Analysis on the prokaryotic microbiome in females and embryonic cell cultures of Rhipicephalus sanguineus tropical and temperate lineages from two specific localities in Brazil

Abstract Two lineages of Rhipicephalus sanguineus are known in Brazil: the temperate or southern and the tropical or northern populations. The distribution patterns of both lineages of R. sanguineus have epidemiological implications that can affect vectorial competence concerning Ehrlichia canis, the agent of canine monocytic ehrlichiosis. Intending to identify the microbiomes of both lineages and compare microorganisms in R. sanguineus, we used the 16S rRNA (V4-V5 region) gene-based metataxonomic approach, through NGS sequencing on the MiSeq Illumina platform. We selected specimens of females from the environment and samples of primary embryonic cell cultures, from both lineages, and this was the first study to investigate the prokaryotic microbiome in tick cell cultures. The results showed that many bacterial taxa detected in the samples were typical members of the host environment. A significant diversity of microorganisms in R. sanguineus females and in embryonic cell cultures from both lineages was found, with emphasis on the presence of Coxiella in all samples, albeit in different proportions. The Coxiella species present in the two lineages of ticks may be different and may have co-evolved with them, thus driving different patterns of interactions between ticks and the pathogens that they can harbor or transmit to vertebrate hosts.

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Initiation of primary cell cultures from embryonic Haemaphysalis bispinosa ticks

Systematic and Applied Acarology ◽

10.11158/saa.22.3.1 ◽

2017 ◽

Vol 22 (3) ◽

pp. 323 ◽

Cited By ~ 1

Author(s):

Fang-shiang Lim ◽

Jing-jing Khoo ◽

Fezshin Chen ◽

Lesley Bell-sakyi ◽

Chee-sieng Khor ◽

...

Keyword(s):

Cell Lines ◽

Cell Cultures ◽

Embryonic Cell ◽

Rrna Gene ◽

Hard Tick ◽

Tick Cell ◽

Haemaphysalis Bispinosa ◽

Ornithodoros Moubata ◽

Floating Cell ◽

Axenic Conditions

Tick cell cultures have been widely used as an important tool for the study of tick-associated microorganisms, specifically for medically important bacteria or viruses that may be difficult to isolate or culture in axenic conditions. In this study, primary embryonic tick cell cultures were initiated separately from each of the egg batches laid by 10 female ticks belonging to the hard tick genus Haemaphysalis. All cultures were maintained at 28°C. After 10 months, 4 healthy cultures were identified with the potential for developing into continuous tick cell lines. These cultures comprise large cells predominantly forming floating cell clumps with multicellular vesicles, which are morphologically similar to cell lines derived from the soft tick Ornithodoros moubata. Subculture has not yet been performed due to the low cell density at the time of writing. Amplification and sequencing of a fragment of the 16S rRNA gene from DNA extracted from the parent ticks showed 99%-100% similarity to published sequences of Haemaphysalis bispinosa. This is the first report of the initiation of embryonic cell cultures from Haemaphysalis ticks found in Malaysia. Such tick cell cultures will be useful for studies of tick-borne pathogens in this region, where recent studies have shown that Haemaphysalis ticks are highly represented and harbor medically important bacteria.

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Prevalence of Ehrlichia canis (Rickettsiales: Ehrlichieae) DNA in Tissues From Rhipicephalus sanguineus (Acari: Ixodidae) Ticks in Areas Endemic for Canine Monocytic Ehrlichiosis in Brazil

Journal of Medical Entomology ◽

10.1093/jme/tjy220 ◽

2018 ◽

Vol 56 (3) ◽

pp. 828-831

Author(s):

Bruno César Miranda Oliveira ◽

Elis Domingos Ferrari ◽

Milena Araúz Viol ◽

Marcos Rogério André ◽

Rosangela Zacarias Machado ◽

...

Keyword(s):

Rhipicephalus Sanguineus ◽

Ehrlichia Canis ◽

Canine Monocytic Ehrlichiosis

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Canine monocytic ehrlichiosis in Buenos Aires, Argentina: comparison of serological and molecular assays

Pesquisa Veterinária Brasileira ◽

10.1590/1678-5150-pvb-6083 ◽

2019 ◽

Vol 39 (8) ◽

pp. 649-654

Author(s):

Paula L. Martin ◽

María N. De Salvo ◽

Gabriel L. Cicuttin ◽

María S. Arauz

Keyword(s):

Buenos Aires ◽

Rhipicephalus Sanguineus ◽

Control Group ◽

Immunochromatographic Test ◽

Serological Tests ◽

Molecular Assays ◽

Worldwide Distribution ◽

Canine Monocytic Ehrlichiosis ◽

Rhipicephalus Sanguineus Sensu Lato ◽

Healthy Dogs

ABSTRACT: Canine monocytic ehrlichiosis (CME) is an infectious disease caused by the bacterium Ehrlichia canis and transmitted by Rhipicephalus sanguineus sensu lato, a tick with worldwide distribution. When not diagnosed and treated early, disease can be severe. Currently, the disease is confirmed by serological or molecular assays. The objective of this study was to compare a serological assay based on immunochromatography (SPEED® EHRLI immunochromatographic test; BVT, France) and a molecular assay (a screening PCR followed by a nested PCR specific for E. canis) for the diagnosis of E. canis in suspected dogs from Buenos Aires city and southern Greater Buenos Aires, Argentina. Blood samples from 20 clinically healthy dogs (Control Group) and from 80 sick dogs suspected of having CME (Groups 1 to 4) were tested in parallel. Neither the immunochromatographic test nor the PCR assay was able to detect the presence of E. canis in the Control Group. In the group which had been previously tested by serology, the agreement between the tests was low (kappa: 0.200), whereas in the group which had been previously tested by PCR, the concordance between the tests was adequate (kappa: 0.650). The concordance between the tests evaluated in the total population studied was moderate (kappa: 0.496). The results of our study suggest that the use of rapid serological tests as a first approach, together with subsequent confirmation by PCR, will improve the diagnosis of CME.

