Tritrichomonas foetus is known to cause chronic diarrhea in the feline species in many different regions of the world. However, there is a paucity of information on T. foetus among cats in Malaysia. This study was conducted to determine the prevalence of Tritrichomonas foetus in the pet and stray cat population in Klang Valley, Malaysia. A total of 201 pet and stray cats' fecal samples were collected in Klang Valley. 24 samples were cultured in the InPouch® TF Feline to observe for motile trophozoites. A nested PCR protocol was used to screen for T. foetus in the collected samples. The prevalence of T. foetus in the cat population in Klang Valley was 33%. There was no association between Tritrichomonas infection and age, sex, breed or management of the cats. However, statistical analysis revealed that stray cats were more likely to be infected with T. foetus compared to pet cats. This study confirmed for the first time the presence of T. foetus among the cat population in Klang Valley, Malaysia.
The study was aimed to investigate the expression of cytosolic and thiolated proteins of Musca domestica larvae under oxidative stress. Proteins from acute treatment of hydrogen peroxide (LC50 = 21.52% (v/v)) on 3rd stage larvae of housefly were extracted and purified using an activated Thiol Sepharose® for thiolated protein purification. Two dimensional gel electrophoresis was used for visualizing and analyzing expression of cytosolic and thiolated proteins. Protein spots with more than 5 fold of expression change were identified using liquid chromatography- tandem mass spectrometry (LC-MS/MS). The cytosolic proteins were actin, tropomyosin, ubiquitin, arginine kinase, pheromone binding protein/general odorant binding protein, and ATP: guanidino phosphotransferase. The thiolated proteins with more than 5 fold change in expression as an effect to the acute treatment were fructose bisphosphate aldolase, short chain dehydrogenase and lactate/malate dehydrogenase. The proteins identified in the study should provide vital information for future reference in oxidative stress defence and response occurring in houseflies.
Helminth infections (HIs) are an important public health problem in tropical countries, and the associated problems have been neglected in rural areas of Thailand. Therefore, this study reports the prevalence and intensity of HIs among inhabitants of the Khon Sawan district, Chaiyaphum province, and Kaeng Samnam Nang district, Nakhon Ratchasima province, which are located near the Chi River and Lahanna water reservoir, northeastern Thailand. A cross-sectional descriptive study was conducted between July 31, 2018, and June 30, 2019, among rural villagers from 40 rural villages in 4 subdistricts. The participants were selected from the village enrolment list after proportional allocation of the total sample size. Faecal samples from 691 inhabitants were prepared using solvent-free faecal parasite concentrator, and helminths were then detected using a light microscope. Statistical analysis included the Chi-square test with Yates correction, and multivariable logistic regression was performed. A P-value of <0.05 was considered statistically significant. The prevalence of HIs was 2.03%. The most prevalent helminths were Opisthorchis viverrini (1.31%), followed by Strongyloides stercoralis (0.44%), Ascaris lumbricoides (0.29%), hookworm (0.15%), Teania spp. (0.15%) and one minute intestinal fluke (0.15%). Coinfections were identified in 2 cases for S. stercoralis and hookworm and 1 case for O. viverrini and S. stercoralis infection. All infected participants had a light intensity of HI. There was no significant difference between general characteristics for all HIs. The prevalence of HIs was not significantly associated with general characteristics. This study indicates that the infections result mainly from foodborne helminths and skin-penetrating nematodes. Therefore, interventions should concentrate on the personal hygiene of the population and improving sanitation to reduce HIs in this area.
Chikungunya virus infection had not been reported by the National Secretary of Health of the State of Baja California Sur, Mexico before 2015, and until now, no information of the disease has been published. Thus, the objective of this study was to assess the tendency that the disease has shown since its introduction. The total number of cases in the state was analyzed, and the incidence rate of infection was evaluated in the general population, age groups, and gender. From 2015-2019, the year with the highest number of reported cases was 2016 with 210 cases, but no cases were reported from 2018-2019. The gender with the highest number of cases was female. The most affected age group was adults 45-64 years of age.
