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Ruminal ciliates are linked to methane production and nitrogen utilization efficiency in ruminants due to their association with other ruminal microorganisms. However, research on the specific interplay between ruminal bacteria and ciliates is still limited, particularly in different dietary conditions. This study examines the effect of the forage-to-concentrate (F:C) ratio on the ruminal bacteriome in vitro , focusing on bacteria associated with Isotricha spp. and small entodinia. The rumen fluid used as the inoculum for this experiment was collected from two cannulated Hanwoo cows. Dietary treatments included high-forage ( HF , F:C of 7:3), high-concentrate ( HC , F:C of 3:7), and control ( CON , F:C of 5:5). After 24-hour incubation, fractions for entodinia-associated bacteria ( EAB ), Isotricha -associated bacteria ( IAB ), and total bacteria ( TB ) were collected for bacteriome analysis using QIIME2 with full-length 16S rRNA gene sequences on the PacBio system. All fermentation parameters, except for NH 3 -N, showed linear changes with increasing F:C ratios (p  ≤  0.05). F:C ratio affected Isotricha spp. and Dasytricha spp. counts. Ciliate-associated bacterial fractions were significantly less diverse than the total bacterial group, as indicated by richness, phylogenetic diversity, and evenness indices. This suggests potential specific associations within ciliate-provided microhabitats. Both diet and ciliate fractions significantly influenced the overall bacteriome (p  ≤  0.05). More bacteriome features were differentially abundant due to the ciliate fraction effect rather than diet (q ≤ 0.05). Our newly proposed washing procedure, using higher ciliate cell counts and minimal bacterial contamination, effectively removed free-living or loosely associated bacteria. This allows focus on ciliate-associated bacterial populations, which may include potential symbionts or engulfed bacteria of host ruminal ciliates. Verifying these associations could provide insights into rumen microbiome dynamics, nitrogen utilization, hydrogen balance, and microbiome variation under different F:C ratios.

Ruminal ciliates are linked to methane production and nitrogen utilization efficiency in ruminants due to their association with other ruminal microorganisms. However, research on the specific interplay between ruminal bacteria and ciliates is still limited, particularly in different dietary conditions. This study examines the effect of the forage-to-concentrate (F:C) ratio on the ruminal bacteriome in vitro, focusing on bacteria associated with Isotricha spp. and small entodinia. The rumen fluid used as the inoculum for this experiment was collected from two cannulated Hanwoo cows. Dietary treatments included high-forage (HF, F:C of 7:3), high-concentrate (HC, F:C of 3:7), and control (CON, F:C of 5:5). After 24-hour incubation, fractions for entodinia-associated bacteria (EAB), Isotricha-associated bacteria (IAB), and total bacteria (TB) were collected for bacteriome analysis using QIIME2 with full-length 16S rRNA gene sequences on the PacBio system. All fermentation parameters, except for NH -N, showed linear changes with increasing F:C ratios (p ≤ 0.05). F:C ratio affected Isotricha spp. and Dasytricha spp. counts. Ciliate-associated bacterial fractions were significantly less diverse than the total bacterial group, as indicated by richness, phylogenetic diversity, and evenness indices. This suggests potential specific associations within ciliate-provided microhabitats. Both diet and ciliate fractions significantly influenced the overall bacteriome (p ≤ 0.05). More bacteriome features were differentially abundant due to the ciliate fraction effect rather than diet (q ≤ 0.05). Our newly proposed washing procedure, using higher ciliate cell counts and minimal bacterial contamination, effectively removed free-living or loosely associated bacteria. This allows focus on ciliate-associated bacterial populations, which may include potential symbionts or engulfed bacteria of host ruminal ciliates. Verifying these associations could provide insights into rumen microbiome dynamics, nitrogen utilization, hydrogen balance, and microbiome variation under different F:C ratios.

