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Infections by gastrointestinal parasites are found in a variety of animals worldwide. For the diagnosis of such infections, the flotation method is commonly used to detect parasitic microorganisms, such as oocysts or eggs, in feces. Instead of adding a flotation solution after the final centrifugation step and using a cover slip to collect the parasites, the method using a wire loop for the recovery of the organisms has been reported as one of alternative methods. However, the recovery rates of microorganisms from the flotation method have not been analysed. In the present study, the utility of a flotation method with the use of a wire loop of 8 mm in diameter (the loop method) was evaluated using different numbers of E. tenella oocysts and Heterakis gallinarum eggs, and chicken fecal samples collected at the farms. Consequently, we found that the oocysts and eggs in tubes could be collected at a ratio of 2.00 to 3.08. Thus, our results indicate that the loop method is a simple and time saving method, implicating the application for the estimated OPG/ EPG (Oocysts/Eggs per gram) of the samples. [ABSTRACT FROM AUTHOR]

RNA activation (RNAa) is a burgeoning area of research in which double-stranded RNAs (dsRNAs) or small activating RNAs mediate the upregulation of specific genes by targeting the promoter sequence and/or AU-rich elements in the 3′- untranslated region (3'-UTR) of mRNA molecules. So far, studies on the phenomenon have been limited to mammals, plants, bacteria, Caenorhabditis elegans, and recently, Aedes aegypti. However, it is yet to be applied in other arthropods, including ticks, despite the ubiquitous presence of argonaute 2 protein, which is an indispensable requirement for the formation of RNA-induced transcriptional activation complex to enable a dsRNA-mediated gene activation. In this study, we demonstrated for the first time the possible presence of RNAa phenomenon in the tick vector, Haemaphysalis longicornis (Asian longhorned tick). We targeted the 3ʹ-UTR of a novel endochitinase-like gene (HlemCHT) identified previously in H. longicornis eggs for dsRNA-mediated gene activation. Our results showed an increased gene expression in eggs of H. longicornis endochitinase-dsRNA-injected (dsHlemCHT) ticks on day-13 post-oviposition. Furthermore, we observed that eggs of dsHlemCHT ticks exhibited relatively early egg development and hatching, suggesting a dsRNA-mediated activation of the HlemCHT gene in the eggs. This is the first attempt to provide evidence of RNAa in ticks. Although further studies are required to elucidate the detailed mechanism by which RNAa occurs in ticks, the outcome of this study provides new opportunities for the use of RNAa as a gene overexpression tool in future studies on tick biology, to reduce the global burden of ticks and tick-borne diseases. [ABSTRACT FROM AUTHOR]

Blood-feeding arthropods, particularly ticks and mosquitoes are considered the most important vectors of arthropod-borne diseases affecting humans and animals. While feeding on blood meals, arthropods are exposed to high levels of reactive oxygen species (ROS) since heme and other blood components can induce oxidative stress. Different ROS have important roles in interactions among the pathogens, vectors, and hosts. ROS influence various metabolic processes of the arthropods and some have detrimental effects. In this review, we investigate the various roles of ROS in these arthropods, including their innate immunity and the homeostasis of their microbiomes, that is, how ROS are utilized to maintain the balance between the natural microbiota and potential pathogens. We elucidate the mechanism of how ROS are utilized to fight off invading pathogens and how the arthropod-borne pathogens use the arthropods' antioxidant mechanism to defend against these ROS attacks and their possible impact on their vector potentials or their ability to acquire and transmit pathogens. In addition, we describe the possible roles of ROS in chemical insecticide/acaricide activity and/or in the development of resistance. Overall, this underscores the importance of the antioxidant system as a potential target for the control of arthropod and arthropod-borne pathogens. [ABSTRACT FROM AUTHOR]

