. Scale bar indicates the nucleotide substitutions per sites

Mentions: Three isolates (F8-2, F8-3 and F6-1) in this study were identified as ST18 which had been reported in B. bovis isolated from cattle in Thailand (B25093). ST23 was represented by two isolates (F1-1 and F8-1) in this study. Other STs (ST24-27) in this study were represented by a single isolate. One of the isolates (F8-6) with unique PCR fingerprint profile (E3) exhibited sequence variation in all the gene fragments except for ssrA gene. The dendrogram constructed based on the concatenated sequences of eight loci demonstrated the segregation of eight B. bovis isolates into lineage II (Fig. 2). The isolate F8-6 (ST27) was placed at a single branch adjacent to those isolates from the lineage II, with a 81 % bootstrap value. The isolate is proposed under a new lineage (designated as lineage IIa).Fig. 2


Prevalence and molecular heterogeneity of Bartonella bovis in cattle and Haemaphysalis bispinosa ticks in Peninsular Malaysia.

Kho KL, Koh FX, Jaafar T, Nizam QN, Tay ST - BMC Vet. Res. (2015)

Phylogenetic placement of Malaysian B. bovis isolates based on the concatenated sequences of ftsZ, gltA, groEL, nuoG, ribC, rpoB, ssrA, and ITS genes. Reference sequences were retrieved from Bai et al. [14] for comparison. Bootstrap analysis was performed with 1000 replications. Numbers in brackets are GenBank accession numbers. Scale bar indicates the nucleotide substitutions per sites
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4502507&req=5

Fig2: Phylogenetic placement of Malaysian B. bovis isolates based on the concatenated sequences of ftsZ, gltA, groEL, nuoG, ribC, rpoB, ssrA, and ITS genes. Reference sequences were retrieved from Bai et al. [14] for comparison. Bootstrap analysis was performed with 1000 replications. Numbers in brackets are GenBank accession numbers. Scale bar indicates the nucleotide substitutions per sites
Mentions: Three isolates (F8-2, F8-3 and F6-1) in this study were identified as ST18 which had been reported in B. bovis isolated from cattle in Thailand (B25093). ST23 was represented by two isolates (F1-1 and F8-1) in this study. Other STs (ST24-27) in this study were represented by a single isolate. One of the isolates (F8-6) with unique PCR fingerprint profile (E3) exhibited sequence variation in all the gene fragments except for ssrA gene. The dendrogram constructed based on the concatenated sequences of eight loci demonstrated the segregation of eight B. bovis isolates into lineage II (Fig. 2). The isolate F8-6 (ST27) was placed at a single branch adjacent to those isolates from the lineage II, with a 81 % bootstrap value. The isolate is proposed under a new lineage (designated as lineage IIa).Fig. 2

Bottom Line: Isolation of B. bovis was successful from all PCR-positive cattle blood samples, except one.At least six genotypes of B. bovis were found circulating in the cattle understudied.New MLST sequence types were identified in Malaysian B. bovis isolates.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Microbiology, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia. khokailing@yahoo.com.

ABSTRACT

Background: Bartonellosis is an emerging zoonotic infection responsible for a variety of clinical syndromes in humans and animals. Members of the genus Bartonella exhibit high degrees of genetic diversity and ecologic plasticity. The infection is usually transmitted to animals and humans through blood-feeding arthropod vectors such as fleas, lice, ticks and sandflies. This study was conducted to investigate the prevalence of Bartonella species in 184 beef cattle, 40 dairy cattle, 40 sheep and 40 goats in eight animal farms across Peninsular Malaysia. Bartonella-specific PCR assays and sequence analysis of partial fragments of the citrate synthase gene were used for detection and identification of B. bovis. Isolation of B. bovis was attempted from PCR-positive blood samples. Molecular heterogeneity of the isolates was investigated based on sequence analysis of gltA, ITS, rpoB genes, ERIC-PCR, as well as using an established multilocus sequence typing (MLST) method. The carriage rate of B. bovis in ticks was also determined in this study.

Results: B. bovis was detected using Bartonella gltA-PCR assays from ten (4.5 %) of 224 cattle blood samples, of which three (1.3 %) were from beef cattle and seven (3.1 %) were from dairy cattle. None of the blood samples from the sheep and goats understudied were positive for B. bovis. Haemaphysalis bispinosa and Rhipicephalus (Boophilus) microplus were the predominant tick species identified in this study. B. bovis was detected from eight of 200 H. bispinosa ticks and none from the R. microplus ticks. Isolation of B. bovis was successful from all PCR-positive cattle blood samples, except one. Strain differentiation of B. bovis isolates was attempted based on sequence analysis of gltA, ITS, rpoB, and ERIC-PCR assay. B. bovis isolates were differentiated into six genotypes using the approach. The genetic heterogeneity of the isolates was confirmed using MLST method. Of the six MLST sequence types identified, five were designated new sequence types (ST23-27), while one (ST18) had been reported previously from Thailand isolates. All except one isolates were segregated into lineage II. A new lineage (IIa) is proposed for a single isolate obtained from a dairy cow.

Conclusions: The current study reported the first detection of B. bovis infection in the cattle and H. bispinosa ticks in Peninsular Malaysia. At least six genotypes of B. bovis were found circulating in the cattle understudied. New MLST sequence types were identified in Malaysian B. bovis isolates. Further study is necessary to explore the zoonotic potential of B. bovis and the vector compatibility of H. bispinosa ticks.

No MeSH data available.


Related in: MedlinePlus