Novel coronavirus disease, the latest world pandemic is one of the most contagious viral infections to date. There has been a lack of uniformity on recognizing this condition clinically because of poorly understood pathophysiology and clinical nature. Also due to ongoing clinical trials, its management is also varied. This is a systematic review from evidence-based studies until March 1st, 2020, covering an update on its clinical features and management. This study shows the multisystem involvement of COVID-19 with dominant respiratory features followed by the musculoskeletal, gastrointestinal system and others. The clinical features varied from asymptomatic to severe forms. Major causes of fatality were acute respiratory distress syndrome, shock, acute cardiac injury, acute kidney injury, rhabdomyolysis, and arrhythmia. Major modalities of management included supportive, antiviral and antibiotic therapy. There was no direct relationship between the specific treatment and the outcome.
To characterize the production and larvicidal activity of Xenorhabdus stockiae KUT6 Petroleum ether extracts from Luria Broth and induced Quorum sensing medium containing N-3- oxododecanoyl Homoserine Lactone inducer against dengue vector Aedes aegypti. The Galleria mellonella larvae were reared for the isolation of Steinernema saimkayi symbiont Xenorhabdus stockiae KUT6 from Cucumber field soil sample in NBTA. Then for the extraction of compounds the KUT6 strains were cultured in Luria Broth and Quorum Sensing optimized media using N-3-oxododecanoyl homoserine lactone inducer. The larvicidal activity of Xenorhabdus stockiae KUT6 of petroleum ether extracts were bioassayed against 4th instar Aedes aegypti dengue vector. The maximum rate of mortality were recorded of the samples A-24h, B-48h, C-72h, A1-24h, B1-48h, C1-72h at different concentrations 50 µg/ml, 100 µg/ml and 150 µg/ml respectively for 24h to 72h of exposure treatment. The morphological characteristics of Xenorhabdus stockiae KUT6 in NBTA were red core colonies with blue background surrounded by zone of inhibition. After 24h exposure maximum rate of 100% mortality of Aedes aegypti 4th instar larvae was attained when treated with sample C1-72h 50 µg/ml of the petroleum ether extracts of quorum sensed medium whereas the sample C 72h petroleum ether extracts of KUT6 cultured in Luria broth recorded 100% mortality at 150 µg on 24h exposure indicates enhancement in the product yield. The study assures the use of Xenorhabdus stockiae KUT6 petroleum ether extracts as biocontrol agent could be beneficial for the control of dengue vectors.
Reduced susceptibility in Burkholderia pseudomallei during carbapenem therapy may lead to treatment failure. We isolated a clinical strain that had developed reduced susceptibility to carbapenems while on treatment. After reviewing the patient's clinical notes, the initial isolate (BUPS01/14) was exposed to carbapenem in vitro to mimic the clinical scenario. The stability of susceptibility of the carbapenem-exposed strain (BUPS01/14R) was examined by serial subculture in antibiotic-free broth. Biochemical and morphological comparison was performed by the VITEK® system and electron microscopy. MICs increased 32-fold following carbapenem exposure and became stable in the antibiotic-free environment. On electron microscopic examination, the BUPS01/14R cells were smoother and less wrinkled compared to BUPS01/14 cells. This report highlights a potential anti-melioidosis treatment failure due to the emergence of resistance while on carbapenem monotherapy. Further study of this strain is necessary to understand the mechanism of resistance at a molecular level.
Canine demodicosis is a common skin disorder with multiple risk factors, including age and breed predisposition. There is relatively limited information about the risk factors for canine demodicosis in large canine populations. This retrospective case-control study was conducted by searching the electronic records of dogs with skin lesions for the presence of Demodex mites in skin scrapings. Diagnosis of demodicosis was based on the presence of skin lesions and mites in skin scrapings. Multivariate analysis was conducted using logistic regression analysis to estimate the relative risk and odds ratio of variables hypothesized to influence the risk of canine demodicosis, such as age, sex, breed, season, and parasitic infection. The results of multivariate logistic regression analysis showed a positive correlation between the dogs' age and demodicosis. Dogs older than three years, as well as puppies, had a high risk of demodicosis (P0.05). Breeds with the greatest association (odds ratio) with demodicosis included the American Staffordshire Terrier (OR=0.9) and Moscow Watchdog (OR=0.2). The presence of intestinal parasites, such as Diphyllobothrium latum, was significantly associated with demodicosis.
