This paper describes a new tick-host record of Haemaphysalis wellingtoni from the helmeted guineafowl (Numida meleagris) in Peninsular Malaysia. This record is also the first report of a tick infestation from N. meleagris in Asia. A checklist of the tick-guineafowl associations has been designed, resulting in 32 tick species including H. wellingtoni recorded as infesting N. meleagris worldwide. A list of pathogens harboured or transmitted by H. wellingtoni is provided. The status of H. wellingtoni as a true parasite of N. meleagris, and the health threat posed to the guineafowls are discussed in this paper.
Haemaphysalis (Rhipistoma) dentipalpis Warburton & Nuttall, 1909 (Acari: Ixodidae) is reinstated here as a valid species and the male is redescribed whereas the female is described for the first time. The adults of H. dentipalpis that we studied were collected from various felid and viverrid carnivorans (Carnivora: Felidae, Viverridae) in Indonesia and Malaysia. For comparative purposes, the male and female of H. (R.) asiatica (Supino, 1897) are redescribed. The adults of H. asiatica that we studied were from various felid and viverrid carnivorans (Carnivora: Felidae, Viverridae) as well as a treeshrew (Scandentia: Tupaiidae) in Thailand and Vietnam. The males and females of both H. dentipalpis and H. asiatica can be differentiated by the pattern of punctations on the conscutum and scutum and the shape and size of the posterodorsal and posteroventral spurs on palpal segment II.
Studies of ticks in Malaysia from past articles were reviewed, resulting in a list of 47 species belonging to seven genera (Argas, Ornithodoros, Amblyomma, Dermacentor, Haemaphysalis, Ixodes, and Rhipicephalus). The most prevalent genus in Malaysia is Haemaphysalis (20 species), followed by Amblyomma (eight species) and Dermacentor (seven species). Out of 47 species, only 28 have bitten humans, mostly belonging to Haemaphysalis. The most researched tick-borne pathogens in Malaysia are Rickettsia and Anaplasma, and most research was focused on the tropical-lineage brown dog ticks, R. sanguineus sensu lato, and the cattle ticks, Haemaphysalis bispinosa and Rhipicephalus microplus. 18 species were excluded from the list due to lack of definite records or dubious findings: Ornithodoros mimon, O. turicata, Amblyomma breviscutatum, A. clypeolatum, A. integrum, A. maculatum, Dermacentor marginatum, D. taiwanensis, Haemaphysalis birmaniae, H. flava, H. humerosa, H. longicornis, H. punctata, H. sulcata, Ixodes holocyclus, Rhipicephalus appendiculatus, R. annulatus and R. bursa. This paper presents the first complete and updated list for Dermacentor and Ixodes tick species in Malaysia since Kohls (1957).
The Asian rodent tick (Ixodes granulatus) occurs throughout much of Asia, it frequently bites humans, and zoonotic pathogens, such as Borrelia burgdorferi (sensu lato) and Rickettsia honei, have been detected within it. Unfortunately, the ecology of I. granulatus remains poorly known, including drivers of its abundance and the interaction ecology with its sylvatic hosts. To elucidate the ecology of this medically important species, the habitat preferences of I. granulatus were assessed in Singapore and Malaysia. Ixodes granulatus showed strong associations with old forest habitats, though across different age classes of old forest there was limited variation in abundance. Ixodes granulatus was absent from other habitats including young forest, scrubland, and parks/gardens. Within its sylvatic rodent hosts, a range of factors were found to be statistically significant predictors of I. granulatus load and/or infestation risk, including sex and body condition index. Male rodents were significantly more likely to be infested and to have higher loads than females, similarly, animals with a lower body condition index were significantly more likely to be infested. Proactive public health efforts targeted at preventing bites by this tick should carefully consider its ecology to minimise ecological overlap between humans and I. granulatus.
Two ticks were collected from a reticulated python (Malayopython reticulatus) caught in Tumpat District, Kelantan, Peninsular Malaysia. The ticks were first identified as Ambylomma sp. through morphological comparison with identification keys. Determination of the tick species was made through PCR and sequencing. However, BLAST analysis revealed 85-88% sequence nucleotide identity with Amblyomma nitidum and Amblyomma geoemydae. Additionally, the morphological features of the ticks collected in this study did not match either A. nitidum or A. geoemydae. Further examination of the ticks confirmed the species as Amblyomma cordiferum. This is the first record of A. cordiferum DNA sequence with morphological support of colour illustrations for adult A. cordiferum. This is also the most recent record of this host association in Peninsular Malaysia. Information from this report can serve as a reference for species identification using the described morphology or molecular sequences.
