Plasmodium Knowlesi Infection Research Articles

First molecular documented case of a rarely reported parasite: Plasmodium knowlesi infection in Denmark in a traveller returning from Malaysian Borneo

Plasmodium knowlesi has been reported as an emerging infection throughout the Southeast Asian region, especially in the Malaysian state of Sabah, where it accounts for the majority of the malaria cases reported. This is in contrast to Europe, where imported P. knowlesi is a rarely reported infection. We present a case of P. knowlesi infection in a Danish woman returning from a short trip to Malaysian Borneo. Microscopy of blood smears revealed 0.8% infected erythrocytes, but due to the atypical morphological presentation, a conclusive species identification was made by molecular methods. Plasmodium knowlesi is a potentially fatal infection and taking the increasing travel activity into consideration after the coronavirus disease 2019 (COVID-19) pandemic, P. knowlesi should be a differential diagnosis in patients with travel-associated illness returning from highly endemic Southeast Asian areas.

SICA-mediated cytoadhesion of Plasmodium knowlesi-infected red blood cells to human umbilical vein endothelial cells

Zoonotic malaria due to Plasmodium knowlesi infection in Southeast Asia is sometimes life-threatening. Post-mortem examination of human knowlesi malaria cases showed sequestration of P. knowlesi-infected red blood cells (iRBCs) in blood vessels, which has been proposed to be linked to disease severity. This sequestration is likely mediated by the cytoadhesion of parasite-iRBCs to vascular endothelial cells; however, the responsible parasite ligands remain undetermined. This study selected P. knowlesi lines with increased iRBC cytoadhesion activity by repeated panning against human umbilical vein endothelial cells (HUVECs). Transcriptome analysis revealed that the transcript level of one gene, encoding a Schizont Infected Cell Agglutination (SICA) protein, herein termed SICA-HUVEC, was more than 100-fold increased after the panning. Transcripts of other P. knowlesi proteins were also significantly increased, such as PIR proteins exported to the iRBC cytosol, suggesting their potential role in increasing cytoadhesion activity. Transgenic P. knowlesi parasites expressing Myc-fused SICA-HUVEC increased cytoadhesion activity following infection of monkey as well as human RBCs, confirming that SICA-HUVEC conveys activity to bind to HUVECs.

Exploring the key anthropological drivers of and barriers to zoonotic malaria preventative behaviour in a community exposed to Plasmodium knowlesi infection in Malaysia: protocol for a qualitative study with a participatory research design

IntroductionPlasmodium knowlesi malaria is a zoonotic mosquito-borne disease with complex epidemiology. According to the WHO, the prevention and control of vector-borne diseases require community participation to increase coherence between malaria...

Risk factor of plasmodium knowlesi infection in Sabah Borneo Malaysia, 2020: A population-based case-control study.

BackgroundIn the Malaysian state of Sabah, P. knowlesi notifications increased from 2% (59/2,741) of total malaria notifications in 2004 to 98% (2030/2,078) in 2017. There was a gap regarding P. knowlesi acquisition risk factors related to practice specifically in working age group. The main objective of this study was to identify the risk factors for acquiring P. knowlesi infection in Sabah among the working age group.Methods and methodsThis retrospective population-based case-control study was conducted in Ranau district to assess sociodemographic, behavioural and medical history risk factors using a pretested questionnaire. The data were entered and analyzed using IBM SPSS version 23. Bivariate analysis was conducted using binary logistic regression whereas multivariate analysis was conducted using multivariable logistic regression. We set a statistical significance at p-value less than or equal to 0.05.ResultsA total of 266 cases and 532 controls were included in the study. Male gender (AOR = 2.71; 95% CI: 1.63–4.50), spending overnight in forest (AOR = 1.92; 95% CI: 1.20–3.06), not using mosquito repellent (AOR = 2.49; 95% CI: 1.36–4.56) and history of previous malaria infection (AOR = 49.34; 95% CI: 39.09–78.32) were found to be independent predictors of P. knowlesi infection.ConclusionsThis study showed the need to strengthen the strategies in preventing and controlling P. knowlesi infection specifically in changing the practice of spending overnight in forest and increasing the usage of personal mosquito repellent.

