Imported case ofLeishmania tropica cutaneous leishmaniasis in a 10-year-old child in Malaysia.

The present paper reported a first imported case of cutaneous leishmaniasis in a 10-year- old child who returned from Saudi Arabia to Malaysia. Six weeks after his travel to Malaysia, two erythematous dermal nodules were developed over his right cheek and chin. Occurrence of intracellular amastigote of Leishmania was observed through examination of skin biopsy with hematoxylin and eosin stain. Furthermore, molecular analysis of ribosomal internal transcribed spacer 1 (ITS1) of Leishmania spp. confirmed the child was infected with Leishmania tropica. The child was given oral fluconazole and he had a 80% recovery before he went back to Saudi Arabia.

Kinetics of immune responses to SARS-CoV-2 proteins in individuals with varying severity of infection and following a single dose of the AZD1222.

To characterize the IgG and IgA responses to different SARS-CoV-2 proteins, we investigated the antibody responses to SARS-CoV-2 following natural infection and following a single dose of AZD1222 (Covishield), in Sri Lankan individuals. The IgG and IgA responses were assessed to S1, S2, RBD, and N proteins in patients at 4 weeks and 12 weeks since the onset of illness or following vaccination. Antibodies to the receptor-binding domain of SARS-CoV-2 wild type (WT), α, ß, and λ and ACE2 (Angiotensin Converting Enzyme 2) receptor blocking antibodies were also assessed in these cohorts. For those with mild illness and in vaccines, the IgG responses to S1, S2, RBD, and N protein increased from 4 weeks to 12 weeks, while it remained unchanged in those with moderate/severe illness. In the vaccines, IgG antibodies to the S2 subunit had the highest significant rise (P < 0.0001). Vaccines had several-fold lower IgA antibodies to all the SARS-CoV-2 proteins tested than those with natural infection. At 12 weeks, the haemagglutination test (HAT) titres were significantly lower to the α in vaccines and significantly lower in those with mild illness and in vaccines to ß and for λ. No such difference was seen in those with moderate/severe illness. Vaccines had significantly less IgA to SARS-CoV-2, but comparable IgG responses those with natural infection. However, following a single dose vaccines had reduced antibody levels to the VOCs, which further declined with time, suggesting the need to reduce the gap between the two doses, in countries experiencing outbreaks due to VOCs.

Seroprevalence of SARS-CoV-2 Infection in the Colombo Municipality Region, Sri Lanka.

Background: As the Municipality Council area in Colombo (CMC) experienced the highest number of cases until the end of January 2021, in Sri Lanka, we carried out a serosurvey prior to initiation of the vaccination program to understand the extent of the SARS-CoV-2 outbreak. Methods: SARS-CoV-2 seropositivity was determined in 2,547 individuals between the ages of 10-86 years, by the Wantai total antibody ELISA. We also compared seroprevalence using the haemagglutination test (HAT) to evaluate its usefulness in carrying out serosurveys. Results: The overall seropositivity rate was 24.46%, while seropositivity by HAT was 18.90%. Although The SARS-CoV-2 infection detection rates by PCR were highest in the population between the ages of 20-60 years of age, there was no statistically significant difference in the seropositivity rates in different age groups. For instance, although the seropositivity rate was highest in the 10-20 age group (34.03%), the PCR positivity rate was 9.80%. Differences in the PCR positivity rates and seropositivity rates were also seen in 60-70-year-olds (8.90 vs. 30.4%) and in individuals >70 years (4.10 vs. 1.20%). The seropositivity rate of the females was 29.70% (290/976), which was significantly higher (p < 0.002) than in males 21.2% (333/1,571). Conclusions: A high seroprevalence rate (24.5%) was seen in all age groups in the CMC suggesting that a high level of transmission was seen during this time. The higher PCR positivity rates between the ages of 20-60 are likely to be due to increased testing carried out in the working population. Therefore, the PCR positivity rates, appear to underestimate the true extent of the outbreak and the age groups which were infected.

Bioefficacy of mosquito mat vaporizers and associated metabolic detoxication mechanisms in Aedes aegypti (Linnaeus) in Selangor, Malaysia: A statewide assessment.

