scholarly journals Phylogenetic study of Theileria ovis and Theileria lestoquardi in sheep from Egypt: Molecular evidence and genetic characterization

2021 ◽  
Vol 14 (3) ◽  
pp. 634-639
Author(s):  
Amira Adel Al-Hosary ◽  
Ahmed ElSify ◽  
Akram A. Salama ◽  
Mohamed Nayel ◽  
Ahmed Elkhtam ◽  
...  
Keyword(s):  
Genetic Characterization ◽  
Small Ruminants ◽  
Rrna Genes ◽  
Effective Control ◽  
Phylogenetic Study ◽  
Local Alignment ◽  
Molecular Evidence ◽  
Blood Samples ◽  
Theileria Lestoquardi ◽  
Theileria Spp

Background and Aim: Ovine theileriosis caused by Theileria ovis and Theileria lestoquardi is an important infectious disease affecting small ruminants in regions of the tropic and subtropic zones. There is limited studies about ovine theileriosis in Egypt; so the present study aims to assess the occurrence of ovine theileriosis in Egypt at the molecular level. Materials and Methods: Blood samples were collected from 115 randomly selected sheep, which were apparently healthy; the ages of the sampled sheep ranged from 1 to 5 years old, from a local breed (barkae and balade), and showed no symptoms indicating infection with Theileria spp. The study was conducted in three governorates representing Lower Egypt (Menoufia and Beheira) and Upper Egypt (El-Wady El-Geded). All blood samples were subjected to polymerase chain reaction (PCR) and semi-nested PCR to target Theileria spp. 18S rRNA genes. Positive samples were sequenced, and these sequences were analyzed using nucleotide basic local alignment search tool (BLAST). Results: Six animals (5.22%) were PCR-positive carriers for ovine theileriosis. Nucleotide BLAST and phylogenetic analyses of the six obtained sequences showed that T. ovis was present in five animals (4.37%) in Menoufia (n=2) and El-Wady El-Geded (n=3), whereas T. lestoquardi was detected in 1 animal (0.87%) in El-Wady El-Geded. Conclusion: This study is the first to provide molecular evidence, genetic characterization, and phylogenetic analysis of ovine Theileria spp. in Egypt. Specifically, T. lestoquardi and T. ovis carrier statuses of sheep were confirmed. These results highlight the importance of developing an effective control strategy against ovine theileriosis carriers that might develop and/or spread theileriosis.

scholarly journals Serological, cultural, and molecular evidence of Brucella infection in small ruminants in Pakistan

2015 ◽  
Vol 9 (05) ◽  
pp. 470-475 ◽  
Author(s):  
Shahzad Ali ◽  
Shamim Akhter ◽  
Heinrich Neubauer ◽  
Falk Melzer ◽  
Iahtasham Khan ◽  
...  
Keyword(s):  
Causative Agent ◽  
Brucella Abortus ◽  
Small Ruminants ◽  
Ring Test ◽  
Effective Control ◽  
Molecular Evidence ◽  
Serum Samples ◽  
Qrt Pcr ◽  
Milk Samples ◽  
Eradication Strategy

Introduction: The objectives of the present study were to determine the seroprevalence and identify the causative agent of brucellosis in small ruminants in Pakistan. Methodology: A total of 278 serum and 212 milk samples were collected from sheep and goats that had close contact with seropositive bovine herds. Data related to age, sex, location, and breed were collected on the sampling day. Serum and milk samples were initially screened using two different Rose Bengal plate test (RBPT) antigens and a milk ring test (MRT). Seropositive samples were subjected to bacterial isolation and PCR analysis using Brucella genus-specific (bcsp31) and Brucella species-specific (IS711 for Brucella abortus and Brucella melitensis) quantitative real-time polymerase chain reactions (qRT-PCR). Results: Twenty-four (8.6%) serum samples were positive by RBPT. Twenty (9.4%) animals were positive for Brucella antibodies using MRT. No Brucella isolates were obtained from the examined blood and milk samples. Of the 24 seropositive serum samples, 18 (75%) were positive in the Brucella genus-specific (bcsp31) and Brucella abortus-specific (IS711) qRT-PCR, respectively. Conclusions: Brucella abortus was identified as causative agent of ovine and caprine brucellosis in Pakistan. Results of this study can be used for the development of an effective control and eradication strategy for brucellosis in livestock, especially small ruminants.

scholarly journals Molecular prevalence, characterization and associated risk factors of Anaplasma spp. and Theileria spp. in small ruminants in Northern Pakistan