The paired review papers in Parts I and II describe the 50-year history of research on the biomechanics of swimming microorganisms and its prospects in the next 50 years: Part I explains the behaviour of individual microorganisms, and Part II explains collective behaviour. Since the discovery of microorganisms by van Leeuwenhoek in the 17th century, many natural scientists have been interested in their motility because it is directly associated with biological function. A research upsurge occurred in the 1970s, with the elucidation of swimming mechanisms among individual microorganisms and the theoretical derivation of swimming speeds. Various swimming strategies of three types of microorganisms, i.e. bacteria, ciliates and microalgae, are explained in this Part I. We show that some of the behaviours of microorganisms can be described by biomechanical equations and are to some extent predictable. Recent researches have revealed the behaviour of microorganisms in more complex environments and more realistic settings, which are also reviewed in the paper. Last, we provide future prospects for research on microbial behaviour.

Green synthesis of metal oxide nanoparticles using plant extracts is a promising alternative to the traditional method of physical and chemical synthesis, as it is a wide research field and environmentally friendly methods. In the current study, iron oxide nanoparticles (α-Fe 2 O 3 -NPs) were bio-synthesized by using Moltkia ciliata plant extract with different concentration of ferric chloride (FeCl 3 ). To improve the yield, we relied on changing the concentration of the saline solution (0.1, 0.05, and 0.025M(. These α-Fe 2 O 3 NPs were analyzed by X-ray diffraction (XRD), scanning electron microscopy (SEM), Fourier transform infrared (FT-IR) spectroscopy, and energy-dispersive X-ray (EDX). UV-Vis analysis shows absorption maximums at the range 200–400nm belonging to Fe-O, and FT-IR shows clear peaks in the range 450–500 cm −1 , which are attributed to the vibration of Fe-O, while the SEM and DRX results are constructed. There are similarities in the structure of the samples in terms of shape and size, despite their different concentrations. Antibacterial studies were used against Gram-positive and -negative bacteria ( Escherichia coli , Pseudomonas aeruginosa , Klebsiella pneumoniae , Staphylococcus aureus ). The sample synthesized at a concentration of 0.05M showed the highest antibacterial activity (inhibition zone) up to 11.7 mm at a concentration of 0.025 M for Klebsiella pneumoniae . This study concluded that the biosynthetic α-Fe 2 O 3 using Moltkia ciliata extract gave significant antibacterial activity.

This study investigated the associations between abundance of rumen ciliate protozoa and the proportion of the main bioactive fatty acids related to rumen biohydrogenation, as 18:0, t10-18:1, t11-18:1, c9,t11-18:2, 18:3n-3 and 18:2 n-6, in rumen and meat of growing lambs, using data derived from 3 production experiments. A global correlation analysis and a linear regression analysis considering the effect of the experiment were performed. Ten of the 86 lambs involved in the experiments did not present ciliate cells in rumen liquor and the remaining lambs presented an average of 1.35 x 106ciliates / ml rumen liquor. From the nine genera of ciliates identified, Entodinium was the most abundant, averaging 1.17 x 106 cells / ml of rumen liquor. A large variation among lambs was observed for both rumen concentration and community structure of ciliates. Rumen t11-18:1 (P< 0.001) and meat deposition of t11-18:1 (P < 0.001) and of c9,t11-18:2 (P < 0.001) increased linearly with total ciliates, whereas the t10/t11 ratio in rumen (P= 0.002) and in meat (P= 0.036) decreased linearly. Entodiniomorphids seems to be strongly related with meat deposition of t11-18:1 and c9,t11-18:2 and with the reduction of the trans-10 shifted pathway. Completeness of RBH decreased linearly with Holotrichs (P = 0.029), Entodiniomorphids (P = 0.029), Isotricha (P = 0.011) and Epidinium (P = 0.027) abundances. Rumen 18:0 also decreased linearly with increasing counts of total ciliates (P = 0.015), Holotrichs (P = 0.020), Entodiniomorphids (P= 0.010) and Isotricha (P= 0.014). Rumen protozoa were positively linked with the deposition of healthy bioactive FA and simultaneously negatively associated with the occurrence of trans-10 shift.

