16S partial gene mitochondrial DNA and internal transcribed spacers ribosomal DNA as differential markers of Trichuris discolor populations

Trichuris suis was isolated from the cecum of two different hosts (Sus scrofa domestica -- swine and Sus scrofa scrofa -- wild boar) and Trichuris vulpis from dogs in Sevilla, Spain. Genomic DNA was isolated and internal transcribed spacers (ITS)1-5.8S-ITS2 segment from the ribosomal DNA (rDNA) was amplified and sequenced using polymerase chain reaction techniques. The sequence of T. suis from both hosts was 1,396 bp in length while that of T. vulpis was 1,044 bp. ITS1 of both populations isolated of T. suis was 661 nucleotides in length, while the ITS2 was 534 nucleotides in length. Furthermore, the ITS1 of T. vulpis was 410 nucleotides in length, while the ITS2 was 433 nucleotides in length. One hundred fifty-four nucleotides were observed along the 5.8S gene of T. suis and T. vulpis. Intraindividual and intraspecific variations were detected in the rDNA of both species. The presence of microsatellites was observed in all the individuals assayed. Sequence analysis of the ITSs and the 5.8S gene has demonstrated no sequence differences between T. suis isolated from both hosts (S. scrofa domestica -- swine and S. scrofa scrofa -- wild boar). Nevertheless, clear differences were detected between the ITS1 and ITS2 of T. suis and T. vulpis. Furthermore, a comparative molecular analysis between both species and the previously published ITS1-5.8S-ITS2 sequence data of Trichuris ovis, Trichuris leporis, Trichuris muris, Trichuris arvicolae, and Trichuris skrjabini was carried out. A common homology zone was detected in the ITS1 sequence of all species of trichurids.

Adults of Trichuris skrjahini have been isolated from the cecum of caprine hosts (Capra hircus), Trichuris ovis and Trichuris globulosa from Ovis aries (sheep) and C. hircus (goats), and Trichuris leporis from Lepus europaeus (rabbits) in Spain. Genomic DNA was isolated and the ITS1-5.8S-ITS2 segment from the ribosomal DNA (rDNA) was amplified and sequenced by polymerase chain reaction (PCR) techniques. The ITS1 of T. skrjabini, T. ovis, T. globulosa, and T. leporis was 495, 757, 757, and 536 nucleotides in length, respectively, and had G + C contents of 59.6, 58.7, 58.7, and 60.8%, respectively. Intraindividual variation was detected in the ITSI sequences of the 4 species. Furthermore, the 5.8S sequences of T. skrjabini, T. ovis, T. globulosa, and T. leporis were compared. A total of 157, 152, 153, and 157 nucleotides in length was observed in the 5.8S sequences of these 4 species, respectively. There were no sequence differences of ITS1 and 5.8S products between T. ovis and T. globulosa. Nevertheless, clear differences were detected between the ITS1 sequences of T. skrjabini, T. ovis, T. leporis, Trichuris muris, and T. arvicolae. The ITS2 fragment from the rDNA of T. skrjabini was sequenced. A comparative study of the ITS2 sequence of T. skrjabini with the previously published ITS2 sequence data of T. ovis, T. leporis, T. muris, and T. arvicolae suggested that the combined use of sequence data from both spacers would be useful in the molecular characterization of trichurid parasites.

Adult worms of Trichuris ovis and T. globulosa were collected from Ovis aries (sheep) and Capra hircus (goats). T. suis was isolated from Sus scrofa domestica (swine) and T. leporis was isolated from Lepus europaeus (rabbits) in Spain. Genomic DNA was isolated and a ribosomal internal transcribed spacer (ITS2) was amplified and sequenced using polymerase-chain-reaction (PCR) techniques. The ITS2 of T. ovis and T. globulosa was 407 nucleotides in length and had a GC content of about 62%. Furthermore, the ITS2 of T. suis and T. leporis was 534 and 418 nucleotides in length and had a GC content of about 64.8% and 62.4%, respectively. There was evidence of slight variation in the sequence within individuals of all species analyzed, indicating intraindividual variation in the sequence of different copies of the ribosomal DNA. Furthermore, low-level intraspecific variation was detected. Sequence analyses of ITS2 products of T. ovis and T. globulosa demonstrated no sequence difference between them. Nevertheless, differences were detected between the ITS2 sequences of T. suis, T. leporis, and T. ovis, indicating that Trichuris species can reliably be differentiated by their ITS2 sequences and PCR-linked restriction-fragment-length polymorphism (RFLP).