Parasite ◽  
2021 ◽  
Vol 28 ◽  
pp. 3
Author(s):  
Sadaf Niaz ◽  
Zia Ur Rahman ◽  
Ijaz Ali ◽  
Raquel Cossío-Bayúgar ◽  
Itzel Amaro-Estrada ◽  
...  
Keyword(s):  
Risk Factors ◽  
Multivariable Analysis ◽  
Small Ruminants ◽  
Rrna Genes ◽  
Infection Prevalence ◽  
Northern Pakistan ◽  
Blood Samples ◽  
Grazing System ◽  
Host Sex ◽  
Associated Risk Factors

This study was conducted in four districts (Malakand, Swat, Bajaur and Shangla) of Northern Pakistan to investigate the prevalence, associated risk factors and phylogenetic analyses of Theileria and Anaplasma species in small ruminants. A total of 800 blood samples, 200 from each district, were collected from apparently healthy animals. PCR assays were performed using generic primers for Anaplasma spp. and Theileria spp. as well as species specific primers for A. ovis and T. ovis. Overall infection prevalence was 361/800 (45.1%). Theileria spp. infection prevalence (187/800, 23.3%) was higher than Anaplasma spp. (174/800, 21.7%). Amplified partial 18S rRNA genes were sequenced and enrolled animals were found to be infected by T. ovis (115/800, 14.3%), and at least two more Theileria species (72/800, 9%) were present (T. lestoquardi and T. annulata). All blood samples that were found to be positive for Anaplasma spp. were also positive for A. ovis. Infection prevalence was higher in sheep (227/361, 28.3%) compared to goats (134/361, 16.6%) (p < 0.005). Univariable analysis of risk factors showed that host, age, grazing system and acaricide treatment were significant determinants (p < 0.05) for both Theileria and Anaplasma infections. Multivariable analysis revealed that host, sex, age, tick infestation and grazing system were significant risk factors (p < 0.005) for both pathogens. Phylogenetic analysis revealed variants among the A. ovis and T. annulata samples analysed, indicating that different genotypes are circulating in the field while T. ovis presented the same genotype for the samples analysed.

scholarly journals Molecular Detection and Differentiation of Theileria lestoquardi, T. ovis and T. annulata in Blood of Goats and Ticks in Kermanshah Province, Iran

2019 ◽  
Author(s):  
Mozhgan Rahmani-Varmale ◽  
Mousa Tavassoli ◽  
Bijan Esmaeilnejad
Keyword(s):  
Nested Pcr ◽  
Molecular Detection ◽  
Ixodid Ticks ◽  
Hyalomma Anatolicum ◽  
Blood Samples ◽  
Theileria Lestoquardi ◽  
Blood Smears ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Theileria Spp

Background: This study was carried out to identify Theileria spp. infections in goats and ticksin Kermanshah Prov­ince, western Iran from May–Sep 2015. Methods: For differentiation of different Theileria spp. both blood and tick samples were examined by nested PCR-RFLP. Results: Light microscopy of blood smears revealed Theileria spp. infection in 22 (5.5%), while 68 (17%) of blood samples were positive using nested PCR. Out of 68 positive samples, 85.3% (58/68) and 11.7% (8/68) were respec­tively positive for Theileria ovis and T. lestoquardi. Mixed infection was detected in 3% (2/68) cases. Overall, 420 ixodid ticks belong to seven different hard ticks species were collected from goats. Rhipicephalus turanicus 112 (26.7%), R. sanguineus 95 (22.6%), R. bursa, 91(21.7%), Hyalomma anatolicum, 55(13.1%), H. excavatum 27(6.4%), H. marginatum, 22(5.3%) and Dermacentor marginatus, 18(4.2%) were the main tick species infesting goats. The PCR products obtained from ticks were subjected to the differentiation of Theileria species. Respectively, 2 and 8 pools of H. marginatum and R. turanicus salivary glands were infected with T. ovis and T. lestoquardi. In addition, T. annulata and T. lestoquardi infection weredetected in three pools of H. anatolicum. Conclusion: This is the first report of goats and collected ticks to Theileria spp infection in Iran. The results suggest that T. ovis has a higher prevalence than T. lestoquardi. It is also postulated H. marginatum, R. turanicus and H. ana­tolicum might play an important role in the field as a vector of Theileria spp in this area.