Malaria is one of the most dangerous infectious diseases due to its high infection and mortality rates, especially in the tropical belt. Plasmodium falciparum (P. falciparum), the most virulent malaria parasite in humans, was recently reported to develop resistance against the final efficient antimalarial drug, artemisinin. Little is known about the resistance mechanisms, which further complicates the problem as a proper counteraction is unable to be taken. Hence, the understanding of drug mode of action and its molecular target is valuable knowledge that needs to be considered to develop the next generation of antimalarial drugs. P. falciparum protein kinase (Pf PK) is an attractive target for antimalarial chemotherapy due to its vital roles in all P. falciparum life stages. Moreover, overall structural differences and the presence of unique Pf PKs that are absent in human kinome, suggesting specific inhibition of Pf PK without affecting human cells is achievable. To date, at least 86 eukaryotic protein kinases have been identified in P. falciparum kinome, by which less than 40 were validated as potential targets at the erythrocytes stage. In this review, recent progress of the furthest validated Pf PKs; Pf Nek-1, Pf CDPK1, Pf CDPK4, Pf PKG, and Pf CLK-3 will be briefly discussed.
The bioactivity of R. nasutus leaf extracts was assessed on Bacillus cereus, Bacillus subtilis, Staphylococcus aureus, Streptococcus pyogenes, Vibrio parahaemolyticus, Enterobacter aerogenes, Proteus mirabilis, and Klebsiella pneumoniae. Crude chloroform, petroleum ether, ethyl acetate, ethanol and methanol extracts were screened by disc diffusion method. Promising crude extract was further subjected to the column fractionation followed by the screening of the antibacterial activity of individual fractions. Biologically active pure fraction was subjected to the advanced analytical studies like HPLC, LC-MS, IR and NMR for characterisation of the bioactive compound. Ethanolic extract exhibited the maximum antibacterial activity against Klebsiella pneumoniae with the maximum of 35±0.42 mm zone of inhibition. The biologically potent column fraction from ethanol extract with 40±0.42 mm zone of inhibition upon subject to the HPLC, LC-MS, IR and NMR revealed that the active compound is rhinacanthin-C, a naphthoquinone.
Ticks are vectors of bacteria, protozoa and viruses capable of causing serious and life threatening diseases in humans and animals. Disease transmission occurs through the transfer of pathogen from tick bites to susceptible humans or animals. Most commonly known tick-borne pathogens are obligate intracellular microorganisms but little is known on the prevalence of culturable pathogenic bacteria from ticks capable of growth on artificial nutrient media. One hundred and forty seven ticks originating from dairy cattle, goats and rodents were collected from nine selected sites in Peninsular Malaysia. The culture of surfacesterilized tick homogenates revealed the isolation of various pathogenic bacteria including, Staphylococcus sp., Corynebacterium sp., Rothia sp., Enterococcus faecalis, Klebsiella pneumoniae, Escherichia coli and Bacillus sp. and its derived genera. These pathogens are among those that affect humans and animals. Findings from this study suggest that in addition to the regular intracellular pathogens, ticks could also harbor extracellular pathogenic bacteria. Further studies, hence, would be needed to determine if these extracellular pathogens could contribute to human or animal infection.
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.
Blastocystis is one of the most common parasites inhabiting the intestinal tract of human and animals. Currently, human Blastocystis isolates are classified into nine subtypes (STs) based on the phylogeny of their small subunit ribosomal RNA (SSU rRNA) gene. Although its pathogenicity remains controversial, the possibility of zoonotic transmission was recognized since eight of the nine STs (except for ST9) have been reported in both humans and animals. A cross-sectional study was conducted to determine the prevalence and subtype distribution of Blastocystis isolated from humans and associated animals in an indigenous community with poor hygiene in Malaysia, where the risk of parasitic infection is high. A total of 275 stool samples were collected, subjected to DNA extraction and amplified by PCR assay. The Blastocystis-positive amplicons were then purified and sequenced. Phylogenetic tree of positive isolates, reference strains and outgroup were constructed using maximum likelihood method based on Hasegawa-KishinoYano+G+I model. The prevalence of Blastocystis infection among humans and domestic animals by PCR assay were 18.5% (45/243) and 6.3% (2/32), respectively. Through molecular phylogeny, 47 isolates were separated into five clusters containing isolates from both hosts. Among human isolates, ST3 (53.3%) was the predominant subtype, followed by ST1 (31.1%) and ST2 (15.6%). Chicken and cattle had lower proportions of ST6 (50%) and ST10 (50%), that were barely seen in humans. The distinct distributions of the most important STs among the host animals as well as humans examined demonstrate that there is various host-specific subtypes in the lifecycle of Blastocystis.
