Prevalence, potential risk factors and genetic diversity of Blastocystis in ruminant livestock animals from Penang, Malaysia.

Blastocystis is a unicellular, anaerobic protist inhabiting the intestinal tract of diverse animal hosts, including human. Information regarding Blastocystis in small ruminants, namely goats and sheep, is limited globally; thus, this study was carried out to investigate the distribution and determinants of Blastocystis in ruminant livestock animals from Penang, Malaysia. Fecal samples from 127 cattle, 149 goats, and 100 sheep were examined for Blastocystis by in vitro cultivation using modified Jones' medium, while DNA barcoding was used for subtyping. Overall, 23.1% (87/376) of animals screened were positive for Blastocystis sp. The prevalence of infection was significantly higher in goats than in cattle and sheep, while the female gender, semi-intensive farming system, and the Northeast Penang Island district were identified as potential risk factors for Blastocystis infection. Blastocystis sp. ST5, ST14, and ST25 were identified in cattle; ST5, ST10, ST13, and ST14 in goats; and ST4, ST5, ST14, and ST15 in sheep. ST5 and ST14 were found to be the most abundant and widespread subtypes in the study area. To the best of our knowledge, this is the first report of ST4 from sheep and ST13 from goats, thus serving as an update to the host range of Blastocystis sp. ST4 and ST13. The isolation of ST4 and ST5 in this study suggests that ruminant livestock animals could serve as reservoirs of human infection.

Performance Evaluation of Nested Polymerase Chain Reaction (Nested PCR), Light Microscopy, and Plasmodium falciparum Histidine-Rich Protein 2 Rapid Diagnostic Test (PfHRP2 RDT) in the Detection of Falciparum Malaria in a High-Transmission Setting in Southwestern Nigeria.

Malaria remains a major public health challenge worldwide. In order to ensure a prompt and accurate malaria diagnosis, the World Health Organization recommended the confirmatory parasitological diagnosis of malaria by microscopy and malaria rapid diagnostic test (RDT) prior to antimalarial administration and treatment. This study was designed to evaluate the performance of nested polymerase chain reaction (nested PCR), light microscopy, and Plasmodium falciparum histidine-rich protein 2 rapid diagnostic test (PfHRP2 RDT) in the detection of falciparum malaria in Akure, Nigeria. A cross-sectional and hospital-based study involving 601 febrile volunteer participants was conducted in Akure, Nigeria. Approximately 2-3 mL venous blood samples were obtained from each study participant for parasitological confirmation by microscopy and PfHRP2-based malaria RDT. Thick and thin films were prepared and viewed under the light microscope for parasite detection, parasite density quantification, and species identification, respectively. Dry blood spot samples were prepared on 3MM Whatman filter paper for nested PCR. The overall prevalence of microscopy, PfHRP2 RDT, and nested PCR were 64.89% (390/601), 65.7% (395/601), and 67.39% (405/601), respectively. The estimates of sensitivity, specificity, positive predictive value, negative predictive value, accuracy, and Youden's j index of microscopy and RDT were 96.30, 100.00, 100.00, 92.89, 97.50, 0.963, and 95.06, 94.90, 97.47, 90.29, 95.01, and 0.899, respectively. Malaria RDT recorded higher false negativity, compared microscopy (4.94% vs. 3.70%). A near perfect agreement was reported between microscopy and nested PCR, and between PfHRP2 RDT and nested PCR with Cohen's kappa (k) values of 0.94 and 0.88, respectively. This study revealed that PfHRP2 RDT and microscopy continues to remain sensitive and specific for falciparum malaria diagnosis in the study area.

First Molecular Identification of Caligus clemensi on Cultured Crimson Snapper Lutjanus erythropterus on Jerejak Island, Penang, Peninsular Malaysia.

