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Copro-molecular study of Entamoeba infection among the indigenous community in Malaysia: A first report on the species-specific prevalence of Entamoeba in dogs

Ngui R, Hassan NA, Nordin NMS, Mohd-Shaharuddin N, Chang LY, Teh CSJ, et al.

Acta Trop, 2020 Apr;204:105334.
PMID: 31926914 DOI: 10.1016/j.actatropica.2020.105334

BACKGROUND: Entamoeba is a free-living protozoan parasitic species that infect a variety of hosts. In humans, Entamoeba histolytica is the causative agent of amoebiasis. Entamoeba species has also been reported in dogs. However, little is known about the molecular epidemiology and the specific species of this parasite in dogs globally, including Malaysia. As dogs are important companion animals for the indigenous community, and close contact with dogs is part of the natural living conditions for this community, this study aims to determine the prevalence and molecular epidemiology of Entamoeba species in human and dogs in Malaysia.
METHOD: The presence of Entamoeba species was examined in 504 fresh fecal samples, collected randomly from 411 humans and 93 dogs using microscopy and polymerase chain reaction (PCR) amplifying 16 s ribosomal RNA (rRNA). Data was analyzed using appropriate statistical analysis.
RESULTS: The microscopy data showed an overall occurrence of Entamoeba species of 26.3% (108/411) and 36.6% (34/93) in humans and dogs respectively. In humans, the most common species was a single infection of E. dispar (26.5%; 13/49), followed by E. histolytica and E. moshkovskii, (20.4% for each species respectively). Double infection of E. dispar + E. moshkovskii was detected at 10.2%, followed by E. dispar + E. histolytica (8.2%) and E. moshkovskii and E. histolytica (6.1%). 8.2% of the samples had triple infection with all three species. In animals, E. moshkovskii (46.7%) was the most common species detected, followed by E. histolytica, and E. dispar, at 20.0% and 13.3% respectively. Double infection with E. moshkovskii + E. histolytica and a triple infection were found in 2 samples (13.3%) and 1 (6.7%) sample respectively. Risk factor analysis showed that members of the community who used untreated water were more prone to be infected with Entamoeba.
CONCLUSION: This study provides information on the species-specific occurrence of Entamoeba infection, the potential risk factors and their zoonotic potential to humans. This is the first report to describe the molecular occurrence of Entamoeba species in dogs in Malaysia. The presence of pathogenic Entamoeba species implies that dogs could be a reservoir or mechanical host for human amoebiasis. Further studies need to be conducted to better understand the transmission dynamics and public health significance of Entamoeba species in human and animal hosts.
Natural infections with larvae of Onchocerca species type I in the human-biting black fly, Simulium nigrogilvum (Diptera: Simuliidae), in western Thailand

Saeung A, Srisuka W, Aupalee K, Fukuda M, Otsuka Y, Taai K, et al.

Acta Trop, 2020 Apr;204:105344.
PMID: 31954685 DOI: 10.1016/j.actatropica.2020.105344

Zoonotic onchocerciasis is a human infection caused by Onchocerca species of animal origins and transmitted by black fly vectors. The reported incidence of this disease has increased throughout the world. This study aims to clarify the vectorial roles of black fly species in zoonotic filarial transmission in Tak province, western Thailand. The integrated approach of morphological and DNA sequence-based analyses was used to identify species of both wild-caught female black flies and infective filarial larvae found in the infected black flies. All of 494 female black flies captured were identified as Simulium nigrogilvum, through scanning electron microscopy (SEM) and DNA sequence analyses based on the cytochrome c oxidase subunit I (COI) and subunit II (COII), and the fast-evolving nuclear elongation complex protein 1 (ECP1) genes. Four females of S. nigrogilvum harbored one to three third-stage larvae (infective larvae) in their thoraces, with an infection rate of 0.81% (4/494). All infective larvae were similar in morphology and size to one another, being identified as Onchocerca species type I (= O. sp. type A), a bovine filaria, originally reported from Japan, and also as O. sp. found in S. nodosum in Thailand, based on their body lengths and widths being 1,068-1,346 µm long by 25-28 µm wide, and morphological characters. Comparisons of cytochrome c oxidase subunit I (COI) and 12S rRNA sequences of mitochondrial DNA (mtDNA) and phylogenetic analyses with those of previous reports strongly supported that all larvae were O. sp. type I. This report is the first indicating the presence of O. sp. type I in Thailand and its vector being S. nigrogilvum.
Geographical variations in king cobra (Ophiophagus hannah) venom from Thailand, Malaysia, Indonesia and China: On venom lethality, antivenom immunoreactivity and in vivo neutralization