Trichuris trichiura is a soil-transmitted helminth infecting human populations globally. Human cases caused by Trichuris suis and Trichuris vulpis have also been reported. Molecular identifications of Trichuris species infecting human populations in Lao PDR and Myanmar are lacking. Here, we explored molecular data obtained from Trichuris eggs recovered from human fecal samples from these countries and compared these with new and existing data from Thailand. Nuclear ribosomal DNA (18S and ITS2) sequences were amplified from Trichuris eggs and sequenced. Forty-one samples showed 99-100% similarity in their 18S sequences to published sequences of T. trichiura and one sample showed 99% similarity to a sequence of T. suis. Similarly, 41 samples showed 92-100% similarity in their ITS2 sequences to published sequences of T. trichiura and one sample showed 94-97% similarity to sequences of T. suis. This study is the first molecular confirmation of human infection with T. suis in northeast Thailand and the first molecular confirmation of the species of Trichuris infecting humans in Lao PDR and Myanmar.

Trichuris suis therapy for active ulcerative colitis: A randomized controlled trial

Zoonotic soil-transmitted helminths (STH), including Ancylostoma ceylanicum, Ancylostoma caninum, Ancylostoma braziliense, Trichuris vulpis, Trichuris suis, and Ascaris suum, are increasingly recognised as potential sources of human infection. Additionally, animals can act as carriers or reservoirs for human STH species. However, the extent of cross-host infection remains poorly understood, primarily due to reliance on morphological diagnostics. This review compiles data on the occurrence of cross-host STH infections, highlighting zoonotic STH in humans and human STH species in domestic and livestock animals. Following PRISMA guidelines, PubMed, Medline, and Web of Science were systematically searched without restriction on publication date, covering records available from inception to December 2024, with the earliest retrieved study published in 1942. Inclusion criteria encompassed studies on cross-host STH infections confirmed by molecular methods. Exclusion criteria included experimental infection studies, studies involving wildlife, and those that did not find cross-host infection. Two independent reviewers assessed bias using Appraisal tool for Cross-sectional studies (AXIS) and Joanna Briggs Institute appraisal tools. The protocol is registered with PROSPERO (CRD42024519067). The review screened 4197 titles and abstracts and included 51 studies. Ancylostoma ceylanicum was the commonest zoonotic STH reported, predominantly in Southeast Asia. Human STH species (Ancylostoma duodenale, Necator americanus, Trichuris trichiura and Ascaris lumbricoides) were found in dogs, cats, and pigs. Studies examining both humans and animals together in shared environments showed STH presence in both populations. Case studies revealed gastrointestinal and dermatological effects in humans particularly infected with zoonotic hookworms. This systematic review highlights STH cross-host species infections underscoring the need for further One health epidemiological investigations of humans and domestic/livestock animals in sympatric environments to better understand the burden and explore the transmission dynamics of cross-host STH infections.

Today, one of the important problems in the cultivation and maintenance of agricultural and domestic animals, both in industrial and private farms, is the spread of ecto- and endoparasites, which cause a significant decline in animal productivity, endanger their health and cause economic losses. The study of the level of distribution and conducting of diagnostic studies on parasitoses of animals in farms with different production orientation is an important and integral part of the overall complex of preventive and health-improving measures. The research was carried out during 2007–2017 in livestock enterprises of Kharkiv Oblast. As a part of the research work, 540 samples were collected from livestock farms, of which 180 were from the premises for keeping pigs, 100 from premises for keeping sheep, 120 from premises for keeping cattle, 80 from premises for dogs and 60 – from the surrounding territories of the livestock enterprises. We found that the objects of livestock rearing (pig complex, sheep farm, dairy farm, cynological center) in Kharkiv Oblast have a significant level of sanitary contamination with exogenous forms of helminths (21.7–45.6%) and the soil of the territories (20.0–36.6%). From samples taken from premises for pigs, eggs of four morphotypes were isolated (Ascaris suum – 5.3%, Trichuris suis – 14.6%, Oesophagostomum dentatum – 60.6%, Strongyloides ransomi – 19.4%), from premises for sheep – three morphotypes (Dicrocoelium lanceatum – 3.8%, Trichuris ovis – 29.3%, Strongylata spp. – 46.9%), from premises for cows – three morphotypes (Neoascaris vitulorum – 2.7%, Trichuris skrjabini – 12.9%, Strongylata spp. – 34.5%), premises for dogs – four morphotypes (Toxocara canis – 6.1%, Toxascaris leonina – 5.4%, Trichuris vulpis – 20.6%, Ancylostoma caninum – 17.5%). It should be noted that the places most contaminated with eggs of helminths were manure gutters (100%) and the floor of livestock buildings (50.0–86.7%). It has also been determined that, in the conditions of the cynological center, the Musca domestica was the source of environmental contamination with exogenous forms of Toxocara canis and Trichuris vulpis, and that the Muscina stabulans and Stomoxys calcitrans can be the source of spread of larvae of Ancylostoma caninum and eggs of Trichuris vulpis respectively.

