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Fulltext Profiling Bartonella infection and its associated risk factors in shelter cats in Malaysia

Alias NNA, Omar S, Ahmad NI, Watanabe M, Tay ST, Aziz NA, et al.

J Vet Sci, 2023 May;24(3):e38.
PMID: 37271506 DOI: 10.4142/jvs.22277

BACKGROUND: Poor disease management and irregular vector control could predispose sheltered animals to disease such as feline Bartonella infection, a vector-borne zoonotic disease primarily caused by Bartonella henselae.
OBJECTIVES: This study investigated the status of Bartonella infection in cats from eight (n = 8) shelters by molecular and serological approaches, profiling the CD4:CD8 ratio and the risk factors associated with Bartonella infection in shelter cats.
METHODS: Bartonella deoxyribonucleic acid (DNA) was detected through polymerase chain reaction (PCR) targeting 16S-23S rRNA internal transcribed spacer gene, followed by DNA sequencing. Bartonella IgM and IgG antibody titre, CD4 and CD8 profiles were detected using indirect immunofluorescence assay and flow cytometric analysis, respectively.
RESULTS: B. henselae was detected through PCR and sequencing in 1.0% (1/101) oral swab and 2.0% (1/50) cat fleas, while another 3/50 cat fleas carried B. clarridgeiae. Only 18/101 cats were seronegative against B. henselae, whereas 30.7% (31/101) cats were positive for both IgM and IgG, 8% (18/101) cats had IgM, and 33.7% (34/101) cats had IgG antibody only. None of the eight shelters sampled had Bartonella antibody-free cats. Although abnormal CD4:CD8 ratio was observed in 48/83 seropositive cats, flea infestation was the only significant risk factor observed in this study.
CONCLUSIONS: The present study provides the first comparison on the Bartonella spp. antigen, antibody status and CD4:CD8 ratio among shelter cats. The high B. henselae seropositivity among shelter cats presumably due to significant flea infestation triggers an alarm of whether the infection could go undetectable and its potential transmission to humans.

Matched MeSH terms: Ctenocephalides*
Flea-borne pathogens in the cat flea Ctenocephalides felis and their association with mtDNA diversity of the flea host

Azrizal-Wahid N, Sofian-Azirun M, Low VL

Comp Immunol Microbiol Infect Dis, 2021 Apr;75:101621.
PMID: 33609991 DOI: 10.1016/j.cimid.2021.101621

Flea-borne pathogens were screened from 100 individual cat fleas using a PCR approach, of which 38 % were infected with at least one bacterium. Overall, 28 % of the flea samples were positive for Bartonella as inferred from ITS DNA region. Of these, 25 % (7/28) were identified as Bartonella clarridgeiae, 42.9 % (12/28) as Bartonella henselae consisted of two different strains, and 32.1 % (9/28) as Bartonella koehlerae, which was detected for the first time in Malaysia. Sequencing of gltA amplicons detected Rickettsia DNA in 14 % of cat flea samples, all of them identified as Rickettsia asembonensis (100 %). None of the flea samples were positive for Mycoplasma DNA in 16S rRNA gene detection. Four fleas were co-infected with Bartonella and Rickettsia DNAs. Statistical analyses reveal no significant association between bacterial infection and mtDNA diversity of the cat flea. Nevertheless, in all types of pathogen infections, infected populations demonstrated lower nucleotide and haplotype diversities compared to uninfected populations. Moreover, lower haplotype numbers were observed in infected populations.

Matched MeSH terms: Ctenocephalides/genetics*; Ctenocephalides/microbiology*
Identification of Rickettsial Infections (Rickettsia sp. TH2014) in Ctenocephalides orientis Fleas (Siphonaptera: Pulicidae)