Tan KY, Ng TS, Bourges A, Ismail AK, Maharani T, Khomvilai S, et al.

Acta Trop, 2020 Mar;203:105311.
PMID: 31862461 DOI: 10.1016/j.actatropica.2019.105311

The wide distribution of king cobra (Ophiophagus hannah), a medically important venomous snake in Asia could be associated with geographical variation in the toxicity and antigenicity of the venom. This study investigated the lethality of king cobra venoms (KCV) from four geographical locales (Malaysia, Thailand, Indonesia, China), and the immunological binding as well as in vivo neutralization activities of three antivenom products (Thai Ophiophagus hannah monovalent antivenom, OHMAV; Indonesian Serum Anti Bisa Ular, SABU; Chinese Naja atra monovalent antivenom, NAMAV) toward the venoms. The Indonesian and Chinese KCV were more lethal (median lethal dose, LD50 ~0.5 μg/g) than those from Malaysia and Thailand (LD50 ~1.0 μg/g). The antivenoms, composed of F(ab)'2, were variably immunoreactive toward the KCV from all locales, with OHMAV exhibited the highest immunological binding activity. In mice, OHMAV neutralized the neurotoxic lethality of Thai KCV most effectively (normalized potency = 118 mg venom neutralized per g antivenom) followed by Malaysian, Indonesian and Chinese KCV. In comparison, the hetero-specific SABU was remarkably less potent by at least 6 to10 folds, whereas NAMAV appeared to be non-effective. The finding supports that a specific king cobra antivenom is needed for the effective treatment of king cobra envenomation in each region.
Genetic diversity and differentiation in the blackflies Simulium cheongi, Simulium jeffreyi and Simulium vanluni (Diptera: Simuliidae) in Peninsular Malaysia

Pavitra SP, Ya'cob Z, Tan TK, Lim YAL, Low VL

Acta Trop, 2020 May;205:105415.
PMID: 32088275 DOI: 10.1016/j.actatropica.2020.105415

The population genetic structures of S. vanluni, S. cheongi and S. jeffreyi were determined from mitochondria-encoded sequences of cytochrome c oxidase subunits I (COI) across different states in Peninsular Malaysia. High levels of genetic diversity and genetic differentiation were observed among three species. All three species revealed an intermediate level of gene flow among the populations. Negative values of Fu's Fs and low values of Raggedness index supported the hypothesis of population expansion in S. vanluni, S. cheongi and S. jeffreyi.
Proteomes of the female salivary glands of Simulium nigrogilvum and Simulium nodosum, the main human-biting black flies in Thailand

Hempolchom C, Reamtong O, Sookrung N, Srisuka W, Sakolvaree Y, Chaicumpa W, et al.

Acta Trop, 2019 Jun;194:82-88.
PMID: 30922801 DOI: 10.1016/j.actatropica.2019.03.026

