House flies were collected from April 2007-April 2008 from two poultry farms (Balik Pulau and Juru) in the state of Penang. The resistance level of the first generation offspring was evaluated against DDT, malathion, propoxur, and permethrin using the topical application method. The resistance ratio (RR) of the Balik Pulau strain house flies for propoxur, malathion and DDT ranged from 10.28 to 99.00, 7.83 to 47.01 and 6.05 to 31.10, respectively. Resistance to propoxur and malathion in house fly was attributed to cross resistance to organophosphate insecticides used in the farm. Increased metabolic detoxification might be the mechanism involved in DDT resistance due to excessive application of cypermethrin formulation. The RR of the Juru strain for propoxur, malathion and DDT was in a decreasing pattern throughout the study period, ranging from 5.58 to 83.38, 15.19 to 27.82, and 10.04 to 22.69, respectively. Permethrin appeared to be the most potent insecticide in controlling house fly in both the Balik Pulau (RR=0.50 to 1.96) and Juru poultry farms (RR=0.64 to 2.40). The fluctuations of insecticides resistance in house fly was also found to correlate with climatic factors due to its rapid breeding. Relative humidity exhibited positive correlation indices with the changes in the resistance level for DDT (r=0.481, p<0.05), malathion (r=0.698, p<0.01), and permethrin (r=0.580, p<0.05) in Balik Pulau. Similarly, relative humidity in Juru also showed strong correlation with the RR for DDT (r=0.900, p<0.01), malathion (r=0.762, p<0.05), permethrin (r=0.760, p<0.05), and propoxur (r=0.897, p<0.01).
Entamoeba gingivalis is present in the oral cavity of humans and is associated with periodontal disease. Consequently, this study aimed to comprehensively investigate the E. gingivalis infection and the associated risk factors among individuals suffering from periodontal conditions. A cross-sectional descriptive study was carried out within a cohort of periodontal patients. Dental plaque specimens were meticulously collected and subsequently subjected to thorough examination using the polymerase chain reaction (PCR)-based technique targeting the small subunit ribosomal RNA (SrRNA) gene of the organism. The occurrence of risk factors for E. gingivalis infection was analyzed by the chi-square test and binary logistic regression. Out of the 230 participants, 60 were clinically diagnosed with periodontitis, while 170 were afflicted with gingivitis. Out of the 230 patients, 25 (10.9%) tested positive for E. gingivalis infections. An in-depth analysis unveiled that a significant majority of infections were recorded within subgroups characterized by a marital status (15.45%), manifestation of periodontitis (25.00%), and concomitant presence of underlying disease (20.83%). Furthermore, the high risk factor associated with E. gingivalis infection was the female (ORadj = 13.65, 95% CI = 1.08-173.21), followed by periodontitis (ORadj = 3.30, 95% CI = 1.21-9.00), respectively. The study employs a molecular diagnostic approach to screen for E. gingivalis enrichment within a subset of periodontal patients with advancing disease. The findings emphasize the necessity for further research to elucidate the pathogenesis of E. gingivalis and advocate for vigilant surveillance within a substantial population of periodontal patients.
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.
Over the last decades, the epidemiology of human brucellosis globally has been subjected to significant changes, with the eradication of many existing endemic hot spots. This paper describes three cases with initial misdiagnosis of brucellosis that were managed during 2011-2017 in Republic of North Macedonia, country that until recently has been declared as endemic region. In spite of the fever, constitutional symptoms, focal disease (spondylitis, pneumonia and orchitis) and previous contact with domestic animals, brucellosis was not initially recognized, and patients were inadequately managed. Brucellosis should be part of differential diagnostic considerations in patients exposed to contacts with animals, with osteoarticular symptoms and signs, constitutional manifestations and different organ involvements in endemic regions where its incidence is diminishing.
