Limited information is available on tropical ticks and tick-borne bacteria affecting the health of humans and animals in the Southeast Asia region. Francisella tularensis is a tick-borne bacterium which causes a potentially life-threatening disease known as tularemia. This study was conducted to determine the occurrence of Francisella spp. in questing ticks collected from Malaysian forest reserve areas. A total of 106 ticks (mainly Dermacentor and Haemaphysalis spp.) were examined for Francisella DNA using a Polymerase chain reaction (PCR) assay targeting the bacterial 16S rDNA. Francisella DNA was detected from 12 Dermacentor ticks. Sequence analysis of the amplified 16S rDNA sequences (1035 bp) show >99% identity with that of Francisella endosymbiont reported in a tick from Thailand. A dendrogram constructed based on the bacterial 16S rDNA shows that the Francisella spp. were distantly related to the pathogenic strains of F. tularensis. Three Francisella-positive ticks were identified as Dermacentor atrosignatus, based on sequence analysis of the tick mitochondrial 16S rRNA gene. Further screening of cattle and sheep ticks (Haemaphysalis bispinosa and Rhipicephalus microplus) and animal samples (cattle, sheep, and goats) did not yield any positive findings. Our findings provide the first molecular data on the occurrence of a Francisella strain with unknown pathogenicity in Dermacentor questing ticks in Malaysia.
The occurrence of Setaria digitata in a horse is reported for the first time in Malaysia. An 8-year-old Thoroughbred cross mare was referred to the University Veterinary Clinic with the primary complaint of corneal opacity and excessive eye discharge. After initial treatment with Terramycin eye ointment, corneal opacity cleared partially to reveal a moving thread-like cylindrical worm in the anterior chamber of the eye. The parasite was successfully removed surgically, and examination under the light microscope revealed that the isolated worm (length = 45 mm) was a 5th stage larva of S. digitata based on morphological criteria. Confirmation of the species of the worm was through molecular methods. The 12S rRNA gene was PCR-amplified, and the purified amplicon was directly sequenced. Phylogenetic analyses revealed that the isolated roundworm showed 100% sequence similarity with that of S. digitata in NCBI GenBank database (Accession no.: KY284626.1). This report is the first confirmed case of equine ocular setariasis by S. digitata in Malaysia. The current study provides evidence that S. digitata is an etiological agent of ocular infection and its presence in Malaysia.
Anaplasma spp. are Gram-negative obligate intracellular, tick-borne bacteria which are of medical and veterinary importance. Little information is available on Anaplasma infection affecting domestic and wildlife animals in Malaysia. This study investigated the presence of Anaplasma spp. in the blood samples of domestic and wildlife animals in Peninsular Malaysia, using polymerase chain reaction (EHR-PCR) assays targeting the 16S rRNA gene of Anaplasmataceae. High detection rates (60.7% and 59.0%, respectively) of Anaplasma DNA were noted in 224 cattle (Bos taurus) and 78 deer (77 Rusa timorensis and one Rusa unicolor) investigated in this study. Of the 60 amplified fragments obtained for sequence analysis, Anaplasma marginale was exclusively detected in cattle while Anaplasma platys/Anaplasma phagocytophilum was predominantly detected in the deer. Based on sequence analyses of the longer fragment of the 16S rRNA gene (approximately 1000 bp), the occurrence of A. marginale, Anaplasma capra and Candidatus Anaplasma camelii in cattle, Candidatus A. camelii in deer and Anaplasma bovis in a goat was identified in this study. To assess whether animals were infected with more than one species of Anaplasma, nested amplification of A. phagocytophilum, A. bovis and Ehrlichia chaffeensis DNA was performed for 33 animal samples initially screened positive for Anaplasmataceae. No amplification of E. chaffeensis DNA was obtained from animals investigated. BLAST analyses of the 16S rDNA sequences from three deer (R. timorensis), a buffalo (Bubalus bubalis) and a cow (B. taurus) reveal similarity with that of Candidatus Anaplasma boleense strain (GenBank accession no.: KX987335). Sequence analyses of the partial gene fragments of major surface protein (msp4) gene from two deer (R. timorensis) and a monitor lizard (Varanus salvator) show the detection of a strain highly similar (99%) to that of A. phagocytophilum strain ZJ-China (EU008082). The findings in this study show the occurrence of various Anaplasma species including those newly reported species in Malaysian domestic and wildlife animals. The role of these animals as reservoirs/maintenance hosts for Anaplasma infection are yet to be determined.
