16S Ribosomal DNA Research Articles

Improved 18S rDNA profiling of parasite communities in salmonid tissues using a host blocking primer

Sensitive screening of eukaryotic communities in aquaculture for research and management is limited by the availability of technologies that can detect invading pathogens in an unbiased manner. Amplicon sequencing of 18S ribosomal DNA (rDNA) provides a potential pan-diagnostic test to overcome these biases; however, this technique is limited by a swamping effect of host DNA on low abundance parasite DNA. In this study, we have adapted a host 18S rDNA blocking assay to amplify eukaryotic DNA from salmonid tissue for amplicon sequencing. We demonstrate that effective salmonid 18S rDNA blocking enables sensitive detection of parasite genera in salmonid gill swabs. Furthermore, 18S rDNA amplicon sequencing with host blocking identified enriched pathogen communities in gill swabs from Atlantic salmon suffering from severe clinical gill infections compared to those exhibiting no clinical signs of gill infection. Application of host 18S rDNA blocking in salmonid samples led to improved detection of the amoebic parasite Neoparamoeba perurans, a parasite of significant threat to the Atlantic salmon aquaculture industry. These results reveal host 18S rDNA blocking as an effective strategy to improve the profiling and detection of parasitic communities in aquaculture species. This assay can be readily adapted to any animal species for improved eukaryotic profiling across agricultural and veterinary industries.

Characterizing living ocular bacterial communities and the effects of antibiotic perturbation in house finches

AbstractDNA‐based methods to measure the abundance and relative abundance of bacterial taxa can be skewed by the presence of dead or transient bacteria. Consequently, the active, functional members of the community may be a small subset of the detected bacterial community. This mismatch can make inferences about the roles of communities in host health difficult and can be particularly problematic for low‐abundance microbiomes, such as those on conjunctival surfaces. In this study, we manipulated bacterial communities on bird conjunctiva with a bacteriostatic antibiotic, reducing bacterial activity while preserving viability, to identify the living and active conjunctival communities using comparisons of 16S ribosomal DNA and RNA in paired samples. DNA amplicons included many more sequence variants than RNA amplicons from the same communities, with consequent differences in diversity. While we found that changes in communities in DNA samples broadly represent shifts in the living (RNA‐amplicon) communities, assessments of community function may be better described by RNA samples, reducing background noise from dead cells. We further used these data to test RNA:DNA ratios, used in other microbiological contexts, to detect shifts in bacterial activity after antibiotic disruption but were unable to detect changes in bacterial activity with this method.

Clinical features and fecal microbiota characteristics of patients with both ulcerative colitis and axial spondyloarthritis

BackgroundThe role of the intestinal microbiota in the pathogenesis of inflammatory bowel disease combined with axial spondyloarthritis (axSpA) is gaining widespread interest.AimsThis study was conducted to investigate the clinical and fecal microbiota characteristics of patients with both ulcerative colitis (UC) and axSpA.MethodsClinical data were collected from patients with UC. Patients were divided into the axSpA and non-axSpA groups according to human leukocyte antigen-B27 serology and sacroiliac joint imaging results. We obtained fecal specimens from 14 axSpA and 26 non-axSpA patients. All samples underwent 16S ribosomal DNA sequencing.ResultsSeventy-three patients with UC were included in this study, and the axSpA incidence was 19.2%. This incidence was significantly higher in patients with C-reactive protein > 10 mg/L. Firmicutes and Faecalibacterium abundances were decreased, and Proteobacteria and Escherichia_Shigella abundances were increased in the axSpA group compared with those of the non-axSpA group. Indicator analysis showed that Escherichia_Shigella was more likely to be an indicator species of axSpA. Additionally, many biosynthetic and metabolic pathways, including glutathione metabolism, fatty acid degradation, geraniol degradation, and biosynthesis of siderophore group nonribosomal peptides, were upregulated in the axSpA group.ConclusionPatients with UC have a high axSpA incidence, which may be related to the relative abundances of Escherichia_Shigella in these patients. The abundances of various biosynthetic and metabolic pathways of the fecal flora were upregulated in patients with axSpA.

Efficacy of Quadruple-coated Probiotics in Patients With Irritable Bowel Syndrome: A Randomized, Double-blind, Placebo-controlled, Parallel-group Study.