This research was undertaken to evaluate the nematicidal activity of various concentrations of aqueous tinctures of 80 plant species towards L1-2 of S. papillosus. For the experiment with larvae of S. papillosus, there were used 0.19%, 0.75% and 3.00% aqueous tinctures of plants. Out of 80 tested species, nematicidal activity against L1-2 of S. papillosus was displayed by 20 plants. The greatest activity (LC50 = 0.060-0.069%) towards larvae of S. papillosus was exerted by Teucrium polium, Achillea millefolium, Genista tinctoria and Ulmus laevis. Less expressed nematicidal activity (LC50 = 0.070-0.079%) was recorded for Thalictrum minus, Stachys recta, Falcaria vulgaris, Lavatera thuringiaca. Even lower effect (LC50 = 0.080-0.089%) was shown by aqueous tinctures of Mentha × piperita, Achillea millefolium, Salvia nutans, Eryngium campestre and Cerasus fruticosa. The following plants could be arranged in declining order of effectiveness of nematicidal activity (LC50 = 0.090-0.165%) Malus sylvestris, Tragopogon orientalis, Erigeron annuus, Grindelia squarrosa, Urtica dioica, Daucus carota, Medicago sativa, Carduus acanthoides, Ulmus minor and Hieracium umbellatum. A far weaker effect on the nematodes was displayed by Bromopsis inermis and Tragopogon podolicus. Aqueous tinctures of 60 other studied species of plants exhibited low nematicidal activity in 3.00% aqueous tincture, while in 0.19% and 0.75% aqueous tinctures, no nematicidal activity was seen. The results of the research suggest that in the conditions of natural ecosystems, some species of plants of the Apiaceae, Asteraceae, Fabaceae, Lamiaceae, Malvaceae, Rosaceae, Ulmaceae and Urticaceae families could reduce vitality of free-living L1-2 larvae of S. papillosus.
Trichuris trichiura, the whipworm of humans, is one of the most prevalent soiltransmitted helminths (STH) reported worldwide. According to a recent study, out of 289 STH studies in Southeast Asia, only three studies used molecular methods. Hence, the genetic assemblages of Trichuris in Southeast Asia are poorly understood. In this study, we used partial mitochondrial DNA (cytochrome c oxidase subunit 1 or COI) sequences for analysis. Trichuris grouped in a same clade with different hosts indicate the potential of cross infection between hosts. Based on COI, the adult Trichuris isolated from a Malaysian patient was most closely related to Trichuris isolated from Papio anubis (olive baboons) from the USA. The Trichuris isolated from the dog from Malaysia was genetically similar to a Trichuris species isolated from Macaca silenus (lion-tailed macaque) from Czech Republic. Both the human and dog isolated Trichuris grouped in clades with different hosts indicating the potential of cross infection between hosts. Specific PCR primers based on the partial COI of T. trichiura isolated from African green monkey and T. serrata were designed and successfully amplified using multiplex PCR of the pooled DNA samples. Our results suggest a complex parasite-host relationship, and support the theory of cross infection of Trichuris between humans and non-human primates as suggested in previous publications.
Dipylidium caninum is a parasite that commonly infects dogs and cats worldwide. The large population of wild and stray dogs and cats may potentially transmit D. caninum to humans via their flea and lice. Humans are an accidental host, and dipylidiasis is more commonly seen in infants and children. There is scant information about human dipylidiasis in Russia. We report nine cases of dipylidiasis - eight in children and one in an adult. The patients were asymptomatic, except for excreting active proglottids in their faeces, which was the most common complaint. The clinical significance of asymptomatic dipylidiasis is not understood, except mothers were anxious because of the continuous appearance of active worms in the faeces of their children. The patients were successfully treated with praziquantel (15 mg/kg). Preventing dipylidiasis in pets and humans requires the control of fleas and lice, avoiding the outdoor defecation of definitive hosts, deworming pets, preventing children from playing with stray animals and spread of information about dipylidiasis among pet owners. Dogs and cats in many places in Russia breed freely, defecate outdoors in any area, and are not subjected to deworming and insect control. These circumstances favour the fact that, although this zoonosis is rare, it is a re-emerging disease and might reach important levels in Russia.
Chloroquine resistance transporter of Plasmodium falciparum (PfCRT) is a food vacuolar transmembrane protein that mediates susceptibility of the parasite to chloroquine. A mutation at K76T of the Pfcrt gene is a key determinant for chloroquine resistance phenotype. In the absence of drug pressure, in vitro growth rate of chloroquine-resistance parasites was outcompeted by wild-type parasites unless intragenic compensatory mutations occurred. Chloroquine-resistant P. falciparum bearing the Cam734 haplotype known to circulate in endemic areas of Cambodia bordering Thailand contains 9 mutations in Pfcrt and exhibits both chloroquine resistance and comparable growth rate to the chloroquine-sensitive 3D7 strain. To analyze the evolution of the Cam734 haplotype, codon-based analysis was performed by using the mixed effects model of evolution (MEME), branch-site random effects likelihood (BR-REL) and other related methods. Results revealed that the Cam734 haplotype has evolved distinctively from other known mutant haplotypes including the most common Dd2 haplotype in Southeast Asia. Evidence of episodic positive selection was detected at codon 144, characterized by c.[430G>T; 431C>T] (p.A144F), known to be indispensable for both chloroquine resistance and restoration of growth rate of the parasites. To survey the prevalence of mutations at codons 76 and 144 in Pfcrt among Thai isolates, restriction fragment analysis of 548 P. falciparum isolates collected from six endemic provinces of Thailand during 1991 and 2016 was performed. The 144F Pfcrt mutant was detected in 7 (1.28%) isolates. All Thai isolates analyzed herein harbored a mutation at codon 76 whilst the wild-type parasite was not found. The low prevalence of isolates bearing the mutation 144F in PfCRT could imply little or lack of survival advantage of this mutant in endemic areas of Thailand where the wild-type parasites seem to be absent or extremely rare.