This study reports the molecular detection of Theileria spp. from six cattle farms, a sheep farm and a goat farm located at different states in Peninsular Malaysia. Animal blood samples were screened for the presence of Theileria DNA using a conventional polymerase chain reaction (PCR) assay. A total of 155 (69.2%) of 224 cattle investigated were PCR-positive for Theileria DNA. The occurrences of Theileria spp. ranged from 17.5% to 100.0% across six cattle farms. Theileria DNA was detected from 90.0% of 40 sheep but none of 40 goats examined in this study. Sequence analyses of amplified 18S rRNA partial fragments (335-338bp) confirmed the identification of Theileria buffeli, Theileria sergenti, and Theileria sinensis in representative samples of cattle and ticks. T. luwenshuni was identified in the infected sheep. The high occurrences of Theileria spp. in our farm animals highlight the needs for appropriate control and preventive measures for theileriosis.
Malaysia is considered a hyperendemic area for canine heartworm (Dirofilaria immitis) due to its favorable climate for the completion of the parasite life cycle. This study provides an updated prevalence data on D. immitis in owned dogs from Kuala Lumpur, Malaysia and compares the trends of D. immitis in Malaysia. In the period between December 2017 and June 2018, 3.85% (5/130) dog blood samples tested positive for the presence of D. immitis antigen. A majority of the tested dogs (122/130) were not on rigorous heartworm prevention. After collating and analyzing information from 10 historical studies (1970-2017), we identified a significant decline in prevalence of D. immitis antigen in Malaysia, after the year 2000. Historically, the prevalence of D. immits antigen in owned dogs was significantly lower than the prevalence seen in stray dogs in Malaysia. This study demonstrates that D. immitis remains active in Kuala Lumpur, implying that accurate compliance of heartworm prevention is essential in Malaysia.
The caligid parasite Caligus coryphaenae has been reported in several marine fishes from Japan. In this study, a total of 223 wild yellowtail (Seriola quinqueradiata) were collected from Kagoshima fish market, separated into three groups according to total length (TL 70 cm), and examined for C. coryphaenae infection. A total of 518 specimens of C. coryphaenae were isolated giving an overall prevalence of infection and mean intensity of 24.2% and 4.7 ± 3.5, respectively. Body surface, fins and operculum of hosts TL > 50 cm were recorded as preferred sites for the attachment of C. coryphaenae. Of female specimens, 56.7% bore egg strings with various egg string lengths (7.1 ± 1.2 mm), carrying from 98 to 140 eggs (0.06 ± 0.005 mm in length) per each string. To our knowledge, this is the first report of C. coryphaenae infection from wild yellowtail in Japanese waters and globally. This parasite has not been reported in yellowtail culture to date, but the finding on wild specimens of yellowtail means that it poses a risk to their culture.
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.
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.