Although several studies have reported pharmacological and immunological activity, as well as the role of black flies in transmitting pathogens to vertebrate hosts through salivary glands (SG) during blood feeding, SG proteomes of the anthropophilic black flies in Thailand have never been reported. Therefore, this study determined the SG proteomes of female S. nigrogilvum and S. nodosum. Sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional (2-DE) gels containing separated SG proteins of individual species were subjected to liquid chromatography-tandem mass spectrometry (LCMS/MS) and an orthologous protein search from eukaryotic organism, nematocera and simuliidae databases for total protein identification. SDS-PAGE and protein staining revealed at least 13 and 9 major protein bands in the SGs of female S. nigrogilvum and S. nodosum, respectively, as well as several minor ones. The 2-DE demonstrated a total of 56 and 41 protein spots for S. nigrogilvum and S. nodosum, respectively. Most of the proteins obtained in both species were enzymes involved in blood feeding, including proteases, apyrases, hyaluronidases, aminopeptidase and elastase. The results obtained in this study provided a new body of knowledge for a better understanding on the role of salivary gland proteins in these black fly species in Thailand.
The electronic song "Scary Monsters and Nice Sprites" reduces host attack and mating success in the dengue vector Aedes aegypti

Dieng H, The CC, Satho T, Miake F, Wydiamala E, Kassim NFA, et al.

Acta Trop, 2019 Jun;194:93-99.
PMID: 30922800 DOI: 10.1016/j.actatropica.2019.03.027

Sound and its reception are crucial for reproduction, survival, and population maintenance of many animals. In insects, low-frequency vibrations facilitate sexual interactions, whereas noise disrupts the perception of signals from conspecifics and hosts. Despite evidence that mosquitoes respond to sound frequencies beyond fundamental ranges, including songs, and that males and females need to struggle to harmonize their flight tones, the behavioral impacts of music as control targets remain unexplored. In this study, we examined the effects of electronic music (Scary Monsters and Nice Sprites by Skrillex) on foraging, host attack, and sexual activities of the dengue vector Aedes aegypti. Adults were presented with two sound environments (music-off or music-on). Discrepancies in visitation, blood feeding, and copulation patterns were compared between environments with and without music. Ae. aegypti females maintained in the music-off environment initiated host visits earlier than those in the music-on environment. They visited the host significantly less often in the music-on than the music-off condition. Females exposed to music attacked hosts much later than their non-exposed peers. The occurrence of blood feeding activity was lower when music was being played. Adults exposed to music copulated far less often than their counterparts kept in an environment where there was no music. In addition to providing insight into the auditory sensitivity of Ae. aegypti to sound, our results indicated the vulnerability of its key vectorial capacity traits to electronic music. The observation that such music can delay host attack, reduce blood feeding, and disrupt mating provides new avenues for the development of music-based personal protective and control measures against Aedes-borne diseases.
Description of the female of Simulium (Gomphostilbia) aziruni (Diptera: Simuliidae) and its genetic relationships with members of the Simulium gombakense species-group

Ya'cob Z, Takaoka H, Low VL, Tan TK, Sofian-Azirun M

Acta Trop, 2019 May;193:66-70.
PMID: 30807749 DOI: 10.1016/j.actatropica.2019.02.023

Simulium (Gomphostilbia) aziruni Takaoka, Hashim & Chen was described initially based only on a pupa and a mature larva collected from Peninsular Malaysia. Herein, we describe the morphological characters of the female of S. aziruni for the first time. It resembles those of the other members of the Simulium gombakense species-group by the genital fork with a distinct projection directed medioposteriorly from each arm and claw with a large basal tooth. Cytochrome c oxidase I (COI) barcoding analysis indicates that S. aziruni is the sister species of S. maleewongae, but both are distantly separated by a genetic distance of 4.9%.
Comparative venom proteomics of banded krait (Bungarus fasciatus) from five geographical locales: Correlation of venom lethality, immunoreactivity and antivenom neutralization

Hia YL, Tan KY, Tan CH

Acta Trop, 2020 Jul;207:105460.
PMID: 32278639 DOI: 10.1016/j.actatropica.2020.105460