Fish parasites such as Caligus clemensi are a serious concern for cultured fish in many regions of the world, including Malaysia. This study was designed to elucidate the parasites' prevalence and intensity coupled with the morphology and molecular identification of C. clemensi on cultured Lutjanus erythropterus in Jerejak Island, Penang, Peninsular Malaysia. The study was carried out on 200 fish specimens of cultured L. erythropterus obtained from the GST group aquaculture farm. Parasites were collected from the infested part of L. erythropterus fish, and their prevalence and intensity were determined. The parasites were identified morphologically using a field emission scanning electron microscope. Molecular studies were performed through PCR amplification and sequencing. MEGA 5 was used to construct a phylogenetic tree using the pairwise distance method. The results showed that only the C. clemensi parasite was found prevalent on L. erythropterus fish with a prevalence and mean intensity (S.D) of 198 (99%) and 36.4 ± 12.2, respectively. The prevalence varied significantly with respect to fish length (p < 0.05). The nucleotide BLAST sequence for 18S ribosomal RNA partial sequences showed 97% with 100% query similarity, E-value 0 with C. clemensi with the accession number DQ123833.1. Conclusively, C. clemensi remains a major parasite of L. erythropterus in the study area.

Analysis of chemical compositions and larvicidal activity of nut extracts from Areca catechu Linn against Aedes (Diptera: Culicidae).

BACKGROUND: There is a growing need to use green alternative larvicidal control for Aedes larvae compared to chemical insecticides. Substantial reliance on chemical insecticides caused insecticide resistance in mosquito populations. Thus, research for alternate chemical compounds from natural products is necessary to control Aedes larvae. This study explores the analysis of chemical compositions from Areca catechu nut as a potential larvicide for Aedes (Diptera: Culicidae). METHODS: The Areca catechu nut collected from Ipoh, Perak, Malaysia was grounded into powder and used for Soxhlet extraction. The chemical analysis of the extracts and their structures were identified using the GCMS-QP2010 Ultra (Shimadzu) system. National Institute of Standards and Technology (NIST) Chemistry WebBook, Standard Reference Database 69 (https://webbook.nist.gov/chemistry/) and PubChem (https://pubchem.ncbi.nlm.nih.gov/), the two databases used to retrieve the synonyms, molecular formula, molecular weight, and 2-dimensional (2D) structure of chemical compounds. Next, following WHO procedures for larval bioassays, the extracts were used to asses larvicidal activity against early 4th instar larvae of Aedes aegypti and Aedes albopictus. RESULTS: The larvicidal activities were observed against early 4th stage larvae with different concentrations in the range from 200 mg/L to 1600 mg/L. The LC50 and LC95 of Aedes aegypti were 621 mg/L and 2264 mg/L respectively; whereas the LC50 and LC95 of Aedes albopictus were 636 mg/L and 2268 mg/L respectively. Mortality was not observed in the non-target organism test. The analysis using gas chromatography and mass spectrometer recovered several chemical compounds such as Arecaidine, Dodecanoic acid, Methyl tetradecanoate, Tetradecanoic acid , and n-Hexadecanoic acid bioactive components. These chemical constituents were used as additive formulations in pesticides, pest control, insect repellent, and insecticidal agents. CONCLUSIONS: Our study showed significant outcomes from the extract of Areca catechu nut and it deserves further investigation in relation to chemical components and larvicidal actions between different species of Aedes mosquitoes. Even though all these findings are fundamental, it may have some interesting potentials to be developed as natural bio-larvicidal products.

Prevalence, Parasite Density and Determinants of Falciparum Malaria Among Febrile Children in Some Peri-Urban Communities in Southwestern Nigeria: A Cross-Sectional Study.