Coccidiosis poses a significant threat to the poultry industry, with synthetic antibiotics and disinfectants being the primary tools for control. This study investigated the potential of Piper betle L essential oil (PBEO) as a natural alternative against Eimeria tenella, one of the most pathogenic Eimeria species affecting poultry. Our findings revealed that PBEO exhibits significant anticoccidial effects through two primary mechanisms: (i) oocysticidal activity by disintegrating oocyst walls and (ii) inhibition of the sporulation process. PBEO demonstrated oocysticidal activities ranging from 8.67 to 95.33% across concentrations from 0.04 to 40%. Notably, at 72 h post-incubation, a 0.04% PBEO concentration significantly reduced the number of sporulated oocysts (P ≤ 0.05) to 71.67%, showing effects comparable to those of formalin. PBEO reduced 50% of oocyst sporulation (IC 50 ) in the concentration of 1.31% at 72 h. Gas chromatography-mass spectrometry (GC-MS) identified the primary constituents of PBEO, including eugenol, beta-caryophyllene, and other key compounds, collectively constituting 96% of the oil. This research underscores the potential of PBEO as a natural anticoccidial agent and lays the groundwork for further studies aimed at identifying, isolating, and developing active compounds that may specifically target the sporogony process in coccidian parasites., (© 2024. The Author(s).)

Background: Extensive attention has been devoted to studies of Trypanosoma lewisi in rodents ever since it became recognised as a zoonotic pathogen known as atypical human trypanosomiasis. Regrettably, although T. lewisi infections of small mammals remain significant public health concerns for humans, there is a lack of comprehensive study in Indonesia., Aim: The aim of the study was to detect T. lewisi from rodents residing in the densely populated residential regions along the coastal areas of Banyuwangi Sub District., Methods: A total of 169 rodents were captured across three villages of Kampung Mandar, Lateng and Kepatihan, using rat single live traps. After being euthanized and identified, the blood samples were collected from each rodent via cardiac puncture. Subsequently, the samples were subjected to native (direct blood microscopic examination), microscopic blood smear examination, and molecular analyses utilizing TRYP1S-TRYP1R (623 bp) and LEW1S-LEW1R (220 bp)., Results: The results demonstrated that two species of rodents were successfully captured: Rattus norvegicus (65.68%) and Rattus tanezumi (34.32%). Based on the native and microscopic blood smear examinations, the prevalence of T. lewisi across three villages was 23.08% and 24.26% for molecular analysis employing both primers, respectively. The highest prevalence was found in Kampung Mandar Village (31.18%), followed by Kepatihan (16.67%) and Lateng Villages (15.71%)., Conclusion: Statistical analysis revealed that T. lewisi was more prevalent in R. tanezumi compared to R. norvegicus . In terms of sex, no statistically significant distinction was observed between female and male infected rodents of either species ( p > 0.05), indicating both species can serve as a source of T. lewisi for humans in the surveyed villages., Competing Interests: The authors declare that there is no conflict of interest.

The Japanese rock ptarmigan ( Lagopus muta japonica ) is an herbivorous species of partridges that inhabits only alpine zones. Alpine plants are their main source of food. These alpine plants contain toxic compounds to deter herbivores from consuming them. A previous analysis of the alpine plants frequently consumed by Japanese rock ptarmigans revealed the presence of a unique mixture of secondary metabolites and a novel compound. Additionally, wild Japanese rock ptarmigans are often infected by two species of Eimeria parasites. When these parasites were experimentally administered to Svalbard rock ptarmigans ( Lagopus muta hyperborean ), which do not feed on alpine plants, the birds exhibited symptoms, such as diarrhea and depression, and in some cases, they died. Although little is known about the pathogenesis of these parasites in wild Japanese rock ptarmigans, it was hypothesized that compounds found in alpine plants, their main food source, may reduce the pathogenicity of Eimeria parasites. In the present study, we evaluated the anticoccidial activity of the compounds derived from alpine plants in vitro using Eimeria tenella , which infects chickens belonging to the same pheasant family, as an experimental model. Twenty-seven natural components were extracted from eight alpine plants. The natural components were added to E. tenella sporozoites and incubated for 24 h to evaluate their direct effect. Additionally, Madin-Darby bovine kidney cells were incubated with sporozoites and natural components for 24 h to evaluate the inhibitory effect of the components on sporozoite cell invasion. Six compounds from four alpine plants decreased sporozoite viability by up to 88.3%, and two compounds inhibited sporozoite invasion into the cells. Although further studies are needed to evaluate the effects of these components against Eimeria infections in vivo , our findings suggest that these alpine plants may reduce the degree of infection by decreasing the number of sporozoites in the intestinal tract., (© 2024 The Authors.)