To evaluate the efficacy of quadruple-coated probiotics (gQlab) in patients with irritable bowel syndrome (IBS), focusing on sex differences and IBS subtypes. One hundred and nine Rome III-diagnosed IBS patients were randomized into either a gQlab or placebo group and received either gQlab or a placebo for 4 weeks. Participants replied to questionnaires assessing compliance, symptoms, and safety. Fecal samples were collected at 0 and 4 weeks to measure the probiotic levels using real-time quantitative polymerase chain reaction (qPCR) and to perform metagenomic analysis via 16S ribosomal DNA sequencing. The primary endpoint was the change in the overall IBS symptoms after 4 weeks of treatment. Ninety-two subjects (47 and 45 in the gQlab and placebo groups, respectively) completed the study protocol. At week 4, there was a higher relief of the overall IBS symptoms in the gQlab group (P = 0.005). The overall IBS symptom improvement was statistically significant (P = 0.017) in female patients of the gQlab group compared with the placebo group. Among the IBS subtypes, constipation-predominant IBS patients showed significant relief of the overall IBS symptoms (P = 0.002). At week 4, the fecal microbiome profiles between the 2 groups did not differ, but the qPCR levels of Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus helveticus, Bifidobacterium longum, and Bifidobacterium breve were increased in the gQlab group (P < 0.05 by repeated measures ANOVA). gQlab administration can improve the overall IBS symptoms, especially in female and constipation-predominant IBS patients. Further research is necessary to clarify the pathophysiology behind sex-related treatment responses in IBS patients.

New insights into the chromosomes of stoneflies: I. Karyotype, C-banding and localization of ribosomal and telomeric DNA markers in Skwalacompacta (McLachlan, 1872) (Polyneoptera, Plecoptera, Perlodidae) from Siberia.

This study provides data on chromosome number (2n♂♀=26), sex determination mechanism (XY♂/XX♀), C-banding pattern, distribution of clusters of telomeric TTAGG repeats and 18S ribosomal DNA in the karyotype of the stonefly Skwalacompacta (McLachlan, 1872). For the first time in the history of stoneflies cytogenetics, we provide photos of the chromosomes of the Plecoptera insects. The karyotype of males and females of S.compacta consists of 12 pairs of autosomes. Three pairs of large autosomes and four pairs of medium-sized autosomes are subacrocentric. The remaining pairs of autosomes are small, with unclear morphology. Pericentromeric C-bands were revealed in all autosomes. The sex chromosomes are also subacrocentric. The short arms of X and Y chromosomes are entirely heterochromatic and are rich in ribosomal DNA sequences. In the X chromosome this arm is larger than in the Y chromosome. It is likely that this arm associated with the nucleolar organizer (NOR). Telomeric DNA (TTAGG)n repeats were detected in the terminal regions of all chromosomes.

P090 Prevotella copri promotes colitis in mice by reducing expression of ATF4 and disturbing the gut microbiota

Abstract Background Colitis is a disease that carries a significant global disease burden, with a growing prevalence. Studies in humans and mice have explored the role of the intestinal microbiota that contribute to inflammatory bowel disease (IBD). Prevotella copri is a common human gut microbe that has been linked to inflammatory disorders. Methods C57BL/6 mice were divided into four groups: normal control group, P. copri group, DSS model group, DSS model+ P. copri group. Mice were gavaged with P. copri or PBS for 14 consecutive days, starting from the first day. Experimental colitis was induced by 2% DSS treatment from day 8. Mouse colon tissues were analyzed by quantitative real-time polymerase chain reaction, immunoblots, immunohistochemistry, and transcriptome. Serum and intestinal epithelial cell (IEC) amino acids were measured by high-performance liquid chromatography-tandem mass spectrometry. The analysis of the microbiome was done through the 16S ribosomal DNA sequencing. Results In the present study, our experiment confirmed that P. copri was significantly associated with the exacerbation of DSS-induced colitis by reducing expression of ATF4 and altering the gut microbiota composition. Leucine concentration of serum and IEC were increased in P. copri-gavaged mice, which clearly suppresses ATF4 expression. The dysbiosis of the intestinal microbiota and impaired α-defensin production by Paneth cells are also the leading causes of why P. copri-gavaged mice are more susceptible to DSS-induced colitis. Conclusion Taken together, the results proposed that P. copri aggravated the severity of colitis in the mice with DSS-induced colitis. This was done by reducing the expression of ATF4 and disturbing the gut microbiota. Thus, targeting P. copri may be an effective strategy in the treatment of IBD.