This study aims to examine the efficacy of mosquito mat vaporizers on Aedes aegypti and their associated metabolic detoxication mechanisms. For this purpose, Aedes aegypti (Linnaeus) was collected from nine districts in Selangor, Malaysia and tested with mosquito vaporizing mat bioassays. The same populations were also subjected to biochemical assays to investigate activities of detoxifying enzymes, namely non-specific esterase (EST), glutathione-S-transferase (GST) and mixed function oxidase (MFO). The efficacy of Ae. aegypti on the active ingredients tested in decreasing order were d- allethrin > dimefluthrin > prallethrin with PBO > prallethrin. The results further indicated significant enhancement mean levels of EST, GST and MFO in pyrethroid-resistant populations. The mortality rate of Ae. aegypti in response to pyrethroid active ingredients was associated with MFO activity, suggesting it is an important detoxification enzyme for the populations tested. In view of the presence of resistance against household insecticide products, pyrethroid efficacy on Ae. aegypti populations needs to be monitored closely to ensure the implementation of an effective vector control program in Malaysia.

Immune responses to a single dose of the AZD1222/Covishield vaccine in health care workers.

Several COVID-19 vaccines have received emergency approval. Here we assess the immunogenicity of a single dose of the AZD1222 vaccine, at one month, in a cohort of health care workers (HCWs) (629 naïve and 26 previously infected). 93.4% of naïve HCWs seroconverted, irrespective of age and gender. Haemagglutination test for antibodies to the receptor binding domain (RBD), surrogate neutralization assay (sVNT) and ex vivo IFNγ ELISpot assays were carried out in a sub-cohort. ACE2 blocking antibodies (measured by sVNT) were detected in 67/69 (97.1%) of naïve HCWs. Antibody levels to the RBD of the wild-type virus were higher than to RBD of B.1.1.7, and titres to B.1.351 were very low. Ex vivo T cell responses were observed in 30.8% to 61.7% in naïve HCWs. Previously infected HCWs, developed significantly higher (p < 0.0001) ACE2 blocking antibodies and antibodies to the RBD for the variants B.1.1.7 and B.1.351. This study shows high seroconversion after one vaccine dose, but also suggests that one vaccine dose may be insufficient to protect against emerging variants.

Comparison of two assays to detect IgG antibodies to the receptor binding domain of SARS­CoV­2 as a surrogate marker for assessing neutralizing antibodies in COVID-19 patients.

BACKGROUND: Neutralizing antibodies (NAbs) are important for protection against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) reinfection. In this study, two assays that are correlated with NAbs were compared: the haemagglutination test (HAT) and the surrogate virus neutralization test (sVNT). METHODS: The specificity of the HAT was compared with the sVNT, and the sensitivity and persistence of antibodies in patients with varying severity of illness was assessed in a cohort of 71 patients at 4-6 weeks and 13-16 weeks. The kinetics were assessed in the first, second, and third weeks in patients with varying severity of acute illness. RESULTS: The specificity of the HAT was >99%, and sensitivity was similar to the sVNT. The levels of HAT were significantly and positively correlated with those of the sVNT (Spearman's r = 0.78, P < 0.0001). Patients with moderate and severe illness had higher HAT titres when compared to those with mild illness. Six of seven patients with severe illness had a titre of >1:640 during the second week of illness, whereas only five of 31 patients with a mild illness had a titre of >1:160 in the second week of illness. CONCLUSIONS: Since the HAT is a simple and very cheap assay to perform, it would be ideal to use as an indicator of NAbs in resource-poor settings.

Serological survey of canine vector-borne diseases in two animal shelters in central Peninsular Malaysia.

Canine vector-borne diseases (CVBDs) are increasingly becoming a cause for global concern because of their high morbidity and mortality rates in dogs. However, information on their occurrence in Malaysia is still scanty. In this study, a total of 103 dog blood samples were collected from two animal shelters in central Peninsular Malaysia and tested for the antibodies against Ehrlichia spp., Anaplasma spp., Borrelia burgdorferi, and the antigen of Dirofilaria immitis. Of the 103 tested dogs, 44.7% (46) were found to be seropositive for Ehrlichia spp., 30.1% (31) for Anaplasma spp. and 13.6% (14) for D. immitis. Co-infections of Anaplasma spp. + Ehrlichia spp. (18.5%, 19) were most prevalent, followed by Anaplasma spp. + D. immitis (1.9%; two) and D. immitis + Ehrlichia spp. (1.0%; one). Furthermore, three dogs (2.9%) were also found to have triple infection, testing seropositive for Ehrlichia spp., Anaplasma spp. and D. immitis. The dogs which were found to be seropositive with at least one pathogen were 66.7% (32/51) at shelter A, and 55.8% (29/52) at shelter B. Serological evidence showed that the exposure of major vector-borne diseases in dogs in shelters was relatively high in the surveyed areas. Routine detection and control of vector-borne diseases are of paramount importance for reducing the risk of CVBDs transmission in dogs and humans.

