Search

ABSTRACTBruno Martorelli Di Genova works in parasitology, focusing on Toxoplasma gondii metabolism. In this mSphere of Influence article, he reflects on how the articles “Metabolic Reprogramming during Purine Stress in the Protozoan Pathogen Leishmania donovani” and “Yeast-Based High-Throughput Screen Identifies Plasmodium falciparum Equilibrative Nucleoside Transporter 1 Inhibitors That Kill Malaria Parasites” impacted him, informing his research strategies and understanding of metabolic flexibility in Toxoplasma gondii.

ABSTRACT: Trypanosoma evansi, the hemoflagellate pathogen that causes “surra,” is a protozoan that infects the widest variety of mammals worldwide, primarily horses, camels, dogs, and wild animals. Cases of canine trypanosomiasis caused by T.evansi have been reported in the Brazilian pantonal region. This study determined the prevalence of T.evansi and investigate risk factors. The study used blood samples from dogs in the municipality of Barão de Melgaço in the pantonal of the state of Mato Grosso, Brazil. The owner was given a structured epidemiological questionnaire containing information about the risk factors. The quantitative polymerase chain reaction technique was used to detect T.evansi DNA using species-specific oligonucleotides TevF and TevR of the gene encoding a 227-bp portion of the glycoprotein of the variant surface (VSG) T.evansi Rode Trypanozoon (RoTat) 1.2. Two (prevalence of 0.5%) genetic materials of T.evansi were reported in the403 samples. Although, no risk factor was associated with infection (P > 0.05), proximity to vegetation and streams, as well as the presence of tabanids, rodents, and marsupials, which are factors in the occurrence of infection, were observed in positive dogs, implying that the municipality of Barão de Melgaço has epidemiological characteristics that allow canine infection by T.evansi.

Parasitoses that are localized to the Central Nervous System (CNS) can cause symptomatic diseases or progress asymptomatically [g1] [AR2]. Any parasites that affect humans could involve the CNS; however, the most common parasitic infection of the CNS is cerebral malaria, followed by neurocysticercosis. Other relatively frequent infections are toxoplasmosis, cystic and alveolar echinococcosis and schistosomiasis [g4] [AR5].

Post COVID-19, there has been renewed interest in understanding the pathogens challenging the human health and evaluate our preparedness towards dealing with health challenges in future. In this endeavour, it is not only the bacteria and the viruses, but a greater community of pathogens. Such pathogenic microorganisms, include protozoa, fungi and worms, which establish a distinct variety of disease-causing agents with the capability to impact the host’s well-being as well as the equity of ecosystem. This review summarises the peculiar characteristics and pathogenic mechanisms utilized by these disease-causing organisms. It features their role in causing infection in the concerned host and emphasizes the need for further research. Understanding the layers of pathogenesis encompassing the concerned infectious microbes will help expand targeted inferences with relation to the cause of the infection. This would strengthen and augment benefit to the host’s health along with the maintenance of ecosystem network, exhibiting host-pathogen interaction cycle. This would be key to discover the layers underlying differential disease severities in response to similar/same pathogen infection.

Abstract The health, growth and fertility of avian species can be negatively affected by parasite infection. This survey assesses the presence, variety and distribution of internal and external parasites among parrots and perching birds in Mashhad, Iran. This study examined 751 caged pet birds from different species and regions in Mashhad for faecal samples and 132 oral swabs for digestive tract parasites. Furthermore, skin scrapings were conducted on 14 canaries displaying dishevelled feathers. During the study, mortalities and moribund birds that had been referred underwent necropsies to examine internal parasites. Following the formol ether faecal examination, only one Malango parrot tested positive for Heterakoidea eggs among 751 faecal samples (0.13%). Further, one cockatiel showed evidence of parasitic infection with Eimeria spp. (0.13%). However, neither Cryptosporidium nor Giardia protozoa were detected in the samples (0%). Oral swabs revealed no evidence of Trichomonas (0%). On the other hand, skin scraping revealed the presence of the mite Dermanyssus in 7 out of 14 canaries with dishevelled feathers (50%). Of 25 moribund and weak budgerigars, 2 were infected with Acuaria in their proventriculus (8%). In addition, 3 out of 14 deceased myna birds carried the nematode Diplotriana in their coelomic cavities (21.42%). In conclusion, the rate of internal parasites has been relatively low in ornamental birds of Mashhad, whereas the prevalence of external parasites has been higher.

Introduction: Soil ciliates, as protozoa, play a crucial role in biogeochemical cycling and the soil food web, yet they are highly sensitive to environmental fluctuations in soil conditions. The diversity and biogeographic characteristics of soil ciliates in the Tibetan Plateau remain poorly understood. As part of a regional survey focused on soil ciliate diversity, we investigated the composition and spatiotemporal variations of soil ciliate communities along the Yarlung Zangbo River, a representative soil habitat in the Tibetan Plateau.Methods: A total of 290 soil samples were collected from four habitat types of grassland, shrubland, forestland and wetland in the middle reaches of the Yarlung Zangbo River during the wet and dry seasons, and 138 species of ciliates were identified.Results: Soil ciliate diversity exhibited greater variation across habitat types than seasons. Moreover, soil ciliate diversity was higher during the wet season compared to the dry season, with the wetland habitat showing the highest diversity and the grassland habitat displaying the lowest. We observed spatiotemporal heterogeneity in the composition of soil ciliate communities across different seasons and habitat types. Notably, Litostomatea, Karyorelictea, and Prostomatea predominated in ciliate communities during the wet season and in grassland habitat. Phyllopharyngers dominated during dry seasons and in forested regions, while Spirotrichea species were prevalent in wetland and forested areas. The co-occurrence network analysis showed that soil ciliate community was more complex in wet season than in dry season, and the stability of soil ciliate community in wet season was higher than that in dry season. The stability of soil ciliate community in wetland was higher than that in forestland, shrubland and grassland, and the anti-interference ability was stronger. Soil temperature (ST), Total nitrogen (TN), Soil organic matter (SOM) and Soil water content (SWC) are important factors affecting the structure of soil ciliate community. By influencing the metabolic rate and nutrient acquisition of soil ciliates, the distribution pattern of soil ciliate community diversity in the middle reaches of Yarlung Zangbo River is shaped.Discussion: In summary, this study revealed the distribution pattern of soil ciliate community diversity in the Yarlung Zangbo River Basin, and the key factors affecting the spatial and temporal differences and stability of the community, enhancing our understanding of how ciliates adapt to environmental conditions in soil habitats across the Tibetan Plateau.

Outbreaks of neurological disease associated with Sarcocystis calchasi have been observed in captive and free-ranging rock pigeons (Columba livia) in Europe and the United States as well as in wild Brandt's cormorants (Phalacrocorax penicillatus) and captive psittacines in California, USA. Experimental and field studies have identified northern goshawks (Accipiter gentilis) and European sparrowhawks (A. nisus) as definitive hosts in Europe while the definitive hosts elsewhere remain unknown. In this study, we aimed to identify the potential definitive host(s) of S. calchasi through molecular analysis of intestinal samples from seven predatory (n = 85) and one omnivorous (n = 11) bird species in California. In total, apicomplexan-generic 28S rRNA PCR products were obtained and sequenced for 42 raptors. Three of 16 (18.8%) Cooper's hawks (A. cooperii) and two of 26 (5.6%) red-tailed hawks (Buteo jamaicensis) also tested positive for the S. calchasi-specific ITS1 PCR and sequencing of the 28S rRNA PCR product was 100% homologous to S. calchasi. In addition to S. calchasi (5.9%; 5/85), other Sarcocystis spp. detected in raptors included: S. jamaicensis (21.2%; 18/85), S. columbae (8.2%; 7/85), S. turdusi (7.1%; 6/85), and S. halieti (4.7; 4/85%). Infections with closely related S. jamaicensis and S. (Frenkelia) microti (9.4%; 8/85) could not be distinguished for eight raptors. Eumonospora henryae (1.2%; 1/85) was detected in one raptor. Our results indicate for the first time that S. calchasi may have a definitive host range in North America that includes at least two raptors, Cooper's hawks and red-tailed hawks, within the family Accipitridae.