DNA barcoding using a partial region (648 bp) of the cytochrome c oxidase I (COI) gene is a powerful tool for species identification and has revealed many cryptic species in various animal taxa. In birds, cryptic species are likely to occur in insular regions like the Japanese Archipelago due to the prevention of gene flow by sea barriers. Using COI sequences of 234 of the 251 Japanese-breeding bird species, we established a DNA barcoding library for species identification and estimated the number of cryptic species candidates. A total of 226 species (96.6%) had unique COI sequences with large genetic divergence among the closest species based on neighbour-joining clusters, genetic distance criterion and diagnostic substitutions. Eleven cryptic species candidates were detected, with distinct intraspecific deep genetic divergences, nine lineages of which were geographically separated by islands and straits within the Japanese Archipelago. To identify Japan-specific cryptic species from trans-Paleartic birds, we investigated the genetic structure of 142 shared species over an extended region covering Japan and Eurasia; 19 of these species formed two or more clades with high bootstrap values. Excluding six duplicated species from the total of 11 species within the Japanese Archipelago and 19 trans-Paleartic species, we identified 24 species that were cryptic species candidates within and surrounding the Japanese Archipelago. Repeated sea level changes during the glacial and interglacial periods may be responsible for the deep genetic divergences of Japanese birds in this insular region, which has led to inconsistencies in traditional taxonomies based on morphology.

The study of the diversity of the parasite fauna in the investigated foxes showed a high level of infestation (100%). The taxonomy of parasitofauna in foxes includes 12 parasitic invasions (Isospora canis – 14,3%, Alaria alata – 51,0%, Mesocestoides lineatus – 21,7%, Taeniidae spp – 27,0%, Syphacia obvelata – 17,0%, Strongyloides stercoralis – 13,3%, Toxocara canis – 59,0%, Toxascaris leonina – 65,5%, Ancylostoma caninum – 8,7%, Trichuris vulpis – 26,1%, Trichuris muris – 4,4%, Capilaria hepatica – 35,0%), which belong to 5 classes, 10 families, 11 genera and about 12 species. The share of species from the Sporozoa class is 8.3%, from the Trematoda class - 8.3%, from the Cestoda class - 16.7%, from the Secernentea class - 41.7% and from the Adenophorea class - 25.0%. Analyzing the parasitic species on epidemiological criterion, it was found that 10 species (83.3%) with large spread have zoonotic impact (A. alata, M. lineatus, Taenia spp., S. obvelata, S. ratti, T. canis, T. leonina, A. caninum, C. hepatica, T. vulpis) with a major risk to public health, and the identified invasions (100%) can parasitize domestic animals, as well as game fauna.

