Isolation and Propagation of Laboratory Strains and a Novel Flea-Derived Field Strain of <i>Wolbachia</i> in Tick Cell Lines
<i>Wolbachia</i> are intracellular endosymbionts of several invertebrate taxa, including insects and nematodes. Although <i>Wolbachia</i> DNA has been detected in ticks, its presence is generally associated with parasitism by insects. To determine whether or not <i>Wolb...
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| Format: | Article |
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MDPI AG
2020-07-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/8/7/988 |
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| author | Jing Jing Khoo Timothy J. Kurtti Nurul Aini Husin Alexandra Beliavskaia Fang Shiang Lim Mulya Mustika Sari Zulkifli Alaa M. Al-Khafaji Catherine Hartley Alistair C. Darby Grant L. Hughes Sazaly AbuBakar Benjamin L. Makepeace Lesley Bell-Sakyi |
| author_facet | Jing Jing Khoo Timothy J. Kurtti Nurul Aini Husin Alexandra Beliavskaia Fang Shiang Lim Mulya Mustika Sari Zulkifli Alaa M. Al-Khafaji Catherine Hartley Alistair C. Darby Grant L. Hughes Sazaly AbuBakar Benjamin L. Makepeace Lesley Bell-Sakyi |
| author_sort | Jing Jing Khoo |
| collection | DOAJ |
| description | <i>Wolbachia</i> are intracellular endosymbionts of several invertebrate taxa, including insects and nematodes. Although <i>Wolbachia</i> DNA has been detected in ticks, its presence is generally associated with parasitism by insects. To determine whether or not <i>Wolbachia</i> can infect and grow in tick cells, cell lines from three tick species, <i>Ixodes scapularis</i>, <i>Ixodes ricinus</i> and <i>Rhipicephalus microplus</i>, were inoculated with <i>Wolbachia</i> strains <i>w</i>Stri and <i>w</i>AlbB isolated from mosquito cell lines. Homogenates prepared from fleas collected from cats in Malaysia were inoculated into an <i>I. scapularis</i> cell line. Bacterial growth and identity were monitored by microscopy and PCR amplification and sequencing of fragments of <i>Wolbachia</i> genes. The <i>w</i>Stri strain infected <i>Ixodes</i> spp. cells and was maintained through 29 passages. The <i>w</i>AlbB strain successfully infected <i>Ixodes</i> spp. and <i>R. microplus</i> cells and was maintained through 2–5 passages. A novel strain of <i>Wolbachia</i> belonging to the supergroup F, designated <i>w</i>CfeF, was isolated in <i>I. scapularis</i> cells from a pool of <i>Ctenocephalides</i> sp. cat fleas and maintained in vitro through two passages over nine months. This is the first confirmed isolation of a <i>Wolbachia</i> strain from a flea and the first isolation of any <i>Wolbachia</i> strain outside the “pandemic” A and B supergroups. The study demonstrates that tick cells can host multiple <i>Wolbachia</i> strains, and can be added to panels of insect cell lines to improve success rates in isolation of field strains of <i>Wolbachia</i>. |
| first_indexed | 2024-03-10T18:45:56Z |
| format | Article |
| id | doaj.art-e05b5287338643a1983ea8e398d5868d |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-10T18:45:56Z |
| publishDate | 2020-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-e05b5287338643a1983ea8e398d5868d2023-11-20T05:32:32ZengMDPI AGMicroorganisms2076-26072020-07-018798810.3390/microorganisms8070988Isolation and Propagation of Laboratory Strains and a Novel Flea-Derived Field Strain of <i>Wolbachia</i> in Tick Cell LinesJing Jing Khoo0Timothy J. Kurtti1Nurul Aini Husin2Alexandra Beliavskaia3Fang Shiang Lim4Mulya Mustika Sari Zulkifli5Alaa M. Al-Khafaji6Catherine Hartley7Alistair C. Darby8Grant L. Hughes9Sazaly AbuBakar10Benjamin L. Makepeace11Lesley Bell-Sakyi12Tropical Infectious Diseases Research and Education Centre, Level 2, High Impact Research Building, University of Malaya, Kuala Lumpur 50603, MalaysiaDepartment of Entomology, University of Minnesota, 219 Hodson Hall, 1980 Folwell Avenue, Saint Paul, MN 55108, USATropical Infectious Diseases Research and Education Centre, Level 2, High Impact Research Building, University of Malaya, Kuala Lumpur 50603, MalaysiaDepartment of Infection Biology and Microbiome, Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool, 146 Brownlow Hill, Liverpool L3 5RF, UKTropical Infectious Diseases Research and Education Centre, Level 2, High Impact Research Building, University of Malaya, Kuala Lumpur 50603, MalaysiaTropical Infectious Diseases Research and Education Centre, Level 2, High