Use of mosquito coils for personal protection against malaria and mosquito nuisance is advocated under mosquito and malaria control programmes. We performed field studies of mosquito coils containing either metofluthrin or esbiothrin in experimental huts situated in Kamhororo village, Gokwe district, Zimbabwe. All tests were performed on 3-5 day old reared female Anopheles gambiae sensu lato mosquitoes. The burning times were 9hr 20min for mosquito coils containing metofluthrin and 8 hr for those containing esbiothrin and the results were significantly different (p = <0.001). The mean knock down rate for mosquito coils containing metofluthrin was 90% and that for esbiothrin was 73.3% and the results were significantly different (p = 0.00). Mosquito coils containing metofluthrin had a mean repellence of 92.7% as compared to 85.4% for esbiothrin and the results were not significantly different (p=0.27). The protection time as required by EPA (1999) was 6 hr for mosquito coils containing metofluthrin and 5 hr for those containing esbiothrin. The mean insecticidal effect of mosquito coils containing metofluthrin was 84% as compared to 83% for those containing esbiothrin and the results were not significantly different (p = 0.56). Both mosquito formulations could not be classified as having insecticidal effect since none of them met the 95% mortality rate criteria.
Japanese encephalitis virus (JEV), a member of the family Flaviviridae, causes severe neurological disorders in humans. JEV infections represent one of the most widely spread mosquito-borne diseases, and therefore, it has been considered as an endemic disease. An effective antiviral drug is still unavailable to treat JEV, and current drugs only provide supportive treatment to alleviate the symptoms and stabilize patients' conditions. This study was designed to evaluate the antiviral activity of the sulphated polysaccharides "Carrageenan," a linear sulphated polysaccharide that is extracted from red edible seaweeds against JEV replication in vitro. Viral inactivation, attachment, and post-infection assays were used to determine the mode of inhibition of Carrageenan. Virus titters after each application were evaluated by plaque formation assay. MTT assay was used to determine the 50% cytotoxic concentration (CC50), and ELISA-like cell-based assay and immunostaining and immunostaining techniques were used to evaluate the 50% effective concentration (EC50). This study showed that Carrageenan inhibited JEV at an EC50 of 15 µg/mL in a dose-dependent manner with CC50 more than 200 µg/mL in healthy human liver cells (WRL68). The mode of inhibition assay showed that the antiviral effects of Carrageenan are mainly due to their ability to inhibit the early stages of virus infection such as the viral attachment and the cellular entry stages. Our investigation showed that Carrageenan could be considered as a potent antiviral agent to JEV infection. Further experimental and clinical studies are needed to investigate the potential applications of Carrageenan for clinical intervention against JEV infection.
Haemonchus spp. or barber's pole worms are one of the most highly pathogenic nematodes of ruminants causing economic losses in livestock worldwide. The current study was a first attempt to identify Haemonchus spp. from goats in Thailand and Lao PDR. Utilizing the inexpensive tools of the discriminant function (DF) combined with synlophe patterns is fundamental for understanding their epidemiological aspects. In total, 255 randomly chosen adult male Haemonchus worms from goats in various areas in each country were identified individually. For both these countries, about 94% based on the DF values, and 99%, 98%, and 97% based on synlophe patterns in the region of the esophageal intestinal junction (EI), 4 mm from the anterior end, and at both these positions, respectively, were identified as H. contortus. Other identified specimens defined as H. placei and hybrids as well as unclassified species based on synlophe patterns were proved using polymerase chain reaction (PCR); this also included some randomly chosen H. contortus by DF and synlophe patterns. All those specimens were confirmed as H. contortus being strongly supported by some genetic evidences and UPGMA analysis. Thus, it was assumed that all specimens in the current study were H. contortus. The morphological differences of this predominant species (H. contortus) in goats between the two countries were: body length, gubernaculum length, and left spicule barb length, while almost all characters of male worms individually measured appeared to overlap, mostly in H. contortus and H. placei, which may lead to misclassification. Therefore, using the DF along with synlophe patterns can assist in increasing the accuracy of Haemonchus spp. identification from goats in some areas where funding is limited, particularly in Lao PDR. The present results revealed that synlophe patterns in the EI region seemed to be promising for the identification of Haemonchus spp., while molecular techniques are also required to address ambiguous identification with some specimens.
