التحليل الجزيئي والتعريف لفصائل بكتيريا البارتونيلا من براغيث تم جمعها من حيوانات حاضنه من مدن فلسطينية Molecular Detection and Identification of Bartonella Species In Fleas (Insecta:Siphonaptera) collected From Animal Hosts In Palestinian Cities

Date
2015-05-06
Authors
احمد 'محمد رشيد' سعدي رشق
Ahmed (Mohd Rasheed) Sadi Rishiq
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Publisher
AL-Quds University
جامعة القدس
Abstract
For an extended period of time, researchers have considered small ectoparasite insects that belong to Siphonaptera order as vectors for many microorganisms as Bartonella and Yersinia bacteria. Bacteria from genus Bartonella are gram-negative, haemotrophic and fastidious were suspected to be transmitted through fleas. However, Bartonella can cause a wide range of diseases according to their species type. No data on Bartonella were ever recorded in Palestine before. This study investigated the prevalence of Bartonella organisms in fleas from cats, dogs, rats, and hyraxes in Palestine and characterized their genetic composition. Collected fleas from different districts in Palestine were characterized. The identified species were subjected to traditional molecular methods (DNA extraction, PCR and RFLP) followed by DNA sequencing for the aim of Bartonella spp identification. Based on the previous steps, dendrogram trees have been constructed using three different genetic loci. Fleas (n=289) were collected from cats (121), dogs (135), hyraxes (26) and rats (7) from northern (n=165), central (n=113), and southern (n=11) regions of Palestine. The prevalent flea species were: Ctenocephalides felis (n=119/289; 41.2%), Ctenocephalides canis (n=159/289; 55%), and Xenopsylla spp. (n=7/289; 2.4%). Targeting the Intergenic Transcribed Spacer (ITS) locus, DNA of Bartonella was detected in 22% (64/289) of all fleas. Fifty percent of the C. felis and 57% of the Xenopsylla spp. contained Bartonella DNA. DNA sequencing showed the presence of Bartonella clarridgeiae (46.7%), Bartonella henselae (25%), and Bartonella koehlerae (3.1%) in C. felis. Xenopsylla spp. collected from Rattus rattus were infected with Bartonella tribocorum, Bartonella elizabethae, and Bartonella rochalimae. By using the 16S-23S ribosomal RNA gene (ITS) for constructing the phylogenetic tree; ITS DNA sequence analysis showed four genetic clusters with unique sublcusters: cluster 1 includes B. henselae and B. koehlerae as its two subclusters, cluster 2 includes B. clarridgeiae. However, B. tribocorum and B. elizabethae formed two more out-group clusters. On the other hand, citrate synthase (gltA) showed two main clusters and RNA polymerase. β subunit (rpoβ) genes displayed two main clusters All confirms the effectiveness of the ITS gene in discriminating between Bartonella spp. These findings showed the important role of cat and rat fleas as vectors of zoonotic Bartonella species in our region. It is anticipated that this study will raise awareness among Palestinian physicians, veterinarians, and other health workers of the highprevalence of Bartonella spp. in fleas and the potential risk of these pathogens to humans and animals in this region. This work has been published in a peer reviewed journal: Journal of Vector Ecology; 2014; 39 (2): 261-270
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Keywords
الكيمياء الحيوية والاحياء الجزيئية, Biochemistry & Molecular Biology
Citation