In this study, Babesia screening was conducted in 55 rodents and 160 tick samples collected from primary forests and an oil palm plantation in Sarawak, Malaysian Borneo. PCR targeting the 18S ribosomal DNA revealed the presence of Babesia spp. DNA detected in two questing male Haemaphysalis shimoga ticks collected from the oil palm plantation. Sequence analysis revealed that both sequences were identical and had 98.6% identity to a Babesia macropus sequence obtained from Eastern grey kangaroos (Macropus giganteus) in Australia. Phylogenetic tree revealed clustering with marsupial-associated Babesia spp. in the Babesia sensu stricto clade. Whether or not H. shimoga is the competent vector and the importance of the Babesia sp. detected in this study warrants more investigation.
We report the presence of a male Haemaphysalis semermis collected from the domestic cat, Felis catus in an aboriginal village located in Pahang, Malaysia. This paper constitutes a new host record of this tick species, and also the first documentation of the infestation of companion animals other than domestic dogs (Canis lupus) by H. semermis in Malaysia. Additionally, we have included an updated host index of the tick species in Southeast Asia.
The close relationship between dogs and humans has led to concerns regarding the transmission of zoonotic pathogens through ectoparasites such as ticks. This study aimed to investigate the occurrence of ticks and the intensity on stray dogs with specific risk factors (size, sex, neutering status, body part). Additionally, identifying the predilection sites of ticks on stray dogs was crucial for developing an effective tick control program in Malaysia. A cross-sectional study was conducted among 64 stray dogs from Kelantan and Selangor States. These dogs were subjected to integumentary examinations, collecting 431 ticks comprising Rhipicephalus sanguineus s.l. "tropical lineage" and Haemaphysalis bispinosa from 53 infested dogs. The overall occurrence of tick infestation was 82.81% (53/64), with an average intensity of 8.13 ticks (range: 1-17) per stray dog. All the potential risk factors considered in this study showed no statistically significant result (P value >0.05). The head, ear, and neck were the most preferred attachment sites for ticks. These findings underscore the importance of implementing tick control programs for stray dogs, which serve as reservoirs of ticks and tick-borne pathogens for owned dogs and humans.
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.
A parasitological survey of 16 pangolins, confiscated from the Department of Wildlife and Nature Park Peninsular Malaysia (DWNP) at Kelantan and Pulau Pinang, Malaysia was conducted in 2011. Amblyomma javanense (family: Ixodidae) was the only ectoparasite found on the pangolins. The prevalence, intensity and life cycle of A. javanense were observed together with the respective pangolins' age and sex. It was found that 68.8% of the pangolins were infected, and significant difference, χ(2)(1, N=16)=4.02, p=0.05 were observed with males higher in infestation (88.9%) as compared to the females (42.9%). However, the mean intensity was higher on females (72) as compared to males (31.6). In addition, significant difference, χ(2) (2, N=16)=6.73, p=0.05 was recorded between adults and juveniles with juveniles found to be 100% infected as compared to adult (63.6%). Nevertheless, the mean intensity was slightly higher on adults (47) than juveniles (35). Adult ticks were found in higher numbers as compared to the nymph and larvae with number of male ticks higher (236) as compared to the females (53). Similarly, a high significant difference χ(2)(2, N=469)=203.47, p=0.05 was recorded in the composition of the tick's life stages with a higher number of adult ticks (61.6%) followed by nymph (30.3%) and larvae (8.1%). As such, the results of this study revealed a picture of the A. javanense life cycle which is related to the age and gender of the Malayan Pangolin.
This paper reports an outbreak of trypanosomiasis due to Trypanosoma evansi in Java deer (Cervus timorensis) on a government deer farm in Lenggong, Perak. Seventeen adult female Java deer were found dead within a week. Symptoms of dullness, inappetence, anaemia, anorexia, respiratory distress and recumbency were seen prior to death in the infected Java deer. Beside trypanosomiasis, other parasitic infections such as theileriosis, helminthiasis and ectoparasite infestation were also recorded. Post mortem results showed generalized anaemia in most animals with isolated cases of jaundice. There was no significant finding with respect to bacteriological and viral investigations.