Pro-and anti-inflammatory cytokines mediate the inflammatory response in sepsis. Therefore, regulation of cytokines with medications in risky situations may protect the patients from sepsis. Hydroxychloroquine and artemisinin are antimalarial drugs with immunomodulatory properties. In this study, we intended to investigate the effects of artemisinin and hydroxychloroquine on the cytokines released during sepsis in the rat model. Twenty-four rats were randomized into four groups. The control group received oral saline, the sepsis group received oral saline and intraperitoneal lipopolysaccharide toxin (LPS), the artemisinin-treated sepsis group received oral 33.33 mg/kg of artemisinin, and the hydroxychloroquinetreated sepsis group received oral 33.33 mg/kg of hydroxychloroquine before LPS injection. Three hours later, serum cytokines were measured. An increase was detected in TNF-a, IL-1, and IL-6 levels in the sepsis group compared to the control (p<0.01). Oral pretreatment with artemisinin resulted in significant downregulation only of IL-1 levels (p<0.01). Cytokines IL-1 and IL-6 were significantly downregulated in the serum of LPS-induced rats pretreated with oral hydroxychloroquine than rats with sepsis (p<0.01). Decreases observed in TNF-a and IL-10 levels were insignificant. These results demonstrated that both artemisinin and hydroxychloroquine attenuate the release of pro-inflammatory cytokines three hours after LPS-induced sepsis in rats. A significant decrease was observed in serum IL-1 and IL-6 levels with hydroxychloroquine and IL-1 levels with artemisinin. Based on our findings, we suggest that the therapeutic potential of artemisinin and hydroxychloroquine may be beneficial in preventing cytokine storm during sepsis, and further research is needed to determine the optimal timing of administration.
This study probes into the prospect of cross-reactivity of HCMV with RCMV which has not been acknowledged to date. We describe the uncovering of a protein with an estimated size of between 61-68 kDa from local RCMV strains which reacted with HCMV positive sera. Our findings are a first disclosure of a plausible immunological cross-reactivity between RCMV with its human counterpart which grounds substantial interest implying existence of conserved determinants between rat and human CMV polypeptides. The cross-reactive protein most likely represents an enveloped glycoprotein, though the precise identification and its degree of similarity needs to be evidently defined and further elucidated in forthcoming experiments.
Chagas disease is endemic to the Americas and is transmitted by blood-feeding kissing bugs. We evaluated the insecticidal potential of a fungus (Beauveria bassiana strain Pr-11) against Triatoma infestans, an important vector in South America. This fungal species was isolated from a locust (Schistocerca piceifrons) that inhabits the Central Andes region of Peru. Ten days post inoculation, this strain induced high insect mortality (97%) at low fungal concentrations (2 × 107 conidia/ml) at 70% relative humidity. The Pr-11 strain outperformed reference strain CCBLE-216 B. bassiana, provided by the Peruvian Ministry of Agriculture. Our results are consistent with previous reports on the virulence of this fungal strain against other insect pests. This is the first study to evaluate an orthopteran-isolated B. bassiana to control Chagas disease vectors. We conclude that strain Beauveria bassiana Pr-11 is effective against Triatoma infestans, resulting in a promising tool to control Chagas disease in Peru and may be used in integrated vector control programs.
Tuberculosis (TB) caused by Mycobacterium tuberculosis remains a serious public health threat with the World Health Organisation (WHO) reporting 5.8 million cases and 1.3 million deaths in the year 2020 due to TB. TB can be diagnosed by imaging, histopathological and bacteriological methods with culture remaining the gold standard. This study was performed to look at the sensitivity and specificity of post-mortem computed tomography (PMCT) imaging when compared to culture in diagnosing pulmonary tuberculosis. This was a retrospective comparative study looking at post mortem cases where lung tissue samples sent for TB culture at Hospital Kuala Lumpur were compared against PMCT imaging. Exclusion criteria included contaminated samples, decomposed cases, immunocompromised subjects and those below 18 years of age. Subjects included 80 medico-legal autopsy cases at the National Institute of Forensic Medicine, Hospital Kuala Lumpur, Malaysia who had whole body PMCT done in accordance with the Institute's protocol and tissue samples sent for bacteriology culture for tuberculosis. PMCT findings were positively associated with acid-fast organisms in 23.5 out of 33 cases (71.2%). Our study also showed that PMCT had a sensitivity of 71.3% and specificity of 54.3% (95% CI: 39.5-68.4) in diagnosing TB based on the protocol set in this study. This study showed that there was relatively good agreement between radiological PMCT findings and bacterial culture, suggesting that radiological examination is a relatively reliable tool for preliminary screening and possible diagnosis of TB prior to a postmortem examination which would be beneficial in reducing the risk of transmission of TB to health workers during autopsy.
