Spotted fever group (SFG) rickettsiae causes a number of diseases in humans worldwide, which can range from mild to highly lethal. Since the clinical presentations of rickettsioses caused by SFG rickettsiae are variable and may be similar to the diseases caused by other rickettsiae, such as Orientia tsutsugamushi (agent for scrub typhus), Coxiella burnetii (agent for Q fever) and the typhus group rickettsiae (agents for epidemic and murine typhus), the accurate diagnosis of infections caused by SFG Rickettsia remains challenging especially in resource-poor settings in developing countries. This review summarizes the various diagnostic and detection tools that are currently available for the confirmation of infections by SFG rickettsiae. The advantages and challenges pertaining to the different serological and molecular detections methods, as well as new assays in development, are discussed. The utility of the detection tools contributing to the surveillance of SFG rickettsiae in arthropods and animals are reviewed.
Durian (Durio zibethinus Murr.) fruits are famous for their unique aroma. This study analysed the Durian fruit transcriptome to discover the expression patterns of genes and to understand their regulation. Three developmental stages of Durian fruit, namely, early [90 days post-anthesis (DPA)], mature (120 DPA), and ripen (127 DPA), were studied. The Illumina HiSeq platform was used for sequencing. The sequence data were analysed using four different mapping aligners and statistical methods: CLC Genomic Workbench, HISAT2+DESeq2, Tophat+Cufflinks, and HISAT2+edgeR. The analyses showed that over 110 million clean reads were mapped to the Durian genome, yielding 19,976, 11,394, 17,833, and 24,351 differentially expressed genes during 90-127 days post-anthesis. Many identified differentially expressed genes were linked to the fruit ripening processes. The data analysis suggests that most genes with increased expression at the ripening stage were primarily involved in the metabolism of cofactors and vitamins, nucleotide metabolism, and carbohydrate metabolism. Significantly expressed genes from the young to mature stage were mainly associated with carbohydrate metabolism, amino acid metabolism, and cofactor and vitamin metabolism. The transcriptome data will serve as a foundation for understanding Durian fruit development-specific genes and could be helpful in fruit's trait improvement.
Durian (Durio zibethinus L.; Family Bombacaceae) is an iconic tropical fruit plant cultivated in Malaysia and the Southeast Asian countries. In Malaysia, durian is recognised as the King of fruits and well known as a rich source of volatile sulphur compounds that make it unique. Fruit pulp of this fruit is an excellent source of nutrients as it contains proteins, dietary fat, fibers, and carbohydrates. Durian leaf and root decoctions are known to have a febrifuge and anti-malarial properties. The understanding of this plant's molecular biology will help breeders to develop a strategy for its further improvements. Hence, there is a need to identify and understand the genes necessary for the quality improvement of the durian fruits. Its genome contains about 46,000 genes which is almost double that of humans (Homo sapiens). The understanding of durian genes will be useful not only in the molecular breeding but also in the microbial production of novel proteins and or enzymes. This review highlights nutritional and medicinal attributes of durian. The molecular studies including the importance of undertaking transcriptomics work and the insights from the most recently reported genome draft are also highlighted.