Assessing the microbiota of the snail intermediate host of trematodes, Galba truncatula

BackgroundThe microbiome is known to play key roles in health and disease, including host susceptibility to parasite infections. The freshwater snail Galba truncatula is the intermediate host for many trematode species, including the liver and rumen flukes Fasciola hepatica and Calicophoron daubneyi, respectively. The snail-parasite system has previously been investigated. However, the specific interaction between the snail-associated microbiota and intra-snail developmental stages of trematodes has yet to be explored.MethodsGalba truncatula snails were collected from farms in Northern Ireland and trematode infection was diagnosed using PCR. High-throughput sequencing analysis of the bacterial 16S ribosomal DNA V3-V4 hypervariable regions was subsequently applied to characterise the microbiota of both uninfected and infected snails.Results We first showed that the snail harboured microbiota that was distinct for its environment. The microbiota of infected snails was found to differ significantly from that of uninfected snails. In particular, the bacterial genera Mycoplasma and Methylotenera were significantly more abundant in infected snails, while genera Sphingomonas and Nocardioides were predominantly associated with uninfected snails. ConclusionThese findings pave the way to future studies on the functional roles of bacteria in host-parasite relationships.Graphical

Non-lethal detection of Eubothrium crassum (Cestoda) in farmed Atlantic salmon, Salmo salar, using anal swabs and real-time PCR.

Detection of intestinal parasites in fish typically requires autopsy, resulting in the sacrifice of the fish. Here, we describe a non-lethal method for detecting the tapeworm Eubothrium crassum in fish using anal swabs and real-time PCR detection. Two assays were developed to detect cytochrome oxidase I (COI) mitochondrial DNA and 18S ribosomal DNA sequences of E. crassum, respectively. The assays were tested on swab samples from confirmed pathogen free Atlantic salmon (Salmo salar L.) and on samples from farmed Atlantic salmon, where the presence and intensity of parasites had been established through autopsy. The COI assay was shown to be specific to E. crassum, while the 18S assay also amplified the closely related E. salvelini, a species infecting Arctic charr (Salvelinus alpinus L.) in freshwater. The COI assay detected E. crassum in all field samples regardless of parasite load while the 18S assay failed to detect the parasite in two samples. The results thus demonstrates that this non-lethal approach can effectively detect E. crassum and can be a valuable tool in assessing the prevalence of infection in farmed salmon, aiding in treatment decisions and evaluating treatment effectiveness.

Microscopic and molecular investigation of vector borne haemoprotozoan diseases in dromedary camel of North Gujarat.

Vector borne haemoprotozoan diseases comprise diverse group of single celled organism transmitted by haematophagus invertebrates. The current study was aimed at identification of major haemoprotozoan (Babesia, Theileria and Trypanosoma) in dromedary camel of North Gujarat region using microscopy and Polymerase Chain Reaction (PCR). A total of 234 blood samples were screened by the microscopic and molecular detection assays. Molecular prevalence studies of Theileria, Trypanosoma spp and Babesia was undertaken using 18s ribosomal DNA, RoTat 1.2 and SS rRNA gene respectively. The data relating to microscopic and molecular prevalence along with associated risk factors were analysed by statistical methods. The overall prevalence of hamoprotozoan disease based on microscopic and molecular investigation was 23.50%. The sensitivity and specificity (95% Confidence Interval) of PCR assay was 100% in comparison to microscopy (45.45% and 100%). The kappa coefficient between PCR and microscopy indicated good level of agreement with a value of 0.704 and SE of 0.159. Despite holding much significance to the animal sector, little work has been undertaken in regional part of India regarding camel parasites. The present paper offers the first preliminary research data investigating haemoprotozoan disease using parasitological and molecular methods in camels in the region.

