BACKGROUND: The encystation of Acanthamoeba castellanii has important ecological and medical significance. Blocking encystation is the key to preventing transmission and curing infections caused by A. castellanii. The formation of autophagosomes is one of the most important changes that occur during the encystation of Acanthamoeba. Our previous studies have shown that the heat shock protein 20 of A. castellanii (Ac-HSP20) is involved in its encystation. This study aimed to determine the role and mechanism of Ac-HSP20 in regulating autophagy involved in the encystation of A. castellanii.
METHODS: Immunofluorescence assay, western blotting and transmission electron microscopy were used to analyze the dynamic changes in autophagy during the initiation and continuation of encystation. The knockdown of Ac-HSP20 was performed to clarify its regulation of encystation and autophagy and to elucidate the molecular mechanism by which Ac-HSP20 participates in autophagy to promote cyst maturation.
RESULTS: The encystation rates and autophagosomes were significantly decreased by treatment with the autophagy inhibitor 3-MA. The autophagy marker LC3B and autophagic lysosomes increased with the induced duration of encystation and reached the maximum at 48 h. The encystation rate, LC3B expression and autophagosomes decreased when Ac-HSP20 was knocked down by siRNA transfection. In addition, the expression levels of Ac-HSP20 and LC3B increased and the expressions of p-AKT and p-mTOR decreased after 48 h of encystation without knockdown. However, the expressions of p-AKT and p-mTOR increased while the expression of LC3B decreased under the knockdown of Ac-HSP20. Furthermore, the protein expression of LC3B increased when the PI3K/AKT/mTOR signaling pathway was inhibited but decreased when the pathway was activated.
CONCLUSIONS: The results demonstrated that autophagy is positively correlated with the encystation of A. castellanii, and Ac-HSP20 regulates autophagy to maintain the homeostasis of A. castellanii by inhibiting the PI3K /AKT /mTOR signaling pathway, thus promoting the maturation and stability of encystation.

Saiga antelope (Saiga tatarica) is a protected species in Kazakhstan. Little is known about the parasitofauna of these mammals. Therefore, the focus of this study was to evaluate the prevalence and species diversity of Eimeria spp. infection in the Volga-Ural Saiga antelope population. In June 2023, 104 Saiga antelope fecal samples collected from the district of Zhanibek, located in the province of West Kazakhstan were evaluated using microscopic and molecular techniques. Based on coprovoscopy results, Eimeria spp. Oocysts were present in 22 samples (21%). The four fecal samples containing the largest numbers of Eimeria spp. Oocysts per 10x field were selected for further genetic analysis. DNA extraction, nested PCR amplification, and sequencing were performed on 91 clones, with 80 clones forming a distinct clade and exhibiting genetic similarity to MT801034 Eimeria sp. Voucher HY3. These clones possibly represent an Eimeria specific to Saiga antelopes and gazelle that has previously been morphologically described as Eimeria elegans (Svanbaev, 1979), underscoring the importance of further research into parasitic infections in this protected species.

The escalating presence of microplastics and heavy metals in marine environments significantly jeopardizes ecological stability and human health. Despite this, research on the combined effects of microplastics/nanoplastics (MPs/NPs) and heavy metals on marine organisms remains limited. This study evaluated the impact of two sizes of polystyrene beads (approximately 2 μm and 200 nm) combined with cadmium (Cd) on the ciliate species Euplotes vannus. Results demonstrated that co-exposure of MPs/NPs and Cd markedly elevated reactive oxygen species (ROS) levels in ciliates while impairing antioxidant enzyme activities, thus enhancing oxidative damage and significantly reducing carbon biomass in ciliates. Transcriptomic profiling indicated that co-exposure of MPs/NPs and Cd potentially caused severe DNA damage and protein oxidation, as evidenced by numerous differentially expressed genes (DEGs) associated with mismatch repair, DNA replication, and proteasome function. Integrated transcriptomic and metabolomic analysis revealed that DEGs and differentially accumulated metabolites (DAMs) were significantly enriched in the TCA cycle, glycolysis, tryptophan metabolism, and glutathione metabolism. This suggests that co-exposure of MPs/NPs and Cd may reduce ciliate abundance and carbon biomass by inhibiting energy metabolism and antioxidant pathways. Additionally, compared to MPs, the co-exposure of NPs and Cd exhibited more severe negative effects due to the larger specific surface area of NPs, which can carry more Cd. These findings provide novel insights into the toxic effects of MPs/NPs and heavy metals on protozoan ciliates, offering foundational data for assessing the ecological risks of heavy metals exacerbated by MPs/NPs.

