A 17-year-old man with acute lymphoblastic leukaemia had fever and diarrhoea during a febrile neutropenic episode. A spiral-shaped, Gram-negative anaerobic bacterium was isolated from blood, and confirmed as Anaerobiospirillum succiniciproducens by 16S rRNA sequencing. The patient responded to imipenem.
Trichuris Dysentery Syndrome (TDS) is a severe persistent trichuriasis associated with heavy worm build-up in the colon that continues to be neglected and underestimated in endemic countries. Trichuriasis is most prevalent in children in tropical countries, and that increases the risk of TDS. We reported a series of four preschool children of both genders chronically having TDS over a period ranging from several months to years presenting with anaemia. The hemoglobin levels ranged from 4.6 to 9.1 g/dl on first admissions. Despite treatment, the cases were reported to have failure to thrive with persistent anaemia. It was concluded that TDS should be considered in endemic areas among children presenting with chronic bloody diarrhea and anaemia.
Parasitic diarrhea among children is a significant health problem worldwide. This cross sectional study described the burden of parasitic diarrhea among children. The objectives of this study were to evaluate the impact of risk factors on the parasitic diarrhea, and to determine the parasitic profile among children in Baghdad-Iraq, during the period extending from September 2003 to June 2004. A total number of 2033 cases were included in the study. The estimated prevalence rate of parasitic diarrhea was 22%. We identified the following major diarrhea determinants were large households size, residential location, water source, low socioeconomic status, and low parent education. Giardia lamblia was found to be the most prevalent parasite with an infection rate of 45.54% followed by Entamoeba histolytica 23.44%, Enterobius vermicularis 12.7%, Hymenolepis nana 9.82%, Trichuris trichiura 5.4%, and Ascaris lumbricoides 2.2%. In conclusion, this study demonstrates that poor sanitation, inadequate environmental conditions, and low socioeconomic status are the main determining factors that predispose children to parasitic diarrhea. Mass deworming programs are recommended for school children, as this population is easily accessible.
Shigellosis is a foodborne illness caused by the genus Shigella and is an important global health issue. The development of effective techniques for rapid detection of this pathogen is essential for breaking the chain of transmission. Therefore, we have developed a novel loop-mediated isothermal amplification (LAMP) assay targeting the invasion plasmid antigen H (ipaH) gene to rapidly detect Shigella species. This assay could be performed in 90 min at an optimal temperature of 64ºC, with endpoint results visualized directly. Notably, the method was found to be more sensitive than conventional PCR. Indeed, the detection limit for the LAMP assay on pure bacterial cultures was 5.9 x 10(5) CFU/ml, while PCR displayed a limit of 5.9 x 10(7) CFU/ml. In spiked lettuce samples, the sensitivity of the LAMP assay was 3.6 x 10(4) CFU/g, whereas PCR was 3.6 x 10(5) CFU/g. Overall, the assay accurately identified 32 Shigella spp. with one enteroinvasive Escherichia coli displaying positive reaction while the remaining 32 non-Shigella strains tested were negative.
The present study aims to develop a system which consists of four pairs of primers that specifically detects Salmonella spp., Salmonella serovar Typhi and Salmonella serovar Paratyphi A with an internal amplification control. The system, when applied in Polymerase Chain Reaction (PCR) under specific conditions, reaction mixture and cycling temperatures produced four bands; 784 bp, 496 bp, 332 bp and 187 bp. The DNA band 784 bp is present in all Salmonella spp., while the bands of 496 bp and 332 bp are only present in S. Paratyphi A and S. Typhi, respectively. An internal amplification control as indicated by the 187 bp shows the system is working in optimum condition in all the tests. This multiplex PCR was evaluated on 241 bacterial cultures and 691 naturally contaminated samples. Overall, this multiplex PCR detection system provides a single step for simultaneous detection of DNAs of Salmonella spp., S. Typhi and S. Paratyphi A.
