Molecular Identification of Trypanosoma evansi Isolated from Arabian Camels (Camelus dromedarius) in Riyadh and Al-Qassim, Saudi Arabia

We analyzed the blood from 400 one-humped camels, Camelus dromedarius (C. dromedarius), in Riyadh and Al-Qassim, Saudi Arabia to determine if they were infected with the parasite Trypanosoma spp. Polymerase chain reaction (PCR) targeting the internal transcribed spacer 1 (ITS1) gene was used to detect the prevalence of Trypanosoma spp. in the camels. Trypanosoma evansi (T. evansi) was detected in 79 of 200 camels in Riyadh, an infection rate of 39.5%, and in 92 of 200 camels in Al-Qassim, an infection rate of 46%. Sequence and phylogenetic analyses revealed that the isolated T. evansi was closely related to the T. evansi that was detected in C. dromedarius in Egypt and the T. evansi strain B15.1 18S ribosomal RNA gene identified from buffalo in Thailand. A BLAST search revealed that the sequences are also similar to those of T. evansi from beef cattle in Thailand and to T. brucei B8/18 18S ribosomal RNA from pigs in Nigeria.

Distinct community structures of soil nematodes from three ecologically different sites revealed by high-throughput amplicon sequencing of four 18S ribosomal RNA gene regions

Quantitative taxonomic compositions of nematode communities help to assess soil environments due to their rich abundance and various feeding habitats. DNA metabarcoding by the 18S ribosomal RNA gene (SSU) regions were preferentially used for analyses of soil nematode communities, but the optimal regions for high-throughput amplicon sequencing have not previously been well investigated. In this work, we performed Illumina-based amplicon sequencing of four SSU regions (regions 1–4) to identify suitable regions for nematode metabarcoding using the taxonomic structures of nematodes from uncultivated field, copse, and cultivated house garden soils. The fewest nematode-derived sequence variants (SVs) were detected in region 3, and the total nematode-derived SVs were comparable in regions 1 and 4. The relative abundances of reads in regions 1 and 4 were consistent in both orders and feeding groups with prior studies, thus suggesting that region 4 is a suitable target for the DNA barcoding of nematode communities. Distinct community structures of nematodes were detected in the taxon, feeding habitat, and life-history strategy of each sample; i.e., Dorylamida- and Rhabditida-derived plant feeders were most abundant in the copse soil, Rhabditida-derived bacteria feeders in the house garden soil, and Mononchida- and Dorylamida-derived omnivores and predators and Rhabditida-derived bacteria feeders in the field soil. Additionally, low- and high-colonizer–persister (cp) groups of nematodes dominated in the house garden and copse soils, respectively, whereas both groups were found in the field soil, suggesting bacteria-rich garden soil, undisturbed and plant-rich copse soil, and a transient status of nematode communities in the field soil. These results were also supported by the maturity indices of the three sampling sites. Finally, the influence of the primer tail sequences was demonstrated to be insignificant on amplification. These findings will be useful for DNA metabarcoding of soil nematode communities by amplicon sequencing.

Morphology and phylogeny of the parasitic nematode Mooleptus rabuka (Machida, Ogawa & Okiyama, 1982) (Rhabditida, Spirurina: Mooleptinae nom. nov.), with notes on taxonomy of the family Gnathostomatidae

The nematode Mooleptus rabuka is recorded in the digestive tract of catshark Apristurus fedorovi caught at the Imperial Ridge (Pacific Ocean). Important morphological features such as the number of cephalic and caudal papillae, the position of amphids and the shape of the gubernaculum are detailed in this parasite species. According to the phylogenetic analyses based on the 18S ribosomal RNA gene sequences, M. rabuka forms a lineage, Mooleptinae nom. nov., which is close to the gnathostomatid genus Echinocephalus (maximum likelihood analysis), or else forms a polytomy with this genus and the lineages of Anguillicola + Spiroxys and Tanqua + ‘Linstowinema’ sp. (Bayesian inference analysis). Overall, our findings do not support the monophyly of the Gnathostomatidae. We elevate spiroxyines to the family status, Spiroxyidae stat. nov., and temporarily consider the Gnathostomatidae to include the following subfamilies: Ancyracanthinae Yorke & Maplestone, 1926, Gnathostomatinae Railliet, 1895 sensu lato and Mooleptinae nom. nov. The name Mooleptinae nom. nov. is suggested instead of the Metaleptinae Moravec & Nagasawa, 2000, which is based on a preoccupied generic name Metaleptus Machida, Ogawa & Okiyama, 1982.