BACKGROUND: Malaria remains a serious public health problem worldwide, particularly in tropical and subtropical regions, including Nigeria. This study investigates the prevalence, parasite density and determinants of malaria among symptomatic children in some peri-urban communities in southwestern Nigeria. METHODS: This was a randomized cross-sectional and hospital-based study. The standard method of microscopy was employed. Thick and thin films were prepared and viewed under a light microscope to identify and quantify malaria parasites. A well-structured and pre-tested questionnaire was used to obtain the subject's information on the demographic, socio-economic and environmental variables. RESULTS: A total of 380 (71.7%) participants were infected with Plasmodium falciparum with a mean parasite density of 1857.11 parasite/µL of blood. Malaria prevalence and mean parasite density were significantly higher among male compared to their female counterparts [80.3% vs 61.4% and 2026.46 vs 1619.63 parasite/µL of blood]. Similarly, age group ≤5 years had the highest malaria prevalence (92.2%) and mean parasite density (2031.66 parasite/µL of blood) than other age groups (AOR 2.281, 95% CI: 1.187-4.384, P < 0.05). The multivariate logistic analysis showed that malaria disease is significantly associated with having mother with no formal education (AOR 12.235, 95% CI: 3.253-46.021, P < 0.05), having well and river as a major source of household water supply (AOR 13.810, 95% CI: 3.012-63.314, P < 0.05 vs AOR 5.639, 95% CI: 1.455-21.853, P < 0.05) and presence of stagnant water around home (AOR 5.22, 95% CI: 2.921-9.332, P < 0.05). Furthermore, protective factors observed include ownership of mosquito bed net (AOR 0.474, 95% CI: 0.223-1.008, P < 0.05) and distance of home to hospital (AOR 0.279, 95% CI: 0.158-0.493, P < 0.05). CONCLUSION: Malaria remains a serious public health problem in the study area. Adopting integrated malaria control measures including educating parents on malaria prevention and control strategies, distributing mosquito bed nets, and establishing larvae source management program is highly imperative.

A cross-sectional study of the prevalence, density, and risk factors associated with malaria transmission in urban communities of Ibadan, Southwestern Nigeria.

BACKGROUND: Malaria is a severe global public health challenge that causes significant morbidity and mortality worldwide, particularly in sub-Saharan Africa. This study was designed to determine the prevalence, parasite density, and risk factors associated with malaria infection transmission among residents of two urban communities of Ibadan, southwestern Nigeria. MATERIALS AND METHODS: A cross-sectional hospital-based study was carried out on 300 participants. Blood samples were obtained. Thick and thin blood films were prepared and viewed using the standard parasitological technique of microscopy. Moreover, data on sociodemographic and environmental variables were obtained using a pre-tested standard questionnaire. RESULTS: Of the 300 participants examined, a total of 165 (55.0%) were found positive for Plasmodium falciparum with a mean (S.D) parasite density of 1814.70 (1829.117) parasite/µL of blood. The prevalence and parasite density of malaria infection vary significantly (P < 0.05) with age group. Children <5 years old were more likely to have malaria infection and high parasite densities than adults (p < 0.05). Similarly, in relation to gender, males significantly (P < 0.05) had a higher prevalence (60.2%) and mean (S.D) parasite density of malaria infection [2157.73 (1659.570) parasite/µL of blood] compared to females. Additionally, those without formal education had the highest prevalence (73.0%) and mean (S.D) parasite density of infection [2626.96 (2442.195) parasite/µL of blood]. The bivariate logistic regression analysis shows that age group 6-10 (Crude Odds Ratio, COR 0.066, 95% CI: 0.007-0.635), presence of streams/rivers (COR 0.225, 95% CI: 0.103-0.492), distance from streams/rivers within ≤1 km (COR 0.283, 95% CI: 0.122-0.654) and travel to rural area (COR 4.689, 95% CI: 2.430-9.049) were the significant risk factors. CONCLUSIONS: Malaria infection is prevalent in the study area and was greatly influenced by traveling activities from the rural areas to urban centers and vice versa. Multifaceted and integrated control strategy should be adopted. Health education on mosquito prevention and chemoprophylaxis before and during travel to rural areas are essential.

Parasitic Gregarine blattarum Found Infecting American Cockroaches, Periplaneta americana, in a Population in Pulau Pinang, Malaysia.

A study on the prevalence of a common endoparasite in the wild population of the American cockroach was conducted in Penang Island using a trapping method at several sampling sites on the island. Gregarine blattarum was found in the digestive tract in 5 out of 115, or 4.35%, of the wild American cockroaches, Periplaneta americana, that were sampled. This is the first report in Malaysia of Gregarine blattarum in local American cockroaches.

The Effect of Macrocyclic Lactones-Ivermectin Exposure on Egg Hatching and Larval Development of Caenorhabditis elegans.