The aim of this study was to document a case of parasite co-infection in a captive Bactrian camel and to supply morphometric data of the found pathogens. It concerned a 20-year-old male animal inhabiting Sofia Zoo, Bulgaria. A decreased appetite and gastrointestinal disorders were observed in it during the summer of 2022. Improvement in the animal’s condition was achieved after the administration of antibiotics, sulfonamides, and other symptomatic medicines. However, two weeks after treatment, clinical symptoms reappeared. Then, a diarrheal fecal sample from the animal was subjected to parasitological examination by direct smear and flotation and sedimentation techniques. Multiple infections by helminths (Trichostrongylus sp., Haemonchus sp., Oesophagostomum sp., Trichuris sp., and Dicrocoelium sp.), ciliates (Buxtonella cameli), and protozoa (Eimeria cameli) were found, with E. cameli being reported for the first time in zoo conditions. Deworming led to the recovery of the general condition and appearance of the animal’s feces, but two weeks later, it died suddenly. We considered that the parasitic infection was not the direct cause of the fatal outcome, and its presence, other health disorders, and the advanced age of the animal were among the contributing factors. This case reveals the need to combine planned preventive deworming with routine parasitological diagnostics to take timely and targeted actions to protect the health of animals inhabiting zoo facilities.

Abstract Background Microbial populations in the rumen play an essential role in the degradation of Cellulosic dietary components and in providing nutrients to the host animal. Objective This study aims to detect the effect of pomegranate seed pulp (PSP) on rumen fermentation, digestibility and methanogens and the protozoa population (by competitive polymerase chain reaction [PCR]) of the camel and goat rumen fluid. Materials and methods PSP was added to the experimental treatments and replaced by wheat bran (0%, 5% and 10%). Rumen fluid was collected from three goats and two camels according to the similarity of sex, breed, origin and time and used for three gas production studies. DNA extraction was performed by the RBB + c method, the ImageJ programme calculated band intensities (target and competing DNA), and line gradients were plotted based on the number of copies and intensity. Results Our result showed that diets did not significantly affect the methanogen and protozoa population. Animal species affected microbial populations so that both populations in camels were less than goats. The production of gas and volatile fatty acids was not affected by diets. These two parameters and NH3 concentration and methane production in goats were higher than in camel. The pH of digested dry matter and microbial protein in camels was higher than in goats. Conclusions Therefore, the competitive PCR technique is an effective method for enumerating rumen microbiota. This supplementation can be considered a strategy to achieve performance and environmental benefits.

The purpose of the research is to review the classical methods of coprological diagnostics, taking into account modern recommendations and proposals. Materials and methods. Literature sources on the issues of coprological methods of diagnosing parasitosis have been studied. Results and discussion. Diagnosis during life is an important element of practical veterinary activity, the success of which largely depends on the correct choice and competent use of suitable laboratory methods. For the diagnosis of parasitic diseases, the study of feces is most often used. Coprological methods are highly informative due to the fact that they can detect not only parasites of the digestive tract, liver, pancreas, but also a number of helminths of the respiratory system and even some tissue parasites that excrete eggs or larvae with feces. Unlike existing indirect immunological and molecular genetic methods, classical research methods are based on the detection and identification of the pathogens themselves at different stages of development. At the same time, the researcher has the opportunity to study morphological features and conduct morphometric analysis. The widespread use of classical coprological methods is also associated with their availability, they are quite simple to perform and relatively inexpensive. A limitation of coprological methods is the inability to diagnose pathogens that are at a non-propagative stage. The effectiveness of diagnostics is influenced by the method of collecting samples, the terms and conditions of transportation, and the choice of an adequate method for examining samples. The article presents the rules for sampling and options for the use of fixatives, a classification of coprological methods is given, and the procedure for performing research is described. Special attention is paid to the choice of flotation solutions, the method of cultivation of strongylid larvae and the method of sporulation of coccidia oocysts. Attention is drawn to the importance of micrometric measurements of detected objects. To ensure the quality of the conducted coprological studies, it is necessary to control the stages of the studies, the technical condition of the equipment and train the personnel. The material presented in the article is intended for both students and practitioners.

Background: Intestinal protozoa are potential diarrhea-causing pathogens and monitored worldwide. The Korea Centers for Disease Control and Prevention has also been monitoring intestinal protozoa causing diarrhea for many years. Recently, the overall protozoa detection rate has decreased to less than 1%, but whether protozoa infection causing diarrhea has declined or is being underestimated has not been studied. This study aimed to investigate the molecular epidemiology of intestinal protozoan pathogens in stool samples collected from multiple Korean centers. Methods: Stool samples were collected from five university hospitals and a commercial laboratory. Direct smear and trichrome staining were performed on all samples. The presence of Cryptosporidium parvum, Giardia lamblia, Entamoeba histolytica, Cyclospora cayetanensis, Dientamoeba fragilis, and Blastocystis hominis were detected using Allplex™ Gastrointestinal Parasite Assays (Seegene Inc., Korea). Microsporidia species and Kudoa septempunctata were detected using PowerChek™ Microsporidia Multiplex and Kudoa Real-time PCR kits (Kogene Biotech, Korea), respectively. Results: The collected samples included 279 diarrheal and 51 non-diarrheal samples. Among the 279 diarrheal samples, nine samples [B. hominis (n=7), C. parvum (n=1), and Microporidia species (n=1)] were positive, but there were no positive samples for K. septempunctata. We could not detect any protozoa by direct smear and trichrome staining. Among the 51 nondiarrheal samples, 10 (19.6%) samples were positive for B. hominis, but no other protozoa were observed. Conclusion: This multicenter study showed that the detection rate of intestinal protozoa is currently low in diarrheal samples from Korea. However, B. hominis was frequently detected in non-diarrheal samples, indicating their low pathogenicity.

Abstract Background Toxoplasma gondii is a protozoan parasite that infects a large spectrum of warm-blooded animals, including humans. Small rodents and insectivores play an important role in the epidemiology of T. gondii and may serve as a source of infection for both, domestic and wild definitive felid hosts. Factors influencing the occurrence of T. gondii in wild small mammals are unknown, despite the fact that many intermediate host species are identified. We have used small mammals (Rodentia and Lipotyphla) captured over two years in various habitats, both in urbanised and in natural landscapes. We assessed the importance of land-use, season and host ecology on T. gondii infection. Results We examined 471 individuals belonging to 20 small mammal species, collected at 63 locations spread over wide altitude, habitat and land-use ranges from Romania. Heart tissue samples were individually analysed by PCR targeting the 529 bp repetitive DNA fragment of T. gondii. The overall prevalence of infection was 7.3%, with nine species of rodents and two species of shrews being found to carry T. gondii DNA. Five species showed high frequency of infection, with the highest prevalence found in Myodes glareolus (35.5%), followed by Spermophilus citellus (33.3%), Sorex minutus (23.1%), S. araneus (21.7%) and Micromys minutus (11.1%). Adults seemed more often infected than young, however when controlling for season, the difference was not significant, as in spring both adults and young showed higher infection rates, but more adults were sampled. Contrary to our expectations, urban/rural areas (with their implicit high density of domestic feline presence) had no effect on infection prevalence. In addition, neither habitat, nor land-use at sampling sites was important as only geographical location and host species were contributing factors to the infection risk. Conclusions High prevalence of T. gondii infection showed a highly localised, patchy occurrence, with long living and higher mobility host species being the most common carriers, especially during autumn.

