Molecular Assay Proves the Presence of Theileria annulata Infection in Camels in Al-Diwaniyah Province, Iraq

Background: Theileria camelensis and T. dromedarii are parasitic protozoans reported by several studies as specific species that infect the one-humped camel (Camelus dromedarius). However, other findings casted significant doubts on the true identity of the causative species of theileriosis in camels. Therefore, the present study was conducted to investigate of T. camelensis and T. dromedarii in one humped camels in Iraq during Apr-Oct 2017. Methods: Blood samples for DNA extraction were obtained from 181 slaughtered camels. Molecular investigation was performed following the amplification of 18S rRNA gene by conventional PCR technique. DNA sequencing was then utilized only for the positive samples to confirm the infection with the Theileria species. Results: Nine (4.97%) out of 181 examined samples showed a positive result to infection with Theileria spp., and all these appeared as a T. annulata when subjected to DNA amplification and sequencing techniques. There was a complete absence of any new sequence outside the known species. Conclusion: Most of Theileria infection in camels in the study area is caused by T. annulata and no other causative agents like T. camelensis or T. dromedarii.

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Genetic Diversity of Bovine Hemoprotozoa in South Korea

Pathogens ◽

10.3390/pathogens9090768 ◽

2020 ◽

Vol 9 (9) ◽

pp. 768

Author(s):

Dongmi Kwak ◽

Min-Goo Seo

Keyword(s):

South Korea ◽

Clinical Symptoms ◽

Surface Protein ◽

18S Rrna Gene ◽

Rrna Gene ◽

Theileria Orientalis ◽

Bovine Theileriosis ◽

Major Piroplasm Surface Protein ◽

Theileria Spp ◽

Epidemiological Information

Tick-borne pathogens cause economically significant diseases in cattle. Theileria spp. are parasitic protozoa and the causative agent of bovine theileriosis. Here we report the distribution and risk factors of bovine Theileria using blood samples taken between 2018 and 2019. Of 737 tested cattle, nine animals (1.2%) were positive for Theileria orientalis infection by 18S rRNA gene amplification. Further analysis of the infected samples using the T. orientalis major piroplasm surface protein (MPSP) gene revealed five different genotypes circulating in the population: Types 1, 2, 3, 7, and N3. To the best of our knowledge, this is the first research to describe the existence of the T. orientalis MPSP genotype N3 in South Korea. Although the prevalence of bovine T. orientalis was low, our study offers data on the geographical distribution and prevalence of bovine Theileria spp. in South Korea. Further studies are warranted to determine the correlation of clinical symptoms with parasite MPSP genotypes. Our data provide epidemiological information to help control bovine theileriosis in South Korea.

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Genetic characterization of Strongyloides spp. from captive, semi-captive and wild Bornean orangutans (Pongo pygmaeus) in Central and East Kalimantan, Borneo, Indonesia

Parasitology ◽

10.1017/s0031182011001284 ◽

2011 ◽

Vol 138 (11) ◽

pp. 1417-1422 ◽

Cited By ~ 17

Author(s):

E. M. LABES ◽

N. WIJAYANTI ◽

P. DEPLAZES ◽

A. MATHIS

Keyword(s):

18S Rrna ◽

Genetic Characterization ◽

18S Rrna Gene ◽

Rdna Locus ◽

Rrna Gene ◽

Fecal Material ◽

East Kalimantan ◽

The One ◽

Bornean Orangutans

SUMMARYOrangutans (Pongo spp.), Asia's only great apes, are threatened in their survival due to habitat loss, hunting and infections. Nematodes of the genus Strongyloides may represent a severe cause of death in wild and captive individuals. In order to better understand which Strongyloides species/subspecies infect orangutans under different conditions, larvae were isolated from fecal material collected in Indonesia from 9 captive, 2 semi-captive and 9 wild individuals, 18 captive groups of Bornean orangutans and from 1 human working with wild orangutans. Genotyping was done at the genomic rDNA locus (part of the 18S rRNA gene and internal transcribed spacer 1, ITS1) by sequencing amplicons. Thirty isolates, including the one from the human, could be identified as S. fuelleborni fuelleborni with 18S rRNA gene identities of 98·5–100%, with a corresponding published sequence. The ITS1 sequences could be determined for 17 of these isolates revealing a huge variability and 2 main clusters without obvious pattern with regard to attributes of the hosts. The ITS1 amplicons of 2 isolates were cloned and sequenced, revealing considerable variability indicative of mixed infections. One isolate from a captive individual was identified as S. stercoralis (18S rRNA) and showed 99% identity (ITS1) with S. stercoralis sequences from geographically distinct locations and host species. The findings are significant with regard to the zoonotic nature of these parasites and might contribute to the conservation of remaining orangutan populations.