Simulium (Simulium) contractum Takaoka from Sulawesi, Indonesia was known only as the pupa. Its female, male and mature larva are described for the first time. The tentative assignment of this species in the Simulium dumogaense species-group is confirmed by the adult characters including the female and male genitalia. The female and male of this species are similar to those of Simulium (Simulium) tumpaense Takaoka and Roberts but are distinguished by the yellowish femora.
Corona virus (COVID-19) infection has been growing as a biggest threat to human society. World Health Organization (WHO) has already declared it as a pandemic for the whole world, with nearly six million positive cases. Highly contagious nature of the virus has challenged the medical facilities of all the developed and developing country health system. Early identification of the infection is very important to provide medical facilities and cease the chain of infection to new persons. The symptoms such as fever, dry cough, breathing issues generally show in patients not before 5-7 days. However, the patients feel the loss of smell or taste (anosmia) as early as second day onwards due to the presence of virus in nose and throat. Low-cost techniques such as SniffIn-sticks ® Smell Test and UPSIT etc. can be used to test anosmia along with medically approved olfactory test leading to identification of COVID-19 infections. With leading researchers findings anosmia test will be effective in breaking the chain infection of COVID-19 virus. In the exit ports, anosmia test kits may be added to thermal testing to identify the infected patients with low symptoms. Additionally, home test kits may be developed at low cost and supplied for large scale testing of the infection.
Third-generation cephalosporin resistant Enterobacteriaceae (TCRE) is a global concern especially in neonatal sepsis. We performed a secondary data analysis in a Thai neonatal intensive care unit to identify the risk factors for acquisition of TCRE sepsis and mortality of Enterobacteriaceae sepsis between 1991 and 2017. Multivariate logistic and Cox proportional regression were used for analysis. Numbers of neonates with TCRE and nonTCRE sepsis were 100 and 41 patients, respectively. Medians (interquartile ranges) of gestational age, birthweight, onset of sepsis and total hospital stay of neonates with Enterobacteriaceae sepsis were 32 (28, 38) weeks, 1670 (1025, 2750) grams, 11 (6, 25) days and 41 (22, 74) days, respectively. Univariate and multivariate analysis, neonates with TCRE sepsis were more likely to have birth asphyxia (adjusted odds ratio [aOR] = 2.6; 95% confidence interval [CI] 1.1-6.0; p = 0.02) and history of aminoglycoside exposure (aOR = 2.9; 95% CI 1.3- 6.7; p = 0.01). In-hospital case fatality rate from Enterobacteriaceae sepsis was 26% (36/141). In Cox regression, neonates with TCRE sepsis was not an independent risk of non-survivors, but septic shock (adjusted hazard ratio = 9.9; 95% CI 5.0-19.7, p < 0.001) increased 30-day mortality in the final model. Asphyxia and previous aminoglycoside consumption were risks of acquisition for neonatal TCRE sepsis while the burden was not a significant difference. Infection prevention and control must be strictly implemented in high multidrug-resistant area.
Pseudomonas aeruginosa is considered an opportunistic pathogen, causing a wide variety of infections in compromised hosts, also frequently develops multi-resistance to antibiotics and can colonize various habitats, including water systems. The main aim of this study was to investigate antibiotics susceptibility pattern, genotypic diversity and detection of resistence genes in P. aeruginosa isolates from clinical and aquatic environment sources. Of the 220 P. aeruginosa isolates examined, 48 were clinical isolates and 172 isolates from wastewater and freshwater. Susceptibility to eight antimicrobial agents was carried out by disk diffusion method. Clinical and environmental isolates were screened for the presence of the genes encoding blaKPC-2, blaCTX-M-9, blaPER-1, blaOXA-10, blaIMP-1, blaVIM-2 and blaampC by polymerase chain reaction (PCR). Isolates were examined with PCR-SSCP analysis of partial DNAr 16S sequence. Isolates were mainly resistant to cefoxitin. Multidrug-resistant P. aeruginosa (MDRPA) strains were found in 70% and 90.3% of the clinical and environmental isolates, respectively. The prevalence rates of â-lactamase genes were recorded (blaKPC-2 41.3%, blaVIM-2 36.8%, blaIMP-1 13.6%, blaCTX-M-9 10.9% and blaampC 10.5%,). The PCR-SSCP analysis showed three conformational patterns. All clinical isolates and most environmental isolates were grouped into a single cluster. In this study, we found that P. aeruginosa strains recovered from city water systems must be considered potential reservoir for ESBL genes, especially blaKPC-2 and blaVIM-2.