High incidence of Plasmodium knowlesi malaria compared to other human malaria species in several hospitals in Malaysia.

Through the regional control programme, Malaysia has been successfully reducing the incidence of Plasmodium falciparum and Plasmodium vivax infections. However, the incidence of zoonotic malaria Plasmodium knowlesi infection is increasing and now has been the major cause of malaria in Malaysia especially Malaysian Borneo. The emergence of knowlesi infection has threatened the malaria elimination programme which the government aims to reduce the overall malaria infections by 2020. Unlike other benign human Plasmodium spp., P. knowlesi can cause fatal infections. The aim of this study was to determine the incidence and distribution of five human malaria parasites including P. knowlesi in Peninsular Malaysia and Malaysian Borneo. A total of 112 blood samples were collected from seven states and district hospitals in Peninsular Malaysia and Malaysian Borneo from year 2015 to 2016. The samples were examined by microscopy and further confirmed by nested PCR assay targeting 18S rRNA gene of Plasmodium spp. Following the nested PCR assays, a total of 54 (48.2%) samples were positive for P. knowlesi infections, 12 (10.7%) cases were positive for P. vivax infections, followed by 7 (6.3%) cases of P. falciparum and 4 (3.5%) cases of P. malariae. There were 3 cases (2.7%) of mixed infections (P. knowlesi/P. vivax). However, no cases were identified as P. ovale. A total of 32 (28.6%) cases were found as negative infections. LoopMediated Isothermal Amplification Assay (LAMP) was performed to confirm inconclusive results produced by microscopy and nested PCR. P. knowlesi showed the highest prevalence in Sarawak (n= 30), Sabah (n=13), Pulau Pinang (n=5) and Pahang (n=6). PCR and LAMP was not able to detect a large number of microscopy positive samples due to DNA degradation during storage and shipping. Among all the states involved in this study, the highest prevalence of P. knowlesi infection was found in Sabah and Sarawak.

Determinants of Parasite Clearance Time among Plasmodium knowlesi Patients in Malaysia

Plasmodium (P.) knowlesi infection was only recently discovered in 2004, and since then the number of cases has increased each year. As such, there is still a need to understand the clinical management of P. knowlesi infection to ensure control of the disease. Therefore, this work was performed to determine factors that affect the clinical outcome of P. knowlesi infected patients. A cross-sectional study from 2012 to 2016 was conducted in Hospital Kuala Lipis, Pahang, Malaysia. The main clinical outcome was parasite clearance. A total of 72 P. knowlesi patients were included, which were mostly male (n=56, 77.80%), with a mean age of 37.39±14.61 years. Fifty-six patients (77.80%) were prescribed with artemisinin-based antimalarial following diagnosis, while the remaining patients (n=16, 22.20%) were treated with chloroquine. The length for parasite clearance of P. knowlesi was 4(4-5) days [(median (IQR)]. Factors that affect the primary outcome of the study (>4 days for parasite clearance) was then determined. A multivariate logistic regression demonstrated that significant predictors of parasite clearance were initial parasite levels and number of symptoms (χ2=11.97, df(69), P=0.003). Those with an initial parasite count of > 2 400/uL were 4.8 times more likely to take > 4 days for parasite clearance (P=0.02). Having < 4 symptoms increased the odds of > 4 days for parasite clearance by 0.14 times (P=0.02). This study shows the need to closely monitor patients with higher initial parasite counts (> 2400/uL) and patients with > 4 symptoms associated with P. knowlesi infection, to ensure appropriate treatment is administered.

Spatial and Temporal Analysis of Plasmodium knowlesi Infection in Peninsular Malaysia, 2011 to 2018.