Fasciolosis is a zoonotic disease, considered an emerging neglected tropical disease threatening ruminant productivity and causing economic losses. Controlling fasciolosis is challenging due to the complex life cycle of Fasciola, which involves snail intermediate hosts. The high rainfall status in Taiping makes it an optimal region for snail abundance, which increases the opportunity to complete Fasciola's life cycle. Previous studies showed that liver condemnation caused by fasciolosis was highly prevalent in the Taiping abattoir compared to other investigated main abattoirs of Peninsular Malaysia. Therefore, the present study determined the prevalence of bovine fasciolosis and risk factors in farms from Larut and Matang (Taiping), Malaysia. Sampling was carried out from February until August 2020. In this cross-sectional study, a total of 371 fecal samples from bovines (dairy cattle, beef cattle, buffalo) were examined from 23 farms selected based on location, farmer consent, and history of anthelmintic usage. Animal's intrinsic and farm management details were recorded, and interview sessions were conducted with farmers to collect information on the potential risk factors. Individual fecal samples were examined for the presence of Fasciola egg using Flukefinder® sedimentation. There was moderate prevalence of bovine fasciolosis in Taiping (36.9%, n = 137/371). Significant risk factors (p 3 years (OR = 5.5, 95% CI: 5.43-5.57), thinner animals with body condition score of 1 to 4 (OR = 1.2-14.9, 95% CI: 1.09-15.08), and larger grazing area (OR = 1.3, 95% CI: 1.30-1.31). Additional risk factors include the presence of more than one ruminant species in the same farm (OR = 2.0-2.1, 95% CI: 2.00-2.22), extensive housing system (OR = 4.0, 95% CI: 3.77-4.23), farm age (OR = 1.2, 95% CI: 1.20-1.21), and also co-infection with Paramphistomes (OR = 1.4, 95% CI: 1.10-1.71). The present study underscores the importance of local bovine fasciolosis epidemiology, which could be used to conduct future veterinary and public health programmes to inform effective parasitic management aimed at reducing the prevalence of fasciolosis.
To shed light on the importance of tick-borne diseases, especially in farm animals that often contact with farm workers, this study aimed to identify ticks and tick-borne pathogens in ruminants in Malaysia. Accordingly, specimen collection was conducted across Peninsular Malaysia yielded a total of 1241 ticks collected from 674 farm ruminants. Among these, four tick species were identified, with Rhipicephalus microplus being the most prevalent, constituting 99.03 % of the total tick population. Analysis of 130 tick pools revealed three positives for Borrelia. BLAST analyses of the flaB and 16S rRNA genes revealed high similarities to Borrelia theileri, ranging from 98.78 to 100 % for flaB and 99.23-99.45 % for 16S rRNA. These results align with the phylogenetic trees, where sequences from both genes clustered together with B. theileri, further supporting this identification. No Rickettsia and Bartonella bacteria were detected. This study represents the first occurrence of B. theileri in R. microplus in Malaysia.
Ticks are the second most important hematophagous ectoparasites after mosquitoes and serve as vectors for various pathogens, transmitting them to wild and domestic animals, as well as humans. Argas persicus are the soft ticks that are known to parasitize domestic poultry. Hard ticks are known to be the reservoir of Toxoplasma (T.) gondii but limited such information is available for soft ticks. This study reports the molecular prevalence and phylogenetic analysis of T. gondii infecting Argas (A.) persicus ticks (N = 70), collected from four domestic poultry breeds in the Kurram District, Khyber Pakhtunkhwa, Pakistan, between June and August 2023. The results showed that 26 out of 70 (37 %) A. persicus samples tested positive for the ITS-1 gene of T. gondii. These findings were confirmed by the DNA sequencing and BLAST analysis of amplified amplicons. Phylogenetic analysis showed that Pakistani isolates were genetically diverse and clustered with isolates that were reported from birds and mammals from Pakistan, Tunisia, Malaysia, Brazil and Iraq. Risk factor analysis indicated that the prevalence of T. gondii was not associated with any specific poultry breed, sampling period, or the body temperature of the fowl hosts. Similarly parasite prevalence was not found associated with tick sex or their developmental stage but parasite infection rate varied with the feeding status of the ticks; highest T. gondii infection rate was observed in fully fed Argas persicus. In conclusion, this is the first report from Pakistan highlighting a high prevalence of T. gondii in A. persicus infesting domestic poultry suggesting a potential zoonotic risk to humans through the consumption of infected poultry. This data is confirming that in addition to oral ingestion of oocysts, T. gondii can be mechanically transmitted through soft ticks.