Molecular detection of vector-borne pathogens in cats tested for FIV and FeLV

Melo, Thyara Bezerra de; Silva, Tatiene Rossana Mota; Almeida, Telga Lucena Alves Craveiro de; Tutija, Juliane Franciele; Silva, Angélica Oliveira da; Lira, Mariana da Silva; Amorim, Débora; Giannelli, Alessio; Ramos, Carlos Alberto do Nascimento; Alves, Lêucio Câmara; Carvalho, Gílcia Aparecida de; Ramos, Rafael Antonio Nascimento

doi:10.1016/j.vprsr.2023.100857

Cited by 3 publications

(3 citation statements)

References 44 publications

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“…Co-occurrence of multiple Bartonella species in their hosts has been reported (Bai et al, 2011 ; Qurollo et al, 2014 ; Melo et al, 2023 ). We mixed DNA from different Bartonella species mimicking the natural occurrence of coinfections.…”

Section: Discussionmentioning

confidence: 98%

“…Because different Bartonella species may co-occur in the same host species or in the same individual (Bai et al, 2011 ; Qurollo et al, 2014 ; Melo et al, 2023 ), we evaluated the quadruplex sequencing assay for its ability to accurately identify co-occurring Bartonella species by testing DNA mixed from different Bartonella species to simulate coinfections. The mixed DNA included (1) B. henselae and B. koehlerae , both of which are cat-associated and can coinfect the same individual (Qurollo et al, 2014 ; Melo et al, 2023 ), and (2) B. henselae, B. koehlerae , and B. grahamii (rodent-associated), which was added to increase the degree of difficulty for species resolution. Three replicates of each mixed DNA template were tested with 10 pg DNA of each Bartonella species.…”

Section: Methodsmentioning

confidence: 99%

“…Many Bartonella species are host-specific, suggesting the maintenance of species in independent enzootic cycles (Vayssier-Taussat et al, 2009 ). On the other hand, coinfections with multiple Bartonella species in the same host species or even in an individual host have been observed (Bai et al, 2011 ; Qurollo et al, 2014 ; Melo et al, 2023 ). Some Bartonella species originating from the same mammalian order are closely related and form a species complex (Kosoy et al, 2012 ).…”

Section: Introductionmentioning

confidence: 99%

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Development of a quadruplex PCR amplicon next generation sequencing assay for detection and differentiation of Bartonella spp.

Bai,

Osikowicz,

Hojgaard

et al. 2023

Front. Microbiol.

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The genus Bartonella includes a group of species that are associated with a wide range of mammalian species, including human. It is challenging to detect all Bartonella species using a single molecular target due to its high genetic diversity. To solve this issue, we developed a quadruplex PCR amplicon sequencing assay using next-generation sequencing (NGS) technology for the detection and differentiation of Bartonella species. Our objective was to obtain the specific sequences of a minimum of two of the four target genes as confirmation of the identity of a particular Bartonella species using the assay. Four pairs of primers targeting specific regions on gltA, groEL, rpoB, and ssrA were evaluated for their capability of differentiating Bartonella species individually and collectively by performing singular PCR amplicon sequencing and quadruplex PCR amplicon sequencing. Using the quadruplex PCR amplicon sequencing, 24 Bartonella reference species were tested, all of which were successfully differentiated by at least two targets. Bartonella species were accurately identified from the artificially mixed DNA templates developed to simulate coinfections. The limit of detection was determined to be 1 fg based on testing a series of 10-fold dilutions of DNA from the Bartonella species. Testing of high DNA concentrations of 19 non-Bartonella species showed high specificity with none of the non-Bartonella species misclassified as Bartonella. Finally, the assay was evaluated by testing DNA extracts from field-collected body lice (Pediculus humanus humanus) and Norway rats (Rattus norvegicus): Bartonella quintana was detected and confirmed by three targets in the lice and Bartonella tribocorum was detected and confirmed by two targets in the rats. These results demonstrated that Bartonella species could be accurately and rapidly detected and differentiated into different tissue types using the quadruplex sequencing assay.

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Section: Discussionmentioning

confidence: 98%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Development of a quadruplex PCR amplicon next generation sequencing assay for detection and differentiation of Bartonella spp.

Bai,

Osikowicz,

Hojgaard

et al. 2023

Front. Microbiol.

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Revolutionizing Veterinary Health with Viral Vector-Based Vaccines

Jogi,

Smaraki,

Rajak

et al. 2024

Indian J Microbiol

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A case of mistaken identity: a systematic review, meta-analysis, and reinvestigation of hemotropic Mycoplasma spp. infection in Ctenocephalides felis (cat flea)

Moore,

Lashnits,

Lappin

et al. 2024

Parasites Vectors

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Background Feline-associated hemotropic Mycoplasma (hemoplasmas) are believed to be transmitted by two primary mechanisms: (1) direct transmission via fighting and (2) vector-borne transmission by the cat flea (Ctenocephalides felis). While the efficiency of transmission by C. felis appears low, most manuscripts focus on the prevalence of hemoplasmas in wild-caught fleas and report either a very low (< 3%) or a high (> 26%) prevalence. Therefore, we aimed to assess the influence of sample processing and PCR methods on C. felis hemoplasma infection prevalence. Methods A systemic review of PubMed articles identified 13 manuscripts (1,531 fleas/flea pools) that met the inclusion criteria (performed PCR for >1 hemoplasma on C. felis collected from cats). Risk of bias was assessed utilizing the ROBINS-E tool. Meta-analysis performed in R of these manuscripts found that not washing samples and a common set of 16S rRNA primers first published in Jensen et al. 2001 were associated with increased hemoplasma prevalence. To evaluate the influence of washing on newly collected fleas, we assessed the hemoplasma status of 20 pools of 5 C. felis each, half of which were washed and half not washed. Results Flea washing did not influence the detection of hemoplasma but instead amplified Spiroplasma. To assess non-specific amplification with the Jensen et al. 2001 primers, 67 C. felis samples (34% previously reported hemoplasma infected) were subject to PCR and sequencing. By this method, hemoplasma was detected in only 3% of samples. In the remaining “hemoplasma infected” fleas, PCR amplified Spiroplasma or other bacteria. Conclusions Therefore, we concluded that hemoplasma infection in C. felis is rare, and future flea prevalence studies should sequence all positive amplicons to validate PCR specificity. Further investigation of alternative methods of feline-associated hemoplasma transmission and the ability of C. felis to maintain hemoplasma infection is necessary. Graphical Abstract

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Molecular detection of vector-borne pathogens in cats tested for FIV and FeLV

Cited by 3 publications

References 44 publications

Development of a quadruplex PCR amplicon next generation sequencing assay for detection and differentiation of Bartonella spp.

Development of a quadruplex PCR amplicon next generation sequencing assay for detection and differentiation of Bartonella spp.

Revolutionizing Veterinary Health with Viral Vector-Based Vaccines

A case of mistaken identity: a systematic review, meta-analysis, and reinvestigation of hemotropic Mycoplasma spp. infection in Ctenocephalides felis (cat flea)

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