The life-threatening zoonotic malaria cases caused by Plasmodium knowlesi in Malaysia has recently been reported to be the highest among all malaria cases; however, previous studies have mainly focused on the transmission of P. knowlesi in Malaysian Borneo (East Malaysia). This study aimed to describe the transmission patterns of P. knowlesi infection in Peninsular Malaysia (West Malaysia). The spatial distribution of P. knowlesi was mapped across Peninsular Malaysia using Geographic Information System techniques. Local indicators of spatial associations were used to evaluate spatial patterns of P. knowlesi incidence. Seasonal autoregressive integrated moving average models were utilized to analyze the monthly incidence of knowlesi malaria in the hotspot region from 2012 to 2017 and to forecast subsequent incidence in 2018. Spatial analysis revealed that hotspots were clustered in the central-northern region of Peninsular Malaysia. Time series analysis revealed the strong seasonality of transmission from January to March. This study provides fundamental information on the spatial distribution and temporal dynamic of P. knowlesi in Peninsular Malaysia from 2011 to 2018. Current control policy should consider different strategies to prevent the transmission of both human and zoonotic malaria, particularly in the hotspot region, to ensure a successful elimination of malaria in the future.

RISK FACTORS FOR PLASMODIUM KNOWLESI IN SOUTHEAST ASIA:A SYSTEMATIC REVIEW Authors

Introduction: Plasmodium knowlesi zoonotic malaria has been identified in Borneo Malaysia in 2004, and has led to an increase in the number of cases throughout Southeast Asia. Many factors have contributed to increasing pattern. Therefore, this systematic review aims to identify risk factors for Plasmodium knowlesi infection that can aid in predicting and controlling the infection. Methods:Systematic search on articles related to risk factors of Plasmodium knowlesi infection was conducted using three databases namely PubMed, Scopus and Science direct. All the articles from year 2015 till May 2019 were selected. Results: A total of three articles were included in this review based on the study protocol. Three common risk factors for Plasmodium knowlesi infection was in identify mainly involved sociodemographic, work-related and behavioural factors. Conclusion: Risk factors for P. Knowlesi ranges not only from individual and community socidemographic background but also by environmental and geographical determinants. The inconsistency of the result was due to the variability and limitations of the studies itself. The research on P. Knowlesi, should be conducted with larger participants and longer length of follow up.

Prevalence of severe Plasmodium knowlesi infection and risk factors related to severe complications compared with non-severe P. knowlesi and severe P. falciparum malaria: a systematic review and meta-analysis

BackgroundPlasmodium knowlesi is a potential cause of severe and fatal malaria, but comprehensive studies of its pooled prevalence and risk factors are lacking. This study aimed to explore the prevalence and risk factors related to severe P. knowlesi infection.MethodsA systematic review was conducted by retrieving all published articles on severe P. knowlesi available in Web of Science (ISI), Scopus, and PubMed (MEDLINE). Titles, abstracts, and full-text articles were screened, and any irrelevant studies were excluded. The random-effects model was used to compute the pooled prevalence estimate of severe P. knowlesi infection by a metaprop command provided in STATA software. Differences in demographic characteristics, clinical characteristics, and laboratory data were analysed using Review Manager Version 5.3 software for patients in the following groups: 1) patients with severe and non-severe P. knowlesi infection and 2) patients with severe P. knowlesi and severe P. falciparum infection.ResultsOut of the 2382 studies retrieved from the three databases, seven studies with a total enrolment of 1124 patients with P. knowlesi infections were eligible to be included in this systematic review and meta-analysis. The pooled prevalence estimate of severe P. knowlesi infection was 19% (95% CI: 11–27%, I2 = 93.7%). Severe acute kidney injuries (AKI) (77 cases, 45.6%), jaundice (71 cases, 42%), and hyperparasitaemia (55 cases, 32.5%) were the common clinical manifestations found among patients with severe complications. In comparison to non-severe P. knowlesi infections, patients with severe P. knowlesi infections had significantly higher age, leucocyte count, and parasitaemia levels (P < 0.05). In comparison to patients with severe P. falciparum infections, patients with severe P. knowlesi infections had significantly higher age, neutrophil count, and creatinine levels (P < 0.05).ConclusionsThis systematic review and meta-analysis demonstrated a high proportion of severe P. knowlesi infections. Patients with severe P. knowlesi infections had higher age, leucocyte count, and parasitaemia levels than those with non-severe P. knowlesi infections. In addition, patients with severe P. knowlesi infections had higher age, neutrophil count, and creatinine levels than those with severe P. falciparum infections.