The correlation between enteric methane emissions (eME) and feed efficiency (FE) in cattle is linked to the anaerobic fermentation of feedstuffs that occurs in the rumen. Several mathematical indices have been developed to predict feed efficiency and identify low methane emitters in herds. To investigate this, the current study aimed to evaluate the rumen microbial composition in the same group of animals ranked according to six different indices (three indices for FE and three for eME). Thirty-three heifers were ranked into three groups, each consisting of 11 animals, based on FE (feed conversion efficiency - FCE, residual weight gain - RG, and residual feed intake - RFI) and eME indices (production, yield, and intensity). Rumen fluids were collected using a stomach tube and analyzed using 16S rRNA and 18S rRNA, targeting rumen bacteria, archaea, and protozoa. The sequencing analysis revealed that the presence of unique microbial species in the rumen varies across animals ranked by the FE and eME indices. The High RG group harbored 17 unique prokaryotic taxa, while the High FCE group contained only seven. Significant differences existed in the microbial profiles of the animals based on the FE and eME indices. For instance, Raoultibacter was more abundant in the Intermediate RFI group but less so in the Intermediate RG and Intermediate FCE groups. The abundance of Entodinium was higher while Diplodinium was lower in the High FCE group, in contrast to the High RG and High RFI groups. Methanobrevibacter exhibited similar abundances across eME indices. However, the heifers did not demonstrate the same production, yield, and intensity of eME. The present findings underscore the importance of standardizing the FE and eME indices. This standardization is crucial for ensuring consistent and reliable assessments of the composition and function of the rumen microbiome across different herds.

Data concerning the sensitivity of various organisms to ultraviolet (UV) radiation exposure are very important in the design of UV disinfection equipment. This review analyzes fluence data from almost 250 studies and organizes the data into a set of recommended fluence values for specific log reductions and an appendix containing all the collected data. This article was sponsored by Dianne L. Poster, Material Measurement Laboratory, and C. Cameron Miller, Physical Measurement Laboratory, National Institute of Standards and Technology (NIST). It is published in collaboration with the International Ultraviolet Association as a complement to the NIST Workshop on Ultraviolet Disinfection Technologies, 14-15 January 2020, Gaithersburg, MD. The views expressed represent those of the authors and not necessarily those of NIST.

Anaerobic protists frequently harbour methanogenic archaea, which apparently contribute to the hosts\' fermentative metabolism by consuming excess H2. However, the ecological properties of endosymbiotic methanogens remain elusive in many cases. Here we investigated the ecology and genome of the endosymbiotic methanogen of the Cononympha protists in the hindgut of the termite Coptotermes formosanus. Microscopic and 16S rRNA amplicon sequencing analyses revealed that a single species, designated here \"Candidatus Methanobrevibacter cononymphae\", is associated with both Cononympha leidyi and Cononympha koidzumii and that its infection rate in Cononympha cells varied from 0.0% to 99.8% among termite colonies. Fine-scale network analysis indicated that multiple 16S rRNA sequence variants coexisted within a single host cell and that identical variants were present in both Cononympha species and also on the gut wall. Thus, \"Ca. Methanobrevibacter cononymphae\" is a facultative endosymbiont, transmitted vertically with frequent exchanges with the gut environment. Indeed, transmission electron microscopy showed escape or uptake of methanogens from/by a Cononympha cell. The genome of \"Ca. Methanobrevibacter cononymphae\" showed features consistent with its facultative lifestyle: i.e., the genome size (2.7 Mbp) comparable to those of free-living relatives; the pseudogenization of the formate dehydrogenase gene fdhA, unnecessary within the non-formate-producing host cell; the dependence on abundant acetate in the host cell as an essential carbon source; and the presence of a catalase gene, required for colonization on the microoxic gut wall. Our study revealed a versatile endosymbiosis between the methanogen and protists, which may be a strategy responding to changing conditions in the termite gut.

Two experiments were conducted to examine the impact of trace mineral (TM) source on in vitro and in vivo solubility characteristics. Experiment 1: Hydroxy TM (HTM) and sulfate TM (STM) sources of Cu, Mn, and Zn were incubated separately in water for 24 h. Immediately after mixing, initial pH of each solution was greater (P < 0.03) for HTM compared to STM for all elements. Final pH tended to be greater for Cu (P = 0.09) and Zn (P = 0.07) from HTM compared to STM. Water solubility of Cu, Mn, and Zn from STM was greater (P < 0.01) than HTM sources. Experiment 2: Eight steers fitted with rumen cannula were blocked by body weight and randomly assigned to treatments. Treatments consisted of 10 mg Cu, 40 mg Mn, and 60 mg Zn/kg DM from either STM or HTM sources. Steers were individually fed a cracked corn-corn silage-based diet. Treatments were top-dressed daily. Rumen contents were collected at 0, 2, and 4 h post-feeding on d 1 and 14. On d 15, strained ruminal fluid and particle-associated microorganisms were obtained. Zinc was more tightly bound (P = 0.01) to the digesta in HTM-supplemented steers compared to STM on d 14. These data indicate that TM source influences pH and solubility of Cu, Mn, and Zn in water and may affect rumen soluble Cu concentrations and binding strength of Zn to solid digesta.