Strongyloides stercoralis genotyping in a human population in southwestern Iran

BackgroundStrongyloidiasis is a neglected tropical disease (NTD) that is caused mainly by Strongyloides stercoralis, with an estimated 600 million people infected worldwide, and in fewer cases by Strongyloides fuelleborni fuelleborni and Strongyloides fuelleborni kellyi. A number of studies have been conducted on the genetic diversity of S. stercoralis in East and Southeast Asia; however, there is very limited corresponding information from West Asian countries, including Iran.MethodsFor Strongyloides worms collected from patients in southwestern Iran, the hypervariable regions I (HVR-I) and IV (HVR-IV) of the nuclear 18S ribosomal DNA (rDNA) locus (SSU) and a fragment of the subunit 1 mitochondrial cytochrome c oxidase gene (cox-1) were sequenced. For a subset of the worms, whole-genome sequencing data were generated.ResultsThe cox-1 sequences of 136 worms isolated from 23 patients indicated that all isolates were S. stercoralis. Among the cox-1 sequences, 33 polymorphic sites and 13 haplotypes were found. The phylogenetic analysis demonstrated that some sequences clustered fairly closely with sequences from humans and dogs from other parts of the world, while others formed a separate, Iran-specific group. Among 64 S. stercoralis analyzed, we found three of the previously described SSU HVR-I haplotypes, with haplotype II being the most frequent haplotype. In contrast to Southeast Asia, where S. stercoralis heterozygous for different haplotypes at the HVR-I locus are rare, we found 20 worms to be heterozygous for two different HVR-I haplotypes, 18 of which fell into the Iran-specific cox-1 cluster. SSU-heterozygous worms also showed elevated heterozygosity at the whole-genome level.ConclusionsWe conclude that the S. stercoralis population from the Khuzestan province shares much of the genetic diversity with the population in Southeast Asia, but there is an indication of additional genetic input. There appears to be some population structure with different subpopulations, which however do interbreed at least occasionally.Graphical

Insight into the chemical compositions of Anhua dark teas derived from identical tea materials: A multi-omics, electronic sensory, and microbial sequencing analysis

Anhua dark teas (DTs), including Tianjian tea, Qianliang tea, Hei brick tea, and Fu brick tea, are unique fermented teas from China’s Anhua County; yet their chemical composition differences remain unclear. Herein, metabolomics, volatolomics, and electronic sensory assessments were employed to analyze and compare chemical compositions and sensory characteristics of five types of Anhua DTs. All of these teas were derived from identical tea materials. Chemical compositions differed significantly among Anhua DTs, with Tianjian tea remarkable. Long-lasting fermentation and complex processing methods led to transformation of multiple compounds, particularly catechins. Eighteen volatile compounds with OVA > 1 were key aroma contributors in Anhua DTs. Internal transcribed spacer and 16S ribosomal DNA sequencing showed that Eurotium, Pseudomonas, and Bacillus are dominant microorganisms in Anhua DTs. Furthermore, this study unveiled notable differences in chemical compositions between Anhua DTs and five other traditional types of tea. This research enhances our understanding of Anhua DTs processing.

Mogroside Alleviates Diabetes Mellitus and Modulates Intestinal Microflora in Type 2 Diabetic Mice.

Mogroside, the main component of Siraitia grosvenorii (Swingle) C. Jeffrey (Cucurbitaceae) is a natural product with hypoglycemic and intestinal microbiota regulating properties. However, whether the alteration of intestinal microbiota is associated with the antidiabetic effect of mogroside remains poorly understood. This study investigated the mechanism underlying the hypoglycemic effect of mogroside in regulating intestinal flora and attenuating metabolic endotoxemia. Kunming mice with type 2 diabetes mellitus (T2DM) induced by high-fat diet and intraperitoneal injection of streptozotocin were randomly divided into model, pioglitazone (2.57 mg/kg) and mogroside (200, 100, and 50 mg/kg) groups. After 28 d of administration, molecular changes related to glucose metabolism and metabolic endotoxemia in mice were evaluated. The levels of insulin receptor substrate-1 (IRS-1), cluster of differentiation 14 (CD14) and toll-like receptor 4 (TLR4) mRNAs were measured, and the composition of intestinal microflora was determined by 16s ribosomal DNA (rDNA) sequencing. The results showed that mogroside treatment significantly improved hepatic glucose metabolism in T2DM mice. More importantly, mogroside treatment considerably reduced plasma endotoxin (inhibition rate 65.93%, high-dose group) and inflammatory factor levels, with a concomitant decrease in CD14 and TLR4 mRNA levels. Moreover, mogroside treatment reduced the relative abundance of Firmicutes and Proteobacteria (the inhibition rate of Proteobacteria was 85.17% in the low-dose group) and increased the relative abundance of Bacteroidetes (growth rate up to 40.57%, high-dose group) in the intestines of diabetic mice. This study reveals that mogroside can relieve T2DM, regulating intestinal flora and improving intestinal mucosal barrier, indicating that mogroside can be a potential therapeutic agent or intestinal microbiota regulator in the treatment of T2DM.

