scholarly journals Molecular and Microscopic Investigation of Sarcocystis Species Isolated from Sheep Muscles in Iran

2021 ◽  
Vol 2021 ◽  
pp. 1-6
Author(s):  
Nader Pestechian ◽  
Hossein Ali Yousefi ◽  
Reza Kalantari ◽  
Rasool Jafari ◽  
Faham Khamesipour ◽  
...  
Keyword(s):  
Distribution Patterns ◽  
18S Rrna Gene ◽  
Microscopic Investigation ◽  
Economic Losses ◽  
Rrna Gene ◽  
Sarcocystis Species ◽  
Digestion Method ◽  
Naked Eye ◽  
Sarcocystis Spp ◽  
Pcr Products

Sarcocystis species is a genus of cyst-forming parasites infecting both humans and animals globally. Some of these species cause clinical and subclinical diseases in the host and may lead to economic losses. This study was carried out to identify the distribution patterns of Sarcocystis spp. in slaughtered sheep based on the digestion method and PCR-RFLP in Isfahan, the center of Iran. In total, 150 fresh muscle samples (30 hearts, 60 esophagi, and 60 diaphragms) were investigated by naked eye observation and then scrutinized based on the digestion method. To this end, pepsin and HCl were used to observe the Sarcocystis parasite via a light microscope. The PCR was carried out to intensify a fragment of the 18S rRNA gene. Afterward, the PCR products were exposed to digestion by endonuclease TaqI, HindII, EcoRI, and AvaI. Consequently, the results of RFLP were confirmed by sequencing, and the phylogenetic placement of all species was analyzed. Through the examination by the naked eye, 5/150 (3.33%) macroscopic cysts were found in the samples. With the tissue digestion and microscopic examination, 116 (77.33%) samples were positive for Sarcocystis spp.; however, 125 (83.33%) samples were positive with PCR. Moreover, the results of sequence analysis on macrocysts and microcysts showed that 4% and 96% of the species belonged to S. gigantea and S. tenella, respectively. According to the results of the current study, sarcocystosis caused by S. tenella are highly prevalent among sheep in the Isfahan region. Due to the high prevalence of Sarcocystis infection in the world and Iran, the development of disease control and prevention policies in sheep would be essential, and changing attitudes in the way of keeping livestock from the traditional type to the industrial method is recommended in this regard.

Molecular approaches in species identification of Sarcocysts in Indian buffalo meat

2018 ◽  
Author(s):  
C. Ramakrishn ◽  
S. Vaithiyanathan ◽  
M. Muthukumar ◽  
L., P. Lavanya and V.V. Kulkarni. R. Chatlod ◽  
P. Lavanya ◽  
...  
Keyword(s):  
Restriction Enzymes ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Restriction Enzyme Digestion ◽  
Pcr Assay ◽  
Naked Eye ◽  
Molecular Approaches ◽  
Pcr Product ◽  
Pcr Products ◽  
Eye Examination

DNA was extracted from sarcocysts (visible on naked eye examination) collected from 168 buffaloes belonging to Mumbai (60), Hyderabad (54) and Kolkata (54) cities of India. They were subjected to PCR assay using 18S rRNA gene primer. All the PCR amplicons of about 900 bp were subjected to restriction enzyme digestion with four different restriction enzymes (BslI, DraI, FokI and RsaI). PCR amplicons showed two different patterns (Pattern A and Pattern B) on RFLP. Twenty one PCR products from Pattern A and one PCR product from Pattern B were subjected to DNA sequencing. S. fusiformis and S. taeniata were identified from Pattern A and S. buffalonis was identified from Pattern B on sequencing.

scholarly journals Sarcocystis gigantea infection associated with granulomatous eosinophilic myositis in a horse

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.

scholarly journals Emergence of Babesia Conradae Infection in Coyote-hunting Greyhounds in Oklahoma, USA

2021 ◽  
Author(s):  
Erin M Stayton ◽  
Megan Lineberry ◽  
Jennifer Thomas ◽  
Tina Bass ◽  
Kelly Allen ◽  
...  
Keyword(s):  
18S Rrna ◽  
Clinical Signs ◽  
18S Rrna Gene ◽  
Post Treatment ◽  
Rrna Gene ◽  
Babesia Gibsoni ◽  
Wide Range ◽  
Pcr Products ◽  
Life Threatening ◽  
Tick Vectors