Digital PCR (dPCR) is an emerging technology that enables the absolute quantification of the targeted nucleic acids. The body of research on the potential applications of this novel tool is growing in human and veterinary medicine. Most of the research on dPCR applications in veterinary parasitology is concentrated on developing and validating new assays to detect and quantify parasites of great financial impact in the food-producing animal industry. Several studies describe the utility of dPCR for individualized medicine in companion animals. Most frequently, dPCR performance is assessed compared to quantitative PCR or Next Generation Sequencing platforms, while others also compare the accuracy of dPCR with traditional parasitological techniques considered gold standard methods. Other researchers describe dPCR assays for surveillance purposes, species identification, and quantification in mixed parasitic infections, the detection of mutations indicative of anthelmintic resistance, and the identification of new targets for drug development. This review provides an overview of the studies that employed dPCR in investigating animal parasites and parasitic diseases from a veterinary perspective and discusses how this novel technology could advance and facilitate diagnosis, surveillance, and the monitoring of response to treatment, or shed light on current gaps in our knowledge of the epidemiology of significant veterinary parasitic diseases.

ABSTRACT Cryptosporidium spp. are important enteropathogen protozoan parasites that infect humans and other animals throughout the world. Cryptosporidiosis in cattle industry leads to considerable economic losses due to diarrhea, dehydration, growth retardation, weight loss, and possibly death, however, data on the occurrence ofCryptosporidiumspp. in cattle in Colombia are limited. Therefore, this study aimed to investigate the occurrence and possible factors associated to the excretion of Cryptosporidium spp. oocyst in pre-weaned calves from dairy farms in Northern Antioquia, Colombia. In addition, Sheather’s sugar floatation (SSF), and Modified Ziehl-Neelsen staining (MZN) methods were compared. A total of 41 fecal samples were collected from calves between 1 and 92 days of age of which 23 were positive (56.1%). Crossbreed calves were nine times less susceptible than purebred dairy cattle to excretion of Cryptosporidium spp. oocysts (OR=0.10). MZN was the best technique for the detection of oocysts in fecal samples, however, the mean number of days to detect cryptosporidial oocysts was lower for the SSF method. Cryptosporidium is widespread among calves under 2 months of age in dairy herds of Northern Antioquia, although further investigations considering a greater number of farms and animals are necessary.

Objective:The aim of the present study was to determine the prevalence of parasitic infections in aquarium fish sold in local pet shops.Methods:Totally 502 fish samples from 8 species were obtained from 7 pet shops. Native preparations were prepared from the gills, fins, skin, intestines, and internal organs of fish and examined under a light microscope.Results:It was detected at the end of the study that 62.7% of the fishes examined were infected with one or more parasite species. Among the fish examined, 28.9% were infected with a single parasite, whereas 33.9% were infected with mixed parasites. Chilodonella sp. identified as the most common species. Other species detected included Trichodina sp., Piscinoodinium sp., Ichthyobodo sp., Ichthyophthirius multifiliis, Apiosoma sp., Epistylis sp., Vorticella sp., Gyrodactylus sp., Dactylogyrus sp., Capillaria sp., Camallanus sp., metacercariae, and nematode larvae, Argulus sp., Philodina sp., Euchlanis sp., Aelosoma sp., and Tetrahymena.Conclusion:The importance of the present study is that highest number of parasite species were detected in aquarium fish sold in pet shops in Türkiye. The aquarium owners should pay attention to the cleanliness of the aquarium water in order to reduce the rate of parasitic infection in fish; when new fish are purchased, they should be taken from reliable sources, and the quarantine process should be observed.

Metronidazole (MTZ) is a clinically important antimicrobial agent that is active against both bacterial and protozoan organisms. MTZ has been used extensively for more than 60 years and until now resistance has been rare. However, a recent and dramatic increase in the number of MTZ resistant bacteria and protozoa is of great concern since there are few alternative drugs with a similarly broad activity spectrum. To identify key factors and mechanisms underlying MTZ resistance, we utilized the protozoan parasite Giardia intestinalis, which is commonly treated with MTZ. We characterized two in vitro selected, metronidazole resistant parasite lines, as well as one revertant, by analyzing fitness aspects associated with increased drug resistance and transcriptomes and proteomes. We also conducted a meta-analysis using already existing data from additional resistant G. intestinalis isolates. The combined data suggest that in vitro generated MTZ resistance has a substantial fitness cost to the parasite, which may partly explain why resistance is not widespread despite decades of heavy use. Mechanistically, MTZ resistance in Giardia is multifactorial and associated with complex changes, yet a core set of pathways involving oxidoreductases, oxidative stress responses and DNA repair proteins, is central to MTZ resistance in both bacteria and protozoa.

Leishmaniasis is a group of tropical diseases caused by protozoa and transmitted by vectors. It is a significant public health issue in many countries. Current chemotherapy options have limitations, and researchers are exploring new therapeutic approaches. Studies have investigated immune responses, B cells, and leukotrienes in the disease and identified potential targets for treatment. The document discusses various studies on the use of natural products, drug repurposing, combination therapies, and alternative treatment protocols. The research is supported by various organizations and funding sources, emphasizing the importance of finding effective treatments for leishmaniasis. [Extracted from the article]

Introduction: According to the guidelines (GL) valid in Germany, persons born or raised in a malaria-endemic area or had continuously stayed in a malaria-endemic area for more than 6 months may only be admitted donating blood if, among other things, validated and quality-assured laboratory diagnostics show that there is no evidence of infectivity. In a statement of the Working Group "Blood" of the Federal Ministry of Health (WGB), a reduction of the deferral period from 4 to 3 years and an antibody test after the deferral period are recommended. Methods: In accordance with the GL, nucleic acid testing (NAT) by means of PCR is carried out at our institution after a retention period of 4 years. In addition to the validated molecular biological testing, an infection serological examination was performed. Case Presentation: In the present cases, Plasmodia genome was detected in the respective single PCR in two blood donors originating from malaria-endemic areas after the expiry of the deferral period. However, one donor tested negative for antibodies against Plasmodia. Discussion/Conclusion: This observation is discussed in the context of a recommendation of the WGB. The question is addressed whether PCR testing is dispensable or whether a combination of infection serological testing and NAT should be favored. [ABSTRACT FROM AUTHOR]

Parasitism by protozoa and monogenean flatworms in freshwater fish from Romania was studied by collecting and examining samples from two major river systems there: 183 fish from 17 species from the Olt River and its tributaries; and 155 fish from 16 species from the Mureș River and its tributary, Târnava Mare. The average rates of parasitism in the samples from the two rivers and their tributaries were as follows: Ichthyiophthirius multifiliis (2%), Trichodina spp. (21%), Apiosoma spp. (18%), Mixobolus spp. (8%), Dactylogyrus spp. (9%), and Gyrodactylus spp. (10%). The number of parasite species varied from one river to another. I. multifiliis was found in only 3 fish species, Trichodina spp. in 13 species, Glosatella spp. in 6 species, and Mixobollus spp., Dactylogyrus spp., and Gyrodactylus spp. in 7 different species each. The highest number of parasite species (six) were identified in the European chub (Squalius cephalus) and schneider (Alburnoides bipunctatus), which seem more susceptible to parasitic infections. The aquatic environment of these rivers may represent a source of parasites for fish from neighboring countries through which these rivers pass.

Intestinal parasites, including helminths and protozoa, account for a significant portion of the global health burden. The gastrointestinal (GI) tract not only serves as the stage for these parasitic infections but also as the residence for millions of microbes. As the intricacies of the GI microbial milieu continue to unfold, it is becoming increasingly apparent that the interactions between host, parasite, and resident microbes help dictate parasite survival and, ultimately, disease outcomes. Across both clinical and experimental models, intestinal parasites have been shown to impact microbial composition and diversity. Reciprocally, microbes can directly influence parasitic survival, colonization and expulsion. The gut microbiota can also indirectly impact parasites through the influence and manipulation of the host. Studying this host–parasite–microbiota axis may help bring about novel therapeutic strategies for intestinal parasitic infection as well as conditions such as inflammatory bowel disease (IBD). In this review, we explore the relationship between intestinal parasites, with a particular focus on common protozoa and helminths, and the gut microbiota, and how these interactions can influence the host defence and intestinal immune response. We will also explore the impact of this tripartite relationship in a clinical setting and its broader implications for human health.

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.