Description of a new species of Pericelis (Polycladida, Diposthidae) from sunken wood in the bathyal zone in Japan

Abstract We describe Pericelis nivea sp. nov. from sunken wood collected 330 m deep, off the coast of Owase, Japan. This is the first record of Pericelis from the bathyal zone. Unlike other congeners, P. nivea sp. nov. is characterized by the absence of eyespots. We provide a partial sequence of the mitochondrial cytochrome c oxidase subunit I gene as a DNA barcode for the new species. Phylogenetic analyses based on concatenated sequences of nuclear 18S and 28S ribosomal DNA showed that P. nivea sp. nov. was nested in the clade of Pericelis with high support; however, the relationship between P. nivea sp. nov. and other Pericelis species was unclear.

Identification of Lepidapedon oregonense as the current world's deepest trematode.

The deepest recorded depth for trematodes currently stands at approximately 6200 m. This depth record was achieved solely through sequence datasets of Lepidapedon sp. obtained from a gastropod. Given that trematodes of this genus typically use fish as definitive hosts, the origin of the trematode sequence was thought to be larval stages. However, the specific species remained unclear owing to the absence of reported adult-stage sequences. In the present study, we definitively identified the deepest trematode as Lepidapedon oregonense by comparing 28S ribosomal DNA sequences from adult worms from the macrourid fish Coelorinchus gilberti with data from the gastropod in the previous study.

Prevalence of Rickettsia spp., Anaplasma spp. and Ehrlichia spp. in Ornithodoros lahorensis from southern Xinjiang, China

Ticks are important vectors for pathogens and transmit a variety of diseases in humans and animals. Numerous studies have reported the presence of Ixodidae ticks and tick-borne pathogens in southern Xinjiang. However, data pertaining to argasid ticks and argasid tick-associated pathogens are sparse. This study collected a total of 407 ticks from southern Xinjiang. The tick species were identified on the basis of their morphological and mitochondrial 16S ribosomal DNA (16S rDNA) characteristics. The citrate synthase gene (gltA), 17-kilodalton antigen gene (17-kDa), and outer membrane protein A and B genes (ompA, ompB) were amplified using four sets of primers. The species of rickettsiae were further identified on the basis of concatenated sequences. To detect Anaplasma and Ehrlichia, nested PCR was used to amplify the Ehrlichia and Anaplasma 16S rRNA genes, respectively. All 407 ticks were Ornithodoros lahorensis. A total of 109 Rickettsia-positive samples and 19 Anaplasma-positive samples were detected in DNA samples from the 407 ticks. R. sibirica, R. massiliae, Candidatus R. barbariae, A. ovis, A. marginale, and A. phagocytophilum were found. All DNA samples from Or. lahorensis were tested for the presence of the Ehrlichia spp. nucleotide, but none of the samples were positive for such pathogens. This is the first time that R. sibirica, R. massiliae, and A. marginale were detected in Or. lahorensis from southern Xinjiang.