The banded krait, Bungarus fasciatus is a medically important venomous snake in Asia. The wide distribution of this species in Southeast Asia and southern China indicates potential geographical variation of the venom which may impact the clinical management of snakebite envenomation. This study investigated the intraspecific venom variation of B. fasciatus from five geographical locales through a venom decomplexing proteomic approach, followed by toxinological and immunological studies. The venom proteomes composed of a total of 9 toxin families, comprising 22 to 31 proteoforms at varying abundances. The predominant proteins were phospholipase A2 (including beta-bungarotoxin), Kunitz-type serine protease inhibitor (KSPI) and three-finger toxins (3FTx), which are toxins that cause neurotoxicity and lethality. The venom lethality varied with geographical origins of the snake, with intravenous median lethal doses (LD50) ranging from 0.45-2.55 µg/g in mice. The Thai Bungarus fasciatus monovalent antivenom (BFMAV) demonstrated a dose-dependent increasing immunological binding activity toward all venoms; however, its in vivo neutralization efficacy varied vastly with normalized potency values ranging from 3 to 28 mg/g, presumably due to the compositional differences of dominant proteins in the different venoms. The findings support that antivenom use should be optimized in different geographical areas. The development of a pan-regional antivenom may be a more sustainable solution for the treatment of snakebite envenomation.
Plasmodium vivax drug resistance markers: Genetic polymorphisms and mutation patterns in isolates from Malaysia

Cheong FW, Dzul S, Fong MY, Lau YL, Ponnampalavanar S

Acta Trop, 2020 Jun;206:105454.
PMID: 32205132 DOI: 10.1016/j.actatropica.2020.105454

Transmission of Plasmodium vivax still persist in Malaysia despite the government's aim to eliminate malaria in 2020. High treatment failure rate of chloroquine monotherapy was reported recently. Hence, parasite drug susceptibility should be kept under close monitoring. Mutation analysis of the drug resistance markers is useful for reconnaissance of anti-malarial drug resistance. Hitherto, information on P. vivax drug resistance marker in Malaysia are limited. This study aims to evaluate the mutations in four P. vivax drug resistance markers pvcrt-o (putative), pvmdr1 (putative), pvdhfr and pvdhps in 44 isolates from Malaysia. Finding indicates that 27.3%, 100%, 47.7%, and 27.3% of the isolates were carrying mutant allele in pvcrt-o, pvmdr1, pvdhfr and pvdhps genes, respectively. Most of the mutant isolates had multiple point mutations rather than single point mutation in pvmdr1 (41/44) and pvdhfr (19/21). One novel point mutation V111I was detected in pvdhfr. Allelic combination analysis shows significant strong association between mutations in pvcrt-o and pvmdr1 (X2 = 9.521, P < 0.05). In the present study, 65.9% of the patients are non-Malaysians, with few of them arrived in Malaysia 1-2 weeks before the onset of clinical manifestations, or had previous history of malaria infection. Besides, few Malaysian patients had travel history to vivax-endemic countries, suggesting that these patients might have acquired the infections during their travel. All these possible imported cases could have placed Malaysia in a risk to have local transmission or outbreak of malaria. Six isolates were found to have mutations in all four drug resistance markers, suggesting that the multiple-drugs resistant P. vivax strains are circulating in Malaysia.
A new species of the Simulium (Gomphostilbia) (Diptera: Simuliidae) from Thailand, with its phylogenetic relationships with related species in the Simulium asakoae species-group

Srisuka W, Takaoka H, Fukuda M, Otsuka Y, Saeung A

Acta Trop, 2019 Sep;197:105043.
PMID: 31153893 DOI: 10.1016/j.actatropica.2019.105043

A new species of black fly, Simulium (Gomphostilbia) rampae, is described, based on adult male, its pupal exuviae and mature larvae collected from Doi Inthanon National Park, northern Thailand. This new species is placed in the Simulium asakoae species-group, and is characterized in the male by the high number of upper-eye facets in 17 vertical columns and 18 horizontal rows, in the pupa by the gill with a long common basal stalk, cone-shaped terminal hook, and cocoon with an anterodorsal projection, and in the larva by the medium-long postgenal cleft. A DNA analysis using COI gene supported its assignment to the S. asakoae species-group and showed its close relationship to S. (G.) udomi Takaoka & Choochote and S. (G.) chiangdaoense Takaoka & Srisuka. This is the fourth member of the S. asakoae species-group recorded from Thailand.
Developing a dengue prediction model based on climate in Tawau, Malaysia