Abstract Ciliated protists are among the oldest unicellular organisms with a heterotrophic lifestyle and share a common ancestor with Plantae. Unlike any other eukaryotes, there are two distinct nuclei in ciliates with separate germline and somatic cell functions. Here, we assembled a near-complete macronuclear genome of Fabrea salina, which belongs to one of the oldest clades of ciliates. Its extremely minimized genome (18.35 Mb) is the smallest among all free-living heterotrophic eukaryotes and exhibits typical streamlined genomic features, including high gene density, tiny introns, and shrinkage of gene paralogs. Gene families involved in hypersaline stress resistance, DNA replication proteins, and mitochondrial biogenesis are expanded, and the accumulation of phosphatidic acid may play an important role in resistance to high osmotic pressure. We further investigated the morphological and transcriptomic changes in the macronucleus during sexual reproduction and highlighted the potential contribution of macronuclear residuals to this process. We believe that the minimized genome generated in this study provides novel insights into the genome streamlining theory and will be an ideal model to study the evolution of eukaryotic heterotrophs.

This research was planned to synthesize a biological potent nanomaterials via an eco-friendly process to combat the diseases causing bacteria and the free radicals generated inside the body. For this purpose, a green synthesis process was employed to prepare SnO2 nanoparticles by utilizing leaf extract of Populus ciliate, and they were characterized via different physico-chemical techniques. The crystallite size of SnO2 nanoparticles was found to be 58.5 nm. The calculated band gap energy of SnO2 nanoparticles was 3.36 eV. The SnO2 nanoparticles showed 38, 49, 57, and 72% antioxidant activity at concentrations of 100, 200, 300, and 400 L with 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonicacid) (ABTS) assays. The antibacterial effects of prepared SnO2 nanoparticles were studied using the agar well diffusion method against Gram-positive bacteria (S. pyogene and S. aureus) and Gram-negative bacteria (K. pneumoniae and E. coli). Both the antioxidant activity and antibacterial activity were seen to increase with increasing the concentration of the nanoparticles.

Changes in the biodiversity of aquatic environments over time and space due to human activities are a topic of theoretical and conservational interest in ecology. Thus, variation in taxonomic beta diversity of the planktonic ciliates community was investigated along a temporal and spatial gradient in two subsystems of a Neotropical floodplain, one impacted by dams (Paraná) and the other free of them along its course (Ivinhema). For the spatial analysis, the Paraná subsystem did not show a significant decrease in beta diversity, presenting a pattern like that observed for the Ivinhema subsystem. Therefore, biotic homogenization was not observed for the ciliate's community downstream of the dams. It was noted that there was a fluctuation in the relevance of the components of beta diversity, regardless of the subsystem analyzed. For the temporal analysis there was a significant change in species composition from the first to the last year investigated, essentially for the subsystem impacted by dams, and that this was determined mainly by species loss. Although spatial beta diversity remained high without a clear process of biotic homogenization, dams promoted remarkable changes in ciliate species composition over the years mainly by continuous loss of species.

Wild aquatic birds are a major reservoir of the influenza A virus in natural ecosystems, facilitating its entry into the aquatic microbial food web through their feces. Free-living protozoa and particularly bacterivorous ciliates are essential players of the microbial food web. This study investigates the interactions between the Influenza A(H1N1)pdm09 virus and the ciliated protozoan Tetrahymena pyriformis at the population and ultrastructural levels. Co-cultivation of Influenza A(H1N1)pdm09 and T. pyriformis resulted in a decline and eventual complete elimination of the viral population. The inactivation of the virus was not mediated by products excreted by T. pyriformis but required A(H1N1)pdm09 endocytosis. Viruses ingested by protozoa lost their virulence within 48 hours post infection (hpi) and, as determined by hemagglutination assays, were entirely inactivated within 72 hpi. When lysates infected with A(H1N1)pdm09 T. pyriformis were applied to MDCK cells 1.5 and 24 hpi the undamaged part of ingested virions caused a cytopathic effect. Confocal laser scanning microscopy (CLSM) and transmission electron microscopy (TEM) of infected T. pyriformis cells revealed large food vacuoles, including multiple undamaged and partly processed virus particles, at 1.5 and 24 hpi. Furthermore, TEM identified coated and half-coated small one-virus endosomes that predominated at 48 hpi. These results demonstrated that A(H1N1)pdm09 inactivation by T. pyriformis includes two types of endosomes that dominated at different periods of interpopulation interactions. The process of A(H1N1)pdm09 inactivation in protozoan cells occurs rapidly, but not instantaneously, that suggesting a dual role of protozoa in the fate of influenza A viruses in natural ecosystems, both as predators and as potential vectors.