Background and Aim: Toxoplasmosis is caused by the parasite Toxoplasma gondii . Cats are the only known hosts that excrete resistant oocysts. Wild rats serve as crucial reservoirs and intermediate hosts for T. gondii's survival and dissemination. Consuming soil and water containing oocysts can lead to illness. This study aimed to estimate the prevalence of toxoplasmosis in wild rats through molecular detection as an indicator of environmental contamination in Surabaya., Materials and Methods: One hundred rats were collected from the three areas (housing, dense settlements, and traditional markets) and distributed into the five zones: West, East, Central, North, and South of Surabaya. Brain tissue samples were extracted using a Geneaid™ (New Taipei City, Taiwan) DNA isolation kit and analyzed through the loop-mediated isothermal amplification (LAMP) method., Results: The study analyzed brain tissue from 100 wild rats, consisting of 77 Rattus tanezumi and 33 Rattus norvegicus , displaying 30% LAMP positivity. The study revealed that 30% (30/100) of wild rats tested were infected with T. gondii . The molecular prevalence rate in male rats was 32.35% (22/68), compared to females with 25% (8/32). 41.9% of the housing population, 33.3% of traditional markets, and 22.6% of dense settlements had the highest molecular prevalence. The high positive molecular rate at the trapping site can be attributed to cats and dense populations., Conclusion: Thirty percentage wild rats were tested positive for toxoplasmosis in Surabaya, East Java, Indonesia using LAMP method. Implementing strict control and monitoring is crucial in preventing the transmission of diseases from wild rats to humans. It is necessary to carry out further research related to genetic analysis of T. gondii to determine the type of T. gondii that infects animals and humans in Surabaya through bioassay and molecular test., Competing Interests: The authors declare that they have no competing interests., (Copyright: © Puspitasari, et al.)

The genus Mansonella Faust, 1929 includes 29 species, mainly parasites of platyrrhine monkeys in South America and anthropoid apes in Africa. In Malaysia, Mansonella ( Tupainema ) dunni (Mullin & Orihel, 1972) was described from the common treeshrew Tupaia glis Diard & Duvaucel (Scandentia). In a recent classification of the genus Mansonella , seven subgenera were proposed, with M . ( Tup .) dunni as a monotypic species in the subgenus Tupainema . In this study, we collected new material of M . ( Tup .) dunni from common treeshrews in Peninsular Malaysia and redescribed the morphological features of this species. We found that M . ( Tup .) dunni differs from M . ( Cutifilaria ) perforata Uni et al., 2004 from sika deer Cervus nippon (Cetartiodactyla) in Japan, with regards to morphological features and predilection sites in their respective hosts. Based on multi-locus sequence analyses, we examined the molecular phylogeny of M . ( Tup .) dunni and its Wolbachia genotype. Species of the genus Mansonella grouped monophyletically in clade ONC5 and M . ( Tup .) dunni was placed in the most derived position within this genus. Mansonella ( Tup .) dunni was closely related to M . ( M .) ozzardi (Manson, 1897) from humans in Central and South America, and most distant from M . ( C .) perforata . The calculated p-distances between the cox 1 gene sequences for M . ( Tup .) dunni and its congeners were 13.09% for M . ( M. ) ozzardi and 15.6-16.15% for M . ( C .) perforata . The molecular phylogeny of Mansonella spp. thus corroborates their morphological differences. We determined that M . ( Tup .) dunni harbours Wolbachia endosymbionts of the supergroup F genotype, in keeping with all other Mansonella species screened to date., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors.)