Ground-based LiDAR also known as Terrestrial Laser Scanning (TLS) technology is an active remote sensing imaging method said to be one of the latest advances and innovations for plant phenotyping. Basal Stem Rot (BSR) is the most destructive disease of oil palm in Malaysia that is caused by white-rot fungus Ganoderma boninense, the symptoms of which include flattening and hanging-down of the canopy, shorter leaves, wilting green fronds and smaller crown size. Therefore, until now there is no critical investigation on the characterisation of canopy architecture related to this disease using TLS method was carried out. This study proposed a novel technique of BSR classification at the oil palm canopy analysis using the point clouds data taken from the TLS. A total of 40 samples of oil palm trees at the age of nine-years-old were selected and 10 trees for each health level were randomly taken from the same plot. The trees were categorised into four health levels - T0, T1, T2 and T3, which represents the healthy, mildly infected, moderately infected and severely infected, respectively. The TLS scanner was mounted at a height of 1 m and each palm was scanned at four scan positions around the tree to get a full 3D image. Five parameters were analysed: S200 (canopy strata at 200 cm from the top), S850 (canopy strata at 850 cm from the top), crown pixel (number of pixels inside the crown), frond angle (degree of angle between fronds) and frond number. The results taken from statistical analysis revealed that frond number was the best single parameter to detect BSR disease as early as T1. In classification models, a linear model with a combination of parameters, ABD - A (frond number), B (frond angle) and D (S200), delivered the highest average accuracy for classification of healthy-unhealthy trees with an accuracy of 86.67 per cent. It also can classify the four severity levels of infection with an accuracy of 80 per cent. This model performed better when compared to the severity classification using frond number. The novelty of this research is therefore on the development of new approach to detect and classify BSR using point clouds data of TLS.
Rickettsia raoultii is one of the causative agents of tick-borne lymphadenopathy in humans. This bacterium was previously isolated and propagated in tick cell lines; however, the growth characteristics have not been investigated. Here, we present the replication kinetics of R. raoultii in cell lines derived from different tick genera (BME/CTVM23, RSE/PILS35, and IDE8). Tick cell cultures were infected in duplicate with cryopreserved R. raoultii prepared from homologous cell lines. By 12-14 days post infection, 100% of the cells were infected, as visualized in Giemsa-stained cytocentrifuge smears. R. raoultii growth curves, determined by rickettsiae-specific gltA qPCR, exhibited lag, exponential, stationary and death phases. Exponential phases of 4-12 days and generation times of 0.9-2.6 days were observed. R. raoultii in BME/CTVM23 and RSE/PILS35 cultures showed, respectively, 39.5- and 37.1-fold increases compared to the inoculum. In contrast, multiplication of R. raoultii in the IDE8 cultures was 110.1-fold greater than the inoculum with a 7-day stationary phase. These findings suggest variation in the growth kinetics of R. raoultii in the different tick cell lines tested, amongst which IDE8 cells could tolerate the highest levels of R. raoultii replication. Further studies of R. raoultii are needed for a better understanding of its persistence within tick populations.
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.
Oil palm is a key agricultural resource in Malaysia. However, palm disease, most prominently basal stem rot caused at least RM 255 million of annual economic loss. Basal stem rot is caused by a fungus known as Ganoderma boninense. An infected tree shows few symptoms during early stage of infection, while potentially suffers an 80% lifetime yield loss and the tree may be dead within 2 years. Early detection of basal stem rot is crucial since disease control efforts can be done. Laboratory BSR detection methods are effective, but the methods have accuracy, biosafety, and cost concerns. This review article consists of scientific articles related to the oil palm tree disease, basal stem rot, Ganoderma Boninense, remote sensors and deep learning that are listed in the Web of Science since year 2012. About 110 scientific articles were found that is related to the index terms mentioned and 60 research articles were found to be related to the objective of this research thus included in this review article. From the review, it was found that the potential use of deep learning methods were rarely explored. Some research showed unsatisfactory results due to limitations on dataset. However, based on studies related to other plant diseases, deep learning in combination with data augmentation techniques showed great potentials, showing remarkable detection accuracy. Therefore, the feasibility of analyzing oil palm remote sensor data using deep learning models together with data augmentation techniques should be studied. On a commercial scale, deep learning used together with remote sensors and unmanned aerial vehicle technologies showed great potential in the detection of basal stem rot disease.