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Failure To Differentiate Cryptosporidium parvum from C. meleagridis Based on PCR Amplification of Eight DNA Sequences

Applied and Environmental Microbiology ◽

10.1128/aem.64.4.1454-1458.1998 ◽

1998 ◽

Vol 64 (4) ◽

pp. 1454-1458 ◽

Cited By ~ 26

Author(s):

Dominique Champliaud ◽

Philippe Gobet ◽

Muriel Naciri ◽

Odile Vagner ◽

José Lopez ◽

...

Keyword(s):

Cryptosporidium Parvum ◽

Dna Sequences ◽

Target Genes ◽

Pcr Amplification ◽

Dna Amplification ◽

Pcr Primers ◽

18S Rrna Gene ◽

Rrna Gene ◽

Restriction Enzyme Digestion ◽

Human Pathogens

ABSTRACT In order to determine the specificities of PCR-based assays used for detecting Cryptosporidium parvum DNA, eight pairs of previously described PCR primers targeting six distinct regions of theCryptosporidium genome were evaluated for the detection ofC. parvum, the agent of human cryptosporidiosis, andC. muris, C. baileyi, and C. meleagridis, three Cryptosporidium species that infect birds or mammals but are not considered to be human pathogens. The four Cryptosporidium species were divided into two groups: C. parvum and C. meleagridis, which gave the same-sized fragments with all the reactions, and C. muris and C. baileyi, which gave positive results with primer pairs targeting the 18S rRNA gene only. In addition to being genetically similar at each of the eight loci analyzed by DNA amplification, C. parvum and C. meleagridiscouldn’t be differentiated even after restriction enzyme digestion of the PCR products obtained from three of the target genes. This study indicates that caution should be exercised in the interpretation of data from water sample analysis performed by these methods, since a positive result does not necessarily reflect a contamination by the human pathogen C. parvum.

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Deep-sea corals provide new insight into the ecology, evolution, and the role of plastids in widespread apicomplexan symbionts of anthozoans

Microbiome ◽

10.1186/s40168-020-00798-w ◽

2020 ◽

Vol 8 (1) ◽

Cited By ~ 2

Author(s):

Samuel A. Vohsen ◽

Kaitlin E. Anderson ◽

Andrea M. Gade ◽

Harald R. Gruber-Vodicka ◽

Richard P. Dannenberg ◽

...

Keyword(s):

Deep Sea ◽

Coral Species ◽

Chlorophyll Biosynthesis ◽

18S Rrna Gene ◽

Rrna Gene ◽

Causative Agents ◽

History Of ◽

Insight Into ◽

Taxonomic Order

Abstract Background Apicomplexans are the causative agents of major human diseases such as malaria and toxoplasmosis. A novel group of apicomplexans, recently named corallicolids, have been detected in corals inhabiting tropical shallow reefs. These apicomplexans may represent a transitional lifestyle between free-living phototrophs and obligate parasites. To shed light on the evolutionary history of apicomplexans and to investigate their ecology in association with corals, we screened scleractinians, antipatharians, alcyonaceans, and zoantharians from shallow, mesophotic, and deep-sea communities. We detected corallicolid plastids using 16S metabarcoding, sequenced the nuclear 18S rRNA gene of corallicolids from selected samples, assembled and annotated the plastid and mitochondrial genomes from a corallicolid that associates with a deep-sea coral, and screened the metagenomes of four coral species for corallicolids. Results We detected 23 corallicolid plastotypes that were associated with 14 coral species from three orders and depths down to 1400 m. Individual plastotypes were restricted to coral hosts within a single depth zone and within a single taxonomic order of corals. Some clusters of closely related corallicolids were revealed that associated with closely related coral species. However, the presence of divergent corallicolid lineages that associated with similar coral species and depths suggests that corallicolid/coral relations are flexible over evolutionary timescales and that a large diversity of apicomplexans may remain undiscovered. The corallicolid plastid genome from a deep-sea coral contained four genes involved in chlorophyll biosynthesis: the three genes of the LIPOR complex and acsF. Conclusions The presence of corallicolid apicomplexans in corals below the photic zone demonstrates that they are not restricted to shallow-water reefs and are more general anthozoan symbionts. The presence of LIPOR genes in the deep-sea corallicolid precludes a role involving photosynthesis and suggests they may be involved in a different function. Thus, these genes may represent another set of genetic tools whose function was adapted from photosynthesis as the ancestors of apicomplexans evolved towards parasitic lifestyles.