Routine surveillance on resistant status of field mosquito populations is important to implement suitable strategies in order to prevent pest outbreaks. WHO test kit bioassay is the most frequent bioassay used to investigate the susceptibility status of field-collected mosquitoes, as it is relatively convenient to be carried out in the field. In contrast, the topical application of active ingredient is less popular in investigating the susceptibility status of mosquitoes. In this study, we accessed the susceptibility status of Aedes albopictus Skuse collected from two dengue hotspots on Penang Island: Sungai Dua and Persiaran Mayang Pasir. Two active ingredients: permethrin and deltamethrin, were used. WHO test kit bioassay showed that both wild strains collected were susceptible to the two active ingredients; while topical application assay showed that they were resistant. This indicated that WHO test kit bioassay less sensitive to low level of resistance compared to topical application assay. Hence, topical application is expected to be more indicative when used in a resistance surveillance programme.
Three genomic regions, VP4 capsid, VP1 capsid and 3D RNA polymerase of human enterovirus 71 (EV-71) and coxsackievirus A16 (CV-A16) were sequenced to understand the evolution of these viruses in Malaysia. A total of 42 EV-71 and 36 CV-A16 isolates from 1997- 2008 were sequenced. Despite the presence of many EV-71 subgenotypes worldwide, only subgenotypes B3, B4, B5, C1 and C2 were present in Malaysia. Importation of other subgenotypes such as C3, C4/D and C5 from other countries was infrequent. For CV-A16, the earlier subgenotype B1 was replaced by subgenotypes B2a and the recent B2c. Subgenotype B2a was present throughout the study while B2c only emerged in 2005. No genetic signatures could be attributed to viral virulence suggesting that host factors have a major role in determining the outcome of infection. Only three EV-71 B3 isolates showed non-consistent phylogeny in the 3D RNA polymerase region which indicated occurrence of recombination in EV-71. High genetic diversity was observed in the Malaysian EV-71 but Malaysian CV-A16 showed low genetic diversity in the three genomic regions sequenced. EV-71 showed strong purifying selection, but that occurred to a lesser extent in CV-A16.
This paper reports the occurrence of helminth and protozoan infections on small ruminants from eight farms situated in Kinta and Perak Tengah district, Perak. The results of this survey indicate that helminthiasis and coccidiosis is rampant in sheep and goat farms. Several anthelmintics have been used for the control of helminths. The smallholders depended on health and extention services from the State Veterinary Department. This survey is part of an ongoing programme by the Department of Veterinary Services to upgrade services and report the current status of parasitic diseases in the state.
Antibodies to the protozoan parasite, Toxoplasma gondii were assayed in sera of 200 goats, 100 pigs, 126 cattle from various states of Malaysia, and 135 dogs and 55 cats around Ipoh region using an indirect fluorescence antibody test (IFAT, cut-off titer 1:200); antibodies were found in 35.5% of goats, 14.5% cats, 9.6% dogs, 7.9% local cattle and 4% yellow cattle but not in pigs. Results indicate that infection is most prevalent in goats.
This study was conducted to evaluate the anthelmintic effect of neem (Azadirachta indica) on nematode parasites of sheep. Twelve Santa Ines cross bred sheep from a government farm were randomly selected and equally divided into control (n = 6) and treated groups (n =6). Faecal egg counts (FEC) using the modified McMaster technique and the FAMACHA score for assessing clinical anaemia were carried out daily and recorded for 6 weeks. At the end of the study all the animals were slaughtered and the total worm count (TWC) was done. The results of FEC showed that there was no significant difference between the control and treated group (p = 0.081). However, worm burden estimations showed that the number of parasites was significantly higher in the control group compared to the treated group (p < 0.05). This result indicated that feeding Neem had an effect on worm numbers in sheep, but was not reflected in their faecal egg counts. Further work is needed to reconfirm the effect of Neem on helminth infections of sheep.
