This is a ten year (1999-2008) retrospective study of amebiasis in patients admitted to UMMC. A total of 34 cases were analyzed. The most common were amebic liver abscess 22(65%) and the rest were amoebic dysentery 12(35%). Majority of the cases occurred among Malaysians 29(85%), with Chinese 14(41%), followed by the Malays 9(26%) and the Indians 6(18%). Foreigners made up of one Indonesian, one Pakistani and three Myanmarese and constituted 5(15%) of the total cases. Males 24(71%) were more commonly affected. Most of the cases occurred between the age group of 40-49 years, 8(23%) and 60 years and above, 8(23%). Age group of 20-50 years constituted 20(60%) of the cases. The most common clinical presentations were fever with chills and rigors 26(76%), diarrhoea 20 (59%), right hypochondrium pain 17(50%), abdominal pain 17(50%), hepatomegaly 16 (47%) and jaundice 7(20%). All were discharged well after treatment except for one case of death in a 69-year-old Chinese male with amebic liver abscess.
Transmission electron microscopy (TEM) can provide high resolution imaging of biological specimens. The study is to establish the effects of a modified glutaraldehyde (GA) compare to the standard GA fixation on Acanthamoeba castellanii from TEM perspectives and thus provide precise and accurate information on the ultrastructure studies of the parasite. By increasing the contrast, the ultrastructures of the parasite were more evident. The TEM images were obtained from parasites fixed with the modified GA and the standard GA and then the area of the nucleus and the gray values of the image of the nucleus of the parasites were measured. The mean areas of the nucleus were found to be significantly reduced in the standard GA fixed parasites (12210.4 nm2) compared to the modified GA fixed parasites (8676.3 nm2) (p < 0.05). The mean gray values of the image were significantly reduced from 2024 in the standard GA fixed parasites (2024) to the modified GA fixed parasites (1636) (p < 0.05). The study shows that the modified GA produced significantly better contrast on TEM images of the A. castellanii compared to the standard GA. This was because the modified GA generated more free water molecules during fixation and the uptake of modified GA by the nucleus of the parasite organizing all protein constituents in the cell into a more closely packed configuration than that of the standard GA. With such properties, the modified GA is a better fixative providing better images for ultrastructures of the parasite.
A total of 10 out of 65 cornea swab samples from cats with eye symptoms showed Acanthamoeba-like morphology after cultivation. By PCR and DNA sequencing of Acanthamoeba diagnostic fragment 3 (DF3), all 10 isolates from the positive samples were categorized into two homologous groups of AfC1 (PM1, PM2, PM3, PF6, KM7, KF8, KMK9) and AfC2 (PM4, PM5, KFK10) due to the presence of bases A(354) and G(354), respectively. Furthermore, DF3 of AfC1 and AfC2 showed 100% similarity with Genbank reference isolates with the accession numbers DQ087314, EU146073 and U07401, GU808323, which were Acanthamoeba castellanii strains genotype T4 originating from human keratitis. This finding suggests that A. castellani strains have the capability to infect cats and human under favorable conditions.
Rhipicephalus (Boophilus) microplus serves as an important ectoparasite of livestock and a vector of several pathogens resulting in diseases, subsequently affecting the agricultural field as well as the economy. The extensive use of synthetic acaricides is known to cause resistance over time and therefore a much safer, effective and environmentally friendly alternative to overcome tick infestation should be implemented. Larval immersion tests (LIT) were done to evaluate the effects of Citrus hystrix (Family: Rutaceae) and Cymbopogon citratus (Family: Poaceae) essential oils (EOs) for their individual and combined (1:1) acaricidal activity against the cattle tick. Results showed that LC50 and LC90 values in 24 and 48 hours for Cit. hystrix EO were 11.98% and 24.84%, and 10.95% and 21.71% respectively. LC50 and LC90 values for Cym. citratus EO were 1.21% and 6.28%, and 1.05% and 6.12% respectively. The mixture of EOs from two plants in 1:1 ratio (Cit. hystrix 50%: Cym. citratus 50%) was found to exhibit antagonistic effect (synergistic factor < 1). The 24 hours and 48 hours LC50 and LC90 values for combined EOs were 1.52% and 2.84%, and 1.50% and 2.76% respectively. Individual and combined essential oils were subjected to qualitative analysis using gas chromatography-mass spectrometry (GC-MS) to screen the chemical components present in EOs. Our results showed that the combination of Cit. hystrix and Cym. citratus at 1:1 ratio resulted in an antagonistic effect and the use of Cym. citratus alone is more toxic to R. (B.) microplus, making it a better alternative to chemical based acaricide.