Wet mount microscopy is the most commonly used diagnostic method for trichomoniasis in clinical diagnostic services all over the world including Sri Lanka due to its availability, simplicity and is relatively inexpensive. However, Trichomonas culture and PCR are the gold standard tests. Unfortunately, neither the culture nor PCR is available for the diagnosis of trichomoniasis in Sri Lanka. Thus, it is important to validate the wet mount microscopy as it is the only available diagnostic test and has not been validated to date in Sri Lanka. The objective was to evaluate the validity and reliability of wet mount microscopy against gold standard Trichomonas culture among clinic based population of reproductive age group women in Western province, Sri Lanka. Women attending hospital and institutional based clinics were enrolled. They were interviewed and high vaginal swabs were taken for laboratory diagnosis by culture and wet mount microscopy. There were 601 participants in the age group of 15-45 years. Wet mount microscopy showed 68% sensitivity, 100% specificity, 100% positive (PPV) and 98% negative predictive values (NPV) (P=0.001, kappa=0.803) respectively against the gold standard culture. The area under the ROC curve was 0.840. Sensitivity of wet mount microscopy is low. However it has high validity and reliability as a specific diagnostic test for trichomoniasis. If it is to be used among women of reproductive age group in Western province, Sri Lanka, a culture method could be adopted as a second test to confirm the negative wet mount for symptomatic patients.
Plasmodium knowlesi has been recently identified as the "fifth human malaria species" following the discovery in Malaysian Borneo of a large focus of this simian malaria parasite in humans. Even though it shares microscopic similarities with Plasmodium malariae, it may cause severe illness with risk of fatality. We describe a case of P. knowlesi infection causing multi-organ failure in a patient who was successfully managed due to early recognition of the infection. Clinicians in this region should be more aware of the infection as it is not as rare as previously thought. This case write up highlight the case of severe malaria infection which presented with multi organ involvement which is caused by P. knowlesi.
A case of human eye infection caused by Brugia pahangi was reported in 2010 in a semi rural village in Selangor, peninsular Malaysia. Our report here reveals results of investigation on the vector and animal host for the transmission of the infection. We conducted entomological survey and cat blood examination in the vicinity of the patient's home. The mosquito species Armigeres subalbatus was incriminated as the vector, whereas cat served as the reservoir host.
Snake bite has been regarded as an important health problem in Myanmar since early 1960's. In the recent years, there has been growing interest in alternative therapies and therapeutic use of natural products, especially those derive from plants. In Myanmar and Indian traditional medicine, various plants have used as a remedy for treating snake bite. The present study was carried out to evaluate the effects of alcohol extract of Tamarind (Tamarindus indica Linn.) seed on some biologic properties of Russell's viper (Daboia russelli siamensis) venom (RVV). The Phospholipase A2 (PLA2) enzyme, coagulase enzyme and caseinolytic enzyme activities of Russell's viper venom (RVV) were reduced when mixed and incubated with the extract. When the RVV and the different amount of extracts were preincubated and injected intramuscularly into mice, all of them survived, but all the mice in the control group died. On the other hand, when RVV were injected first followed by the extract into mice, all of them died. If the extract was injected near the site where Russell's viper venom was injected, all the mice survived for more than 24 hours and the survival time prolonged but they all died within 96 hours. In conclusion, according to the results obtained, the extract neutralizes some biologic properties of the Russell's viper venom and prolonged the survival time if the extract was injected near the site where the Russell's viper venom was injected.
Battling malaria will be a persistent struggle without the proper means to diagnose the parasitic infection. However, the inherent limitations of microscopy, the conventional method of diagnosing malaria, affect the accuracy of diagnosis. The present study aimed to compare the accuracy of two different set of primers targeting the small subunit ribosomal RNA (ssRNA) and the dihydrofolate reductase-thymidylate synthase linker region (dhfr-ts) in detecting species specific malaria infections by nested PCR. The sensitivity and specificity of nested PCR assay using the two primers were calculated with reference to microscopy as the 'gold standard'. The results show that 18S rRNA primers had 91.9% sensitivity and 100% specificity in detecting human Plasmodium species as opposed to dhfr-ts primers which had 51.4% sensitivity and 100% specificity. The higher sensitivity of 18S rRNA primers suggests that it may be a better diagnostic tool for detecting human malaria.
