Molecular detection of Bartonella spp. in Malaysian small flying foxes (Pteropus hypomelanus).

Bartonella spp. are emerging zoonotic pathogens responsible for a wide variety of clinical syndromes in humans. Bats have been increasingly reported as reservoirs for Bartonella spp. In this study, molecular investigation showed the presence of Bartonella DNA in two of 30 blood samples of Malaysian small flying foxes (Pteropus hypomelanus). Two strains (Bartonella sp. KS013a and KS013b) were isolated from a PCR-positive blood sample after five days of incubation on blood agar. Based on the dendrogram constructed from 16S rRNA gene sequences, the two strains were genetically most closely related to ruminant associated Bartonella spp. Both strains are regarded as potentially novel Bartonella species as their citrate synthase (gltA) sequences exhibit less than 96% similarities to all previously identified Bartonella spp. Additionally, high gltA sequence similarity was observed between the strains with that reported from a bat fly (Cyclopodia horsfieldi) collected from P. hypomelanus. Possible transmission of Bartonella infection through bat flies and the impact of the infection in P. hypomelanus are yet to be investigated.

Molecular evidence of hemoplasmas in Malaysian cattle and ticks.

Hemotrophic mycoplasma (hemoplasmas) is a cell wall-less bacterium causing infectious anemia in animals. As data on hemoplasmas infecting cattle in Malaysia is scarce, specific polymerase chain reaction assays were used for detection of hemoplasmas from blood samples of cattle and ticks in this study. Hemoplasma DNA was detected in 69 (69.0%) of 100 cattle blood samples obtained from different breeds. A total of 50.0% of the cattle in this study were infected with only Mycoplasma wenyonii, while 2.0% were infected with only Candidatus Mycoplasma haemobos and 17% were infected with both species. Based on sequence analysis of the partial or nearly full length sequences of hemoplasma 16S rRNA gene, the presence of M. wenyonii and Candidatus M. haemobos was confirmed. Hemoplasmapositive cattle of less than three years appeared to have higher infection rate compared to other age groups. M. wenyonii was identified for the first time in approximately 30% of cattle ticks (Rhipicephalus microplus and Haemaphysalis sp.) in this study. This study presents the first molecular evidence of hemoplasmas in Malaysian cattle and ticks.

Molecular Detection of Coxiella burnetii from Farm Animals and Ticks in Malaysia.

Coxiella burnetii, the causative agent of Q fever, is an intracellular bacterium of medical and veterinary importance. The reservoirs of C. burnetii are extensive which include mammals and arthropods, particularly ticks. As the organism is difficult to culture, this study was aimed to detect C. burnetii DNA in animal (mainly blood and vaginal samples of cattle, goats and sheep) and tick samples obtained from farm animals, wild rodents and vegetation. Two polymerase chain reaction (PCR) assays targeting IS1111 transposon-like gene (TransPCR) and com1 gene (OMP-PCR) were used for C. burnetii detection. Sequence determination of the amplified fragments and a real-time PCR assay were used to confirm PCR findings. C. burnetii DNA was detected from 9.1% of cattle blood and 4.2% vaginal samples, respectively. A small percentage (5.8%) of ticks (including Amblyomma, Dermacentor, Rhipicephalus and Haemaphysalis spp.) haboring C. burnetii were identified in this study. This study provides molecular evidence on the presence of C. burnetii in cattle and ticks. The possible zoonotic transmission of C. burnetii is yet to be investigated.

Molecular evidence of hemoplasmas in Malaysian cattle and ticks

Hemotrophic mycoplasma (hemoplasmas) is a cell wall-less bacterium causing infectious anemia in animals. As data on hemoplasmas infecting cattle in Malaysia is scarce, specific polymerase chain reaction assays were used for detection of hemoplasmas from blood samples of cattle and ticks in this study. Hemoplasma DNA was detected in 69 (69.0%) of 100 cattle blood samples obtained from different breeds. A total of 50.0% of the cattle in this study were infected with only Mycoplasma wenyonii, while 2.0% were infected with only Candidatus Mycoplasma haemobos and 17% were infected with both species. Based on sequence analysis of the partial or nearly full length sequences of hemoplasma 16S rRNA gene, the presence of M. wenyonii and Candidatus M. haemobos was confirmed. Hemoplasmapositive cattle of less than three years appeared to have higher infection rate compared to other age groups. M. wenyonii was identified for the first time in approximately 30% of cattle ticks (Rhipicephalus microplus and Haemaphysalis sp.) in this study. This study presents the first molecular evidence of hemoplasmas in Malaysian cattle and ticks.

Molecular survey and sequence analysis of Anaplasma spp. in cattle and ticks in a Malaysian farm.

This study was conducted to determine the occurrence of Anaplasma spp. in the blood samples of cattle, goats, deer and ticks in a Malaysian farm. Using polymerase chain reaction (PCR) and sequencing approach, Anaplasma spp. was detected from 81(84.4%) of 96 cattle blood samples. All blood samples from 23 goats and 22 deer tested were negative. Based on the analysis of the Anaplasma partial 16S ribosomal RNA gene, four sequence types (genotypes 1 to 4) were identified in this study. Genotypes 1-3 showed high sequence similarity to those of Anaplasma platys/ Anaplasma phagocytophilum, whilst genotype 4 was identical to those of Anaplasma marginale/ Anaplasma centrale/ Anaplasma ovis. Anaplasma DNA was detected from six (5.5%) of 109 ticks which were identified as Rhipicephalus (formely known as Boophilus) microplus ticks collected from the cattle. This study reported for the first time the detection of four Anaplasma sequence types circulating in the cattle population in a farm in Malaysia. The detection of Anaplasma DNA in R. microplus ticks in this study provides evidence that the ticks are one of the potential vectors for transmission of anaplasmosis in the cattle.

Molecular survey and sequence analysis of Anaplasma spp. in cattle and ticks in a Malaysian farm

This study was conducted to determine the occurrence of Anaplasma spp. in the blood samples of cattle, goats, deer and ticks in a Malaysian farm. Using polymerase chain reaction (PCR) and sequencing approach, Anaplasma spp. was detected from 81(84.4%) of 96 cattle blood samples. All blood samples from 23 goats and 22 deer tested were negative. Based on the analysis of the Anaplasma partial 16S ribosomal RNA gene, four sequence types (genotypes 1 to 4) were identified in this study. Genotypes 1-3 showed high sequence similarity to those of Anaplasma platys/ Anaplasma phagocytophilum, whilst genotype 4 was identical to those of Anaplasma marginale/ Anaplasma centrale/ Anaplasma ovis. Anaplasma DNA was detected from six (5.5%) of 109 ticks which were identified as Rhipicephalus (formely known as Boophilus) microplus ticks collected from the cattle. This study reported for the first time the detection of four Anaplasma sequence types circulating in the cattle population in a farm in Malaysia. The detection of Anaplasma DNA in R. microplus ticks in this study provides evidence that the ticks are one of the potential vectors for transmission of anaplasmosis in the cattle.