Jayaraj VJ, Avoi R, Gopalakrishnan N, Raja DB, Umasa Y

Acta Trop, 2019 Sep;197:105055.
PMID: 31185224 DOI: 10.1016/j.actatropica.2019.105055

Dengue is fast becoming the most urgent health issue in Malaysia, recording close to a 10-fold increase in cases over the last decade. With much uncertainty hovering over the recently introduced tetravalent vaccine and no effective antiviral drugs, vector control remains the most important strategy in combating dengue. This study analyses the relationship between weather predictors including its lagged terms, and dengue incidence in the District of Tawau over a period of 12 years, from 2006 to 2017. A forecasting model purposed to predict future outbreaks in Tawau was then developed using this data. Monthly dengue incidence data, mean temperature, maximum temperature, minimum temperature, mean relative humidity and mean rainfall over a period of 12 years from 2006 to 2017 in Tawau were retrieved from Tawau District Health Office and the Malaysian Meteorological Department. Cross-correlation analysis between weather predictors, lagged terms of weather predictors and dengue incidences established statistically significant cross-correlation between lagged periods of weather predictors-namely maximum temperature, mean relative humidity and mean rainfall with dengue incidence at time lags of 4-6 months. These variables were then employed into 3 different methods: a multivariate Poisson regression model, a Seasonal Autoregressive Integrated Moving Average (SARIMA) model and a SARIMA with external regressors for selection. Three models were selected but the SARIMA with external regressors model utilising maximum temperature at a lag of 6 months (p-value:0.001), minimum temperature at a lag of 4 months (p-value:0.01), mean relative humidity at a lag of 2 months (p-value:0.001), and mean rainfall at a lag of 6 months (p-value:0.001) produced an AIC of 841.94, and a log-likelihood score of -413.97 establishing it as the best fitting model of the methodologies utilised. In validating the models, they were utilised to develop forecasts with the model selected with the highest accuracy of predictions being the SARIMA model predicting 1 month in advance (MAE: 7.032, MSE: 83.977). This study establishes the effect of weather on the intensity and magnitude of dengue incidence as has been previously studied. A prediction model remains a novel method of evidence-based forecasting in Tawau, Sabah. The model developed in this study, demonstrated an ability to forecast potential dengue outbreaks 1 to 4 months in advance. These findings are not dissimilar to what has been previously studied in many different countries- with temperature and humidity consistently being established as powerful predictors of dengue incidence magnitude. When used in prognostication, it can enhance- decision making and allow judicious use of resources in public health setting. Nevertheless, the model remains a work in progress- requiring larger and more diverse data.
Ticks and tick-borne pathogens in animals and humans in the island nations of Southeast Asia: A review

Sharifah N, Heo CC, Ehlers J, Houssaini J, Tappe D

Acta Trop, 2020 Sep;209:105527.
PMID: 32447029 DOI: 10.1016/j.actatropica.2020.105527

Ticks are blood-feeding ectoparasites and major vectors of pathogens that cause infectious diseases in humans and animals worldwide including mammals, birds and reptiles. Despite the growing scientific effort in the 20th century, there is still limited information on ticks and tick-borne pathogens in Southeast Asia, especially concerning medical, veterinary, socioeconomic and agricultural aspects in the island nations. This review provides an overview of the current state of knowledge of ticks and their pathogens in the island nations of Southeast Asia and peninsular Malaysia. We aim to stimulate further research studies on ticks and tick-borne pathogens of human and veterinary importance in this geographical region.
Proteomics and preclinical antivenom neutralization of the mangrove pit viper (Trimeresurus purpureomaculatus, Malaysia) and white-lipped pit viper (Trimeresurus albolabris, Thailand) venoms

Liew JL, Tan NH, Tan CH

Acta Trop, 2020 09;209:105528.
PMID: 32442435 DOI: 10.1016/j.actatropica.2020.105528
Vector competence of Malaysian Aedes aegypti to Zika virus and impact of sequential arbovirus infections

Ong NH, Chua CL, Liew JWK, Wan Sulaiman WY, Chan YF, Sam IC, et al.