Observations of scrotal mass, liver mass, haemolytic jaundice, and central vestibular disorder in Brugia pahangi-infected dogs.

Brugia pahangi is known to infect humans and dogs. Its associated symptoms and complications, however, have not been fully understood in dogs. Herein, we reported the observations of B. pahangi infections in dogs with scrotal mass, liver mass, haemolytic jaundice, and central vestibular disorder.

Nematode control failure due to anthelmintic resistance in a sheep farm in Malaysia: First identification of the F200Y mutation in the isotype 1 ß-tubulin gene.

This paper reports total nematode anthelmintic resistance towards albendazole, fenbendazole, levamisole and ivermectin in a commercial sheep farm located in Terengganu, Malaysia. Faecal Egg Count Reduction Test (FECRT) was conducted on 25 sheep, where five sheep in each group were treated with the respective four anthelmintics based on live bodyweight. The balance of five sheep placed in the control group were not treated with any anthelmintics. At day 13 post-treatment, faecal egg count was conducted and nematode worm egg count reduction percentage was calculated to determine the resistance status towards the respective anthelmintics tested. Results showed that nematodes were resistant to all the anthelmintics tested, namely albendazole, fenbendazole, levamisole and ivermectin with reduction percentage of 87%, 46%, 94% and 68%, respectively. Subsequently, the third stage larvae of Haemonchus contortus and Trichostrongylus colubriformis recovered from post-treatment faecal cultures were subjected to allele-specific polymerase chain reaction (AS-PCR) assay to determine the presence of the benzimidazole resistance gene. This study reports the occurrence of the classical F200Y mutation in the isotype 1 ßtubulin gene, for the first time in Malaysia.

Occurrence of gastro-intestinal parasites among small ruminants in Malaysia: highlighting Dicrocoelium infection in goats.

The aim of the present study was to determine the gastro-intestinal (GI) parasitic infections among small ruminants (i.e., goats, sheep, deer) in Malaysia through formalin-ether concentration technique. Overall, 70.9% or 302 out of 426 small ruminants (79.4% or 251/316 goats; 87.5% or 35/40 sheep; 22.9% or 16/70 deer) were infected with at least one species of GI parasites. Overall, ten types of GI parasites [Helminth: strongyle (57.7%), Moniezia spp. (5.4%), Paramphistomum spp. (4.5%), Strongyloides spp. (4.2%), Dicrocoelium spp. (2.3%), Trichuris spp. (2.3%); Protozoa: Eimeria spp. (23.7%), Entamoeba spp. (18.8%), Giardia spp. (1.9%), Cryptosporidium spp. (0.2%)] were detected in this study. Among the studied animals, goats harboured the highest diversity of GI parasites (ten types), followed by sheep (six types) and deer (two types). Polyparasitism was observed in goats (43.7% or 138 of 316) and sheep (15.0% or 6 of 40). Cumulatively, a total of 32 combinations of coinfections (Helminth+Helminth: 8 combinations; Helminth+Protozoa: 20 combinations; Protozoa+Protozoa: 4 combinations) between detected parasites with up to quintuple infections were reported. Among these parasites, "strongyle + Eimeria spp." and "Moniezia spp. + strongyle" were the commonest infections in goats (13.5% or 34 of 251) and sheep (5.7% or 2 of 6), respectively. This study is a comprehensive documentation on multiple GI parasitisms among small ruminant in Malaysia, and the findings are crucial for effective farm management, especially for the formulation of parasitic control and elimination strategies.

First molecular genotyping of A302S mutation in the gamma aminobutyric acid (GABA) receptor in Aedes albopictus from Malaysia.

Given the lack of molecular evidence in altered target-site insecticide resistance mechanism in Aedes albopictus (Skuse) worldwide, the present study aims to detect the presence of A302S mutation in the gene encoding the gamma aminobutyric acid receptor resistant to dieldrin (Rdl) in Ae. albopictus for the first time from its native range of South East Asia, namely Malaysia. World Health Organization (WHO) adult susceptibility bioassay indicated a relatively low level of dieldrin resistance (two-fold) in Ae. albopictus from Petaling Jaya, Selangor. However, PCR-RFLP and direct sequencing methods revealed the presence of the A302S mutation with the predomination of heterozygous genotype (40 out of 82 individuals), followed by the resistant genotype with 11 individuals. This study represents the first field evolved instance of A302S mutation in Malaysian insect species.