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Efficacy of a Doxycycline Treatment Regimen Initiated during Three Different Phases of Experimental Ehrlichiosis

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01622-09 ◽

2010 ◽

Vol 54 (12) ◽

pp. 5012-5020 ◽

Cited By ~ 26

Author(s):

Jennifer C. McClure ◽

Michelle L. Crothers ◽

John J. Schaefer ◽

Patrick D. Stanley ◽

Glen R. Needham ◽

...

Keyword(s):

Antibiotic Therapy ◽

Treatment Regimen ◽

Rhipicephalus Sanguineus ◽

Ehrlichia Canis ◽

Clinical Parameters ◽

Blood Samples ◽

Persistently Infected ◽

Doxycycline Treatment ◽

Canine Monocytic Ehrlichiosis ◽

Three Phases

ABSTRACT Doxycycline is the treatment of choice for canine monocytic ehrlichiosis (CME), a well-characterized disease and valuable model for tick-borne zoonoses. Conflicting reports of clearance of Ehrlichia canis after treatment with doxycycline suggested that the disease phase during which treatment is initiated influences outcomes of these treatments. The purpose of this study was to evaluate the efficacy of a 28-day doxycycline regimen for clearance of experimental E. canis infections from dogs treated during three phases of the disease. Ten dogs were inoculated with blood from E. canis carriers and treated with doxycycline during acute, subclinical, or chronic phases of CME. Daily rectal temperatures and semiweekly blood samples were monitored from each dog, and Rhipicephalus sanguineus ticks were acquisition fed on each dog for xenodiagnosis. Blood collected from dogs treated during acute or subclinical CME became PCR negative for E. canis as clinical parameters improved, but blood samples collected from dogs treated during chronic CME remained intermittently PCR positive. R. sanguineus ticks fed on dogs after doxycycline treatments became PCR positive for E. canis, regardless of when treatment was initiated. However, fewer ticks became PCR positive after feeding on two persistently infected dogs treated with doxycycline followed by rifampin, suggesting that antibiotic therapy can reduce tick acquisition of E. canis.

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Encephalomyelitis vaccines: a Vero-derived cell culture alternative to primary duck embryonic cell cultures

Vaccine ◽

10.1016/0264-410x(88)90003-5 ◽

1988 ◽

Vol 6 (1) ◽

pp. 6

Author(s):

J.B. Katz ◽

S.K. Hanson

Keyword(s):

Cell Culture ◽

Cell Cultures ◽

Embryonic Cell

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Effects of Glycerol on Chicken Spermatozoa Incubated In Vitro at 41 C in Oviducal and Embryonic Cell Cultures

Poultry Science ◽

10.3382/ps.0670135 ◽

1988 ◽

Vol 67 (1) ◽

pp. 135-140 ◽

Cited By ~ 5

Author(s):

J.R. LATORRE ◽

G.C. HARRIS ◽

J.K. SKEELES ◽

Z.B. JOHNSON

Keyword(s):

Cell Cultures ◽

Embryonic Cell

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Developmental modulation of protein synthesis in Drosophila primary embryonic cell cultures

Developmental Genetics ◽

10.1002/dvg.1020020303 ◽

1981 ◽

Vol 2 (3) ◽

pp. 237-252 ◽

Cited By ~ 11

Author(s):

Carolyn H. Buzin ◽

Robert L. Seecof

Keyword(s):

Protein Synthesis ◽

Cell Cultures ◽

Embryonic Cell ◽

Developmental Modulation

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Contamination of primary embryonic bovine kidney cell cultures with parainfluenza type 2 simian virus 5 and infectious bovine rhinotracheitis virus

Journal of Clinical Microbiology ◽

10.1128/jcm.7.2.214-218.1978 ◽

1978 ◽

Vol 7 (2) ◽

pp. 214-218

Author(s):

R A Crandell ◽

J C Hierholzer ◽

J W Krebs ◽

S S Drysdale

Keyword(s):

Cell Cultures ◽

Simian Virus ◽

Embryonic Cell ◽

Cross Contamination ◽

Primary Cells ◽

Infectious Bovine Rhinotracheitis ◽

Pig Kidney ◽

Simian Virus 5 ◽

Infectious Bovine Rhinotracheitis Virus

Two different viruses were isolated from bovine embryonic cell cultures after two subcultures from the primary cells. One virus was identified as parainfluenza type 2 simian virus 5 (SV-5), and the other was identified as infectious bovine rhinotracheitis virus. Six months later, stock cultures of pig kidney (PK-15) cells were found to be contaminated with SV-5 virus. We believe that the source of the SV-5 virus in the bovine cells was a cross-contamination from monkey kidneys during preparation of the cell cultures. The infectious bovine rhinotracheitis contamination was probably of endogenous origin. The bovine embryonic cell cultures were the probable source of contamination of the PK-15 cells with SV-5 virus.

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