Ticks serve as vectors of a wide range of infectious agents deleterious to humans and animals. Tick bite prevention is based to a large extent on the use of chemical repellents and acaricides. However, development of resistance in targeted ticks, environmental pollution, and contamination of livestock meat and milk are major concerns. Recently, metal, metal oxide and carbon nanoparticles, particularly those obtained through green fabrication routes, were found to be highly effective against a wide array of arthropod pests and vectors. We summarize current knowledge on the toxicity of nanoparticles against tick vectors of medical and veterinary importance. We also discuss the toxicity of products from botanical- and bacterial-based as well as classic chemical nanosynthesis routes, showing differences in bioactivity against ticks based on the products used for the fabrication of nanoparticles. Further research is needed, to validate the efficacy of nanoparticle-based acaricides in the field and clarify mechanisms of action of nanoparticles against ticks. From a technical point of view, the literature analyzed here showed little standardization of size and weight of tested ticks, a lack of uniform methods to assess toxicity and concerns related to data analysis. Finally, an important challenge for future research is the need for ecotoxicology studies to evaluate potential negative effects on non-target organisms and site contamination arising from nanoparticle-based treatments in close proximity of livestock and farmers.
Spirochetes from the Borrelia genus are known to cause diseases in humans, namely Lyme disease and relapsing fever. These organisms are commonly transmitted to humans by arthropod vectors including ticks, mite, and lice. Here, we report the molecular detection of a Borrelia sp. from a Haemaphysalis hystricis Supino tick collected from wildlife in an Orang Asli settlement in Selangor, Malaysia. Phylogenetic analyses of partial 16s rRNA and flaB gene sequences revealed that the Borrelia sp. is closely related to the relapsing fever group borreliae, Borrelia lonestari, Borrelia miyamotoi, and Borrelia theileri, as well as a number of uncharacterized Borrelia sp. from ticks in Portugal and Japan. To our knowledge, this is the first report of a Borrelia sp. detected in H. hystricis, and in Malaysia. The zoonotic potential of this Borrelia sp. merits further investigation.
Babesia bigemina is a tick-borne protozoan that affects cattle in almost all regions of the world. Despite its importance, there is no report of its prevalence in cattle using molecular detection methods in Peninsular Malaysia. This study describes the prevalence, distribution, and risk factors associated with B. bigemina infection using molecular diagnostic methods. Also, the species of ticks infesting cattle and the attitude of cattle farmers towards tick control in Peninsular Malaysia were studied. Blood samples were collected from 1045 cattle from 43 herds throughout the country, and were subjected to molecular studies to detect B. bigemina. Tick samples for entomological studies were also collected and identified. Epidemiological information of each cattle and farm were obtained using a well-structured questionnaire containing open-ended and closed-ended questions. Data were statistically analyzed using Univariate and Multivariate models. The 211-base pair of AMA-1 gene of B. bigemina was amplified and confirmed in 30.5 % (319/1045; 95 % CI = 27.8-33.4) of the sampled population, with the haemoprotozoan detected in all the sampled herds. Breed, age, physiological status, management type, rate of de-ticking, and closeness to human settlement were the risk factors significantly (p < 0.05) associated with the prevalence of B. bigemina in cattle. Rhipicephalus (Boophilus) microplus and Haemaphysalis bispinosa were the species of ticks collected from cattle, with the former been more prevalent. A large number of cattle farmers (12/43; 28 %) do not control ticks in their herds. The findings of this study will create baseline data on the epidemiology of the haemoprotozoan and control patterns of its tick vectors that will guide the government in enacting policies that will improve food security and the economy of the nation.
Blood meal analysis (BMA) from ticks allows for the identification of natural hosts of ticks (Acari: Ixodidae). The aim of this study is to identify the blood meal sources of field collected on-host ticks using PCR analysis. DNA of four genera of ticks was isolated and their cytochrome b (Cyt b) gene was amplified to identify host blood meals. A phylogenetic tree was constructed based on data of Cyt b sequences using Neighbor Joining (NJ) and Maximum Parsimony (MP) analysis using MEGA 5.05 for the clustering of hosts of tick species. Twenty out of 27 samples showed maximum similarity (99%) with GenBank sequences through a Basic Local Alignment Search Tool (BLAST) while 7 samples only showed a similarity range of between 91-98%. The phylogenetic trees showed that the blood meal samples were derived from small rodents (Leopoldamyssabanus, Rattustiomanicus and Sundamysmuelleri), shrews (Tupaiaglis) and mammals (Tapirusindicus and Prionailurusbengalensis), supported by 82-88% bootstrap values. In this study, Cyt b gene as a molecular target produced reliable results and was very significant for the effective identification of ticks' blood meal. The assay can be used as a tool for identifying unknown blood meals of field collected on-host ticks.