Using metal nanoparticles (ZnO and Ag) to enhance DNA isolation and amplification

Recently, there have been significant improvements in the methods used to isolate and purify genomic DNA. The implementation of high-throughput facilities has accelerated the advancement of technology, enabling more rapid and effective DNA processing than ever before. The study's goal was to develop a cost-effective and efficient technique to enhance the characterization of isolated DNA that is suitable for subsequent molecular downstream applications. The DNA used as study object obtained from suspensions of pathogenic bacteria, Klebsiella pneumoniae (K. pneumoniae) and the Enterobacter cloacae complex (E. cloacae) using a manual salting out method. The method includes utilizing ZnO and Ag nanoparticles (Nps), to enhance isolation of genomic DNA. The study showed that using Nps at concentrations of 0.02 mg/ml for Ag and 0.004 mg/ml for ZnO led to better DNA quality and quantity compared to the control group without Nps, resulting in ideal outcomes. In addition, the presence of these Nps greatly increased the efficiency of PCR amplification for detecting the 16S ribosomal DNA (rDNA) of K. pneumoniae and E. cloacae. This resulted in the effective amplification of a 709 base pair segment in both type of Nps. The incorporation of nanoparticles technology into DNA extraction methods is in line with the current demands of the fast-progressing fields of molecular biology and biotechnology.

Comparative 16s metagenomic analysis of prokaryotic diversity in freshwater and permanent snow-line glacial lakes in Türkiye

Freshwater lakes are critical to healthy ecosystems, providing vital services like drinking water and recreation for surrounding communities. Microorganisms within these ecosystems play essential roles, driving biogeochemical cycles for elements like carbon, nitrogen, and sulfur. This study utilized a metagenomic approach to examine the prokaryotic communities of three freshwater lakes in T?rkiye: the Eber and Bey?ehir lakes, located at close altitudes (967 m and 1,115 m, respectively), which serve as primary water sources for nearby communities, and Lake Uludag Buzlu (2,390 m) that lies at the permanent snow border within the Uludag glacial lake system. Metagenomics allowed us to identify species, genetic structures, and the functional roles of microorganisms. Employing high-throughput next-generation sequencing (NGS) technology, we analyzed 16S ribosomal DNA (rDNA) sequences (V3-V4 regions) from the lake samples. EzBioCloud software facilitated the analysis of prokaryotic diversity obtained using Illumina NovaSeq technology. While Eber and Bey?ehir lakes had similar diversity, Bacillota dominated in the higher-altitude Lake Uludag Buzlu. Genus-level analysis revealed Parabacteroides as the most prevalent in Lake Uludag Buzlu, contrasting with Limnohabitans dominance in Lake Eber; Lake Bey?ehir exhibited co-dominance of Limnohabitans and Planktophila.

Comparative evaluation of enriched formula milk powder with OPO and MFGM vs. breastfeeding and regular formula milk powder in full-term infants: a comprehensive study on gut microbiota, neurodevelopment, and growth.

This study investigated the non-inferiority of feeding term healthy infants with enriched formula milk powder containing 1,3-dioleoyl-2-palmitoylglycerol (OPO) and milk fat globular membrane (MFGM), compared to breast milk, in terms of the formation of gut microbiota, neurodevelopment and growth. Infants were divided into three groups: breast milk group (BMG, N = 50), fortified formula group (FFG, N = 17), and regular formula group (RFG, N = 12), based on the feeding pattern. Growth and development information was collected from the infants at one month, four months, and six months after the intervention. Fecal samples were collected from infants and analyzed for gut microbiota using 16S ribosomal DNA identification. The study found that at the three time points, the predominant bacterial phyla in FFG and BMG were Proteobacteria, Firmicutes, and Bacteroidetes, which differed from RFG. The abundance of Bifidobacterium in the RFG was lower than the FFG (one month, p = 0.019) and BMG (four months, p = 0.007). The abundance of Methanoprebacteria and so on (genus level) are positively correlated with bone mineral density (BMD) of term infants, and have the potential to be biomarkers for predicting BMD. The abundance of beta-galactosidase, a protein that regulates lactose metabolism and sphingoid metabolism, was higher in FFG (six months, p = 0.0033) and BMG (one month, p = 0.0089; four months, p = 0.0005; six months, p = 0.0005) than in the RFG group, which may be related to the superior bone mineral density and neurodevelopment of infants in the FFG and BMG groups than in the RFG group. Our findings suggest that formula milk powder supplemented with OPO and MFGM is a viable alternative to breastfeeding, providing a practical alternative for infants who cannot be breastfed for various reasons.