We aimed at determination of acaricidal, larvacidal, and repellent activities of green synthesized silver nanoparticles (SNP) against Hyalomma dromedarii as one of the most common ticks in camels. SNP were green synthesized by reducing Lupinus albus extract through the precipitation technique. The acaricidal, larvicidal, and repellent activity of SNP against H. dromedarii was studied through the adult immersion test (AIT), the larval packet test (LPT), the vertical movement behavior of tick's larvae method, anti-acetylcholinesterase (AChE) activity, and oxidative enzyme activity. The green synthesized SNP displayed a spherical form with a size ranging from 25-90 nm; whereas the most distribution of particles size was reported at 50-65 nm. SNP dose-dependently (p<0.001) increased the mortality rate of H. dromedarii adult; whereas at 16 and 32 µg/mL completely killed the adult females. Treatment of exposure of H. dromedarii adult to SNP markedly (p<0.001) declined the mean number, weight, and hatchability of eggs. Treatment of H. dromedarii larvae with SNP reduced the viability rate of larvae with the LC50 and LC90 values of 3.1 and 6.9 µg/mL, respectively. Exposure of H. dromedarii larvae to SNP, especially at ½ LC50 and LC50, markedly (p<0.001) increased the oxidative stress and declined the level of antioxidant enzymes in H. dromedarii larvae; whereas, markedly suppressed the AChE activity of the larvae stage of H. dromedarii in comparison to the control group. These results showed that SNP green synthesized by L. albus extract had promising acaricidal, larvicidal and repellent activity against H. dromedarii adults and larvae as a dose-dependent response. SNP also considrably decreased the level of acetylcholinesterase and antioxidant activity and also provokes oxidative stress in H. dromedarii larvae. However, more investigation must be designed to clear the accurate mechanisms and the efficacy of SNP in practical use.
The study was aimed to evaluate the effectiveness of maggot therapy in healing of cutaneous infected wound in streptozotocin (STZ) induced diabetic Wistar rat. For live maggots, the sterilized eggs of Lucilia sericata were obtained from colonies established in laboratory. Diabetes model was established in 48 male Wister rat by intra-peritoneal injection of STZ at the dose of 60 mg/kg body-weight. Cutaneous wounds exposed with mixed colonies of bacteria like Staphylococcus aureus, E. coli and Pseudomonas aeruginosa were prepared in all rat. The animals equally divided in 4 groups with 12 rats each being presented as treatment group of control, antibiotic, maggot and maggot with antibiotic in combination. All treatments were done once and hold for 24 hours. Wound kinetics and bacterial bio burden were measured at weekly interval to till complete healing. Significant reduction in wound area with maximum contraction was found (>95%) in maggot treated group when compared to antibiotic treated (79%) and control (72%). In maggot as well as maggot and antibiotic in combination group showed early elimination of bacterial bio-burden 7.88±0.03log CFU/ml to 1.12±0.65log CFU/ml and 7.86±0.04) log CFU/ml to 1.54±0.52log CFU/ml respectively in three weeks of time. Early healing indication was also experienced on histomorphological examination of wounded tissue of maggot treated groups by early and better epithelialization, collagenation and neovascularization with complete healing of wound in three weeks in comparison to antibiotic and control respectively. However, the present study did not show any difference in healing of wound with use of maggot alone or in antibiotic combination. Live maggot of Lucilia sericata effectively lower bacterial bioburden and and accelerate healing of infected cutaneous wound in diabetic conditions.