Mitochondrial DNA analyses reveal low genetic diversity in Culex quinquefasciatus from residential areas in Malaysia.

The present study explored the intraspecific genetic diversity, dispersal patterns and phylogeographic relationships of Culex quinquefasciatus Say (Diptera: Culicidae) in Malaysia using reference data available in GenBank in order to reveal this species' phylogenetic relationships. A statistical parsimony network of 70 taxa aligned as 624 characters of the cytochrome c oxidase subunit I (COI) gene and 685 characters of the cytochrome c oxidase subunit II (COII) gene revealed three haplotypes (A1-A3) and four haplotypes (B1-B4), respectively. The concatenated sequences of both COI and COII genes with a total of 1309 characters revealed seven haplotypes (AB1-AB7). Analysis using tcs indicated that haplotype AB1 was the common ancestor and the most widespread haplotype in Malaysia. The genetic distance based on concatenated sequences of both COI and COII genes ranged from 0.00076 to 0.00229. Sequence alignment of Cx. quinquefasciatus from Malaysia and other countries revealed four haplotypes (AA1-AA4) by the COI gene and nine haplotypes (BB1-BB9) by the COII gene. Phylogenetic analyses demonstrated that Malaysian Cx. quinquefasciatus share the same genetic lineage as East African and Asian Cx. quinquefasciatus. This study has inferred the genetic lineages, dispersal patterns and hypothetical ancestral genotypes of Cx. quinquefasciatus.

First molecular genotyping of voltage gated sodium channel alleles in Culex quinquefasciatus populations in Malaysia.

A nationwide investigation was performed to detect the presence of 1014 mutation(s) in voltage gated sodium channel (kdr) gene of Culex quinquefasciatus from 14 residential areas across 13 states and a federal territory in Malaysia. Molecular genotyping of kdr mutation was performed via a modified three tubes allele-specific-polymerase chain reaction (AS-PCR) and direct sequencing of kdr gene. Based on the results of AS-PCR, homozygous susceptible (SS) genotype was found in nine out of 14 populations with 38 individuals from a total sample size of 140. Heterozygous (RS) genotype was most predominant (99 individuals) and distributed across all study sites. Homozygous resistance (RR) genotype was detected in Perak (one individual) and Selangor (two individuals). The resistance kdr allele frequencies ranged from 0.1 to 0.55, with the highest being detected in Cx. quinquefasciatus population from Selangor. This study has documented the first field-evolved instance of 1014F mutation in Malaysian mosquitoes and the findings of this study could be utilized in the implementation of strategic measures in vector control programs in Malaysia.

Subdural haematoma due to dural metastases from bronchogenic carcinoma in a previously well patient: an unusual cause of non-traumatic recurrent intracranial haematomata.

Subdural haematomata (SDH) are usually traumatic in aetiology. Non-traumatic instances of SDH are uncommon, and can rarely be due to metastases involving the dura. Computed tomography or magnetic resonance imaging can be misleading, as the underlying aetiology may be masked by the SDH, or the appearance can simulate meningiomas. A high index of suspicion for SDH is thus required. Under such circumstances, when no overt cause is identified, dural tissue should be sent for histological analysis and blood clot for cytology, even if the appearances are grossly normal at surgery. We present a rare case of a 42-year-old woman who was previously well, but presented with progressive weakness due to acute spontaneous SDH. She required repeated surgical evacuations for SDH and for subsequent recurrent extradural haematomata. After extensive investigations, the cause was identified to be secondary dural metastases from a primary lung carcinoma.

The anaesthetist's role in the setting up of an intraoperative MR imaging facility.