The fox is the most common wild carnivore, showing a synanthropic tendency, being widespread on all continents. They have a primary role in the biology of the development cycles of a large number of parasitic species, including those with zoonotic impact. The aim of the paper is the eco-parasitological study regarding the determination of the epidemiological characteristics of the parasite fauna in Vulpes vulpes (Linnaeus, 1758) from various natural and anthropogenic biotopes of the Republic of Moldova. The parasitological investigations were carried out according to the methods of Popova, Baermann, Fuileborn, Darling and through successive washings, in the laboratory of Parasitology and Helminthology of the Institute of Zoology. The results of the investigations show an increased level of infestation (100%) in the investigated foxes. Taxonomically, the parasite species fall into 5 classes, 10 families, 11 genera and about 12 species (Isospora canis - 14,3%, Alaria allata - 51,0%, Mesocestoides lineatus - 21,7%, Taeniidae spp - 27,0%, Syphacia obvelata - 17,0%, Strongyloides stercoralis - 13,3%, Toxocara canis - 59,0%, Toxascaris leonine - 65,5%, Ancylostoma caninum - 8,7%, Trichuris vulpis - 26,1%, Trichuris muris - 4,4%, Capilaria hepatica - 35,0%. The share of species from class Sporozoais of 8,3%, from class Trematoda - 8,3%, from class Cestoda - 16,7%, from class Secernentea - 41,7%, and from class Adenophorea - 25,0%. Analyzing the parasitic species according to epidemiological criteria, we found that 10 species (83.3%) with increased spread have a zoonotic impact (A. allata, M. lineatus, Taenidae, S. obvelata, S. ratti, T. canis, T. leonina, A. caninum, C. hepatica, T. vulpis), with a major risk for public health and the identified invasions, can also totally (100%) parasitize domestic animals and hunting fauna.

ABSTRACTThe dinoflagellate order Peridiniales encompasses several well circumscribed families. However, the family level of some genera, such as Bysmatrum and Vulcanodinium, has remained elusive for many years. Four Peridinium-like strains were established from the Atlantic coast of France and North Sulawesi, Indonesia through cyst germination or isolation of single cells. The cyst-theca relationship was established on specimens from the French Atlantic. Their morphologies were examined using light, scanning and transmission electron microscopy. The cells were characterized by a much larger epitheca relative to the hypotheca, a large anterior sulcal (Sa) plate deeply intruding the epitheca and a small first anterior intercalary plate. The plate formula was identified as Po, cp, X, 4′, 3a, 7′′, 6C, 5S, 5′′′, 2′′′′, shared by Apocalathium, Chimonodinium, Fusiperidinium and Scrippsiella of the family Thoracosphaeraceae but the configuration of Sa plate and anterior intercalary plates is different. Transmission electron microscopy showed that the eyespot was located within a chloroplast comprising two rows of lipid globules and thus belongs to type A. All four strains were classified within a new genus Caladoa as C. arcachonensis gen. et sp. nov. Small subunit ribosomal DNA (SSU rDNA), partial large subunit ribosomal DNA (LSU rDNA) and internal transcribed spacer ribosomal DNA (ITS rDNA) sequences were obtained from all strains. Genetic distance based on ITS rDNA sequences between French and Indonesian strains reached 0.17, suggesting cryptic speciation in C. arcachonensis. The maximum likelihood and Bayesian inference analysis based on concatenated data from SSU and LSU rDNA sequences revealed that Caladoa is monophyletic and closest to Bysmatrum. Our results supported that Caladoa and Bysmatrum are members of the order Peridiniales but their family level remains to be determined. Our results also support that Vulcanodinium is closest to the family Peridiniaceae.

To identify the species of trematodes isolated from laying ducks in Nanchang City using morphological and molecular approaches. Trematodes were isolated from the hepatobiliary duct, gallbladder and large intestine of market-sold laying ducks in Nanchang City. Following morphological characterization, total DNA was extracted from all trematode specimens, and internal transcribed spacer region (ITS) and cytochrome C oxidase subunit 1 (Cox1) genes were amplified using PCR assay and sequenced. Sequence alignment was performed using the Blast software, and homology and phylogenetic analyses were done in the trematode isolates based on ITS and Cox1 gene sequences. The morphological characteristics of two trematode isolates from the large intestine of laying ducks were similar to those of Echinostoma revolutum and E. miyagawai, and the morphological characteristics of eight trematode samples isolated from the hepatobiliary duct and gallbladder of laying ducks were similar to those of Amphimerus anatis. The ITS and Cox1 gene sequences of the two trematode isolates from the large intestine of laying ducks had 99.3% and 98.9%-99.4% homology with E. miyagawai, and the phylogenetic analysis showed that two trematode isolates had the closest genetic relationship with E. miyagawai based on ITS and Cox1 gene sequences. The ITS gene sequences of eight trematode isolates from the hepatobiliary duct and gallbladder of laying ducks shared 95.1%-95.5% with Opisthorchis sudarikovi and Clonorchis sinensis, while the Cox1 gene sequences of eight trematode isolates from the hepatobiliary duct and gallbladder of laying ducks shared 86.3%-86.4% and 85.5%-85.7% with O. viverrini and O. sudarikovi. ITS gene sequence-based phylogenetic analysis showed that the duck-derived trematode isolates had the closest genetic relationship with C. sinensis, and Cox1 gene sequence-based phylogenetic analysis showed that the duck-derived trematode isolates had the closest genetic relationship with Metorchis orientalis and O. viverrini. The trematode isolates from the large intestine of laying ducts in Nanchang City may be E. miyagawai, and the trematode isolates from the hepatobiliary duct and gallbladder may be an unidentified trematode species of the family Opisthorchiidae.