Apart from occasional reports of clinical disease affecting horses, there is no information about Trypanosoma evansi in horses in Peninsula Malaysia. Thus, a cross-sectional study was conducted in eight states in Peninsula Malaysia to determine the active presence of T. evansi in horses. A total of 527 blood samples were obtained and examined by haematocrit centrifugation technique (HCT), Giemsa-stained thin blood smear (GSS), morphometric measurements, polymerase chain reaction (PCR) and cloning of PCR products. The results showed an overall parasitological prevalence of 0.57% (3/527, CI: 1.6-0.19%) with both HCT and GSS. Morphometric study revealed the mean total length of the trypanosomes including the free flagellum was 27.94 ± 2.63 μm. PCR successfully amplified a trypanosome specific 257 bp in 1.14% of samples (6/527, CI: 2.4-0.52%) and was confirmed by nucleotide sequences. The mean packed cell volume (PCV) for the positive cases detected by HCT was lower (23% ± 7.00) compared to the positive cases detected by PCR alone in the state of Terengganu (35% ± 4.73). In conclusion, this study showed T. evansi infection occurred in low frequency in horses in Peninsula Malaysia, and anaemia coincided with parasitaemic animals. PCR is considered as a sensitive diagnostic tool when parasitaemia is undetectable. The slight lengthier mean of parasite and anaemia may indicate a virulent strain of T. evansi circulating throughout the country. Thus, it's highly recommended to shed light on host-parasite relationship for better epidemiological understanding.
The susceptibility of Culex quinquefasciatus to chemical insecticides in two field sites in Kuala Lumpur was evaluated using the WHO standard susceptibility test. Less then 7 days old female mosquitos, reared from wild caught females were exposed to discriminating dosages of insecticides at recommended exposure periods. The larval bioassay were conducted using the multiple concentrations and the LC50 value was determined. The results indicated that cyfluthrin is the most effective among all the insecticides tested with LT50 value of 29.95 min and 28.59 min, for the strain from Ampang Hill and Pantai Dalam, respectively. It was surprisingly to note that both these field strains showed 0% mortality when tested against malathion and DDT. The LC50 value indicated that both strains were highly resistant to malathion with resistance ratio of 17,988 folds and 14,053 folds, respectively. This concludes that resistance at larval stages is extremely high compared to adult stages.
Decomposing carrion provides a temporary microhabitat and food source for a distinct community of organisms. Arthropods constitute a major part of this community and can be utilized to estimate the postmortem interval (PMI) of cadavers during criminal investigations. However, in Malaysia, knowledge of carrion arthropod assemblages and their succession is superficial. Therefore, a study on three types of forensic entomology animal model was conducted from 27 September 2010 to 28 October 2010 in a tropical rainforest at National University of Malaysia, Bangi, Selangor, Malaysia. Over one month collections of arthropods were made on nine animal carcasses: three laboratory rats (Rattus norvegicus, mean weight: 0.508 ± 0.027 kg), three rabbits (Oryctolagus cuniculus, mean weight: 2.538 ± 0.109 kg) and three long tailed macaque (Macaca fascicularis, mean weight: 5.750 ± 0.551 kg). A total of 31,433 arthropods belonging to eight orders and twenty-eight families were collected from all carcasses. Among 2924 of adults flies collected, approximately 19% were calliphorids with Chrysomya megacephala (Fabricius, 1794) being the most abundant. Arthropod taxon richness was lower on rat carcasses compared to that of rabbit and monkey carcasses, and this was more apparent during the first week of decomposition. However, there were no significant differences in Shannon-Weiner index (H'), Simpson dominance index (C) and Pielou's Evenness index (J) between different animal model. The arthropod assemblages associated to animal model were different significantly (p<0.05) while decomposition stage was a significant factor influencing insect assemblages (p<0.05). Analysis on the arthropods succession indicated that some taxa have a clear visitation period while the others, particularly Coleoptera, did not show a clear successional pattern thus require futher insect succession study. Although human bodies were not possible for the succession study, most of the arthropods collected are necrophagous, and will also possibly colonize human cadaver, and potentially be useful in assisting in estimates of PMI in future forensic cases in Malaysia.
Nasopharyngeal diphtheria is an acute infectious upper respiratory tract disease caused by toxigenic strains of Corynebacterium diphtheriae. We report a case of a young adult who presented to us with a short history of fever, sore throat, hoarseness of voice and neck swelling. He claimed to have received all his childhood vaccinations and had no known medical illnesses. During laryngoscopy, a white slough (or membrane) was seen at the base of his tongue. The epiglottis was also bulky and the arytenoids were swollen bilaterally. The membrane was sent to the microbiology laboratory for culture. A diagnosis of nasopharyngeal diphtheria was made clinically and the patient was treated with an antitoxin together with erythromycin, while awaiting the culture result. Nevertheless, the patient's condition deteriorated swiftly and although the laboratory eventually confirmed an infection by toxin-producing C. diphtheriae, the patient had already succumbed to the infection.
