Detection and genetic characterization of "Candidatus Mycoplasma haemomacaque" infection among long-tailed macaques (Macaca fascicularis) in Thailand using broad-range nested polymerase chain reaction assay

Background and Aim: Hemoplasmas are defined as small, epicellular parasitic bacteria that can infect the red blood cells of several mammalian species. Diseases caused by these bacteria range from asymptomatic infections to acute hemolytic anemia. However, data on hemoplasmas in non-human primates in Thailand remain to be limited. Therefore, this study aims to determine the occurrence and genetic diversity of hemoplasmas among long-tailed macaques in Thailand. Materials and Methods: Blood samples were collected from 339 long-tailed macaques in three provinces of Thailand. DNA was then extracted from the blood samples and tested for hemoplasma using broad-range nested polymerase chain reaction (PCR) based on the 16S rRNA gene. PCR-positive samples were sequenced, and phylogenetic analysis for species identification was conducted. Results: In total, 38 (11.2%) out of the 339 samples were found to be positive for hemoplasmas, based on the broad-range nested PCR assay of the 16S rRNA gene. The 16S rRNA sequences of Mycoplasma spp. were highly similar (98-99% identity) to "Candidatus Mycoplasma haemomacaque." Furthermore, phylogenetic analysis using maximum likelihood demonstrated that the sequences were located in the same cluster of "Ca. M. haemomacaque." Conclusion: The detection of hemoplasmas among long-tailed macaques in Thailand is reported. Genetic characterization confirmed that these hemoplasmas are closely related to "Ca. M. haemomacaque." These results indicate that long-tailed macaques in several locations in Thailand may be infected and serve as reservoirs for this parasite.

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Molecular detection of Campylobacter species from human and cattle faecal samples in Kilosa district, Tanzania

10.21203/rs.3.rs-46746/v1 ◽

2020 ◽

Author(s):

Noel Gahamanyi ◽

Leonard E.G. Mboera ◽

Mecky I. Matee ◽

Dieudonné Mutangana ◽

Raghavendra G. Amachawadi ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Phylogenetic Analysis ◽

16S Rrna ◽

Rrna Gene ◽

Chain Reaction ◽

Faecal Samples ◽

Rrna Sequencing ◽

Polymerase Chain ◽

Campylobacter Species ◽

The 16S Rrna Gene

Abstract Background: A growing number of Campylobacter species other than C. jejuni and C. coli have been considered as emerging human and animal pathogens. However, the contribution of these species to human gastroenteritis is poorly documented. This study aimed at detecting Campylobacter species from human and cattle faecal samples in Kilosa district, Tanzania using Polymerase Chain Reaction (PCR) amplification of the 16S rRNA gene, and Sanger sequencing . Methods: A total number of 100 faecal samples (70 from human and 30 from cattle) were collected from diarrheic and non-diarrheic patients and healthy cattle in Kilosa district, Tanzania from July to October 2019. Species identification was conducted by PCR and 16S rRNA sequencing. The phylogenetic analysis was carried out by comparison of the 16S rRNA gene sequences to reference strains by the Neighbor-Joining method in MEGA X. Results: Campylobacter species detection rate by PCR was 65.7% (46/70) and 20% (6/30) in humans and cattle, respectively. There were five human diarrheic cases, four showed Campylobacter presence and two were from children ≤15 years of age. In humans, the 16S rRNA sequencing revealed that C. concisus was the most predominant species occurring at a frequency of 37.8% (14/37), followed by uncultured Campylobacter spp. 24.3% (9/37) and C. hominis 21.6% (8/37). The least represented species were C. jejuni and C. lanienae all occurring at 2.7% (1/37). In cattle, five (100%) sequenced PCR products matched with C. lanienae . Phylogenetic analysis revealed that Campylobacter 16S rRNA sequences were closely related to C. concisus , uncultured Campylobacter sp., C. hominis , and C. gracilis . Conclusion: The non- C. jejuni / C. coli species are present in human and cattle faecal samples and their true occurrence is probably under-reported due to shortcomings of conventional techniques used in most diagnostic microbiology laboratories. Based on our findings, we recommend that molecular techniques be adopted for direct detection of Campylobacter species during routine laboratory screening and surveillance studies. Keywords: Campylobacter , molecular diagnostics, polymerase chain reaction, sequencing, gastroenteritis, Tanzania

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