First Molecular Evidence of Theileria Lestoquardi in Small Ruminants in Northern Iran

2021 ◽  
Author(s):  
Seyed Mousa Motavalli Haghi ◽  
Mahdi Fakhar ◽  
Mitra Sharbatkhori ◽  
Abdol Sattar Paghe
Keyword(s):  
Staining Method ◽  
Small Ruminants ◽  
Molecular Study ◽  
Common Species ◽  
Giemsa Staining ◽  
Molecular Evidence ◽  
Northern Iran ◽  
Theileria Lestoquardi ◽  
High Prevalence ◽  
Peripheral Blood Smears

Abstract Ovine theileriosis as a critical agent in small ruminant production, can cause lethal infections. Different species of Theileria have been reported in various parts of the world, and each species causes different diseases in the host. This is the first molecular study to investigate the prevalence of ovine theileriosis and identify the dominant Theileria species in northern Iran. A number of 220 small ruminants, including sheep and goats, were randomly sampled from 22 flocks. Peripheral blood smears are stained by the Giemsa staining method. As well as for species identification, all samples were examined by molecular method. From 220 samples, 160 and 60 were sheep and goat, respectively. By the Giemsa staining method, Theileria parasite was observed in 20 (9%) samples. But by polymerase chain reaction (PCR) method, 30 (13.6%) samples were positive for Theileria species. Theileria lestoquardi was the most common species found in these animals. The high prevalence of theileriosis in small ruminants demonstrates the emergence of ovine theileriosis in Mazandaran and Golestan provinces in northern Iran.

scholarly journals Development of a Multiplex PCR and Magnetic DNA Capture Assay for Detecting Six Species Pathogens of the Genera Anaplasma and Ehrlichia in Canine, Bovine, Caprine and Ovine Blood Samples from Grenada, West Indies

Pathogens ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 192
Author(s):  
Bhumika Sharma ◽  
Roman R. Ganta ◽  
Diana Stone ◽  
Andy Alhassan ◽  
Marta Lanza-Perea ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Clinical Signs ◽  
Small Ruminants ◽  
High Sensitivity ◽  
Molecular Evidence ◽  
Blood Samples ◽  
Persistent Problem ◽  
Magnetic Capture ◽  
Low Sensitivity ◽  
Species Specific

Infections with tick-borne pathogens belonging to Anaplasma/Ehrlichia in various vertebrate hosts are a persistent problem resulting in nonspecific clinical signs during early infection. Diagnosis of single and multi-infections with these pathogens, causing diseases in companion/agricultural animals and people, remains a challenge. Traditional methods of diagnosis, such as microscopy and serology, have low sensitivity and specificity. Polymerase chain reaction (PCR) assays are widely used to detect early-phase infections, since these have high sensitivity and specificity. We report the development and validation of an assay involving PCR followed by magnetic capture method using species-specific oligonucleotides to detect six Anaplasma/Ehrlichia species pathogens in canine, bovine, caprine, and ovine blood samples. Overall, the assay application to 455 samples detected 30.1% (137/455) positives for one or more out of six screened pathogens. Single-pathogen infections were observed in 94.9% (130/137) of the positive samples, while co-infections were detected in 5.1% (7/137). Anaplasma marginale infection in cattle had the highest detection rate (34.4%), followed by canines positive for Anaplasma platys (16.4%) and Ehrlichia canis (13.9%). The assay aided in documenting the first molecular evidence for A. marginale in cattle and small ruminants and Ehrlichia chaffeensis and Ehrlichia ewingii in dogs in the Caribbean island of Grenada.

scholarly journals Genetic characterization and phylogenetic study of Indonesian indigenous catfish based on mitochondrial cytochrome B gene

2020 ◽  
Vol 13 (1) ◽  
pp. 96-103
Author(s):  
Dorothea Vera Megarani ◽  
Herjuno Ari Nugroho ◽  
Zahrah Prawita Andarini ◽  
Yura Dwi Risa B. R. Surbakti ◽  
Rini Widayanti
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Cytochrome B ◽  
Genetic Characterization ◽  
Phylogenetic Study ◽  
Cyt B ◽  
Phylogenetic Structure ◽  
Mitochondrial Cytochrome B ◽  
Sperata Seenghala ◽  
Cyt B Gene