Reactive oxygen species (ROS) contain at least one oxygen atom and one or more unpaired electrons and include singlet oxygen, superoxide anion radical, hydroxyl radical, hydroperoxyl radical, and free nitrogen radicals. Intracellular ROS can be formed as a consequence of several factors, including ultra-violet (UV) radiation, electron leakage during aerobic respiration, inflammatory responses mediated by macrophages, and other external stimuli or stress. The enhanced production of ROS is termed oxidative stress and this leads to cellular damage, such as protein carbonylation, lipid peroxidation, deoxyribonucleic acid (DNA) damage, and base modifications. This damage may manifest in various pathological states, including ageing, cancer, neurological diseases, and metabolic disorders like diabetes. On the other hand, the optimum levels of ROS have been implicated in the regulation of many important physiological processes. For example, the ROS generated in the mitochondria (mitochondrial ROS or mt-ROS), as a byproduct of the electron transport chain (ETC), participate in a plethora of physiological functions, which include ageing, cell growth, cell proliferation, and immune response and regulation. In this current review, we will focus on the mechanisms by which mt-ROS regulate different pathways of host immune responses in the context of infection by bacteria, protozoan parasites, viruses, and fungi. We will also discuss how these pathogens, in turn, modulate mt-ROS to evade host immunity. We will conclude by briefly giving an overview of the potential therapeutic approaches involving mt-ROS in infectious diseases.

Saiga antelope and Turkmenian kulans are considered critically endangered and near threatened, respectively, by the International Union for Conservation of Nature (IUCN). Due to these species’ fragile status, it is important to understand the pathogens infecting their remaining populations. A total of 496 faecal samples were collected from Ural saiga antelope in western Kazakhstan during June, September, and November of 2021 and May and August of 2022 and 149 faecal samples were collected from kulans in the Altyn-Emel nature reserve in south-eastern Kazakhstan from June to August of 2021. Additionally, endo- and ecto-parasites were collected from 17 saiga that were found deceased due to natural causes. Nine helminths (3 cestodes, 6 nematodes) and two protozoans were found in Ural saiga antelope. In addition to intestinal parasites, one case of cystic echinococcosis due to Echinococcus granulosus infection and one case of cerebral coenurosis due to Taenia multiceps infection was identified on necropsy. None of the collected ticks (all Hyalomma scupense) were found positive for Theileria annulate (enolase gene) or Babesia spp. (18 S ribosomal RNA gene) via PCR. Three intestinal parasites (Parascaris equorum, Strongylus sp., and Oxyuris equi) were found in kulans. All identified parasites, in both saiga and kulans, are also found in domesticated livestock, suggesting a need for better understanding of how parasites are maintained within and between regional wild and domestic ungulate populations.

Background and Aim: Herbal plants have the potential to reduce the population of metagonic bacteria and protozoa due to the bioactive compound contained in herbal plants. This study aimed to evaluate the effect of herbal plant supplementation on rumen fermentation characteristics, methane (CH4) gas emissions, in vitro nutrient digestibility, and protozoan populations. Materials and Methods: This study consisted of two stages. Stage I involved determining the potential of herbal plants to increase total gas production (Orskov and McDonald methods) and reduce the protozoan population (Hristov method). Three potential herbs were selected at this stage and used in Stage II as supplements in the palm kernel cake (PKC)-based diet (30% herbal plants + 70% PKC). Proximate and Van Soest analyses were used to determine the chemical composition. In vitro dry matter digestibility (IVDMD), organic matter (IVOMD), and rumen fermentation characteristics were determined using Theodorous method. Conway microdiffusion was used to determine ammonia concentration (NH3). Gas chromatography was used to determine the total and partial volatile fatty acid production. Results: The results of the first stage showed that seven herbal plants (Moringa oleifera, Rhodomyrtus tomentosa, Clerodendron serratum, Curcuma longa Linn., Urena lobata, Uncaria, and Parkia timoriana) significantly differed in terms of total gas production (p < 0.05). Herbal plants can increase gas production and reduce protozoan populations. The highest total gas production was observed using P. timoriana, M. oleifera, and C. longa Linn. Moringa oleifera plants were the most effective in lowering protozoa population. In Stage 2, the supplementation of herbal plants in PKC-based-diet significantly increased IVDMD, that was ranged from 56.72% to 65.77%, IVOMD that was ranged from 52.10% to 59.54%, and NH3, that was ranged from 13.20 mM to 17.91 mM. Volatile fatty acid partial and total gas production potential and CH4> gas emissions were also significantly different from those of the control (p < 0.05). Conclusion: Supplementation of M. oleifera, C. longa Linn., and P. timoriana in ruminant diet effectively increased total gas production, IVDMD percentage, and IVOMD, and reduced CH4 gas emissions and protozoa populations during rumen fermentation.

Cooks have an important role in the dissemination and transmission of enteropathogenic microorganisms, including intestinal parasites such as Blastocystis sp. responsible for blastocystosis.

ABSTRACT This study investigated the change in the microbiome of tomato rhizosphere soils after the invasion of Ralstonia solanacearum and analyzed the correlation between microbes and soil physicochemical properties. Diversity analyses of the bacteria in healthy and diseased rhizosphere soil samples (HRS and DRS) revealed that HRS had a higher species diversity and were compositionally different from DRS (P ≤ 0.05). Substantial differences in the relative abundance of Actinobacteria (37.52% vs 28.96%, P ≤ 0.05) and Proteobacteria (29.20% vs 35.59%, P ≤ 0.05) were identified in HRS and DRS, respectively. Taxonomic composition analysis showed ten differentially abundant genera, and seven of them (Gaiella, Roseisolibacter, Solirubrobacter, Kribbella, Acidibacter, Actinomarinicola, and Marmoricola) are more abundant in HRS. Soil pH and enzyme activities were negatively correlated with the abundance of R. solanacearum. The contents of total nitrogen (TN), total phosphorus (TP), total potassium (TK), alkaline nitrogen (alkaline N), available phosphorus (AP), available potassium (AK), NO3—N(NN), NH4+-N (AN), and organic matter (OM) were all significantly increased in DRS. The composition and richness of protozoa in the samples show significant differences. Cephalobus, Acrobeles, Heteromita, norank_Tylenchida, and Rotylenchulus were enriched in DRS. Microbial interaction networks revealed that the HRS networks were more complex than the DRS networks. Overall, the results of this study demonstrate that healthy soil has a more complex microbial community structure and higher enzyme activity, and the invasion of R. solanacearum damages the soil microbial system.IMPORTANCEHow does the invasion of Ralstonia solanacearum affect tomato rhizosphere bacteria and protozoa? Which microbial changes can affect the growth of R. solanacearum? To date, most research studies focus on bacteria, with little research on protozoa, and even less on the synergistic effects between protozoa and bacteria. Here, we analyzed the correlation between tomato rhizosphere bacterial and protozoan communities and soil physicochemical properties during the invasion of R. solanacearum. We found that the diversity and abundance of rhizosphere microorganisms in healthy rhizosphere soil samples (HRS) were significantly higher than those in diseased rhizosphere soil samples (DRS), and there were significant changes in soil pH and enzyme activity. Overall, in this study, the analysis of microbial changes during the invasion of R. solanacearum provides a theoretical basis for the prevention and control of bacterial wilt.