Intestinal parasitic infections (IPIs) pose a serious public health threat across the globe, particularly in children residing in poor and most deprived communities like Ethiopia. Many published articles were available separately, and a detailed nationwide review was essential to combine all the results to draw a conclusion and avoid any informational conflicts, ambiguities, or misunderstandings. Therefore, this systematic review and meta-analysis aimed to provide pooled estimates for the individually available data on IPIs and its determinant factors among preschool-aged children (PSAC) in Ethiopia. Published and unpublished articles from various electronic databases were accessed using MeSH terms and keywords. I 2 and sensitivity analysis tests were used to assess potential sources of heterogeneity across studies. Funnel plot, Begg\'s, and Egger\'s regression tests were used to check publication bias. A random-effects model with a 95% confidence interval (CI) was used to calculate the pooled estimate of IPI. In this meta-analysis, a total of 14,994 PSAC were included in the 32 eligible studies. The pooled national prevalence of IPIs among PSAC was 32.52% (95% CI: 26.24, 38.80). Of these, single and mixed infections contribute 31.08% and 1.44%, respectively. According to the subgroup analysis, the highest prevalence was observed in simple random studies (39.61%; 95% CI: 29.19, 50.03), the Tigray region (58.00%; 95% CI: 54.10, 61.90), studies conducted in >384 (39.47%; 95% CI: 27.73, 51.20) sample sizes, cross-sectional studies (32.76%), community-based studies (42.33%; 95% CI: 31.93, 52.74), and from 2005 to 2016 (34.53%; 95% CI: 20.13, 48.92) study periods. Intestinal parasites were significantly associated with eating raw fruits and vegetables (aOR = 3.21; 95%CI: 1.11-5.31). The high prevalence of STHs observed in this systematic review and meta-analysis underscores the need for appropriate control and prevention strategies suitable for PSAC in Ethiopia.

BACKGROUND: Toxoplasmosis has been of public health concern due to direct associations with socioeconomic vulnerability and inadequate living conditions.
METHODS: Accordingly, the present study aimed to assess antibodies against T. gondii, historical reported toxoplasmosis cases and associated socio-environmental risk factors in Pinhais, a full urban area of Curitiba, currently the eighth biggest metropolitan area of Brazil. Anti-Toxoplasma gondii antibodies were assessed by an indirect immunofluorescence reaction (RIFI). Owner and dog samples were also tested by IFAT to anti-Leishmania spp. and anti-Trypanosoma cruzi antibodies.
RESULTS: Overall, 20/135 (14.8%) persons and 13/133 (9.8%) dogs from 25 different households were considered seropositive to T. gondii. All samples were seronegative to Leishmania spp. and Trypanosoma cruzi.
CONCLUSIONS: Although no significant covariates were found in the regression model, statistically associated risk factors in the bivariate analysis included no public water use (p = 0.016) and drinking raw milk (p = 0.041) for owners, and obesity (p = 0.028) and tick infestation (p = 0.03) for dogs. In addition, a spatial cluster of T. gondii seropositivity for both owners and their dogs overlapped the location of historic reported cases of human acquired, gestational and congenital toxoplasmosis. Finally, the results herein showed tick infestation as an indicator of socio-environmental risk for T. gondii exposure in the household environment, and dogs may be used as sentinels for human toxoplasmosis cases.

The relationship between the protozoan communities and environmental variables was studied in the Nile River to evaluate their potential as water quality indicators. Protozoans were sampled monthly at six sampling sites in the Nile\'s Damietta Branch across a spatial gradient of environmental conditions during a 1-year cycle (February 2016-January 2017). The Protozoa community was comprised of 54 species belonging to six main heterotrophic Protozoa phyla. The abundance (average, 1089 ± 576.18 individuals L-1) and biomass (average, 86.60 ± 106.13 μg L-1) were comparable between sites. Ciliates comprised the majority of protozoan species richness (30 species), abundance (79.72%), and biomass (82.90%). Cluster analysis resulted in the distribution of protozoan species into three groups, with the most dominant species being the omnivorous ciliate Paradileptus elephantinus. Aluminium, fluoride, and turbidity negatively affected abundance and biomass, while dissolved oxygen and potassium positively impacted biomass. Of the dominant species recorded over the study area, the amoebozoa Centropyxis aculeata was associated with runoff variables, while the bacterivorous ciliates Colpidium colpoda, Glaucoma scintillans, and Vorticella convallaria were related to the abundance of heterotrophic bacteria, phytoplankton biomass, and total organic carbon. Total dissolved salts, PO4, NH3, NO2, dissolved oxygen, and total organic carbon were the strongest causative factors for protozoa distribution. The α-Mesosaprobic environment at site VI confirmed a high load of agricultural runoffs compared to other sites. This study demonstrates that protozoans can be a potential bioindicator of water quality status in this subtropical freshwater river system.