Aim: This study aimed to determine the genetic characterization and phylogenetic structure of Indonesian indigenous catfish using cytochrome B (Cyt B) sequences. Materials and Methods: The genomes of 26 catfishes caught from nine rivers from nine different geographical locations around Indonesia were analyzed. The tissue isolation method was used to isolate the total genome of the fishes. Furthermore, polymerase chain reaction was done to amplify the mtDNA Cyt B using the CytBF and CytBR primers. Following sequencing, the analysis of genetic variation and the phylogenetic relationship was performed using MEGA version X software. Results: Cyt B gene sequencing attained a total of 1139 nucleotides encrypting 379 amino acids for all samples. The ClustalW alignment program using MEGA X software revealed 395 substituted nucleotides, which then translated into 63 amino acid variation sites among all 26 samples. No amino acids in catfish BB were different compared to catfish PM, MP, and KR2,3. Catfish MS had one modified amino acid; KR1 and KS had two different amino acids; BF had 38 different amino acids; EM had 31 different amino acids; and BSBJ had 26 different amino acids compared to catfish BB. The most significant alteration of amino acids was between catfish EM and BF (49 amino acids). Conclusion: Indonesian catfish were divided into five clades based on the Cyt B gene. Samples KR and MP (Sumatra); MS and BB (Kalimantan); and PM (Java) were clustered with Hemibagrus nemurus and Hemibagrus wyckioides (Bagridae family). Samples from Kalimantan (KS) and one sample of KR (KR1) from Sumatra were clustered with Sperata seenghala and Hemibagrus spilopterus (Bagridae family). Samples from Java (BSBJ) were clustered with Pseudolais pleurotaenia (Pangasiidae family). Samples EM (Java) were together with Mystus cavasius (Bagridae family). Samples from West Papua were clustered with Potamosilurus latirostris (Ariidae family).

scholarly journals Microscopic And Molecular Identification of Anaplasma Ovis in Small Ruminants in Duhok Province in Kurdistan Region of Iraq

2020 ◽  
Vol 23 (2) ◽  
pp. 228-235
Author(s):  
Adnan Ahmed ◽  
Jassim M Abdo
Keyword(s):  
Microscopic Examination ◽  
Prevalence Rate ◽  
Small Ruminants ◽  
Surface Protein ◽  
Positive Sample ◽  
Pcr Analysis ◽  
Anaplasma Ovis ◽  
Blood Samples ◽  
Major Surface Protein ◽  
Major Surface

In last ten years, there has been a developing enthusiasm for microscopic organisms from the genus Anaplasma, particularly the species A. ovis. It is associated with the pathogenic action of these microscopic organisms in livestock. Anaplasma ovis is a tick-borne obligate intracellular rickettsial bacterium that causes anaplasmosis in domestic and wild small ruminants. The samples of the present study were collected from small ruminants from inside seven distinct regions (Akre, Simele, Zummar, Feshchapoor, Deraboon, Bajed Kandal,Karoda)of Duhok province, 389 (goats 75 and sheep 314) during the period of April and May 2018, blood sample were taken and thin smear was formed, after Giemsa’s staining the slide is observed under microscope. In this study used Giemsa stain for microscopic examination out of 389 animals 250 were found positive for Anaplasma ovis infection with a prevalence rate of 64.26 % and 139 of them were negative with a prevalence rate of 35.73 %. According to the species of animals, the highest prevalence of A. ovis infection in animals by using microscopic examination was 67.83 %, 213 positive sample from total 314 blood samples from sheep and lowest prevalence was 49.33 %, 37 positive sample from total 75 blood samples from goats. PCR analysis of 100 blood samples obtained from total 250 positive blood samples after DNA extraction and measure of concentration and purity we used 2 primers that target major surface protein 4 (MSP4) in A. ovis genomic DNA. The results of PCR test with major surface protein 4 primer was 83 samples positive from total 100 samples, According to the species of animals, the highest prevalence of A. ovis was 83.7 %, 72 positive sample from total 86 blood samples from sheep and lowest prevalence was 78.5 %, 11 positive sample from total 14 blood samples from goats.

scholarly journals Epizootic Situation on Anaplasmosis of Small Ruminants in the Irkutsk Region

2021 ◽  
Vol 6 (1) ◽  
pp. 60-68
Author(s):  
O. V. Suntsova ◽  
V. A. Rar ◽  
O. V. Lisak ◽  
I. V. Meltsov ◽  
E. K. Doroschenko ◽  
...  
Keyword(s):  
Nucleotide Substitution ◽  
Small Ruminants ◽  
Wide Distribution ◽  
Anaplasma Ovis ◽  
Single Nucleotide ◽  
Blood Samples ◽  
Sheep Blood ◽  
Irkutsk Region ◽  
Probable Role