Dynamic gut microbiome-metabolome in cationic bovine serum albumin induced experimental immune-complex glomerulonephritis and effect of losartan and mycophenolate mofetil on microbiota modulation

Dynamic changes in gut dysbiosis and metabolomic dysregulation are associated with immune-complex glomerulonephritis (ICGN). However, an in-depth study on this topic is currently lacking. Herein, we report an ICGN model to address this gap. ICGN was induced via the intravenous injection of cationized bovine serum albumin (c-BSA) into Sprague-Dawley (SD) rats for two weeks, after which mycophenolate mofetil (MMF) and losartan were administered orally. Two and six weeks after ICGN establishment, fecal samples were collected and 16S ribosomal DNA (rDNA) sequencing and untargeted metabolomic were conducted. Fecal microbiota transplantation (FMT) was conducted to determine whether gut normalization caused by MMF and losartan contributed to their renal protective effects. A gradual decline in microbial diversity and richness was accompanied by a loss of renal function. Approximately 18 genera were found to have significantly different relative abundances between the early and later stages, and Marvinbryantia and Allobaculum were markedly upregulated in both stages. Untargeted metabolomics indicated that the tryptophan metabolism was enhanced in ICGN, characterized by the overproduction of indole and kynurenic acid, while the serotonin pathway was reduced. Administration of losartan and MMF ameliorated microbial dysbiosis and reduced the accumulation of indoxyl conjugates in feces. FMT using feces from animals administered MMF and losartan improved gut dysbiosis by decreasing the Firmicutes/Bacteroidetes (F/B) ratio but did not improve renal function. These findings indicate that ICGN induces serous gut dysbiosis, wherein an altered tryptophan metabolism may contribute to its progression. MMF and losartan significantly reversed the gut microbial and metabolomic dysbiosis, which partially contributed to their renoprotective effects.

A NEW EIMERIA SPECIES (APICOMPLEXA: EIMERIIDAE) PARASITIZING COMMERCIAL CHUKAR PARTRIDGES (ALECTORIS CHUKAR) IN SOUTHERN ONTARIO.

A commercial producer hatching and rearing chukar partridges (Alectoris chukar) in Ontario, Canada had flocks experiencing coccidiosis. Microscopic analysis of Eimeria species isolated from a field sample indicated the presence of 2 distinct oocyst morphotypes; the most abundant species was determined to be Eimeria chapmani, based on oocyst morphology and sequence-based genotyping, and the less abundant, second Eimeria sp. was an undescribed parasite. Oocysts of the unknown Eimeria sp. were large and oval-shaped; dimensions averaged 27.9 μm by 17.0 μm (shape index = 1.65 μm). Oocysts contained at least 1 polar granule and 4 almond-shaped sporocysts with average dimensions measuring 12.5 μm by 6.9 μm (shape index = 1.83). Each sporocyst featured a Stieda body, sub-Stieda body, and sporocyst residuum; a sporocyst contained 2 sporozoites that each possessed a small anterior refractile body and a larger posterior refractile body. Virtually all oocysts sporulated after 24 hr when suspended in potassium dichromate at room temperature (22 C) on a rotary platform. Experimental infections with various doses of oocysts demonstrated elevated parasite shedding from birds gavaged with higher challenge doses; fecundity generally decreased in heavier infections. The approximate prepatent period of the parasite was 4-5 days (unsporulated oocysts observed histologically at 90 hr postinfection and in feces by day 5) and patency lasted until day 12 postinfection. To characterize the endogenous development of the Eimeria sp., tissues were collected at 8 regions along the intestinal tract (including the ceca and rectum) every 6 hr throughout the estimated prepatent period. Parasites were observed to infect the descending and ascending duodenum, midjejunum, proximal and distal ileum, and the ceca. The endogenous stages identified included intracellular sporozoites, 3 generations of merogony, and gametogonic stages. Sequences of the mitochondrial genome (GenBank MW934555) and nuclear 18S ribosomal DNA (GenBank MW934259) were obtained using polymerase chain reaction amplification for Sanger sequencing, and these were unique from all published sequences on GenBank. Molecular data, in conjunction with the unique biology of the Eimeria sp. isolated from the chukar partridge flock, support that this coccidium is new to science.