BackgroundApples are important for human nutrition because these provide vital nutrients, including vitamins and minerals, that are needed for a balanced diet. A suitable environment for the growth and survival of various microorganisms is also provided by multiple nutrients, such as carbohydrates, minerals, vitamins, and amino acids. Penicillium spp. are the cause of blue mold, one of the most common postharvest apple fruit diseases. Using morphological description and ITS gene sequencing, the current study aimed to identify Penicillium isolates associated with infected apple. High-performance liquid chromatography (HPLC) was used to demonstrate these isolates' capability for producing mycotoxins.ResultsThe initial identification of fungi was based on micromorphological characteristics, growth texture, and colony patterns. Penicillium (35.01%) and Talaromyces (15.62%) were the most prevalent fungal genera. These isolates' morphological identity was validated by ITS gene amplification and sequencing. The generated fragments' ITS gene sequencing values were 477, 480, 510, 478, and 478 bps for Penicillium expansum strain AP1, Penicillium crustosum strain AP2, Talaromyces atroroseus strain AP3, Penicillium expansum strain AP4, and Penicillium expansum strain AP5, respectively. It was examined whether the five isolates could produce citrinin and patulin. The highest value of toxins was recorded for patulin (24180 ppb), that produced by P. expansum AP5, followed by P. crustosum AP2 (19360 ppb). However, the highest value for citrinin was 24890 ppb detected for P. expansum AP4, followed by P. expansum AP5 (19320 ppb).ConclusionBlue mold caused by Penicillium expansum, Penicillium crustosum, and Talaromyces atroroseus. These isolates have the ability to produce citrinin and patulin with different degrees. So blue mold is one of the most harmful diseases in post-harvest apple fruits.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12870-025-07205-2.

Development of a PCR‐based diagnostic test for <i>Spongospora subterranea</i> f.sp. <i>nasturtii</i>, the causal agent of crook root of watercress (<i>Rorippa nasturtium‐aquaticum</i>)

Seven isolates of Cryptocaryon irritans from different host species and geographical locations in Mainland China were characterized by the first (ITS-1) and second (ITS-2) internal transcribed spacers (ITS) of nuclear ribosomal DNA (rDNA) using two isolates of Ichthyophthirius multifiliis for comparative purposes. The rDNA region including the ITS-1, 5.8S, ITS-2, and flanking 18S and 28S sequences were amplified by polymerase chain reaction and the amplicons were sequenced directly. The ITS-1, 5.8S, and ITS-2 sequences were 129, 160, and 190 bp in length, respectively, for all seven C. irritans isolates, whereas the corresponding sequences for the two I. multifiliis isolates were 142, 153, and 194 bp, respectively. While sequence variation among the seven C. irritans isolates ranged from 0 to 1.6% in both the ITS-1 and ITS-2, and the two I. multifiliis isolates differed by 1.4% in the ITS-1 and 1.0% in the ITS-2; C. irritans differed from I. multifiliis by 57.1-60.9% in the ITS-1 and 79.4-83.0% in the ITS-2, indicating that ITS sequences provide reliable genetic markers for the identification and differentiation of the two species. Phylogenetic analysis using the sequence pairwise-distance data using the neighbor-joining method inferred that the seven C. irritans isolates from Mainland China and two other isolates (T.A and Aus.C) from other countries clustered together to show monophyly, which could be readily distinguished from the other monophyletic group all from other regions. Therefore, ITS sequence data and phylogenetic analysis provided strong support that C. irritans isolates from Mainland China represent a single species. The definition of genetic markers in the ITS rDNA provide opportunities for studying the ecology and population genetic structures of the C. irritans from Mainland China and elsewhere and is also relevant to the diagnosis and control of fish diseases they cause.