To date, the ivermectin resistance in nematode parasites has been reported and many studies are carried out to determine the causes of this problem. A free-living Caenorhabditis elegans is used as a model system for this study to investigate the response of C. elegans to ivermectin exposure by using larval development assay. Worms were exposed to ivermectin at concentration from 1 ng/mL to 10 ng/mL and dimethyl sulphoxide (DMSO) as a control. The developments of the worms were monitored for 24, 48, 72, and 96 hours until the worms become adults. Results indicated that worms' growth began to be affected by ivermectin at a concentration of 5 ng/mL, while at the concentration of 6, 7, 8, 9, and 10 ng/mL, the growth of worms were inhibited compared to control worms. Further study of the protein expression in C. elegans should be done to investigate the up-regulated and down-regulated proteins involve in ivermectin resistance.

Neobenedenia melleni Parasite of Red Snapper, Lutjanus erythropterus, with Regression Statistical Analysis between Fish Length, Temperature, and Parasitic Intensity in Infected Fish, Cultured at Jerejak Island, Penang, Malaysia.

The fish parasites collected from Lutjanus erythropterus fish species showed a correlation with parasitic intensity, fish size, and temperature, and statistical model summary was produced using SPSS version 20, statistical software. Statistical model summary concluded that among the variables which significantly predict the prevalence of Neobenedenia melleni parasites are fish length and water temperature, both significant at 1% and 5%. Furthermore, the increase in one unit of fish length, holding other variables constant, increases the prevalence of parasite by approximately 1 (0.7≈1) unit. Also, increasing the temperature from 32°C to 33°C will positively increase the number of parasites by approximately 0.32 units, holding other variables constant. The model can be summarized as estimated number of Neobenedenia melleni parasites = 8.2 + 0.7 ⁎ (fish length) + 0.32 ⁎ (water temperature). Next, this study has also shown the DNA sequence and parasitic morphology of Neobenedenia melleni. Nucleotide sequence for 18s ribosomal gene RNA in this study showed 99% similarity with N. melleni EU707804.1 from GenBank. Finally, all the sequence of Neobenedenia melleni in this study was deposited in GenBank with accession numbers of KU843501, KU843502, KU843503, and KU843504.

Oviposition deterring and oviciding potentials of Ipomoea cairica L. leaf extract against dengue vectors.

Bioprospecting of plant-based insecticides for vector control has become an area of interest within the last two decades. Due to drawbacks of chemical insecticides, phytochemicals of plant origin with mosquito control potential are being utilized as alternative sources in integrated vector control. In this regard, the present study aimed to investigate oviposition deterring and oviciding potentials of Ipomoea cairica (L.) (Family: Convolvulaceae) crude leaf extract against dengue vectors, Aedes aegypti and Aedes albopictus. Ipomoea cairica is an indigenous plant that has demonstrated marked toxicity towards larvae of Ae. aegypti and Ae. albopictus. Leaves of I. cairica were extracted using Soxhlet apparatus with acetone as solvent. Oviposition deterrent activity and ovicidal assay was carried out in oviposition site choice tests with three different concentrations (50, 100, 450 ppm). Acetone extract of I. cairica leaf strongly inhibited oviposition with 100% repellence to Ae. aegypti at lower concentration of 100 ppm, while for Ae. albopictus was at 450 ppm. The oviposition activity index (OAI) values which ranged from -0.69 to -1.00 revealed that I. cairica demonstrated deterrent effect. In ovicidal assay, similar trend was observed whereby zero hatchability was recorded for Ae. aegypti and Ae. albopictus eggs at 100 and 450 ppm, respectively. It is noteworthy that I. cairica leaf extract had significantly elicited dual properties as oviposition deterrent and oviciding agent in both Aedes species. Reduction in egg number through oviposition deterring activity, reduction in hatching percentage and survival rates, suggested an additional hallmark of this plant to be integrated in Aedes mosquito control. Ipomoea cairica deserved to be considered as one of the potential alternative sources for the new development of novel plant based insecticides in future.