Burkholderia pseudomallei the causative agent of melioidosis, is being increasingly recognized as an important cause of morbidity and mortality in South East Asia. Biofilm formation of B. pseudomallei may be responsible for dormancy, latency and relapse of melioidosis. Based on the colonial morphology of the bacteria on B. pseudomallei selective agar medium, seven distinct morphotypes were identified. This study was conducted to assess the in vitro biofilm produced by B. pseudomallei and to investigate possible correlation between B. pseudomallei morphotypes with biofilm forming abilities of the isolates. Using a standard biofilm crystal violet staining assay, comparison was made between the biofilm forming ability of 76 isolates of B. pseudomallei and Burkholderia thailandensis ATCC 700388. Amongst the blood isolates, 30.2% were considered as high biofilm producers and 27.9% were low producers, 33.3% of the pus isolates were considered as high and 16% low biofilm producers. Most of the isolates were identified as morphotype group 1 which displayed a rough centre with irregular circumference on the agar medium. However, we did not find any correlation of B. pseudomallei morphotypes with biofilm forming abilities (p > 0.05). Additional studies are needed to identify internal and external factors which contribute to the high and low biofilm formation of B. pseudomallei.
Trichuris dysentery syndrome is caused by Trichuris trichiura which contributes to one of the most common helminthic infections in the world. It is associated with heavy colonic infection that manifests as mucoid diarrhoea, rectal bleeding, rectal prolapse, iron deficiency anaemia, and finger clubbing. Here, we report a case of trichuris dysentery syndrome complicated with severe chronic iron deficiency anaemia in a 4-year-old girl who required blood transfusion. The nematode was visualized on stool microscopic and colonoscopic examination. A longer duration of anti-helminthic treatment is required to achieve effective and better outcome.
A study was carried out to isolate Brucella melitensis using established bacteriological and PCR techniques in Brucella seropositive goats in farms in Selangor, Negeri Sembilan, Melaka and Pulau Pinang. Brucella melitensis was isolated from 7 of 134 reactors with the highest isolation from the vaginal swabs (57.14%) followed by the spleen (28.57%), uterine fluid (14.29%). No Brucella was isolated from the lymph nodes. PCR confirmed all the seven isolates as B. melitensis and isolates were phylogenetically related to other isolates from India, Iran, and Israel but most closely related to isolates from Singapore.
In this study we have cloned unreported gene fragments of Toxoplasma gondii GRA7 and SAG1 and expressed the corresponding recombinant proteins, followed by evaluation of their usefulness for the serological diagnosis of toxoplasmosis. Both recombinant proteins were expressed efficiently in insoluble form, purified by single step Ni-NTA affinity chromatography and their antigenicity to detect toxoplasma specific IgG antibodies were determined by immunoblotting. A total of 60 serum samples from three groups of individuals based on their anti-toxoplasma antibody profiles were tested, namely (I) IgM+, IgG+ (n=20), (II) IgM-, IgG+ (n=20) and (III) IgM-, IgG- (n=20). Both recombinant proteins exhibited high sensitivity (100%) with sera from Group I. rGRA7 and rSAG1 reacted 40% and 80% respectively with Group II sera. The specificity of the recombinant proteins based on reactivities with Group III sera were 100% and 80% with rGRA7 and rSAG1 respectively. Thus rGRA7 was found to be better at discriminating probable acute from chronic phases of toxoplasmosis, and it also showed higher specificity.
Oxyspiruriasis caused by the bird eyeworm, Oxyspirura mansoni, a thelaziid nematode, in three species of pheasants, 3 Chrysolophus pictus (golden pheasant), 7 Lophura nycthemera (silver pheasant) and 9 Phasianus colchicus (common pheasant) in Zoo Negara Malaysia are reported. Birds with the disease were treated with a solution of 0.5% iodine or 0.5% lysol. Antistress powder for 4 days in water and non-strep vitamin powder in water was also provided. Control measures included removal of the cockroach intermediate host, Pycnoscelus surinamensis (Surinam cockroach) from the vicinity of the birds. The golden pheasant is a new host for O. mansoni in peninsular Malaysia.
Penicilliosis is a rare occurrence among non human immunodeficiency virus (HIV) infected patients. We report here two cases of Penicillium marneffei infection in patients with systemic lupus erythematosus (SLE). Both patients had a recent flare of lupus and were on immunosuppressive drugs when they presented with prolonged fever without an obvious foci of infection, unresponsive to broad-spectrum antibiotics. They were leucopaenic upon admission, with rapid deterioration during the course of the illness. Diagnosis of penicilliosis via fungal isolation from blood culture was delayed resulting in the late initiation of antifungal agents. While both patients ultimately recovered, the delay in diagnosis led to a prolonged hospital stay with increased morbidity. Clinicians should be aware of this uncommon but emerging fungal pathogen in SLE patients and maintain a high index of suspicion in diagnosing this potentially fatal but treatable disease.