Waterborne diseases such as cryptosporidiosis and giardiasis are among the leading public health problems worldwide because these protozoa are more resistant to water disinfection methods. This study aimed to evaluate the presence of oocysts of Cryptosporidium spp. and cysts of Giardia spp. in samples of raw and treated water. During one year, 24 raw and treated water samples were collected from the public water supply system in Canavieiras, Bahia, Brazil. Cysts and oocysts were concentrated using membrane filtration and visualized through Direct immunofluorescence assay (DFA). DNA isolated with a commercial kit was subjected to nested PCR (nPCR). The DFA obtained a better result than the nPCR in detecting both protozoa. By DFA, Cryptosporidium spp. was present in 25% (6/24) of the raw water and 4.2% (1/24) of the treated water samples. Giardia spp. was detected in 41.6% (10/24) of the raw water and 16.6% (4/24) of the treated water samples. In nPCR, Giardia spp. was detected in treated and raw water at 16.6% (4/24) and 16.6% (4/24), respectively. Cryptosporidium spp. was detected in raw water at 8.3% (2/24). The DFA and nPCR results differed despite the molecular analysis having greater sensitivity. Many inhibitors in the samples may have influenced the result. The physicochemical parameters pH and turbidity were not under the standards established by the legislation. In raw water, microbiological parameters were above the required legal standard. Precipitation and pH correlated with Cryptosporidium nPCR results in raw water. The presence of these protozoa in the water samples constitutes a potential risk for transmitting diarrheal diseases to this municipality's human and animal populations.

Abstract Highly diverse exoenzymes mediate the energy flow from substrates to the multitrophic microbiota within the soil decomposer micro‐food web. Here, we used a “soil enzyme profile analysis” approach to establish a series of enzyme profile indices; those indices were hypothesized to reflect micro‐food web features. We systematically evaluated the shifts in enzyme profile indices in relation to the micro‐food web features in the restoration of an abandoned cropland to a natural area. We found that enzymatic C:N stoichiometry and decomposability index were significantly associated with substrate availability. Furthermore, the higher Shannon diversity index in the exoenzyme profile, especially for the C‐degrading hydrolase, corresponded to a greater microbiota community diversity. The increased complexity and stability of the exoenzyme network reflected similar changes with the micro‐food web networks. In addition, the gross activity of the enzyme profile as a parameter for soil multifunctionality, effectively predicted the substrate content, microbiota community size, diversity, and network complexity. Ultimately, the proposed enzymic channel index was closely associated with the traditional decomposition channel indices derived from microorganisms and nematodes. Our results showed that soil enzyme profile analysis reflected very well the decomposer food web features. Our study has important implications for projecting future climate change or anthropogenic disturbance impacts on soil decomposer micro‐food web features by using soil enzyme profile analysis.

ABSTRACT The zoonotic Cryptosporidium parvum is a global contributor to infantile diarrheal diseases and opportunistic infections in immunocompromised or weakened individuals. Like other apicomplexans, it possesses several specialized secretory organelles, including micronemes, rhoptry, and dense granules. However, the understanding of cryptosporidial micronemal composition and secretory pathway remains limited. Here, we report a new micronemal protein in C. parvum, namely, thrombospondin (TSP)-repeat domain-containing protein-4 (CpTSP4), providing insights into these ambiguities. Immunostaining and enzyme-linked assays show that CpTSP4 is prestored in the micronemes of unexcysted sporozoites but secreted during sporozoite excystation, gliding, and invasion. In excysted sporozoites, CpTSP4 is also distributed on the two central microtubules unique to Cryptosporidium. The secretion and microtubular distribution could be completely blocked by the selective kinesin-5 inhibitors SB-743921 and SB-715992, resulting in the accumulation of CpTSP4 in micronemes. These support the kinesin-dependent microtubular trafficking of CpTSP4 for secretion. We also localize γ-tubulin, consistent with kinesin-dependent anterograde trafficking. Additionally, recombinant CpTSP4 displays nanomolar binding affinity to the host cell surface, for which heparin acts as one of the host ligands. A novel heparin-binding motif is identified and validated biochemically for its contribution to the adhesive property of CpTSP4 by peptide competition assays and site-directed mutagenesis. These findings shed light on the mechanisms of intracellular trafficking and secretion of a cryptosporidial micronemal protein and the interaction of a TSP-family protein with host cells.IMPORTANCECryptosporidium parvum is a globally distributed apicomplexan parasite infecting humans and/or animals. Like other apicomplexans, it possesses specialized secretory organelles in the zoites, in which micronemes discharge molecules to facilitate the movement and invasion of zoites. Although past and recent studies have identified several proteins in cryptosporidial micronemes, our understanding of the composition, secretory pathways, and domain-ligand interactions of micronemal proteins remains limited. This study identifies a new micronemal protein, namely, CpTSP4, that is discharged during excystation, gliding, and invasion of C. parvum sporozoites. The CpTSP4 secretion depends on the intracellular trafficking on the two Cryptosporidium-unique microtubes that could be blocked by kinesin-5/Eg5 inhibitors. Additionally, a novel heparin-binding motif is identified and biochemically validated, which contributes to the nanomolar binding affinity of CpTSP4 to host cells. These findings indicate that kinesin-dependent microtubular trafficking is critical to CpTSP4 secretion, and heparin/heparan sulfate is one of the ligands for this micronemal protein.

Cancer remains a significant global health issue, despite advances in screening and treatment. While existing tumor treatment protocols such as surgery, chemotherapy, radiotherapy, targeted therapy, and immunotherapy have proven effective in enhancing the prognosis for some patients, these treatments do not benefit all patients. Consequently, certain types of cancer continue to exhibit a relatively low 5-year survival rate. Therefore, the pursuit of novel tumor intervention strategies may help improve the current effectiveness of tumor treatment. Over the past few decades, numerous species of protozoa and their components have exhibited anti-tumor potential via immune and non-immune mechanisms. This discovery introduces a new research direction for the development of new and effective cancer treatments. Through in vitro experiments and studies involving tumor-bearing mice, the anti-tumor ability of Toxoplasma gondii, Plasmodium, Trypanosoma cruzi, and other protozoa have unveiled diverse mechanisms by which protozoa combat cancer, demonstrating encouraging prospects for their application. In this review, we summarize the anti-tumor ability and anti-tumor mechanisms of various protozoa and explore the potential for their clinical development and application.

Soil bacterial and fungal communities play key roles in the degradation of organic contaminants, and their structure and function are regulated by bottom-up and top-down factors. Microbial ecological effects of polycyclic aromatic hydrocarbons (PAHs) and trophic interactions among protozoa and bacteria/fungi in PAH-polluted soils have yet to be determined. We investigated the trophic interactions and structure of the microbiome in PAH-contaminated wasteland and farmland soils. The results indicated that the total concentration of the 16 PAHs (∑PAHs) was significantly correlated with the Shannon index, NMDS1 and the relative abundances of bacteria, fungi and protozoa (e.g., Pseudofungi) in the microbiome. Structural equation modelling and linear fitting demonstrated cascading relationships among PAHs, protozoan and bacterial/fungal communities in terms of abundance and diversity. Notably, individual PAHs were significantly correlated with microbe-grazing protozoa at the genus level, and the abundances of these organisms were significantly correlated with those of PAH-degrading bacteria and fungi. Bipartite networks and linear fitting indicated that protozoa indirectly modulate PAH degradation by regulating PAH-degrading bacterial and fungal communities. Therefore, protozoa might be involved in regulating the microbial degradation of PAHs by predation in contaminated soil.

IntroductionThis study explores if promoting a complex rumen microbiota represents an advantage or a handicap in the current dairy production systems in which ruminants are artificially reared in absence of contact with adult animals and fed preserved monophyte forage.MethodsIn order to promote a different rumen microbial diversity, a total of 36 newborn goat kids were artificially reared, divided in 4 groups and daily inoculated during 10 weeks with autoclaved rumen fluid (AUT), fresh rumen fluid from adult goats adapted to forage (RFF) or concentrate (RFC) diets, or absence of inoculation (CTL). At 6 months of age all animals were shifted to an oats hay diet to determine their ability to digest a low quality forage.Results and discussionEarly life inoculation with fresh rumen fluid promoted an increase in the rumen overall microbial diversity which was detected later in life. As a result, at 6 months of age RFF and RFC animals had higher bacterial (+50 OTUs) and methanogens diversity (+4 OTUs) and the presence of a complex rumen protozoal community (+32 OTUs), whereas CTL animals remained protozoa-free. This superior rumen diversity and presence of rumen protozoa had beneficial effects on the energy metabolism allowing a faster adaptation to the forage diet, a higher forage digestion (+21% NDF digestibility) and an energetically favourable shift of the rumen fermentation pattern from acetate to butyrate (+92%) and propionate (+19%) production. These effects were associated with the presence of certain rumen bacterial taxa and a diverse protozoal community. On the contrary, the presence of rumen protozoa (mostly Entodinium) had a negative impact on the N metabolism leading to a higher bacterial protein breakdown in the rumen and lower microbial protein flow to the host based on purine derivatives urinary excretion (-17% to -54%). The inoculation with autoclaved rumen fluid, as source of fermentation products but not viable microbes, had smaller effects than using fresh inoculum. These findings suggest that enhancing rumen microbial diversity represents a desirable attribute when ruminants are fed forages in which the N supply does not represent a limiting factor for the rumen microbiota.