Acta Trop, 2020 Aug;208:105472.
PMID: 32389451 DOI: 10.1016/j.actatropica.2020.105472

Zika virus (ZIKV) is a mosquito-borne flavivirus with global impact since 2015. Although ZIKV was first isolated from Aedes aegypti in Malaysia in 1965, not much is known about the competency of Malaysian Ae. aegypti to ZIKV. To date only 9 cases of ZIKV have been reported in Malaysia despite the abundance of mosquito vectors. This study aimed to determine the susceptibility of Ae. aegypti to ZIKV, and the impact of sequential infections in Ae. aegypti mosquitoes with DENV serotype 2 (DENV-2) followed by ZIKV. Field-caught urban Ae. aegypti were orally challenged with a Martinique strain of ZIKV, and midgut, head/thorax and saliva were collected at 3, 7 and 14 days post-infection (dpi). At 14 dpi, ZIKV-exposed mosquitoes had infection and dissemination rates of 59% (n=10/17) and 90% (n=9/10), respectively. Average titres of 3.9 and 4.4 log pfu infectious ZIKV were recovered in midgut and head/thorax, respectively. In sequential infection, prior exposure of Ae. aegypti to DENV did not affect the subsequent ZIKV infection in head/thorax albeit with a low sample size. In conclusion, Malaysian urban Ae. aegypti is susceptible to the contemporary Asian lineage of ZIKV. The established and continuous DENV circulation in Ae. aegypti did not suppress ZIKV emergence in Malaysia. Other factors contributing to low level of ZIKV circulation in Malaysia remain to be explored.
Soft ticks (Acari: Argasidae) in the island nations of Southeast Asia: A review on their distribution, associated hosts and potential pathogens

Kazim AR, Houssaini J, Ehlers J, Tappe D, Heo CC

Acta Trop, 2021 Nov;223:106085.
PMID: 34389326 DOI: 10.1016/j.actatropica.2021.106085

Soft ticks (Acari: Argasidae) are the second major family of the blood feeding metastriates and vectors of a number of viral and bacterial pathogens for both humans and animals. Despite the growing effort on tick surveillance and studies worldwide, there is still limited information on the soft tick distribution in the island nations of Southeast Asia, especially species that are medically and veterinarily important. With the aim to provide an overview of the current status of knowledge on soft tick distribution in the island nations of Southeast Asia (Malaysia, Singapore, Brunei, Indonesia, the Philippines and Timor-Leste), this article reviews the species of soft ticks (Acari: Argasidae) and their associated hosts and pathogens, with the addition of a pictorial summary and list of tick species discovered in this region. The most prevalent soft tick genus is Carios, and the host species most associated with findings of soft ticks in this region are bats, particularly of the Pteropodidae and Vespertilionidae families. Furthermore, the only known pathogen originating from soft ticks in the island nations of Southeast Asia was the Keterah virus, which was isolated from Argas pusillus tick in Malaysia.
Two new black fly species of the Simulium (Gomphostilbia) asakoae species-group (Diptera: Simuliidae) from Taiwan

Takaoka H, Low VL, Huang YT, Fukuda M, Ya'cob Z

Acta Trop, 2021 Oct;222:106023.
PMID: 34161814 DOI: 10.1016/j.actatropica.2021.106023