Abstract Background: Babesia species are intraerythrocytic Apicomplexan parasites that infect a wide range of vertebrate hosts. These pathogens are typically transmitted either by tick vectors or by direct blood-to-blood contact, and may cause life-threatening clinical disease such as thrombocytopenia, hemolytic anemia, and acute renal failure in canine hosts. While Babesia vogeli and Babesia gibsoni infections have both been reported in Oklahoma, reports of B. conradae infections have been limited to California. Methods: Whole blood samples were collected in EDTA tubes from all dogs in four separate kennels in Oklahoma. DNA was extracted from each blood sample and a nested PCR was performed using general Apicomplexan primers for the partial 18S rRNA gene. PCR products were electrophoresed in agarose matrix and appropriately sized amplicons were sequenced. Sequences were compared to reference 18S rRNA sequences available in GenBank, and samples with >98% homology to B. conradae (GenBank MK256976) were considered positive. B. conradae positive dogs were then treated with atovaquone (13.5 mg/kg TID) and azithromycin (10 mg/kg SID) for 10 days and retested at 30 and 60 days post treatment by PCR. Results: Fifteen of 40 dogs tested positive for B. conradae with 98–100% sequence homology to B. conradae from California. All positive cases were coyote-hunting Greyhounds. Treatment of clinically ill dogs with atovaquone and azithromycin resulted in complete clinical recovery in clinically ill dogs and all treated dogs had negative follow-up PCR at 30 and 60 days post treatment. Conclusions: Collectively, this study (i) documents the occurrence of B. conradae in Oklahoma, (ii) highlights this pathogen as a differential to be considered when clinical signs are present, and (iii) supports the use of atovaquone and azithromycin as effective treatment in these cases.

scholarly journals Stenamoeba dejonckheerei sp. nov., a Free-Living Amoeba Isolated from a Thermal Spring

Pathogens ◽  
2020 ◽  
Vol 9 (7) ◽  
pp. 586
Author(s):  
Manuel Alejandro Borquez-Román ◽  
Luis Fernando Lares-Jiménez ◽  
Libia Zulema Rodriguez-Anaya ◽  
Jose Reyes Gonzalez-Galaviz ◽  
Paul A. Fuerst ◽  
...  
Keyword(s):  
Thermal Spring ◽  
Small Subunit ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Likelihood Analysis ◽  
Ribosomal Ribonucleic Acid ◽  
Pcr Products ◽  
Northwestern Mexico ◽  
Rrna Gene Sequencing ◽  
A New Species

Two amoeboid organisms were obtained from water samples taken from a thermal spring, "Agua Caliente", in Northwestern Mexico. The isolates were obtained when samples were cultivated at 37 °C on non-nutrient agar coated with Escherichia coli. The initial identification of the isolates was performed morphologically using light microscopy. The samples were found to have trophozoite morphology consistent with members of the genus Stenamoeba, a genus derived in 2007 from within the abolished polyphyletic genus Platyamoeba. Further analysis was performed by sequencing PCR products obtained using universal eukaryotic primers for the small subunit ribosomal ribonucleic acid (SSU rRNA) gene. Sequencing primers were designed to allow the comparison of the 18S rRNA gene sequences of the new isolates with previous sequences reported for Stenamoeba. Phylogenetic relationships among sequences from Stenamoeba were determined using Maximum Likelihood analysis. The results showed the two "Agua Caliente" sequences to be closely related, while clearly separating them from those of other Stenamoeba taxa. The degrees of sequence differentiation from other taxa were considered sufficient to allow us to propose that the Mexican isolates represent a new species.

scholarly journals Phylogenetic information of freshwater copepod (Diaptomus sicilis) with special reference to 18S rRNA

2016 ◽  
Vol 4 (1) ◽  
pp. 25 ◽  
Author(s):  
Gomathi Jeyam Mookkaiah ◽  
Ramanibai Ravichandran
Keyword(s):  
Tamil Nadu ◽  
18S Rrna ◽  
Molecular Data ◽  
Small Subunit ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Calanoid Copepods ◽  
Universal Primer ◽  
Pcr Products ◽  
The Individual