While fleas are often perceived simply as a biting nuisance and a cause of allergic dermatitis, they represent important disease vectors worldwide, especially for bacterial zoonoses such as plague (transmitted by rodent fleas) and some of the rickettsioses and bartonelloses. The cosmopolitan cat (Ctenocephalides felis) and dog (Ctenocephalides canis) fleas, as well as Ctenocephalides orientis (restricted to tropical and subtropical Asia), breed in human dwellings and are vectors of cat-scratch fever (caused by Bartonella spp.) and Rickettsia spp., including Rickettsia felis (agent of flea-borne spotted fever) and Rickettsia asembonensis , a suspected pathogen. These Rickettsia spp. are members of a phylogenetic clade known as the ‘transitional group’, which includes both human pathogens and arthropod-specific endosymbionts. The relatively depauperate flea microbiome can also contain other endosymbionts, including a diverse range of Wolbachia strains. Here, we present circularized genome assemblies for two C. orientis-derived pathogens ( Bartonella clarridgeiae and R. asembonensis ) from Malaysia, a novel Wolbachia strain (wCori), and the C. orientis mitochondrion; all were obtained by direct metagenomic sequencing of flea tissues. Moreover, we isolated two Wolbachia strains from Malaysian C. felis into tick cell culture and recovered circularized genome assemblies for both, one of which (wCfeF) is newly sequenced. We demonstrate that the three Wolbachia strains are representatives of different major clades (‘supergroups’), two of which appear to be flea-specific. These Wolbachia genomes exhibit unique combinations of features associated with reproductive parasitism or mutualism, including prophage WO, cytoplasmic incompatibility factors and the biotin operon of obligate intracellular microbes. The first circularized assembly for R. asembonensis includes a plasmid with a markedly different structure and gene content compared to the published plasmid; moreover, this novel plasmid was also detected in cat flea metagenomes from the USA. Analysis of loci under positive selection in the transitional group revealed genes involved in host–pathogen interactions that may facilitate host switching. Finally, the first B. clarridgeiae genome from Asia exhibited large-scale genome stability compared to isolates from other continents, except for SNPs in regions predicted to mediate interactions with the vertebrate host. These findings highlight the paucity of data on the genomic diversity of Ctenocephalides-associated bacteria and raise questions regarding how interactions between members of the flea microbiome might influence vector competence.
Rat bocavirus (RBoV) and rodent bocavirus (RoBoV) have previously been detected in Rattus norvegicus; however, these viruses have not been reported in rodent populations in Malaysia. We investigated the presence of RBoV and RoBoV in archived rodent specimens. DNA barcoding of the rodent cytochrome c oxidase gene identified five different species: Rattus tanezumi R3 mitotype, Rattus tiomanicus, Rattus exulans, Rattus argentiventer, and Rattus tanezumi sensu stricto. Three spleens were positive for RBoV (1.84%; 3/163), but no RoBoV was detected. Phylogenetic analyzes of the partial non-structural protein 1 gene grouped Malaysian RBoV strains with RBoV strains from China. Further studies among rats from different geographical locations are warranted for this relatively new virus.
Chigger mites are vectors of the bacterial disease scrub typhus, caused by Orientia spp. The bacterium is vertically transmitted in the vector and horizontally transmitted to terrestrial vertebrates (primarily wild small mammals), with humans as incidental hosts. Previous studies have shown that the size of the chigger populations is correlated with the density of small mammals in scrub typhus-endemic regions. Here, we explore interactions between the small mammals and chiggers in two oil palm plantations located in the Perak and Johor states of Peninsular Malaysia. The location in Perak also contained an aboriginal (Orang Asli) settlement. A ~5% sub-sample from 40,736 chigger specimens was identified from five species of small mammals (n = 217), revealing 14 chigger species, including two new records for Malaysia. The abundance and species richness of chiggers were significantly affected by habitat type (highest in forest border), state (highest in Perak), and season (highest in dry). The overall prevalence of Orientia tsutsugamushi DNA in small-mammal tissues was 11.7% and was not significantly affected by host or habitat characteristics, but in Johor, was positively associated with infestation by Leptotrombidium arenicola. These findings highlight the risk of contracting scrub typhus in oil palm plantations and associated human settlements.