Molecular Detection and Genetic Analysis of Theileria equi Detected in Apparently Healthy Horses in Nigeria

Equine theileriosis, an apicomplexan debilitating tick-borne parasitic disease of horses has caused considerable havoc to equine production all over the world. There is a dearth of information on the molecular characteristic of the parasites, Theileria equi Laveran, 1901, in Nigeria. Thus, in this study microscopy techniques and PCR were used to detect the T. equi of horses in Ogun, Oyo and Lagos States of Nigeria. We also characterized the partial region of 18S ribosomal RNA gene by sequencing and sequences analysis. One hundred and two horses consisting of Argentine 34 (33.3 %), Sudanese 21 (20.6 %) and local breeds 47 (46.1 %) including 2 females and 100 males were randomly sampled from the Polo Clubs in Ibadan, Lagos and from privately owned horse stables in Abeokuta. Blood samples were collected from the jugular vein, thin smears were prepared and stained with a field stain. The DNA was extracted from the blood and a partial region of the 18S ribosomal RNA gene was amplified. The amplified products were sequenced unidirectionally and subjected to phylogenetic analysis with those sequences obtained from the Gen-Bank. Of the 102 horses tested, 12 (11.7 %) were positive for T. equi by microscopy which included 9 (19.1 %) local breeds, 2 (5.8 %) Argentine breed and 1 (4.8 %) Sudanese breed. In contrast, 7 (6.8 %) were positive by the PCR method; out of which 5 (10.6 %) of these samples were from the local breed of horses while the remaining 2 (5.8 %) were from the Argentine breed. The Packed Cell Volume (PCV) of the infected and non-infected horses did not show any significant (P < 0.05) difference. The sequences lengths obtained were 311 bp and they had 97.43—98.07 % homologies with available sequences in the GenBank. The phylogenetic analysis of the sequences suggested that the strain of T. equi detected in the study area formed a new genotype different from the established genotypes around the world. In conclusion, the prevalence of T. equi was very low in the study area and one strain of the parasite may be in circulation among the studied horses.

Using Fomitopsis pinicola for bioinspired synthesis of titanium dioxide and silver nanoparticles, targeting biomedical applications

This study proposes a bio-directed approach for the formation of titanium oxide and silver nanoparticles (TiO2 and Ag NPs), using a wild mushroom, Fomitopsis pinicola, identified by 18S ribosomal RNA gene sequencing (gene accession no. MK635350) and phenotypic examination.

Molecular detection of Moniezia spp. (Cestoda) in Pergalumna persica (Acari: Oribatida) in Iran

Some oribatid mites are intermediate hosts of anoplocephalid cestodes, where the cestodes complete their larval development in the bodies of these oribatid mites. In order to investigate the potential of oribatid mites as vectors of anoplocephalid cestodes, 64 species of these mites were collected from different pastures of Shiraz County (Fars Province, Iran) during 2010–12. Two predominant species, Pergalumna persica and Acrogalumna lanceolata, were selected for study of both laboratory and natural infections. For artificial infection, these two species were exposed separately to eggs of Moniezia expansa and Moniezia benedeni, which are the common anoplocephalid cestodes of sheep and cattle in the Shiraz region. In order to develop a PCR-based technique for identification and detection of these cestodes, species-specific primers were designed to amplify the 18S ribosomal RNA gene. We demonstrate that the mite P. persica can be infested by M. expansa and M. benedeni in the laboratory and it can be naturally infected by the former species; however cestodes were not detected in the mite A. lanceolata. This is the first record of P. persica as the intermediate host of anoplocephalid tapeworms.

First molecular identification of Strongyloides fuelleborni in long-tailed macaques in Thailand and Lao People's Democratic Republic reveals considerable genetic diversity

Strongyloides fuelleborni is a soil-transmitted nematode parasite of non-human primates. The worm is prevalent also in human populations in Africa and South-East Asia. In this study, we amplified and sequenced a portion of the 18S ribosomal RNA gene (rRNA) and of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene of Strongyloides adult males recovered from faecal samples from long-tailed macaques (Macaca fascicularis) in Thailand and Lao PDR. The prevalence in Thailand was 31.1% (55/177) and in Lao PDR it was 62.1% (41/66), with an overall prevalence of 39.5% (96/243). All 18S rRNA sequences that we obtained (n = 96) showed 100% identity with published S. fuelleborni sequences. The 96 cox1 sequences that we obtained represented 32 new haplotypes. When included with the 17 previously known haplotypes from S. fuelleborni, the cox1 sequences fell into four clusters, which had clear geographical structure. This is the first molecular confirmation of S. fuelleborni in long-tailed macaques in Thailand and Lao PDR. Clearly, awareness needs to be raised of the zoonotic potential of S. fuelleborni. A monitoring programme should be organized, taking into account the role of reservoir hosts (i.e. monkeys) in the natural background of human strongyloidiasis caused by S. fuelleborni.