The adulticidal activity of methanol extracts from three Malaysian plants namely Acorus calamus Linn., Litsea elliptica Blume and Piper aduncum Linn. against adult of Aedes aegypti (L.) were studied. Standard WHO bioassay tests were used to evaluate the effectiveness of these plant extracts. The hexane fraction from methanol extract of Acorus calamus rhizome was the most effective, exhibiting LC50 and LC90 values of 0.04 mgcm(-2) and 0.09 mgcm(-2) respectively. For L. elliptica, the methanol fraction also displayed good adulticidal property with the LC50 and LC90 values of 0.11 mgcm(-2) and 6.08 mgcm(-2) respectively. It is found that hexane fraction of the P. aduncum crude extract was the least effective among the three plants showing LC50 and LC90 values of 0.20 mgcm(-2) and 5.32 mgcm(-2), respectively. However, although A. calamus showed lowest LC values, the LT50 results indicated that the methanol fraction of L. elliptica was most potent extract among the extracts tested.
Early detection of a dengue outbreak is an important first step towards implementing effective dengue interventions resulting in reduced mortality and morbidity. A dengue mathematical model would be useful for the prediction of an outbreak and evaluation of control measures. However, such a model must be carefully parameterized and validated with epidemiological, ecological and entomological data. A field study was conducted to collect and analyse various parameters to model dengue transmission and outbreak. Dengue prone areas in Kuala Lumpur, Pahang, Kedah and Johor were chosen for this study. Ovitraps were placed outdoor and used to determine the effects of meteorological parameters on vector breeding. Vector population in each area was monitored weekly for 87 weeks. Weather stations, consisting of a temperature and relative humidity data logger and an automated rain gauge, were installed at key locations in each study site. Correlation and Autoregressive Distributed Lag (ADL) model were used to study the relationship among the variables. Previous week rainfall plays a significant role in increasing the mosquito population, followed by maximum humidity and temperature. The secondary data of rainfall, temperature and humidity provided by the meteorological department showed an insignificant relationship with the mosquito population compared to the primary data recorded by the researchers. A well fit model was obtained for each locality to be used as a predictive model to foretell possible outbreak.
African swine fever (ASF) is a transboundary haemorrhagic viral disease that affected domestic and wild pigs of all ages. The disease is caused by African swine fever virus (ASFV) and was introduced to China in 2018 before spreading rapidly to neighbouring Asian countries. As such, putting countries free from ASF like Malaysia at risk. ASF is highly lethal with no vaccine or treatment available. In February 2021, we confirmed backyard pigs from various locations in Sabah were infected with ASF using real time polymerase chain reaction (real- time PCR). Further characterization of the Sabah ASFVs indicated that they were of p72 genotype II with intergenic region (IGR) variant II that displayed an addition tandem repeat sequence (TRS) insertion, similar to ASFV from Indonesia, V ietnam and China. These results indicate and support the transboundary expansion of a homogenotypic ASFV (p72 genotype II and IGR variant II) in the Europe and Asia-Pacific, emphasizing the need for a holistic international collaboration in control and preventing further spreading of the current ASF pandemic. Importantly, our results informed the first detection and characterization of ASF, a disease previously not detected in Malaysia. This information is crucial for further mitigation and preventive measures.
Accurate estimation of the minimum post-mortem interval (minPMI) is important in the investigation of forensic cases. Various thanatological methods are being used to estimate this interval. However, entomology approach is the most reliable method for this minPMI estimation especially when death has occurred over 72 hours and involved insects or other arthropods evidence at the death scene. The current methods of age estimation are daunting and destructive especially when dealing with pupal stage. The aims of this study were to characterize the morphological changes during intra-puparial period of Chrysomya megacephala (Fabricius) (Diptera: Calliphoridae) and their relation with minPMI estimation by using a high resolution micro-Computed Tomography (micro-CT). Gravid C. megacephala were collected from a rural area in Sungai Buloh, Selangor and cultured in the laboratory at 23.83±0.25°C with light: dark hour of 12:12 to initiate oviposition. The resulting larvae were reared until pupal stage. A pupa was collected at first (24 hours), second (48 hours), third (72 hours), and fourth quarter (96 hours) of the intra-puparial period. The pupal samples were placed directly into 70% ethanol for preservation. Micro-CT scanning was employed to acquire microstructural information following pupal sample staining for contrast enhancement. Eight age-informative internal morphological landmarks were mapped from the micro-CT scanning. The present study enhanced the potential value of micro-CT for the estimation of minPMI based on the internal morphological changes of C. megacephala pupae. This novel method is a promising tool for improving medico-legal investigations in forensic entomology.