Malaria cross-sectional surveys identified asymptomatic infections of Plasmodium falciparum, Plasmodium vivax and Plasmodium knowlesi in Surat Thani, a southern province of Thailand

ObjectivesMalaria cross-sectional surveys are rarely conducted in very low transmission settings. This study aimed to determine the prevalence and risk factors of Plasmodium infection in a near-elimination setting in southern Thailand. MethodsTwo cross-sectional surveys were conducted in areas of active transmission in the Surat Thani province of Thailand in January and May 2019. PCR was used to detect Plasmodium infection. ResultsThe prevalence of Plasmodium blood infection was 0.45% and 0.61% in January and May 2019, respectively. The major parasite species was Plasmodium falciparum in January and Plasmodium vivax in May. Unexpectedly, Plasmodium knowlesi infections were also detected. Most infections, including those of Plasmodium knowlesi, were asymptomatic. Being male and staying outdoors at night-time were the only significant identified risk factors. Of people infected in January 28.0% were positive in May for the same parasite species, suggesting persistent asymptomatic infections. ConclusionsDespite the very low incidence rate in Surat Thani, most malaria infections were asymptomatic. Outdoor mosquito biting at night-time is likely an important mode of malaria transmission. Unexpectedly, asymptomatic Plasmodium knowlesi infection was found, confirming previous reports of such infection in mainland Southeast Asia.

Vector compositions change across forested to deforested ecotones in emerging areas of zoonotic malaria transmission in Malaysia

In lowland areas of Malaysia, Plasmodium knowlesi infection is associated with land use change and high proportions of the vector Anopheles balabacensis. We conducted a 15-month study in two Malaysian villages to determine the effect of habitat on vector populations in understudied high-altitude, high-incidence districts. Anopheles mosquitoes were sampled in human settlements, plantations and forest edges, and screened for Plasmodium species by PCR. We report the first An. donaldi positive for P. knowlesi. This potential vector was associated with habitat fragmentation measured as disturbed forest edge:area ratio, while An. balabacensis was not, indicating fragmented land use could favour An. donaldi. Anopheline species richness and diversity decreased from forest edge, to plantation, to human settlement. Greater numbers of An. balabacensis and An. donaldi were found in forest edges compared to human settlements, suggesting exposure to vectors and associated zoonoses may be greater for people entering this habitat.

Concurrent juvenile myelomonocytic leukemia with thalassemia in a case with Plasmodium knowlesi infection from Sabah, Malaysian Borneo.

A 3-year-old male child was presented with worsening abdominal pain, abdominal distension, lethargy, pallor and hepatosplenomegaly. The patient had multiple outpatient visits in the past and was treated with oral antibiotics, oral anthelmintic agents, albeit with minimal benefit. The patient also had non-neutropenic pyrexia spikes and oral ulcers. The patient was an adopted child; hence details about his biological parents’ previous history were unclear. Differential diagnosis of Chronic Myelomonocytic Leukemia (CMML), Juvenile Myelomonocytic Leukemia (JMML), Gaucher’s disease, Thalassemia and discrete pancreatic pathology was considered. Hemoglobin electrophoresis was indicative of thalassemia. Also, molecular detection method by polymerase chain reaction confirms a concurrent infection with Plasmodium knowlesi malaria. The BCR-ABL fusion gene was found to be negative. Correlating with peripheral monocytosis, bone marrow aspiration and trephine biopsy with blasts only 3-4% and hepatosplenomegaly, a diagnosis of JMML was established. We present a rare phenomenon with an overlap of signs and symptoms between JMML, underlying thalassemia, and Plasmodium knowlesi, posing a diagnostic challenge to physicians.

Occupationally Acquired Plasmodium knowlesi Malaria in Brunei Darussalam.