Two new black fly species, Simulium (Gomphostilbia) kaohsiungense and S. (G.) shoufengense, are described from females, males and pupae in Taiwan. These new species are placed in the S. asakoae species-group. Simulium (G.) kaohsiungense sp. nov. is characterized by a combination of the elongate female sensory vesicle, male upper-eye (large) facets in 15 vertical columns and 15 or 16 horizontal rows, male hind basitarsus 1.02-1.15 times the width of the hind femur, and widened pupal terminal hooks. These characteristics distinguish this new species from most species of the S. asakoae species-group. Simulium (G.) shoufengense sp. nov. is similar in the female and male to S. (G.) asakoae Takaoka & Davies originally described from Peninsular Malaysia but is distinguished from the latter species in the pupa by the combined length of the primary and secondary stalks of the dorsal triplet of gill filaments as long as or longer than the common basal stalk of the gill, and dorsum of abdominal segments 1 and 2 not darkened. The species previously regarded in Taiwan as S. (G.) metatarsale Brunetti, originally described from India, is morphologically and molecularly identified as S. (G.) asakoae.
A point-of-care cassette test for detection of Strongyloides stercoralis

Noordin R, Osman E, Kalantari N, Anuar NS, Gorgani-Firouzjaee T, Sithithaworn P, et al.

Acta Trop, 2022 Feb;226:106251.
PMID: 34808116 DOI: 10.1016/j.actatropica.2021.106251

Strongyloides stercoralis is a parasite that causes strongyloidiasis worldwide. It may lead to a life-long infection in immunocompetent people and hyperinfection in immunosuppressed patients. A point-of-care (POC) rapid test is helpful for patient diagnosis in resource-limited settings and as a detection tool in elimination/control programs. Previously, we reported a rapid IgG4 dipstick test (Ss Rapid®) for Strongyloides suitable for a laboratory setting. A POC cassette format of the test, which is field-applicable, has since been developed. Here, we report on a laboratory-based evaluation of the Ss Rapid® cas sette test on 285 sera. We assessed the diagnostic sensitivity of the Ss Rapid® cas sette with 32 sera, comprising samples from larval and/or DNA positive individuals from three countries. Additionally, we also tested samples from 33 seropositive endemic areas residents. We evaluated the diagnostic specificity of the test using 220 samples, comprising sera from other infections (n = 101), allergy cases with high IgE antibodies (n = 4), and blood donors (n = 115). The test showed high diagnostic sensitivity (97%, 31/32), and all sera of the seropositive endemic residents were reactive. It also showed high diagnostic specificity (94.5%, 208/220), and all false-positive samples tested negative after sera adsorption using recombinant NIE-coated microsphere beads. Additionally, we showed that the test worked with spiked whole blood samples. The study results showed that the SsRapid® cas sette test merits further laboratory and field evaluations.
Coexistence and intraguild competition of Chrysomya albiceps and Lucilia sericata larvae: case reports and experimental studies applied to forensic entomology

Ivorra T, Martínez-Sánchez A, Rojo S

Acta Trop, 2022 Feb;226:106233.
PMID: 34808117 DOI: 10.1016/j.actatropica.2021.106233

The larval development of Chrysomya albiceps and Lucilia sericata is a well-known and valuable tool for estimating the postmortem interval (PMI). The third instar larvae of Ch. albiceps are facultative predators of the larvae of other necrophagous dipterans, and this behaviour is undoubtedly an important factor to consider. Both species are found together during human autopsies in Spain and other parts of Europe, where they cohabit, so a precise knowledge of their larval coexistence data is essential. The aim of this study is to investigate their coexistence and the intraguild predation of Ch. albiceps on L. sericata under experimental conditions and in real case reports. To analyse intra- and interspecific competition, four densities were used under controlled abiotic conditions [25ºC, 60-70% RH and 12:12 (D:N)]. The experimental data were compared with data corresponding to their coexistence in natural conditions, so annual activity and forensic case reports for both species were studied. The results indicate that the mortality of immature specimens in both species, the preimaginal developmental time, and adult size were affected by competition. In natural conditions, adult coexistence during the spring-summer period was confirmed. The simultaneity of the two species colonising human corpses is frequent in the studied area, with Ch. albiceps being the dominant species, and L. sericata the relevant species for estimating the minimum postmortem interval.
Phytochemical, anti-Acanthamoeba, and anti-adhesion properties of Garcinia mangostana flower as preventive contact lens solution

Sangkanu S, Mitsuwan W, Mahboob T, Mahabusarakam W, Chewchanwuttiwong S, Siphakdi P, et al.