Schistosomiasis is the second most common parasitic disease post Malaria around the world. Praziquantel (PZQ) is known as the most efficient anti- schistosomal drug but has no anti-fibrotic effect. Metformin (Met) is a well-known drug for type 2 diabetes. This study aimed to evaluate the role of Met as anti-schistosomal and anti-fibrotic agents alone or in combination with PZQ treatment. Forty male CD1 mice were divided into four groups (n=10 mice) as following; the first group (Gp1) was served as a negative control. Gp2, Gp3, Gp4, and Gp5 were infected with (60-80) S. mansoni cercariae. After a month of infection, Gp3 was administered orally with PZQ (500 mg/Kg) for 2 consecutive days. Gp4 was administered orally with Met (150 mg/Kg) for 15 consecutive days, and Gp5 was orally administered with PZQ followed by Met for 15 consecutive days at the same doses as in Gp 3 and 4. The results showed that PZQ had potent worms and egg reduction in liver and intestine tissues with no anti-fibrotic effect of the granuloma formation. However, Met or PZQ/Met treatment postinfection led to a reduction in egg count in both liver and intestine tissues with a significant reduction in granuloma site. Treatment of S. mansoni-infected mice with Met or PZQ/Met ameliorated the hematological and biochemical alterations induced by S. mansoni infection. Collectively, Met has no anti-schistosomal activity but led to a reduction in egg deposition and showed an anti-fibrotic effect on granulomatous development either when used alone or in combination with PZQ treatment. This study shed light on the possible role of Met as an anti-fibrotic agent when administered with PZQ for S. mansoni infected humans.
Timely and rapid diagnosis is crucial for faster and proper malaria treatment planning. Microscopic examination is the gold standard for malaria diagnosis, where hundreds of millions of blood films are examined annually. However, this method's effectiveness depends on the trained microscopist's skills. With the increasing interest in applying deep learning in malaria diagnosis, this study aims to determine the most suitable deep-learning object detection architecture and their applicability to detect and distinguish red blood cells as either malaria-infected or non-infected cells. The object detectors Yolov4, Faster R-CNN, and SSD 300 are trained with images infected by all five malaria parasites and from four stages of infection with 80/20 train and test data partition. The performance of object detectors is evaluated, and hyperparameters are optimized to select the best-performing model. The best-performing model was also assessed with an independent dataset to verify the models' ability to generalize in different domains. The results show that upon training, the Yolov4 model achieves a precision of 83%, recall of 95%, F1-score of 89%, and mean average precision of 93.87% at a threshold of 0.5. Conclusively, Yolov4 can act as an alternative in detecting the infected cells from whole thin blood smear images. Object detectors can complement a deep learning classification model in detecting infected cells since they eliminate the need to train on single-cell images and have been demonstrated to be more feasible for a different target domain.
An understanding of common pathogens and their antibiotic sensitivity patterns is critical for proper management of sepsis in Emergency Department (ED). The goal of the study was to identify common organisms isolated from blood cultures of patients attended to ED and their antimicrobial susceptibility. Beginning from 2002, all cases of positive blood culture collected by the ED, Hospital Universiti Sains Malaysia (HUSM) were recorded and analysed. Over the period of eight years, we documented 995 cases of positive blood cultures. Of these samples, 549 (55.2%) were Gram-negative bacteria; 419 (42.1%) were Gram-positive bacteria; 10 (1.0%) were anaerobic organisms; 10 (1.0%) were fungus; and 7 (0.7%) cases were mixed organisms. Gram-negative bacteria were observed to develop more resistance to antimicrobial agents, especially those commonly used in an outpatient setting with less than 80% sensitivity to ampicillin, cotrimoxazole and ciprofloxacin. By contrast, there has been no marked change in the sensitivity trends of Gram-positive bacteria over the same period. In conclusion, ED physicians are more equipped to initiate empirical antimicrobial therapy especially when dealing with possibility of Gram-negative sepsis.