Plasmodium knowlesi is a simian malaria parasite and is recently recognized as the fifth malaria parasite infecting humans. Manifestation of the infection may resemble other infection particularly dengue fever leading to inappropriate management and delay in treatment. We reported three cases of naturally acquired P. knowlesi in Hospital Universiti Sains Malaysia, Kubang Kerian, Kelantan, Malaysia. Clinical manifestations were quite similar in those cases. Microscopically, the diagnosis might be challenging. These cases were confirmed by polymerase chain reaction method which serves as a gold standard.
The biodiversity and the killer activity of yeasts isolated from various types of fermented food in Malaysia were investigated in this study. Of 252 yeasts isolated from 48 fermented food samples in this study, 19 yeast species were identified based on sequence analysis of the ITS1-5.8S-ITS2 partial fragments of the yeasts. A total of 29 (11.5%) of the yeast isolates demonstrated killer activity to at least one Candida species tested in this study; including 22 isolates of Trichosporon asahii, 4 isolates of Pichia anomala, and one isolate each of Pichia norvegensis, Pichia fermentans and Issatchenkia orientalis, respectively. The presence of killer yeasts reflects antagonism that occurs during microbial interaction in the fermented food, whereby certain yeasts produce killer toxins and possibly other toxic substances in competition for limited nutrients and space. The anti-Candida activity demonstrated by killer yeasts in this study should be further explored for development of alternative therapy against candidiasis.
Human immunodeficiency virus (HIV)-infected individuals have greater susceptibility to infections by a myriad of microorganisms which can cause significant morbidity and mortality compared to immunocompetent individuals. Of these microbial infections, intestinal parasitic infections (IPIs) however are receiving less attention than bacterial and viral infections, hence, the lack of information of parasitic infections in HIV individuals. Prevalence of IPIs among 346 HIV-infected individuals in Malaysia was determined in this study. The overall prevalence of intestinal parasitic infections (IPIs) was 37.9% (131 of 346) with protozoa infections (18.8%) being more common compared to helminth infections (7.5%). Observed protozoa include Entamoeba histolytica/dispar (16.8%), Cryptosporidium parvum (12.4%), Isospora belli (10.1%), Cyclospora cayetanensis (4.9%) and Giardia duodenalis (intestinalis) (3.2%) whilst helminthes which were detected comprised of Ascaris lumbricoides (13.9%), Trichuris trichiura (6.4%) and hookworms (0.6%). Among those 131 infected, 50.4% had multiple infections and 48.9% had single parasitic infection. The CD4 counts were significantly lower (i.e., 200 cells/mm³) in patients harbouring IPIs. Of those individuals infected with intestinal parasites, 49% were intravenous drug users and 58% were not on any antiretroviral therapy. Most were asymptomatic and had concurrent opportunistic infections (OIs) mainly with Mycobacterium tuberculosis infection. These results confirmed that IPIs are ubiquitous among HIV-infected individuals, especially those presenting with low CD4 T cells counts, and provide useful insights into the epidemiology of these infections among HIV-infected patients in Malaysia. It is therefore recommended, that diagnosis of these intestinal parasitic pathogens should be conducted on a routine basis for better management of gastrointestinal illnesses among HIV individuals.
The cytochrome oxidase subunit I (COXI) gene sequences of three recent (2007-2008) clinical Plasmodium knowlesi isolates from Klang Valley, peninsular Malaysia, were determined and compared with those of older (1960's) peninsular Malaysia, recent isolates from Sarawak (on Borneo Island), and an isolate from Thailand. Multiple alignment of the sequences showed that the three clinical isolates were more similar to the older peninsular Malaysia isolates than to those from Sarawak and Thailand. Phylogenetic tree based on the COXI sequences revealed three distinct clusters of P. knowlesi. The first cluster consisted of isolates from peninsular Malaysia, the second consisted of Sarawak isolates and the third composed of the Thailand isolate. The findings of this study highlight the usefulness of mitochondrial COXI gene as a suitable marker for phylogeographic studies of P. knowlesi.