Simian malaria is a zoonotic disease caused by Plasmodium knowlesi infection. The common natural reservoir of the parasite is the macaque monkey and the vector is the Anopheles mosquito. Human cases of P. knowlesi infection has been reported in all South East Asian countries in the last decade, and it is currently the most common type of malaria seen in Malaysia and Brunei. Between 2007–2017, 73 cases of P. knowlesi infection were notified and confirmed to the Ministry of Health in Brunei. Of these, 15 cases (21%) were documented as work-related, and 28 other cases (38%) were classified as probably related to work (due to incomplete history). The occupations of those with probable and confirmed work related infections were border patrol officers, Armed Forces and security personnel, Department of Forestry officers, boatmen and researchers. The remaining cases classified as most likely not related to work were possibly acquired via peri-domestic transmission. The risk of this zoonotic infection extends to tourists and overseas visitors who have to travel to the jungle in the course of their work. It can be minimised with the recommended use of prophylaxis for those going on duty into the jungles, application of mosquito/insect repellants, and use of repellant impregnated uniforms and bed nets in jungle camp sites.

Plasmodium knowlesi infection in a returning German traveller from Thailand: a case report on an emerging malaria pathogen in a popular low-risk travel destination

BackgroundThailand is a major destination for German travellers with more than 760,000 arrivals in 2015. At the same time, malaria is a concern in travel recommendations with regard to this destination. The World Malaria Report of 2016 mentions only P. falciparum and P. vivax as prevalent species for Thailand, however, P. knowlesi infections have been occasionally reported in Thailand. In German travellers, only five cases of P. knowlesi infections have been reported to date.Case presentationA 45-year-old German male tourist travelled to Thailand from 25 December 2016 to 13 January 2017. On 14 January he developed fever with no other symptoms, and presented on 17 January at the Division for Tropical Medicine and Infectious Diseases in Munich, Germany.Malaria was diagnosed, primarily based on a single parasite in the thin smear microscopy, while commercial rapid diagnostic testing remained negative. Only the result of a differential PCR assay revealed P. knowlesi infection.ConclusionsP. knowlesi has to be considered in travellers returning from Thailand. Cases may present with an extremely low parasitaemia. This is in contrast to the assumption that P. knowlesi was likely to cause high parasitaemia due to its short replication cycle.

A fatal malaria caused by Plasmodium knowlesi infection in a healthy man, Betong, Yala, Thailand April, 2016

A fatal malaria caused by Plasmodium knowlesi infection in a healthy man, Betong, Yala, Thailand April, 2016

Neglected tropical diseases among the Association of Southeast Asian Nations (ASEAN): overview and update.

The ten member states of the Association of Southeast Asian Nations (ASEAN) constitute an economic powerhouse, yet these countries also harbor a mostly hidden burden of poverty and neglected tropical diseases (NTDs). Almost 200 million people live in extreme poverty in ASEAN countries, mostly in the low or lower middle-income countries of Indonesia, the Philippines, Myanmar, Viet Nam, and Cambodia, and many of them are affected by at least one NTD. However, NTDs are prevalent even among upper middle-income ASEAN countries such as Malaysia and Thailand, especially among the indigenous populations. The three major intestinal helminth infections are the most common NTDs; each helminthiasis is associated with approximately 100 million infections in the region. In addition, more than 10 million people suffer from either liver or intestinal fluke infections, as well as schistosomiasis and lymphatic filariasis (LF). Intestinal protozoan infections are widespread, while leishmaniasis has emerged in Thailand, and zoonotic malaria (Plasmodium knowlesi infection) causes severe morbidity in Malaysia. Melioidosis has emerged as an important bacterial NTD, as have selected rickettsial infections, and leptospirosis. Leprosy, yaws, and trachoma are still endemic in focal areas. Almost 70 million cases of dengue fever occur annually in ASEAN countries, such that this arboviral infection is now one of the most common and economically important NTDs in the region. A number of other arboviral and zoonotic viral infections have also emerged, including Japanese encephalitis; tick-borne viral infections; Nipah virus, a zoonosis present in fruit bats; and enterovirus 71 infection. There are urgent needs to expand surveillance activities in ASEAN countries, as well as to ensure mass drug administration is provided to populations at risk for intestinal helminth and fluke infections, LF, trachoma, and yaws. An ASEAN Network for Drugs, Diagnostics, Vaccines, and Traditional Medicines Innovation provides a policy framework for the development of new control and elimination tools. Together with prominent research institutions and universities, the World Health Organization (WHO), and its regional offices, these organizations could implement important public health improvements through NTD control and elimination in the coming decade.

Parasite accumulation in placenta of non-immune baboons during Plasmodium knowlesi infection.