Kho KL, Tay ST

J Med Entomol, 2019 02 25;56(2):526-532.
PMID: 30312440 DOI: 10.1093/jme/tjy169

Rickettsia felis (Rickettsiales: Rickettsiaceae) is an emergent human pathogen that causes febrile illnesses in various parts of the world. This study describes the identification and growth characteristics of a R. felis-like organism (designated as Rickettsia sp. TH2014) cultured from Ctenocephalides orientis fleas in rural Malaysia. In this study, culturing of rickettsiae from filtered triturated flea lysates was performed in Aedes albopictus C6/36 cells. Cytopathic effects were observed from one of the samples 4 d post-inoculation. Electron microscopy revealed actively replicating intracytosolic coccobacillary organisms in the rickettsia-infected cells. Sequence analysis of amplified citrate synthase (gltA) gene fragment shows complete match of the rickettsia with Rickettsia sp. Rf31 in Southeast Asia, and 'Candidatus Rickettsia senegalensis' strain PU01-02 in Africa. The whole-genome sequence of Rickettsia sp. TH2014 was determined and assembled. The estimated genome size and guanine + cytosine content of the rickettsia are 1.37 Mb and 32.9%, respectively. The high values of average nucleotide identity and tetra-nucleotide signature correlation index obtained from pairwise genome comparison study suggest the identification of the rickettsia as R. felis. The whole-genome single-nucleotide polymorphism analysis demonstrates close genetic relatedness of the rickettsia with R. felis and Rickettsia asemboensis. However, based on sequence analyses of rickettsial genes (16S rDNA, gltA, ompB, and sca4), Rickettsia sp. TH2014 is found to be distinct from R. felis and R. asemboensis. The sequence analyses reveal that Rickettsia sp. TH2014 is highly similar to 'Ca. Rickettsia senegalensis' detected in fleas from Africa, Asia, and North America. Further investigation to provide insights on pathogenic potential and transmission dynamics of the rickettsia is warranted.

Matched MeSH terms: Ctenocephalides/microbiology*
New insights into the haplotype diversity of the cosmopolitan cat flea Ctenocephalides felis (Siphonaptera: Pulicidae)

Azrizal-Wahid N, Sofian-Azirun M, Low VL

Vet Parasitol, 2020 May;281:109102.
PMID: 32289653 DOI: 10.1016/j.vetpar.2020.109102

The present study investigated the genetic profile of the cosmopolitan cat flea, Ctenocephalides felis (Siphonaptera: Pulicidae) from Malaysia and the reference data available in the National Center for Biotechnology Information (NCBI) GenBank. A set of sequences of 100 Malaysian samples aligned as 550 characters of the cytochrome c oxidase subunit I (cox1) and 706 characters of the II (cox2) genes revealed ten haplotypes (A1-A10) and eight haplotypes (B1-B8), respectively. The concatenated sequences of cox1 and cox2 genes with a total of 1256 characters revealed 15 haplotypes (AB1-AB15). Analyses indicated that haplotype AB1 was the most frequent and the most widespread haplotype in Malaysia. Overall haplotype and nucleotide diversities of the concatenated sequences were 0.52909 and 0.00424, respectively, with moderate genetic differentiation (FST = 0.17522) and high gene flow (Nm = 1.18). The western population presented the highest genetic diversity (Hd = 0.78333, Pi = 0.01269, Nh = 9), whereas the southern population demonstrated the lowest diversity (Hd = 0.15667, Pi = 0.00019, Nh = 3). The concatenated sequences showed genetic distances ranged from 0.08 % to 4.39 %. There were three aberrant haplotypes in cox2 sequences that highly divergent, suggesting the presence of cryptic species or occurrence of introgression. In the global point of view, the aligned sequences of C. felis revealed 65 haplotypes (AA1-AA65) by the cox1 gene (n = 586), and 27 haplotypes (BB1-BB27) by the cox2 gene (n = 204). Mapping of the haplotype network showed that Malaysian C. felis possesses seven unique haplotypes in both genes with the common haplotypes demonstrated genetic affinity with C. felis from Southeast Asia for cox1 and South America for cox2. The topologies of cox1 and cox2 phylogenetic trees were concordant with relevant grouping pattern of haplotypes in the network but revealed two major lineages by which Malaysian haplotypes were closely related with haplotypes from the tropical region.

Matched MeSH terms: Ctenocephalides/genetics*
Fulltext Wolbachia endosymbionts, Rickettsia felis and Bartonella species, in Ctenocephalides felis fleas in a tropical region

Tay ST

J Vector Ecol, 2013 Jun;38(1):200-2.
PMID: 23701629 DOI: 10.1111/j.1948-7134.2013.12030.x
Matched MeSH terms: Ctenocephalides/microbiology*
Molecular detection of pathogens from ectoparasites recovered from peri-domestic animals, and the first description of a Candidatus Midichloria sp. from Haemaphysalis wellingtoni from rural communities in Malaysia

Khoo JJ, Husin NA, Lim FS, Oslan SNH, Mohd Azami SNI, To SW, et al.