Bats carry many zoonotic pathogens without showing pronounced pathology, with a few exceptions. The underlying immune tolerance mechanisms in bats remain poorly understood, although information-rich omics tools hold promise for identifying a wide range of immune markers and their relationship with infection. To evaluate the generality of immune responses to infection, we assessed the differences and similarities in serum proteomes of wild vampire bats (Desmodus rotundus) across infection status with five taxonomically distinct pathogens: bacteria (Bartonella spp., hemoplasmas), protozoa (Trypanosoma cruzi), and DNA (herpesviruses) and RNA (alphacoronaviruses) viruses. From 19 bats sampled in 2019 in Belize, we evaluated the up- and downregulated immune responses of infected versus uninfected individuals for each pathogen. Using a high-quality genome annotation for vampire bats, we identified 586 serum proteins but found no evidence for differential abundance nor differences in composition between infected and uninfected bats. However, using receiver operating characteristic curves, we identified four to 48 candidate biomarkers of infection depending on the pathogen, including seven overlapping biomarkers (DSG2, PCBP1, MGAM, APOA4, DPEP1, GOT1, and IGFALS). Enrichment analysis of these proteins revealed that our viral pathogens, but not the bacteria or protozoa studied, were associated with upregulation of extracellular and cytoplasmatic secretory vesicles (indicative of viral replication) and downregulation of complement activation and coagulation cascades. Additionally, herpesvirus infection elicited a downregulation of leukocyte-mediated immunity and defense response but an upregulation of an inflammatory and humoral immune response. In contrast to our two viral infections, we found downregulation of lipid and cholesterol homeostasis and metabolism with Bartonella spp. infection, of platelet-dense and secretory granules with hemoplasma infection, and of blood coagulation pathways with T. cruzi infection. Despite the small sample size, our results suggest that vampire bats have a similar suite of immune mechanisms for viruses distinct from responses to the other pathogen taxa, and we identify potential biomarkers that can expand our understanding of pathogenesis of these infections in bats. By applying a proteomic approach to a multi-pathogen system in wild animals, our study provides a distinct framework that could be expanded across bat species to increase our understanding of how bats tolerate pathogens.

ABSTRACT: The aim was to describe the incidence and lesions caused by metazoan and protozoan parasites in Didelphis virginiana opossums in Mexico. From 2019 to 2021, twenty D. virginiana were collected from the state of Colima, Mexico. Post mortem, parasitological, and histopathological studies were performed to identify and describe their lesions and findings. All opossums(20/20) presented at least one type of parasite of the twelve found; the most abundant were nematodes 58.33% (7/12) Didelphostrongylus hayesi, Turgida turgida, Cruzia sp., Viannaia sp., Trichuris didelphis, Trichostrongylidae and Gnathostoma sp. Protozoa 25% (3/12), including Sarcocystis sp., Besnoitia sp., and Sarcocystis sp. A single specimen of Mathevotaenia sp., 8.3% (1/12), and Paragonimus mexicanus 8.3%, were identified (1/12). The most affected systems were the digestive, respiratory, and musculoskeletal. The most abundant parasites were Turgida turgida (20/20) 100%, Cruzia sp., (16/20) 80%, and Viannaia sp. (6/20) 30%, followed by Sarcocystis sp. cysts, (6/20) 30%. The associated parasitic lesions were: D. hayesi produce eosinophilic granulomatous bronchopneumonia, P. mexicanus eosinophilic focal granulomatous pneumonia, and Besnoitia sp., focal eosinophilic lymphoplasmacytic interstitial pneumonia. Gnathostoma and T. turgida caused severe eosinophilic granulomatous focal gastritis. Viannaia sp., Sarcocystis sp., Mathevotaenia sp., and Trichostrongylidae; caused diffuse eosinophilic mucosal enteritis. Cruzia sp., and T. didelphis, induced diffuse eosinophilic mucosal typhlitis. Sarcocystis sp. cysts were detected in the cytoplasm of muscle fibers without histological changes.

The protozoan parasites Cryptosporidium parvum, Cyclospora cayetanensis, and Toxoplasma gondii are major causes of waterborne and foodborne diseases worldwide. The assessment of their removal or inactivation during water treatment and food processing remains challenging, partly because research on these parasites is hindered by various economical, ethical, methodological, and biological constraints. To address public health concerns and gain new knowledge, researchers are increasingly seeking alternatives to the use of such pathogenic parasites. Over the past few decades, several non-pathogenic microorganisms and manufactured microparticles have been evaluated as potential surrogates of waterborne and foodborne protozoan parasites. Here, we review the surrogates that have been reported for C. parvum, C. cayetanensis, and T. gondii oocysts, and discuss their use and relevance to assess the transport, removal, and inactivation of these parasites in food and water matrices. Biological surrogates including non-human pathogenic Eimeria parasites, microorganisms found in water sources (anaerobic and aerobic spore-forming bacteria, algae), and non-biological surrogates (i.e. manufactured microparticles) have been identified. We emphasize that such surrogates have to be carefully selected and implemented depending on the parasite and the targeted application. Eimeria oocysts appear as promising surrogates to investigate in the future the pathogenic coccidian parasites C. cayetanensis and T. gondii that are the most challenging to work with.

Introduction. Trichomonas vaginalis genome is among the largest genome size and coding capacities. Combinations of gene duplications, transposon, repeated sequences, and lateral gene transfers (LGTs) have contributed to the unexpected large genomic size and diversity. This study is aimed at investigating genomic exchange and seeking for presence of the CRISPR CAS system as one of the possible mechanisms for some level of genetic exchange. Material and Methods. In this comparative analysis, 398 publicly available Trichomonas vaginalis complete genomes were investigated for the presence of CRISPR CAS. Spacer sequences were also analyzed for their origin using BLAST. Results. We identified a CRISPR CAS (Cas3). CRISPR spacers are highly similar to transposable genetic elements such as viruses of protozoan parasites, especially megavirals, some transposons, and, interestingly, papillomavirus and HIV-1 in a few cases. Discussion. There is a striking similarity between the prokaryotes/Archaean CRISPR and what we find as eukaryotic CRISPR. About 5-10% of the 398 T. vaginalis possess a CRISPR structure. Conclusion. According to sequences and their organization, we assume that these repeated sequences and spacer, along with their mentioned features, could be the eukaryotic homolog of prokaryotes and Archaean CRISPR systems and may involve in a process similar to the CRISPR function. [ABSTRACT FROM AUTHOR]