Hard ticks taken from the Japanese serow, Capricornis crispus, in Yamagata Prefecture, Honshu, harboured infective larvae of onchocercid filariae after incubation from the 22nd to the 158th day. Haemaphysalis flava and H. japonica contained one to eight filarial larvae; females, males and a nymph of the ticks were infected. The 44 infective larvae recovered were 612-1,370 μm long, and 11 of them, 930-1,340 μm long, were studied in detail. The larvae possessed the morphologic characteristics of the larvae of the genus Cercopithifilaria, namely an oesophagus with a posterior glandular part, no buccal capsule and a long tail with three terminal lappets. Five types (A to E) of infective larvae were identified based on the morphologic characteristics. While to date five species of Cercopithifilaria have been described from the Japanese serow, a specific identification of the larvae found in this study was generally not possible. Only type E larvae could be tentatively assigned to Cercopithifilaria tumidicervicata, as they had a cervical swelling similar to that of the adults of this species. A key for the identification of the five larval types is presented. The study presents circumstantial evidences indicating that H. flava and H. japonica may transmit Cercopithifilaria spp. to Japanese serows. It also suggests the possibility that such filarial larvae will be found in hard ticks anywhere, because Cercopithifilaria is distributed worldwide, though this genus generally goes unnoticed, as its microfilariae occur in the skin, not in the blood, of host animals.
Amblyomma ticks parasitize a wide range of animals in tropical regions. This study describes the identification of Amblyomma ticks from wild snakes in Malaysia and the detection of potential human pathogens such as Rickettsia, Anaplasma, Ehrlichia and bartonellae in the ticks.
Bartonellosis is an emerging zoonotic infection responsible for a variety of clinical syndromes in humans and animals. Members of the genus Bartonella exhibit high degrees of genetic diversity and ecologic plasticity. The infection is usually transmitted to animals and humans through blood-feeding arthropod vectors such as fleas, lice, ticks and sandflies. This study was conducted to investigate the prevalence of Bartonella species in 184 beef cattle, 40 dairy cattle, 40 sheep and 40 goats in eight animal farms across Peninsular Malaysia. Bartonella-specific PCR assays and sequence analysis of partial fragments of the citrate synthase gene were used for detection and identification of B. bovis. Isolation of B. bovis was attempted from PCR-positive blood samples. Molecular heterogeneity of the isolates was investigated based on sequence analysis of gltA, ITS, rpoB genes, ERIC-PCR, as well as using an established multilocus sequence typing (MLST) method. The carriage rate of B. bovis in ticks was also determined in this study.
Ticks are vectors in the transmission of many important infectious diseases in human and animals. Ticks can be readily found in the semi-forested areas such as the settlements of the indigenous people in Malaysia, the Orang Asli. There is still minimal information available on the bacterial agents associated with ticks found in Malaysia. We performed a survey of the bacterial communities associated with ticks collected from domestic animals found in two Orang Asli villages in Malaysia. We collected 62 ticks, microscopically and molecularly identified as related to Haemaphysalis wellingtoni, Haemaphysalis hystricis and Haemaphysalis bispinosa. Bacterial 16s rRNA hypervariable region (V6) amplicon libraries prepared from the tick samples were sequenced on the Ion Torrent PGM platform. We detected a total of 392 possible bacterial genera after pooling and sequencing 20 samples, indicating a diverse bacterial community profile. Dominant taxa include the potential tick endosymbiont, Coxiella. Other dominant taxa include the tick-associated pathogen, Rickettsia, and environmental bacteria such as Bacillus, Mycobacterium, Sphingomonas and Pseudomonas. Other known tick-associated bacteria were also detected, including Anaplasma, Ehrlichia, Rickettsiella and Wolbachia, albeit at very low abundance. Specific PCR was performed on selected samples to identify Rickettsia and Coxiella. Sequence of Rickettsia felis, which causes spotted fever in human and cats, was identified in one sample. Coxiella endosymbionts were detected in three samples. This study provides the baseline knowledge of the microbiome of ticks in Malaysia, focusing on tick-associated bacteria affecting the Orang Asli communities. The role of the herein found Coxiella and Rickettsia in tick physiology or disease transmission merits further investigation.