Parasitol Int, 2021 Feb;80:102202.
PMID: 33038482 DOI: 10.1016/j.parint.2020.102202

Rural communities in Malaysia have been shown to be exposed to Coxiella, Borrelia and rickettsial infections in previous seroprevalence studies. Further research is necessary to identify the actual causative agents and the potential vectors of these infections. The arthropods parasitizing peri-domestic animals in these communities may serve as the vector in transmitting arthropod-borne and zoonotic agents to the humans. Molecular screening of bacterial and zoonotic pathogens from ticks and fleas collected from dogs, cats and chickens from six rural communities in Malaysia was undertaken. These communities were made up of mainly the indigenous people of Malaysia, known as the Orang Asli, as well as settlers in oil palm plantations. The presence of Coxiella burnetii, Borrelia, and rickettsial agents, including Rickettsia and Anaplasma, was investigated by performing polymerase chain reaction (PCR) and DNA sequencing. Candidatus Rickettsia senegalensis was detected in one out of eight pools of Ctenocephalides felis fleas. A relapsing fever group Borrelia sp. was identified from one of seven Haemaphysalis hystricis ticks tested. The results from the PCR screening for Anaplasma unexpectedly revealed the presence of Candidatus Midichloria sp., a potential tick endosymbiont, in two out of fourteen Haemaphysalis wellingtoni ticks tested. C. burnetii was not detected in any of the samples tested. The findings here provide evidence for the presence of potentially novel strains of rickettsial and borrelial agents in which their impact on public health risks among the rural communities in Malaysia merit further investigation. The detection of a potential endosymbiont of ticks also suggest that the presence of tick endosymbionts in the region is not fully explored.

Matched MeSH terms: Ctenocephalides/microbiology*; Ctenocephalides/parasitology*
Fulltext Cat flea infestation in a hospital: a case report

Muthupalaniappen L, Norhayati M, Lee YY

Korean J Parasitol, 2012 Mar;50(1):79-82.
PMID: 22451739 DOI: 10.3347/kjp.2012.50.1.79

Cat flea bite in humans results in extremely pruritic skin lesions. It has been reported to occur among those living in domiciliary accommodation. However, nosocomial infestation with cat flea has not been reported. We hereby report a case of nosocomial infestation of cat flea in a hospital facility. Identification of the parasite, its appropriate eradication, and adequate medical management of the patients resulted in a satisfactory outcome.

Matched MeSH terms: Ctenocephalides/physiology*
Infestation with the cat flea, Ctenocephalides felis felis (Siphonaptera: Pulicidae) among students in Kuala Lumpur, Malaysia

Chin HC, Ahmad NW, Lim LH, Jeffery J, Hadi AA, Othman H, et al.

Southeast Asian J Trop Med Public Health, 2010 Nov;41(6):1331-4.
PMID: 21329307

This paper reports six male undergraduate students living at a local university hostel who were infested with cat fleas (Siphonaptera: Pulicidae: Ctenocephalides felis felis) in February 2009. All of them suffered from maculopapular rashes and severe pruritus after the bites. Investigation revealed the presence of a stray cat in the hostel building; five of the students had a history of contact with the cat. Six cat fleas were collected at the hostel and identified as C. felis felis. Most of the students were not aware of this infestation and did not seek medical treatment.

Matched MeSH terms: Ctenocephalides*
Fulltext Evidence for a specific host-endosymbiont relationship between 'Rickettsia sp. genotype RF2125' and Ctenocephalides felis orientis infesting dogs in India

Hii SF, Lawrence AL, Cuttell L, Tynas R, Abd Rani PA, Šlapeta J, et al.

Parasit Vectors, 2015;8:169.
PMID: 25884425 DOI: 10.1186/s13071-015-0781-x

Fleas of the genus Ctenocephalides serve as vectors for a number of rickettsial zoonoses, including Rickettsia felis. There are currently no published reports of the presence and distribution of R. felis in India, however, the ubiquitous distribution of its vector Ctenocephalides felis, makes it possible that the pathogen is endemic to the region. This study investigates the occurrence of Rickettsia spp. infection in various subspecies of C. felis infesting dogs from urban areas of Mumbai, Delhi and Rajasthan in India.

Matched MeSH terms: Ctenocephalides/classification; Ctenocephalides/microbiology*
Risk factors associated with flea infestation on cats

Azrizal-Wahid N, Sofian-Azirun M, Low VL

Trop Biomed, 2019 Dec 01;36(4):810-821.
PMID: 33597453

Fleas are the common cause of skin disorders in cats. They are well-known for transmitting various pathogens to both cats and humans. Accordingly, this study was conducted to gain insights on the risk factors associated with flea infestation on cats. Flea combing conducted on 426 cats from four distinct regions in Peninsular Malaysia revealed a relatively high rate of flea infestation on 306 cats (71.83%). A total of 651 fleas were collected, all of them were identified as Ctenocephalides felis with the total intensity of 2.13 and abundance of 1.53. The sex ratio of fleas was female-biased at 2.5:1 (♀=464, ♂=187). Statistical analysis of the data revealed that flea infestation was significantly (P<0.05) associated with several risk factors including region, age, weight, status (stray, sheltered, pet), body condition, and hair length. Higher flea prevalence was also observed in female cats (77.99%), big-sized cats (91.76%), stray cats (84.94%), cats with clean body condition (73.35%), and cats with long hairs (78.38%) as compared to their contemporaries within the same comparison variables. The high infestation of fleas in this study is indicative of cats as a flea reservoir particularly C. felis. Thus the findings of this study and the knowledge gained on the risk factors can be used to develop and improve control measures and management of flea infestations.