Background: Apicomplexan protozoa are a diverse group of obligate intracellular parasites causing many diseases that affect humans and animals, such as malaria, toxoplasmosis, and cryptosporidiosis. Apicomplexan protozoa possess unique thioredoxins (Trxs) that have been shown to regulate various cellular processes including metabolic redox regulation, parasite survival, and host immune evasion. However, it is still unknown how synonymous codons are used by apicomplexan protozoa Trxs. Methods: Codon usage bias (CUB) is the unequal usage of synonymous codons during translation which leads to the over- or underrepresentation of certain nucleotide patterns. This imbalance in CUB can impact a variety of cellular processes including protein expression levels and genetic variation. This study analyzed the CUB of 32 Trx coding sequences (CDS) from 11 apicomplexan protozoa. Results: The results showed that both codon base composition and relative synonymous codon usage (RSCU) analysis revealed that AT-ended codons were more frequently used in Cryptosporidium spp. and Plasmodium spp., while the Eimeria spp., Babesia spp., Hammondia hammondi, Neospora caninum, and Toxoplasma gondii tended to end in G/C. The average effective number of codon (ENC) value of these apicomplexan protozoa is 46.59, which is > 35, indicating a weak codon preference among apicomplexan protozoa Trxs. Furthermore, the correlation analysis among codon base composition (GC1, GC2, GC3, GCs), codon adaptation index (CAI), codon bias index (CBI), frequency of optimal codons (FOP), ENC, general average hydropathicity (GRAVY), aromaticity (AROMO), length of synonymous codons (L_sym), and length of amino acids (L_aa) indicated the influence of base composition and codon usage indices on CUB. Additionally, the neutrality plot analysis, PR2-bias plot analysis, and ENC-GC3 plot analysis further demonstrated that natural selection plays an important role in apicomplexan protozoa Trxs codon bias. Conclusions: In conclusion, this study increased the understanding of codon usage characteristics and genetic evolution of apicomplexan protozoa Trxs, which expanded new ideas for vaccine and drug research. [ABSTRACT FROM AUTHOR]

Parasitic infestations are one of the most concerning problems limiting ornamental fish farming. In addition to the direct economic losses due to the major mortalities, parasites may significantly negatively impact the body shape, coloration, growth rate, and health condition of the fish. The results of the study highlight the importance of comprehensive parasitological analysis in the diagnosis and treatment of parasitic infections in ornamental fish farms. The presence of multiple parasites in the affected fish emphasizes the need for effective biosecurity measures, such as extending the quarantine period for newly imported fish, closely monitoring fish populations, and implementing isolation units to prevent the spread of infections. By implementing these preventative measures, ornamental fish farmers can reduce the risk of parasitic infections and ensure the health and well-being of their fish populations. This, in turn, can lead to increased profitability and sustainability for their business. Overall, the current study aimed to conduct a clinical, histopathological, and phylogenetic analysis of the epibiont ciliated protozoan Epistylis wuhanensis and the copepod crustacean Lernaea cyprinacea in a freshwater ornamental fish farm in Iran. Furthermore, it provides valuable insights into the prevalence and impact of parasitic infections in ornamental fish farms and underscores the need for continued research and the development of effective preventative measures to address this issue. A total of 60 symptomatic freshwater ornamental fish, including 30 guppy (Poecilia reticulata) and 30 sailfin molly (Poecilia latipinna), were packed in polyethylene bags filled with oxygenated pond water and transported to the Ornamental Fish Clinic, Faculty of Veterinary Medicine, University of Tehran, for parasitological analysis. Following the clinical examination, histopathological analysis was performed on 10% NBF (neutral buffered formalin)-fixed samples from affected tissues, including the skin, skeletal muscle, and liver, to identify any pathological changes associated with the parasitic infections. Furthermore, the DNA was extracted from the 99% ethanol-fixed samples using a commercial DNA extraction tissue kit (SinaPure DNA, Iran), and PCR was performed using Peri18S-F1 (5′-ACC TGG TTG ATC CTG CCA GT-3′) and Peri18S-R1 (5′-TGC AGG TTC ACC TAC GGA AA-3′) (first reaction), and Peri18S-F2 (5′-CCG CGG TAA TTC CAG CTC-3′) and Peri18S-R2 (5′-GAT CCC CTA ACT TTC GTT CTT GA-3′) (second round) primers for the identified parasites. Finally, the PCR products were sequenced using Sanger dideoxy sequencing methods, and the resulting sequences were compared to sequences in the BLAST search program to provide a comprehensive picture of the current parasite-based disorder. The crustacean L. cyprinacea and the epibiont sessilid E. wuhanensis were identified in the examined ornamental guppy (6/30) and sailfin molly (6/30), with an overall parasitic prevalence of 20.00% (12/60). Ciliates were found in all tissue lesions but not in fish without lesions. A great number of the ciliated protozoan E. wuhanensis were found attached to the integumentary area of L. cyprinacea. Microscopically, oval to round granulomatous lesions were observed in cutaneous and skeletal muscles. Lymphoplasmacytic dermatitis and myositis were also observed. The crustacean L. cyprinacea serves as a mechanical vector for E. wuhanensis infection and spreads the disease in ornamental fish farming operations. For the first time in Iran, we successfully presented diagnostic morphological and molecular data on sessilids isolated from L. cyprinacea. Based on the findings of the current study, such parasitic infections may cause significant economic losses following invasion of the integument area of fish, eventually leading to death, if treatment is neglected or inadequate. Furthermore, the findings of the analysis were used to develop effective diagnostic approaches for the affected fish, as well as recommendations for improved health conditions to prevent future outbreaks of parasitic infections. However, further research is needed to determine the precise mechanisms of crustacean attachment and host-crustacean-peritrich protozoan interactions. Furthermore, the direct and indirect effects of various environmental factors on the emergence and spread of the current disease should be considered. [ABSTRACT FROM AUTHOR]

Trypanosoma cruzi is a parasitic protozoa causative of Chagas disease. As part of our interest in studying the basic biology of this microorganism, this work reports our observations related to the characterization of motifs and structural domains present in two fibrillarin isoforms (TcFib1 and TcFib2) that were found to be necessary for the nuclear targeting of these nucleolar proteins. Previous characterization of these proteins indicated that they share 68.67% of identical amino acids and are both expressed as nucleolar proteins in T. cruzi epimastigotes. Using an approach based on the transfection of recombinant genes encoding fluorescent fibrillarin-EGFP fusion proteins, this study found evidence for the presence of 4 motifs or protein domains that help target these proteins to the nucleus: The GAR domain and carboxyl terminus in both TcFibs, as well as two lysines and a computationally predicted cNLS in TcFib1. As a distinctive feature, the GAR domain of TcFib2 proved to be essential for the nuclear localization of this protein paralog. Such a difference between TcFib1 and Tcfib2 nuclear localization signals can be explained as the presence of two partially related nuclear import pathways for the two fibrillarin homologues in this organism. • Fibrillarins are essential nucleolar proteins required during ribosomal biogenesis in eukaryotic cells. • Trypanosoma cruzi , the causative agent of Chagas disease, expresses two paralogous fibrillarins. • The two types of TcFibrillarin present non identical signals for nuclear transport. • TcFibrillarins may be translocated to T. cruzi nucleus using different molecular pathways. [ABSTRACT FROM AUTHOR]

Innovative technologies are needed to enhance access to clean water and avoid waterborne diseases. We investigated the performance of cold atmospheric plasma (CAP), a clean and sustainable approach for microbial inactivation and total organic carbon (TOC) degradation in environmental water. Water matrices played a crucial role in the performance of CAP efficacy; for example, complete removal of ɸX174 from dH2O required 1 min of treatment, while ɸX174 reductions of ~ 2log10 and 4log10 were obtained after 10 min of CAP exposure in river water and wastewater samples, respectively. Similarly, after 10 min of CAP treatment, bacterial concentrations decreased by 3 log10 and 4 log10, in river and wastewater samples, respectively. In contrast, after 30 s of contact time, a 4 log10 reduction of bacteria was accomplished in dH2O. Complete removal of Acanthamoeba from dH2O was found after 30 min of CAP treatment, whereas it was not removed from surface water or wastewater at the same exposure time. Additionally, the approach successfully reduced TOC, and the degradation kinetics of TOC were represented by pseudo-first-order. CAP showed higher rates of TOC degradation in the final effluent of the wastewater treatment plant compared to surface water. The difference in CAP performance between river water and wastewater could be attributed to the bulk structure of humic acids in river water compared to small organic byproducts in the final effluent of WWTP. Overall, the findings reported here support the idea that CAP holds promise as a sustainable solution for controlling pathogens, removing organic water pollution, and integrating with traditional purification processes. Low-cost systems may advance CAP technology and increase its widespread use. [ABSTRACT FROM AUTHOR]