The clinical and economic advantages of intraoperative magnetic resonance (MR) imaging for neurosurgery are apparent. Hence, more medical institutions are setting up such services. Establishing such a service can be daunting because of the cost and specifications, its highly technical nature and many safety considerations. The anaesthetists have an integral part to play during this process because of their stake as key users and their vital role in conducting anaesthesia for complex neurosurgery within this hostile, and sometimes remote, environment. Moreover, their experiences with efficient workflows, patient screening and concern about safety make them eminently qualified members of the planning and building process. They are also no strangers to budgets and equipment appraisals. The complex interactions between conducting anaesthesia in a hostile environment and in a remote site conspire to make this a challenging undertaking. This article describes the role, the practical considerations and the difficulties experienced by the neurosurgical anaesthetists in setting up an intraoperative MR imaging operating theatre at Singapore General Hospital, from planning to equipment procurement and streamlining the workflow. Safety concerns and training are vital aspects of this article. It is hoped that our experiences will be of help to others who will be called upon at some point in a similar undertaking.

How severe acute respiratory syndrome (SARS) affected the department of anaesthesia at Singapore General Hospital.

The effects of the outbreak of severe acute respiratory syndrome (SARS) on the Singapore General Hospital's Department of Anaesthesia are described. Urgent measures to protect staff and patients were implemented. Clear direction in administration issues and clinical pathways with good logistic support were essential. Anaesthetists were at risk and anaesthetic practice had to change in view of strict infection control provisions. Contingency planning for future infectious diseases outbreaks is required. Lessons can be learnt by hospitals yet to be visited by this virulent infection.

Somatosensory evoked potential monitoring during carotid endarterectomy in patients with a stroke.

UNLABELLED: The aim of our study was to assess the characteristics and feasibility of somatosensory evoked potential (SSEP) monitoring in patients who have had a stroke undergoing carotid endarterectomy. We retrospectively reviewed the medical and SSEP records of 204 patients. The patients were divided into two groups: Stroke (n = 65) and No-Stroke (n = 139). The amplitude and latency of the N20-P25 cortical complex on the ipsilateral side (surgical) were compared with the contralateral side in each group and between groups. Stroke patients showed asymmetry of their cortical waveforms; the ipsilateral N20-P25 baseline amplitude was 1.5 +/- 1.0 microv versus 1.9 +/- 1.2 microv for the contralateral (P = 0.001), for No-Stroke patients 2.0 +/- 1.1 microv versus 2.1 +/- 1.1 microv (P = 0.2). Forty-eight percent of Stroke patients had a ratio (ipsilateral/contralateral amplitude) of <1.0 +/- 0.2 compared with 26% for No-Stroke patients (P = 0.01). There were no differences in latency measurements, in the incidences of significant SSEP changes (four Stroke, six No-Stroke) and immediate postoperative neurological deficits (two Stroke, six No-Stroke) between the two groups. Nine patients (three Stroke, six No-Stroke) had a decrease in ipsilateral N20-P25 amplitude >50% after cross-clamping, and had a shunt inserted. In conclusion, patients with a history of a stroke before surgery had a decrease in the amplitude of the ipsilateral cortical peak. There were no differences in the incidences of SSEP changes or neurological deficits. IMPLICATIONS: Patients who have had a preoperative stroke may show asymmetry of their cortical baseline somatosensory evoked potential waveforms; however, this does not interfere with the ability to use somatosensory evoked potential as a monitor during surgery.

Concurrent production of chitin from shrimp shells and fungi.

Crustacean shells constitute the traditional and current commercial source of chitin. Conversely, the control of fungal fermentation processes to produce quality chitin makes fungal mycelia an attractive alternative source. Therefore, the exploitation of both of these sources to produce chitin in a concurrent process should be advantageous and is reported here. Three proteolytic Aspergillus niger (strains 0576, 0307 and 0474) were selected from a screening for protease activity from among 34 zygomycete and deuteromycete strains. When fungi and shrimp shell powder were combined in a single reactor, the release of protease by the fungi facilitated the deproteinization of shrimp-shell powder and the release of hydrolyzed proteins. The hydrolyzed proteins in turn were utilized as a nitrogen source for fungal growth, leading to a lowering of the pH of the fermentation medium, thereby further enhancing the demineralization of the shrimp-shell powder. The shrimp-shell powders and fungal mycelia were separated after fermentation and extracted for chitin with 5% LiCl/DMAc solvent. Chitin isolates from the shells were found to have a protein content of less than 5%, while chitin isolates from the three fungal mycelia strains had protein content in the range of 10-15%. The relative molecular weights as estimated by GPC for all chitin samples were in the 10(5) dalton range. All samples displayed characteristic profiles for chitin in their FTIR and solid-state NMR spectra. All chitin samples evaluated with MTT and Neutral Red assays with three commercial cell lines did not display cytotoxic effects.