Emerging cases of Fasciola and Paramphistomes co-infection have been reported, especially in tropical regions. Thisis due to Fasciola and Paramphistomes sharing biological factors which influence the pattern of transmission, especially in faecal egg shedding due to interaction and competition in the definitive host. Most reports surveyed the occurrence of fasciolosis in ruminants with a lack of observation of faecal egg distribution. Therefore, present study is aimed to assess the distribution of Fasciola and Paramphistomes faecal egg count (fec) in co-infected large ruminants in Larut, Matang, and Selama areas (Taiping). A total of 371 faecal samples were collected at random from 23 ruminant herds. Flukefinder® sedimentation was used to quantify the Fasciola and Paramphistomes eggs. Descriptive analyses were performed to determine the prevalence of co-infections, and Spearman correlation analysis was used to correlate the fec. Overall, the prevalence of Fasciola and Paramphistomes co-infection was 23.7% (n=89/371) in Taiping. Prevalence of paramphistomosis was always higher than fasciolosis in overall and single infection, with 46.9% (n=174/371) and 22.9% (n=85/371) compared to 36.9% (n=137/371) and 12.9% (n=48/371) respectively. Egg per gram (epg) of both parasites were positively skewed with a median of 1.5 epg in fasciolosis and 10.5 epg in paramphistomosis. Spearman correlation analysis of the epg in co-infected bovine was found to have a moderately positive correlation with rs=0.39 (p-value<0.01). The recent study observed a moderate prevalence of Fasciola and Paramphistomes coinfection in a large ruminant population from Taiping, with the prevalence of paramphistomosis being higher than fasciolosis. Hence, this suggests that infection with one of these parasites increases the chance of infection with another. There is a need to integrate fec in parasite surveillance to monitor the trend of parasite transmission. Findings in the present study could tailor control strategies, especially for fasciolosis to limit the economic loss and prevent zoonotic transmission.
The objective of this study was to elucidate the association of various risk factors with dengue cases reported in Lundu district, Sarawak, by analyzing the interaction between environmental, entomological, socio-demographic factors. Besides conventional entomological, serological and house surveys, this study also used GIS technology to generate geographic and environmental data on Aedes albopictus and dengue transmission. Seven villages were chosen based on the high number of dengue cases reported. A total of 551 households were surveyed. An overall description of the socio-demographic background and basic facilities was presented together with entomological and geographical profiles. For serological and ovitrap studies, systematic random sampling was used. Serological tests indicated that 23.7% of the 215 samples had a history of dengue, either recent or previous infections. Two samples (0.9%) were confirmed by IgM ELISA and 49 samples (22.8%) had IgG responses. A total of 32,838 Aedes albopictus eggs were collected in 56 days of trapping. Cluster sampling was also done to determine whether any of the risk factors (entomological or geographical) were influenced by geographical location. These clusters were defined as border villages with East Kalimantan and roadside villages along Lundu/Biawas trunk road. The data collected were analyzed using SPSS version 10.01. Descriptive analysis using frequency, means, and median were used. To determine the association between variables and dengue cases reported, and to describe the differences between the two clusters of villages, two-sample t-test, and Pearson's Chi-Square were used. Accurate maps were produced with overlay and density function, which facilitates the map visualization and report generating phases. This study also highlights the use of differential Global Positioning System in mapping sites of 1m accuracy. Analysis of the data revealed there are significant differences in clusters of villages attributable to container density, house density, distance of the house from the main road, and number of Ae. albopictus eggs from ovitraps set indoor, outdoor and in dumping sites (Person's Chi-Square = 6.111, df = 1, p < 0.01). Further analysis using t-test showed that house density, container density, indoor mosquitoes egg count, outdoor mosquitoes egg count, and dumping sites mosquitoes egg count were higher at the roadside villages compared to border villages. A number of potential risk factors including those generated from GIS were investigated. None of the factors investigated in this study were associated with the dengue cases reported.
Malaysia first reported H5N1 poultry case in 2004 and subsequently outbreak in poultry population in 2007. Here, a recombinant gene encoding of peptide epitopes, consisting fragments of HA1, HA2 and a polybasic cleavage site of H5N1 strain Malaysia, was amplified and cloned into pET-47b(+) bacterial expression vector. DNA sequencing and alignment analysis confirmed that the gene had no alteration and in-frame to the vector. Then, His-tagged truncated HA protein was expressed in Escherichia coli BL21 (DE3) under 1 mM IPTG induction. The protein expression was optimized under a time-course induction study and further purified using Ni-NTA agarose under reducing condition. Migration size of protein was detected at 15 kDa by Western blot using anti-His tag monoclonal antibody and demonstrated no discrepancy compared to its calculated molecular weight.