New colonies of Formosan subterranean termites are founded by monogamous pairs. During swarming season, alates (winged reproductives) leave their parental colony. After swarming, they drop to the ground, shed their wings, and male and female dealates find suitable nesting sites where they mate and become kings and queens of new colonies. The first generation of offspring is entirely dependent on the nutritional resources of the founder pair consisting of the fat and protein reserves of the dealates and their microbiota, which include the cellulose-digesting protozoa and diverse bacteria. Since termite kings and queens can live for decades, mate for life and colony success is linked to those initial resources, we hypothesized that gut microbiota of founders affect pair formation. To test this hypothesis, we collected pairs found in nest chambers and single male and female dealates from four swarm populations. The association of three factors (pairing status, sex of the dealates and population) with dealate weights, total protozoa, and protozoa Pseudotrichonympha grassii numbers in dealate hindguts was determined. In addition, Illumina 16S rRNA gene sequencing and the QIIME2 pipeline were used to determine the impact of those three factors on gut bacteria diversity of dealates. Here we report that pairing status was significantly affected by weight and total protozoa numbers, but not by P. grassii numbers and bacteria diversity. Weight and total protozoa numbers were higher in paired compared to single dealates. Males contained significantly higher P. grassii numbers and bacteria richness and marginally higher phylogenetic diversity despite having lower weights than females. In conclusion, this study showed that dealates with high body weight and protozoa numbers are more likely to pair and become colony founders, probably because of competitive advantage. The combined nutritional resources provided by body weight and protozoa symbionts of the parents are important for successful colony foundation and development. [ABSTRACT FROM AUTHOR]

Euglena gracilis (Euglena) is a microalgae found in most freshwater environments that produces paramylon, an insoluble β-1,3-glucan linked to human immunity. We hypothesized that Euglena powder has effects on immune function in apparently healthy adults. The study included male or female volunteers between the ages of 20 and 70 years who had white blood cell counts ranging from 4 × 103/µL to 10 × 103/µL, a "severe" rating on the stress questionnaire from the Korea National Health and Nutrition Examination Survey, and at least 2 upper respiratory infections with cold-like symptoms in the previous year. Participants received either a placebo or 700 mg of Euglena powder daily for 8 weeks. The study measured natural killer cell activity, cytokine concentrations, and blood lipid profiles to confirm the immune effect of Euglena consumption. In conclusion, Euglena improved immunological function through natural killer cell activity. Safety assessment showed no significant changes in vital signs or clinical chemistry indicators, and there were no adverse events associated with Euglena consumption. Euglena supplementation may help boost the immune systems of healthy individuals. Euglena gracilis is a freshwater microalgae that produces paramylon, which benefits human immunity. This study examined the effects of Euglena on immune function in apparently healthy subjects. Participants received either a placebo or 700 mg of Euglena powder daily for 8 weeks. Results showed improved immune function through natural killer (NK) cell activity, with no adverse events or safety concerns. Euglena supplementation may enhance the immune systems of apparently healthy individuals. [Display omitted] [ABSTRACT FROM AUTHOR]

This study simulates in vitro the effects of (i) rumen acidity and (ii) change in rumen protozoa numbers on the recovery of aflatoxins (AFs). Two 24‐h fermentation experiments were carried out using the same batch in vitro fermentation systems and substrate (dried corn meal) containing 11.42, 2.42, 7.65 and 1.70 µg/kg of AFB1, AFB2, AFG1 and AFG2 respectively. In Experiment 1, two buffer concentrations (normal salts dosage or lowered to 25%) were tested. Buffer reduction decreased gas production (730 vs. 1101 mL, p < 0.05), volatile fatty acids (VFA) and NH3 concentrations in the fermentation liquid (39.8 vs. 46.3 mmol/L, and 31.7 vs. 46.5 mg/dL respectively, p < 0.01). Recovery of all four AFs types was higher (p < 0.01) in the reduced buffer fermentation fluid, both as a percentage of total AF incubated (73.6% vs. 62.5%, 45.9% vs. 38.1%, 33.6% vs. 17.9% and 18.9% vs. 6.24% for AFB1, AFB2, AFG1 and AFG2 respectively) and as amounts relative to VFA production (163.4 vs. 123.5, 22.1 vs. 15.7, 48.8 vs. 22.5 and 6.16 vs. 1.86 ng/100 mmol of VFA, for AFB1, AFB2, AFG1 and AFG2 respectively). In Experiment 2, Stevia rebaudiana Bertoni extracts (S) or a Camphor essential oil (Cam) were added to fermenters and compared to the control (no additives, C). S and Cam addition resulted in a 25% reduction (p < 0.05) and a 15% increase (p < 0.05) in protozoa counts respectively, when compared to C. Both plant additives slightly reduced (p < 0.05) AFB1 recovery as a percentage of total AFB1 incubated (68.5% and 67.7% vs. 74.9% for S, Cam and C respectively). Recoveries of all other AFs were unaffected by the additives. In conclusion, the rumen in vitro AFB1 recovery (63%–75%) was higher than other AFs (3%–46%) and the acidic fermentation environment increased it. In our conditions, changes in protozoa numbers did not affect AFs recovery. [ABSTRACT FROM AUTHOR]

Permafrost soils store a substantial part of the global soil carbon and nitrogen. However, global warming causes abrupt erosion and gradual thaw, which make these stocks vulnerable to microbial decomposition into greenhouse gases. Here, we investigated the microbial response to abrupt in situ permafrost thaw. We sequenced the total RNA of a 1 m deep soil core consisting of up to 26 500-year-old permafrost material from an active abrupt erosion site. We analysed the microbial community in the active layer soil, the recently thawed, and the intact permafrost, and found maximum RNA:DNA ratios in recently thawed permafrost indicating a high microbial activity. In thawed permafrost, potentially copiotrophic Burkholderiales and Sphingobacteriales, but also microbiome predators dominated the community. Overall, both thaw-dependent and long-term soil properties significantly correlated with changes in community composition, as did microbiome predator abundance. Bacterial predators were dominated in shallower depths by Myxococcota, while protozoa, especially Cercozoa and Ciliophora, almost tripled in relative abundance in thawed layers. Our findings highlight the ecological importance of a diverse interkingdom and active microbial community highly abundant in abruptly thawing permafrost, as well as predation as potential biological control mechanism. [ABSTRACT FROM AUTHOR]

Gastrointestinal nematode parasites and gastrointestinal protozoan parasites are considered detrimental to the livestock population and manifest production-limiting effects. Small and large ruminants (cattle, buffalo, goats, and sheep) are important components of the rural economy of northern India. However, the epidemiology of gastrointestinal parasites in this agro-climatic region has not been studied extensively. In this study, the prevalence of gastrointestinal parasites was determined in 163 animals, including cattle (n = 86), buffalo (n = 11), goats (n = 48), and sheep (n = 18) from 26 sampling sites by copro-parasitological analysis. The prevalence values of 94.47% and 66.87% were recorded for the nematodes and protozoa, respectively. The group-wise prevalence of gastrointestinal nematode parasites was 95.3%, 90.9%, 93.7%, and 94.4% in cattle, buffalo, goats, and sheep, respectively, whereas for gastrointestinal protozoan parasites, the respective values were 70.9%, 54.5%, 60.4%, and 72.2%. Copromicroscopy revealed ten genera of nematodes—Ascaris, Capillaria, Cooperia, Haemonchus, Nematodirus, Oesophagostomum, Ostertagia, Strongyloides, Trichostrongylus, Trichuris, and one protozoan genus—Eimeria. The prevalence of Trichostrongylus spp. was highest in buffaloes, whereas in cattle, Ascaris spp. were predominant. In both goats and sheep, Haemonchus contortus was found to be predominant. The highest prevalence of gastrointestinal parasites was recorded in the rainy season. These findings indicate the prevalence of gastrointestinal parasites in the ruminant population in this region and necessitate the implementation of preventive and control strategies for effective animal health management. [ABSTRACT FROM AUTHOR]