Study site: Emergency department, Hospital Universiti Sains Malaysia (HUSM), Kelantan, Malaysia
Time series modelling and forecasting plays an important role in various domains. The objective of this paper is to construct a simple average ensemble method to forecast the number of cases for infectious diseases like dengue and typhoid and compare it by applying models for forecasting. In this paper we have also evaluated the correlation between the number of typhoid and dengue cases with the ecological variables. The monthly data of dengue and typhoid cases from 2014 to 2017 were taken from integrated diseases surveillance programme, Government of India. This data was analysed by three models namely support vector regression, neural network and linear regression. The proposed simple average ensemble model was constructed by ensemble of three applied regression models i.e. SVR, NN and LR. We combine the regression models based upon the error metrics such as Mean Square Error, Root Mean Square Error and Mean Absolute Error. It was found that proposed ensemble method performed better in terms of forecast measures. The finding demonstrates that the proposed model outperforms as compared to already available applied models on the basis of forecast accuracy.
This is a cross-sectional study conducted from January to September 2004 in a group of school children aged 8 -10 years old. The schools and study subjects were selected using stratified systematic sampling technique. A total of 44 schools and 1100 subjects were selected from schools with iodinator and schools without iodinator. Samples collected were spot urine and drinking water. Dietary and iodised water consumption data were obtained from interviews. A total of 931 subjects (84.6%) responded; 558 (50.7%) from schools with iodinator and 373 (33.9%) from schools without iodinator. Results showed that in more than half (53.8%) of the schools with iodinators, mean water iodine level was below 25 microg/L. The study population in Terengganu was found to be mildly iodine deficient with an overall median urine iodine concentration (uIC) of 74 microg/L. Based on WHO criteria, 4.1% with uIC <20 microg/L (severe), 19.5% with uIC between 20-49 microg/L (moderate), 49.2% with uIC between 50-99 microg/L (mild) and 27.2% was iodine sufficient with uIC >100 microg/L. Majority of the study subjects were found to have high seafood intake (> 90%) and low in goitrogen food intake. This study suggests water iodinator system may not be a suitable method of supplying iodine and an alternative is needed in order to eradicate the iodine deficiency problem seen in some parts of Malaysia.
This study was conducted in Samsun Province of Turkey to investigate the serological status of domesticated water buffaloes for both Crimean-Congo Hemorrhagic Fever (CCHF) and Lumpy Skin Disease (LSD). Serum was collected from a total of 272 water buffaloes from different age groups and both genders; of the total, 48.1% had been vaccinated against LSD with heterologous sheep-goat pox vaccine. The serum samples were individually assessed by using a commercial ID screen enzyme-linked immune-sorbent assay (ELISA) to detect neutralizing antibodies against both CCHF virus and LSD virus. All 272 buffaloes were negative for antibodies against the CCHF virus. All the unvaccinated buffaloes (141) were seronegative for LSD virus but of the 131 vaccinated buffaloes, 10 (7.6%) were seropositive for the LSD virus. In addition, 8.6% of vaccinated animals age >1 year old were seropositive for LSD, whereas the seropositivity was 5.1% for the animals age <= 1 year old. There was no significant difference for seropositivity between male and female animals in the >1 year old or <= 1 year old age groups. When seroprevalances for LSD in the tested water buffaloes are evaluated by gender, there was a significant difference between females (8.6%) and males (0%) in the <1 year old water buffaloes (X2=20.24; P<0.001). Separately, the results of this study indicate that Bafra district water buffaloes are not infected by CCHFV and LSDV and some of the buffaloes that vaccinated with LSDV did not develop sufficient antibodies to protect them after they were vaccinated for the LSD virus. Furthermore, the authors of this study conclude that both the commercially produced vaccine that is currently administered and the vaccination strategy have to be urgently evaluated by the veterinary authorities in Turkey. This is essential in order to combat the spread of LSD virus infection with an effective vaccine and a comprehensive management strategy across Turkey.