BackgroundPlacental malaria (PM) causes adverse pregnancy outcomes in the mother and her foetus. It is difficult to study PM directly in humans due to ethical challenges. This study set out to bridge this gap by determining the outcome of PM in non-immune baboons in order to develop a non-human primate model for the disease.MethodsTen pregnant baboons were acquired late in their third trimester (day 150) and randomly grouped as seven infected and three non-infected. Another group of four nulligravidae (non-pregnant) infected was also included in the analysis of clinical outcome. Malaria infection was intravenously initiated by Plasmodium knowlesi blood-stage parasites through the femoral vein on 160th day of gestation (for pregnant baboons). Peripheral smear, placental smear, haematological samples, and histological samples were collected during the study period. Median values of clinical and haematological changes were analysed using Kruskal-Wallis and Dunn’s Multiple Comparison Test. Parasitaemia profiles were analysed using Mann Whitney U test. A Spearman’s rank correlation was run to determine the relationship between the different variables of severity scores. Probability values of P <0.05 were considered significant.ResultsLevels of white blood cells increased significantly in pregnant infected (34%) than in nulligravidae infected baboons (8%). Placental parasitaemia levels was on average 19-fold higher than peripheral parasitaemia in the same animal. Infiltration of parasitized erythrocytes and inflammatory cells were also observed in baboon placenta. Malaria parasite score increased with increase in total placental damage score (rs = 0.7650, P <0.05) and inflammatory score (rs = 0.8590, P <0.05). Although the sample size was small, absence of parasitized erythrocytes in cord blood and foetal placental region suggested lack of congenital malaria in non-immune baboons.ConclusionThis study has demonstrated accumulation of parasitized red blood cells and infiltration of inflammatory cells in the placental intravillous space (IVS) of baboons that are non-immune to malaria. This is a key feature of placental falciparum malaria in humans. This presents the baboon as a new model for the characterization of malaria during pregnancy.

Genotyping of the Duffy blood group among Plasmodium knowlesi-infected patients in Malaysia.

The Duffy blood group is of major interest in clinical medicine as it plays an important role in Plasmodium knowlesi and Plasmodium vivax infection. In the present study, the distribution of Duffy blood group genotypes and allelic frequencies among P. knowlesi infected patients as well as healthy individuals in Peninsular Malaysia were determined. The blood group of 60 healthy blood donors and 51 P. knowlesi malaria patients were genotyped using allele specific polymerase chain reaction (ASP-PCR). The data was analyzed using Fisher's exact test in order to assess the significance of the variables. Our results show a high proportion of the FY*A/FY*A genotype (>85% for both groups) and a high frequency of the FY*A allele (>90% for both groups). The FY*A/FY*A genotype was the most predominant genotype in both infected and healthy blood samples. The genotype frequency did not differ significantly between the donor blood and the malaria patient groups. Also, there was no significant correlation between susceptibility to P. knowlesi infection with any Duffy blood genotype.

Recent advances in the management of Plasmodium knowlesi infection.

Plasmodium knowlesi (P. knowlesi) has been detected to be the fifth malarial parasite that can cause malaria in human beings. The parasite is known to commonly infect macaque monkeys. The infection is highly prevalent in South-East Asia. It has morphologic similarities to Plasmodium malariae and Plasmodium falciparum. P. knowlesi is known to replicate every 24 h in the human host and hence, causes “quotidian malaria.” It causes a wide spectrum of clinical manifestations and sometimes can cause fatal illness. Chloroquine is effective in the treatment of uncomplicated P. knowlesi infection. Severe and complicated P. knowlesi malaria can be managed with artemisinin combination therapy.

Plasmodium knowlesi infection imported to Germany, January 2013

To the editor: We read with great interest the article by Orth et al. [1] in Eurosurveillance on a recent case of imported Plasmodium knowlesi infection in Germany. This case nicely illustrates the pivotal role of microscopy on thick and thin blood films by experienced microscopists for malaria diagnosis. The statement in the discussion section that only five cases imported to Europe have been published so far, underestimates the occurrence of this infection. Two more cases imported to the Netherlands have been described previously [2,3].