<p>In the present investigation to isolate freshwater calanoid copepods (<em>Diaptomus sicilis</em>) was characterized and identify the organisms by 18S rRNA sequencing. Plankton samples containing <em>D. sicilis</em> were collected during January 2014 (Post-monsoon) from Madippakkam Lake (12°57'41"N80°11'27"E) Chennai, Tamil Nadu. Immediately after sampling, specimens for genetic analyses were fixed in 95% ethyl alcohol. The total DNA was extracted from the individual copepod <em>D. sicilis</em> using Qiagen Blood tissue kit. The nuclear small subunit 18S rRNA gene was amplified using the Universal primer LCO —1490 (5’-GGTCAACAAATCATAAAGATATTGG-3’) and HCO-2198 (5’-TAAACTTCAGGGTGACCAAAAAATCA-3’). PCR products were loaded onto a 1% TAE agarose gel. Sequences were carried out an automated sequencer. The nucleotide sequence of 1282 base pair region of 18S rRNA was determined for D. sicilis. The similarity of sequences of <em>D. sicilis</em> was retrieved by BLASTn pro­gram and maximum identity and E-value was 76% and 0.00, respectively. The PCR products of <em>D. sicilis</em> individuals showed 80% similarity with the partial nuclear small subunit 18S rRNA gene region of other calanoid copepods. Based on molecular data the freshwater Calanoid copepods showed different algorithms and similar types of topologies useful for designing molecular analyses using phylogeny tree construction.Present molecular stud­ies on the relationship of D. sicilis with other freshwater calanoid copepods indicate that this species is close to <em>D. castor</em> followed by <em>D. keniraensis</em><em>.</em></p>

Molecular Survey of Babesia microti (Aconoidasida: Piroplasmida) in Wild Rodents in Turkey

2019 ◽  
Vol 56 (6) ◽  
pp. 1605-1609 ◽  
Author(s):  
Selma Usluca ◽  
Bekir Celebi ◽  
Djursun Karasartova ◽  
A Semra Gureser ◽  
Ferhat Matur ◽  
...  
Keyword(s):  
Sequence Data ◽  
18S Rrna Gene ◽  
Babesia Microti ◽  
Myodes Glareolus ◽  
Rrna Gene ◽  
Wild Rodents ◽  
Sarcocystis Spp ◽  
Zoonotic Parasite ◽  
Reservoir Hosts ◽  
Babesia Spp

Abstract Babesia microti (Aconoidasida: Piroplasmida) (Franca, 1910) is an important tick-borne zoonotic parasite with rodents serving as reservoir hosts. In the present study, 536 rodents were captured from Burdur, Bartin, Giresun, and Yozgat provinces of Turkey between the years 2010 and 2012, and blood samples were examined for the presence of Babesia spp. using conventional PCR which targeted the 18S rRNA gene. The sequence analysis of PCR amplicons was tested for B. microti as well as for Hepatozoon spp., and Sarcocystis spp. Overall, 5.8% of the rodents were positive for B. microti: 41% in Myodes glareolus, 7.7% in Chionomys roberti, and 2% in Apodemus spp., whereas no Babesia DNA was detected in Mus macedonicus and Microtus spp. Six rodents were positive for Hepatozoon spp. and one rodent was positive for Sarcocystis spp. Overall, 14.9 and 4.5% of rodents captured from Bartin and Giresun provinces, respectively, were PCR positive for B. microti, whereas none of rodents captured in Burdur and Yozgat were positive for Babesia spp. The sequence data of B. microti from rodents revealed that all sequences belonged to the zoonotic genotype. Sequences of B. microti obtained from rodents of the Bartin province were genotypically closer to European isolates, whereas those obtained from rodents of the Giresun province were closer to Russian and Mongolian isolates.

scholarly journals Distribution and diversity of eukaryotic microalgae in Kuwait waters assessed using 18S rRNA gene sequencing

PLoS ONE ◽  
2021 ◽  
Vol 16 (4) ◽  
pp. e0250645
Author(s):  
Vinod Kumar ◽  
Sabah Al Momin ◽  
Vanitha V. Kumar ◽  
Jasim Ahmed ◽  
Lamya Al-Musallam ◽  
...  
Keyword(s):  
Microbial Community ◽  
Microbial Communities ◽  
18S Rrna ◽  
Biological Diversity ◽  
Marine Ecosystem ◽  
Distribution Patterns ◽  
Arid Environment ◽  
18S Rrna Gene ◽  
Anthropogenic Factors ◽  
Rrna Gene