Impact Research Building, University of Malaya, Kuala Lumpur 50603, MalaysiaDepartment of Infection Biology and Microbiome, Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool, 146 Brownlow Hill, Liverpool L3 5RF, UKDepartment of Infection Biology and Microbiome, Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool, 146 Brownlow Hill, Liverpool L3 5RF, UKDepartment of Infection Biology and Microbiome, Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool, 146 Brownlow Hill, Liverpool L3 5RF, UKDepartments of Vector Biology and Tropical Disease Biology, Centre for Neglected Tropical Disease, Liverpool School of Tropical Medicine, Pembroke Place, Liverpool L3 5QA, UKTropical Infectious Diseases Research and Education Centre, Level 2, High Impact Research Building, University of Malaya, Kuala Lumpur 50603, MalaysiaDepartment of Infection Biology and Microbiome, Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool, 146 Brownlow Hill, Liverpool L3 5RF, UKDepartment of Infection Biology and Microbiome, Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool, 146 Brownlow Hill, Liverpool L3 5RF, UK<i>Wolbachia</i> are intracellular endosymbionts of several invertebrate taxa, including insects and nematodes. Although <i>Wolbachia</i> DNA has been detected in ticks, its presence is generally associated with parasitism by insects. To determine whether or not <i>Wolbachia</i> can infect and grow in tick cells, cell lines from three tick species, <i>Ixodes scapularis</i>, <i>Ixodes ricinus</i> and <i>Rhipicephalus microplus</i>, were inoculated with <i>Wolbachia</i> strains <i>w</i>Stri and <i>w</i>AlbB isolated from mosquito cell lines. Homogenates prepared from fleas collected from cats in Malaysia were inoculated into an <i>I. scapularis</i> cell line. Bacterial growth and identity were monitored by microscopy and PCR amplification and sequencing of fragments of <i>Wolbachia</i> genes. The <i>w</i>Stri strain infected <i>Ixodes</i> spp. cells and was maintained through 29 passages. The <i>w</i>AlbB strain successfully infected <i>Ixodes</i> spp. and <i>R. microplus</i> cells and was maintained through 2–5 passages. A novel strain of <i>Wolbachia</i> belonging to the supergroup F, designated <i>w</i>CfeF, was isolated in <i>I. scapularis</i> cells from a pool of <i>Ctenocephalides</i> sp. cat fleas and maintained in vitro through two passages over nine months. This is the first confirmed isolation of a <i>Wolbachia</i> strain from a flea and the first isolation of any <i>Wolbachia</i> strain outside the “pandemic” A and B supergroups. The study demonstrates that tick cells can host multiple <i>Wolbachia</i> strains, and can be added to panels of insect cell lines to improve success rates in isolation of field strains of <i>Wolbachia</i>.https://www.mdpi.com/2076-2607/8/7/988<i>Wolbachia</i>tick cell line<i>Ctenocephalides</i>fleaMalaysiain vitro culture |
| spellingShingle | Jing Jing Khoo Timothy J. Kurtti Nurul Aini Husin Alexandra Beliavskaia Fang Shiang Lim Mulya Mustika Sari Zulkifli Alaa M. Al-Khafaji Catherine Hartley Alistair C. Darby Grant L. Hughes Sazaly AbuBakar Benjamin L. Makepeace Lesley Bell-Sakyi Isolation and Propagation of Laboratory Strains and a Novel Flea-Derived Field Strain of <i>Wolbachia</i> in Tick Cell Lines Microorganisms <i>Wolbachia</i> tick cell line <i>Ctenocephalides</i> flea Malaysia in vitro culture |
| title | Isolation and Propagation of Laboratory Strains and a Novel Flea-Derived Field Strain of <i>Wolbachia</i> in Tick Cell Lines |
| title_full | Isolation and Propagation of Laboratory Strains and a Novel Flea-Derived Field Strain of <i>Wolbachia</i> in Tick Cell Lines |
| title_fullStr | Isolation and Propagation of Laboratory Strains and a Novel Flea-Derived Field Strain of <i>Wolbachia</i> in Tick Cell Lines |
| title_full_unstemmed | Isolation and Propagation of Laboratory Strains and a Novel Flea-Derived Field Strain of <i>Wolbachia</i> in Tick Cell Lines |
| title_short | Isolation and Propagation of Laboratory Strains and a Novel Flea-Derived Field Strain of <i>Wolbachia</i> in Tick Cell Lines |
| title_sort | isolation and propagation of laboratory strains and a novel flea derived field strain of i wolbachia i in tick cell lines |
| topic | <i>Wolbachia</i> tick cell line <i>Ctenocephalides</i> flea Malaysia in vitro culture |
| url | https://www.mdpi.com/2076-2607/8/7/988 |
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