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Molecular characterization of Theileria spp. in livestock and the first report on the occurrence of Theileria sp. OT3 in Iran

Acta Parasitologica ◽

10.1515/ap-2018-0061 ◽

2018 ◽

Vol 63 (3) ◽

pp. 515-521 ◽

Cited By ~ 1

Author(s):

Seyyed Javad Seyyed Tabaei ◽

Adel Spotin ◽

Ramin Pouriran ◽

Abbas Shahbazi ◽

Amirreza Javadi Mamaghani

Keyword(s):

Distance Matrix ◽

18S Rrna Gene ◽

Pairwise Distance ◽

Rrna Gene ◽

Sequencing Data ◽

Nested Polymerase Chain Reaction ◽

Blood Samples ◽

First Report ◽

Theileria Lestoquardi ◽

Causative Agents

Abstract Theileria lestoquardi, T. ovis, and T. annulata are recognized as major causative agents for ovine and cattle theileriosis in Iran, respectively. Recently, there have been reports on the presence of Theileria spp. (Theileria sp. OT1, Theileria sp. OT3, and Theileria sp.). In this study, 37 blood samples were collected from sheep and cattle with clinically suspected Theileria infection in the Northwest of Iran. The samples were analyzed using a light microscope. DNA samples were amplified via nested-polymerase chain reaction (PCR) of 18S rRNA gene. The amplicons were digested with HpaII, following restriction fragment length polymorphism (RFLP) and sequenced to reconfirm Theileria species. The microscopic examination indicated that 4 out of 37 (10.8%) blood samples were infected with Theileria spp. Based on the nested PCR-RFLP and sequencing data, 5.4%, 13.5%, and 27% of blood samples were infected with Theileria sp. OT3, T. ovis, and T. annulata, respectively. The pairwise distance matrix of Theileria sp. OT3 showed 99.8–100% identity and 0–0.2% divergence in comparison with the registered sequences. The present study is the first report of Theileria sp. OT3 in Iran. To the evaluate evolution of Theileria spp. and providing resultant genetic data, further research with a larger sample is necessary in the region.

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Cross-Genera PCR Amplification of DNA from Apicomplexan Parasites

Journal of Arthropod-Borne Diseases ◽

10.18502/jad.v12i3.83 ◽

2018 ◽

Author(s):

Philippe Gil de Mendonça

Keyword(s):

Pathogenic Bacteria ◽

Hypervariable Region ◽

Pcr Amplification ◽

18S Rrna Gene ◽

Rrna Gene ◽

Apicomplexan Parasites ◽

Diagnostic Samples ◽

Successful Amplification ◽

Babesia Spp ◽

Theileria Spp

Background: The discovery of an unexpected genetic sequence raised doubts about the specificity of a primer pair targeting Babesia spp. and Theileria spp. This study aimed to check the specificity of this primer pair. Methods: Conventional end-point PCR and real-time PCR protocols using primers targeting the 18S rRNA gene V4 hypervariable region of Babesia spp. and Theileria spp. were tested for potential cross-genera amplification using DNA from a palette of parasitic protists and pathogenic bacteria as a template. These investigations took place at the Ludwig Maximilian University of Munich (Germany) in 2010 as part of the EDEN project. Results: Successful amplification was obtained with DNA from five apicomplexan genera: Babesia, Theileria, Hepatozoon, Toxoplasma, and Hammondia. No amplicons were obtained when DNA from Leishmania infantum or bacteria within the genera Borrelia, Leptospira or Anaplasma was used as a template. Conclusion: This cross-genera amplification ability is useful for the quick exclusion of many parasite species from PCR negative diagnostic samples. Accurate species identification from PCR positive samples requires genetic sequencing of the amplicon.