Acta Trop, 2022 Feb;226:106266.
PMID: 34890540 DOI: 10.1016/j.actatropica.2021.106266

Acanthamoeba keratitis infection extends due to the growing number of contact lens users. Indigenous plants including Garcinia mangostana play a vital role in human health and well being. Many species of this plant have been reported with myriads of potent medicinal properties. However, the aims of this study were, for the first time, to isolate compounds from the flower of G. mangostana and to test their anti-Acanthamoeba and anti-adhesion activity against Acanthamoeba triangularis. Powdered flowers of G. mangostana were extracted and chromatographed on a silica gel column. The structures of the compounds were established with the aid of 1H NMR. More so, the anti-Acanthamoeba and anti-adhesion properties were tested on a 96-well polystyrene microtiter plate and soft contact lenses. Scanning electron microscope (SEM) was used to determine the features of A. triangularis on contact lenses. Eight pure compounds were obtained, namely 9-hydroxycalabaxanthone, tovophillin A, garcinone E, garcinone B, α-mangostin, gartinin, 8-deoxygartinin and γ-mangostin. The extract and pure compounds exhibited anti-Acanthamoeba activity with MIC values in the range of 0.25-1 mg/mL. In addition, the extract and α-mangostin displayed significant activity against the adhesion of A. triangularis trophozoites both in polystyrene plate and in contact lenses at 0.5 × MIC (0.25 mg/mL). Furthermore, α-mangostin has the potential to remove A. triangularis adhesion in contact lenses similar to a commercial multipurpose solution (MPS). SEM study confirmed that crude extract and α-mangostin are effective as solutions for contact lenses, which removed A. triangularis trophozoites within 24 h. Alpha-mangostin was non-toxic to Vero cells at a concentration below 39 μM in 24 h. Crude extract of G. mangostana flower and its α-mangostin serve as candidate compounds in the treatment of Acanthamoeba infection or as lens care solution, since they can be used as a source of natural products against Acanthamoeba and virulence factor associated with the adhesion of A. triangularis.
Report of rare genotypes of Acanthamoeba from soil source of the Payeh Maga Highland forest, North-eastern Sarawak, Malaysia

Chan LL, Toh HC, Jaikwang K, Loo EM, Wong JH, Liew YK, et al.

Acta Trop, 2022 Feb 16.
PMID: 35182491 DOI: 10.1016/j.actatropica.2022.106372

Pathogenic Acanthamoeba species are the causative agents of Acanthamoeba keratitis and granulomatous amoebic encephalitis. Members of this amoeba genus are ubiquitous in the environments. In Malaysia, most environmental studies performed to date have targeted the detection and characterisation of Acanthamoeba sp. in different water sources, dust and soil samples collected near human habitats. However, no local study has yet to examine these amoebae in a forest, an isolated terrestrial environment, where human activity is relatively scarce. Further, there are also limited studies to investigate the same globally. The current study reported the isolation, morphological and genotypic characterisations of eleven Acanthamoeba sp. isolated from soils of the Payeh Maga Highland forest, Sarawak, Malaysia. Morphological analysis revealed that nine isolates belonged to Group II, whereas two isolates belonged to Group III as defined by the criteria of Pussard and Pons. The phylogenetic analysis based on complete 18S rRNA gene sequences showed that the isolates belonged to the rare T1 (six isolates), T6 (two isolates) and T13 (three isolates) genotypes. To the best of our knowledge, this is the first report about the detection of T6 Acanthamoeba sp. in this country. Overall, the current findings have enriched the knowledge pertaining to the occurrence, morphological and genotypic characteristics of Acanthamoeba sp. in an isolated terrestrial environment of Malaysia.

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