Matched MeSH terms: Ctenocephalides
Fulltext The dog louse Heterodoxus spiniger from stray cats in Penang, Malaysia

Norhidayu S, Mohd Zain SN, Jeffery J, Lewis JW

Trop Biomed, 2012 Jun;29(2):301-3.
PMID: 22735853 MyJurnal

Stray cats collected from Georgetown, Penang from 2008 to 2010 were screened for ectoparasites via fine-tooth combing. Two cats from a total 102 examined were infested with the dog louse, Heterodoxus spiniger. Both cats, a juvenile male and female were found in close contact with each other prior to capture. The number of lice ranged from 5 and 14 in the male and female cat respectively. Other ectoparasites recovered included the common cat flea, Ctenocephalides felis, one louse species Felicola subrostratus, one tick species Haemaphysalis bispinosa and one mite species of Listrophoridae. The present study reports for the first time the finding of H. spiniger on cats from peninsular Malaysia.

Matched MeSH terms: Ctenocephalides/pathogenicity
Identification of rickettsiae from wild rats and cat fleas in Malaysia

Tay ST, Mokhtar AS, Low KC, Mohd Zain SN, Jeffery J, Abdul Aziz N, et al.

Med Vet Entomol, 2014 Aug;28 Suppl 1:104-8.
PMID: 25171613 DOI: 10.1111/mve.12075

Rickettsioses are emerging zoonotic diseases reported worldwide. In spite of the serological evidence of spotted fever group rickettsioses in febrile patients in Malaysia, limited studies have been conducted to identify the animal reservoirs and vectors of rickettsioses. This study investigated the presence of rickettsiae in the tissue homogenates of 95 wild rats and 589 animal ectoparasites. Using PCR assays targeting the citrate synthase gene (gltA), rickettsial DNA was detected in the tissue homogenates of 13 (13.7%) wild rats. Sequence analysis of the gltA amplicons showed 98.6-100% similarity with those of Rickettsia honei/R. conorii/R. raoultii (Rickettsiales: Rickettsiaceae). Sequence analysis of outer membrane protein A gene (ompA) identified Rickettsia sp. TCM1 strain from two rats. No rickettsia was detected from Laelaps mites, Rhipicephalus sanguineus and Haemaphysalis bispinosa ticks, and Felicola subrostratus lice in this study. R. felis was identified from 32.2% of 177 Ctenocephalides felis fleas. Sequence analysis of the gltA amplicons revealed two genotypes of R. felis (Rf31 and RF2125) in the fleas. As wild rats and cat fleas play an important role in the enzoonotic maintenance of rickettsiae, control of rodent and flea populations may be able to reduce transmission of rickettsioses in the local setting.

Matched MeSH terms: Ctenocephalides/microbiology*
Fulltext Cat flea outbreak in Kuching District, Sarawak

Lau, S.M., Jamail, M., Sarina, S., Bahtiar, R., Asmad, M.

Malaysian Journal of Public Health Medicine, 2010;10(1):57-61.
MyJurnal

Cat fleas were reported to attack human in RPR Batu Kawa, a housing area about 3 km from Kuching town, resulting in an outbreak. A total of 19 people (10 adults and 9 children) were attacked by fleas. They presented with red spots, slightly raised (swollen) and irritation of skin, mostly found on the ankles and legs. The first 4 cases were reported on 29 September 2007 and the last case was on 17 November 2007. The remaining 12 cases which represent the majority of cases reported on 4th October 2007. The study conducted based mainly on field investigation and flea sampling from animals on field at that moment to find out the causes of the disease spread. Flea samples from human and cats were found to be Ctenocephalides felis; which is the most prevalent species in the world. However, no fleas were found on dog, rabbit and rat. This is the first reported case in Kuching; the study was carried out to determine the cause and the epidemiological pattern of the disease. This is important, because cat flea might attack human especially if house owners fail to monitor their pets and practice proper sanitation method to avoid the presence of cat flea larvae at home.

Matched MeSH terms: Ctenocephalides

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