The microbial communities play a crucial role in ecosystem functioning through interactions among individuals and taxonomic groups in a highly dynamic marine ecosystem. The structure and functioning of the microbial communities are often influenced by the changes in the surrounding environment. Monitoring the microbial diversity of the marine ecosystem helps to understand spatial patterns of microbial community and changes due to season, climate, and various drivers of biological diversity. Kuwait is characterized by an arid environment with a high degree of temperature variation during summer and winter. Our understanding of spatial distribution patterns of microbial communities, their diversity, and the influence of human activities on the degree of changes in the diversity of the microbial community in Kuwait territorial waters remain unclear. In this study, we employed 18S rRNA sequencing to explore marine microalgal community composition and dynamics in seawater samples collected from Kuwait waters over two seasonal cycles across six locations. A total of 448,184 sequences across 36 replicates corresponding to 12 samples from six stations were obtained. The quality-filtered sequences were clustered into 1,293 representative sequences, which were then classified into different eukaryotic taxa. This study reveals that the phytoplankton community in Kuwait waters is diverse and shows significant variations among different taxa during summer and winter. Dinoflagellates and diatoms were the most abundant season-dependent microalgae taxa in Kuwait waters. Alexandrium and Pyrophacus were abundant in summer, whereas Gonyaulax was abundant during the winter. The abundance of Coscinodiscus and Navicula, of the diatom genera, were also dependent upon both seasonal and possible anthropogenic factors. Our results demonstrate the effectiveness of a sequencing-based approach, which could be used to improve the accuracy of quantitative eukaryotic microbial community profiles.

scholarly journals Design of primer pairs for species-specific diagnosis of Leishmania (Leishmania) infantum chagasi using PCR

2012 ◽  
Vol 21 (3) ◽  
pp. 304-307 ◽  
Author(s):  
Osvaldo José da Silveira Neto ◽  
Sabrina Castilho Duarte ◽  
Hérika Xavier da Costa ◽  
Guido Fontgalland Coelho Linhares
Keyword(s):  
18S Rrna ◽  
18S Rrna Gene ◽  
Reference Sequence ◽  
Rrna Gene ◽  
Analytical Sensitivity ◽  
Specific Detection ◽  
Pcr Products ◽  
Specific Amplification ◽  
Pcr Assays ◽  
Species Specific

The objective of this study was to design and evaluate new primers for species-specific detection of L. infantum chagasi using PCR. Two combinations of primer pairs were established with the aim of obtaining specific amplification products from the L. infantum chagasi 18S rRNA gene. The combinations of the primer pairs and the respective sizes of the PCR products, based on the U422465 GenBank reference sequence of L. infantum chagasi, were: LCS1/LCS3 (259 bp) and LCS2/LCS3 (820 bp). It was concluded that the new PCR assays optimized using the primer pairs LCS1/LCS3 and LCS2/LCS3 were effective for specific detection of L. infantum chagasi, with analytical sensitivity to detect 1 pg/µL of DNA.

scholarly journals Phylogenetic Analysis of the Hypervariable Region of the 18S rRNA Gene of Cryptosporidium Oocysts in Feces of Canada Geese (Branta canadensis): Evidence for Five Novel Genotypes

2004 ◽  
Vol 70 (1) ◽  
pp. 452-458 ◽  
Author(s):  
Kristen L. Jellison ◽  
Daniel L. Distel ◽  
Harold F. Hemond ◽  
David B. Schauer
Keyword(s):  
Genetic Diversity ◽  
Phylogenetic Analysis ◽  
18S Rrna ◽  
Hypervariable Region ◽  
Pcr Amplification ◽  
The United States ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Canada Geese ◽  
Pcr Products

ABSTRACT To assess genetic diversity in Cryptosporidium oocysts from Canada geese, 161 fecal samples from Canada geese in the United States were analyzed. Eleven (6.8%) were positive for Cryptosporidium spp. following nested PCR amplification of the hypervariable region of the 18S rRNA gene. Nine PCR products from geese were cloned and sequenced, and all nine diverged from previously reported Cryptosporidium 18S rRNA gene sequences. Five sequences were very similar or identical to each other but genetically distinct from that of Cryptosporidium baileyi; two were most closely related to, but genetically distinct from, the first five; and two were distinct from any other sequence analyzed. One additional sequence in the hypervariable region of the 18S rRNA gene isolated from a cormorant was identical to that of C. baileyi. Phylogenetic analysis provided evidence for new genotypes of Cryptosporidium species in Canada geese. Results of this study suggest that the taxonomy of Cryptosporidium species in geese is complex and that a more complete understanding of genetic diversity among these parasites will facilitate our understanding of oocyst sources and species in the environment.

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