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Molecular Detection of Tick-Borne Agents in Cats from Southeastern and Northern Brazil

Pathogens ◽

10.3390/pathogens11010106 ◽

2022 ◽

Vol 11 (1) ◽

pp. 106

Author(s):

Marcos Rogério André ◽

Ana Cláudia Calchi ◽

Maria Eduarda Chiaradia Furquim ◽

Isabela de Andrade ◽

Paulo Vitor Cadina Arantes ◽

...

Keyword(s):

18S Rrna ◽

Clinical Signs ◽

18S Rrna Gene ◽

Molecular Techniques ◽

Rrna Gene ◽

Molecular Identity ◽

Northern Brazil ◽

Rrna Gene Sequencing ◽

First Time ◽

Theileria Spp

Even though the epidemiology of tick-borne agents (TBA) in dogs has been extensively investigated around the world, the occurrence, vectors involved, and molecular identity of these agents in cats remains elusive in many regions. Among TBA, Ehrlichia, Anaplasma, Babesia, Cytauxzoon, and Hepatozoon are responsible for diseases with non-specific clinical signs in cats, making essential the use of molecular techniques for accurate diagnosis and proper treatment. The present work aimed to investigate the occurrence and molecular identity of tick-borne agents (Ehrlichia, Anaplasma, Babesia/Theileria, Cytauxzoon, and Hepatozoon) in cats from southeastern (states of São Paulo (SP) and Minas Gerais (MG)) and northern (state of Rondônia (RO)) Brazil. For this purpose, 390 blood samples were collected from domiciled cats in MG (n = 155), SP (n = 151), and RO(n = 84) states, submitted to DNA extraction and PCR assays for Ehrlichia spp. (dsb gene), Anaplasma spp. (rrs gene), piroplasmids (18S rRNA gene), and Hepatozoon spp. (18S rRNA gene), sequencing, and phylogenetic inferences. The overall positivity for Anaplasma spp., Ehrlichia spp., Babesia/Theileria spp., Cytauxzoon spp., and Hepatozoon spp. were 7.4% (12.3% (MG) and 6.6% (SP)), 2% (4.5% (MG) and 0.6% (SP)), 0.7% (0.6% (MG), 0.6% (SP) and 1.2% (RO)), 27.2% (41.9% (MG), 24.5% (SP) and 4.8% (RO), and 0%, respectively. The phylogenetic analysis grouped the obtained sequences with ‘Candidatus Anaplasma amazonensis’, A. platys, B. vogeli, and Cytauxzoon sp. previously detected in wild felids from Brazil. qPCR specific for E. canis based on the dsb gene confirmed the molecular identity of the detected ehrlichial agent. The present study expanded the list and geographical distribution of hemoparasites in cats. ‘Candidatus Anaplasma amazonensis’, recently detected in sloths from northern Brazil, was described for the first time in cats. This is the first report of piroplasmids infecting cats in northern Brazil. Coinfection by Cytauxzoon and other TBA (Ehrlichia, Anaplasma, and B. vogeli) reported in the present study raises the need for veterinary practitioners’ awareness of cats parasitized by multiple TBA.

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Sarcocystis gigantea infection associated with granulomatous eosinophilic myositis in a horse

Journal of Veterinary Diagnostic Investigation ◽

10.1177/1040638720935847 ◽

2020 ◽

Vol 32 (4) ◽

pp. 611-615

Author(s):

Fabrizia Veronesi ◽

Stefano Di Palma ◽

Simona Gabrielli ◽

Giulia Morganti ◽

Giovanni L. Milardi ◽

...

Keyword(s):

Incidental Finding ◽

Pectoral Muscle ◽

18S Rrna Gene ◽

Rrna Gene ◽

Sarcocystis Species ◽

Sarcocystis Spp ◽

Formalin Fixed Paraffin Embedded ◽

Veterinary Surgeon ◽

Specific Species ◽

The Right

The only Sarcocystis species currently known to inhabit the fibers of skeletal and cardiac muscles in horses are S. fayeri, S. bertrami, and S. asinus. We describe herein the invasion of myofibers in a horse by S. gigantea, a sheep-specific species with low virulence in the original host. A hunter gelding was referred to a veterinary surgeon in Newmarket (UK). The anamnestic data reported that the horse had an initial history of swelling of the right forelimb with fluid on the front of the carpus and edema spreading up the forearm. Subsequently, 2 firm lumps were found on the left pectoral muscle adjacent to the axilla of the left forelimb. Histologic examination of biopsies from the lumps revealed multifocal granulomatous eosinophilic myositis associated with intact and degenerate encysted parasites, consistent with Sarcocystis spp. Based on amplification and DNA sequencing of the 18S rRNA gene obtained from formalin-fixed, paraffin-embedded tissue blocks, S. gigantea was identified. The presence of sarcocysts in equine skeletal muscles has been considered an incidental finding, and there are only sporadic associated reports of myositis. Our finding suggests that some Sarcocystis spp. have a wider intermediate host range than believed previously, and that Sarcocystis of other species (not considered horse-associated) can invade the muscle fibers of equids, leading to myositis.

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Plankton Microorganisms Coinciding with Two Consecutive Mass Fish Kills in a Newly Reconstructed Lake

The Scientific World JOURNAL ◽

10.1100/2012/504135 ◽

2012 ◽

Vol 2012 ◽

pp. 1-14 ◽

Cited By ~ 39

Author(s):

Andreas Oikonomou ◽

Matina Katsiapi ◽

Hera Karayanni ◽

Maria Moustaka-Gouni ◽

Konstantinos Ar. Kormas

Keyword(s):

Gene Diversity ◽

18S Rrna Gene ◽

Rrna Gene ◽

Prymnesium Parvum ◽

Fish Kill ◽

Aquatic Life ◽

Unicellular Eukaryotes ◽

Toxic Cyanobacteria ◽

Causative Agents ◽

Lake Karla

Lake Karla, Greece, was dried up in 1962 and its refilling started in 2009. We examined the Cyanobacteria and unicellular eukaryotes found during two fish kill incidents, in March and April 2010, in order to detect possible causative agents. Both microscopic and molecular (16S/18S rRNA gene diversity) identification were applied. Potentially toxic Cyanobacteria included representatives of thePlanktothrixandAnabaenagroups. Known toxic eukaryotes or parasites related to fish kill events werePrymnesium parvumandPfiesteriacf.piscicida, the latter being reported in an inland lake for the second time. Other potentially harmful microorganisms, for fish and other aquatic life, included representatives of Fungi, Mesomycetozoa, Alveolata, and Heterokontophyta (stramenopiles). In addition, Euglenophyta, Chlorophyta, and diatoms were represented by species indicative of hypertrophic conditions. The pioneers of L. Karla’s plankton during the first months of its water refilling process included species that could cause the two observed fish kill events.

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Species-specific PCR assay for the detection of Babesia odocoilei

Journal of Veterinary Diagnostic Investigation ◽

10.1177/10406387211032927 ◽

2021 ◽

pp. 104063872110329

Author(s):

Hilary J. Burgess ◽

Betty P. Lockerbie ◽

Lisanework E. Ayalew ◽

Antonia Dibernardo ◽

Kristýna Hrazdilová ◽

...

Keyword(s):

Dna Amplification ◽

18S Rrna Gene ◽

Rrna Gene ◽

Species Sensitivity ◽

Pcr Assay ◽

Banding Patterns ◽

Specific Pcr ◽

Pcr Product ◽

Specific Pcr Assay ◽

Species Specific

We developed a PCR assay for the detection of Babesia odocoilei based on the 18S rRNA gene. Multiple specimens of B. odocoilei were examined, and the assay consistently produced a small specific PCR product of 306 bp. The PCR assay was also challenged with DNA from 13 other Babesia species and 2 Theileria species, originating from 10 different host species; however, nonspecific DNA amplification and multiple banding patterns were observed, and the amplicon banding patterns varied between different isolates of the same species. Sensitivity was determined to be 6.4 pg of DNA, and an estimated 0.0001% parasitism. This assay can be utilized for species-specific differential detection of B. odocoilei.

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