Anaplasmosis of ruminants is a group of natural focal infections caused by bacteria from the genus Anaplasma of the Anaplasmataceae family. The main etiological agent of anaplasmosis in sheep, goats, and wild ruminants is Anaplasma ovis, which parasitizes in the erythrocytes of these animals. The purpose of this study was the finding and identification of Anaplasma spp. in the blood of small ruminants using genetic methods and obtaining data on the distribution of anaplasmosis in the Irkutsk region. 20 goat blood samples, 611 sheep blood samples and 209 Dermacentor nuttalli ticks from 12 districts of the Irkutsk region were examined for the presence of Anaplasma spp. Only one type of anaplasma, A. ovis, was found among the genotyped samples. A. ovis was found in the blood of sheep and goats in all of the studied districts of the Irkutsk region. The proportion of sheep blood samples containing anaplasma DNA varied from 30 % to 85 %, in goats – from 10 % to 100 % in different districts, and averaged 57.8 % in sheep and 55,0 % in goats. Frequency of infection of D. nuttalli ticks with A. ovis was 5.7 %. The nucleotide sequences of the samples detected in the blood of small ruminants on the territory of the Irkutsk region differed from each other by a single nucleotide substitution and were identical to the sequences of the type strain Haibei, as well as the sequences of A. ovis previously found in the blood of sheep from Mongolia, deer from China, and Dermacentor niveus and Dermacentor nuttalli ticks from China. These sequences were also identical to the sequences previously found in the blood of sheep from Altai and in Dermacentor nuttalli ticks from Tuva, which indicates the wide distribution of these A. ovis genovariants in Siberia and the probable role of D. nuttalli as a carrier of the agent of anaplasmosis of small ruminants in the Irkutsk region.

scholarly journals Molecular identification of the Trypanosoma (Herpetosoma) lewisi clade in black rats (Rattus rattus) from Australia

2019 ◽  
Author(s):  
Siobhon L. Egan ◽  
Casey L. Taylor ◽  
Jill M. Austen ◽  
Peter B. Banks ◽  
Liisa A. Ahlstrom ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Molecular Identification ◽  
Ribosomal Rna ◽  
Rattus Rattus ◽  
Molecular Evidence ◽  
Diverse Range ◽  
Blood Samples ◽  
Trypanosoma Lewisi ◽  
Sydney Australia ◽  
Black Rats

AbstractInvasive rodent species are known hosts for a diverse range of infectious microorganisms and have long been associated with the spread of disease globally. The present study describes molecular evidence for the presence of a Trypanosoma sp. from black rats (Rattus rattus) in northern Sydney, Australia. Sequences of the 18S ribosomal RNA (rRNA) locus were obtained in two out of eleven (18%) blood samples with subsequent phylogenetic analysis confirming the identity within the Trypanosoma lewisi clade.

scholarly journals An Assessment of the Molecular Diversity of Ticks and Tick-Borne Microorganisms of Small Ruminants in Pakistan

2020 ◽  
Vol 8 (9) ◽  
pp. 1428 ◽  
Author(s):  
Abdul Ghafar ◽  
Adil Khan ◽  
Alejandro Cabezas-Cruz ◽  
Charles G. Gauci ◽  
Sadaf Niaz ◽  
...  
Keyword(s):  
Sequence Data ◽  
Phylogenetic Analyses ◽  
Strong Support ◽  
Small Ruminants ◽  
Ixodid Ticks ◽  
Nucleotide Sequence Data ◽  
Health Significance ◽  
Second Internal Transcribed Spacer ◽  
Theileria Spp

This study investigated ticks and tick-borne microorganisms of small ruminants from five districts of the Federally Administered Tribal Area (FATA) of Pakistan. Morphological (n = 104) and molecular (n = 54) characterization of the ticks revealed the presence of six ixodid ticks: Rhipicephalus (Rh.) haemaphysaloides, Rh. microplus, Rh. turanicus, Haemaphysalis (Hs.) punctata, Hs. sulcata and Hyalomma anatolicum. Phylogenetic analyses of nucleotide sequence data for two mitochondrial (16S and cytochrome c oxidase 1) and one nuclear (second internal transcribed spacer) DNA regions provided strong support for the grouping of the six tick species identified in this study. Microfluidic real-time PCR, employing multiple pre-validated nuclear and mitochondrial genetic markers, detected 11 potential pathogens and endosymbionts in 72.2% of the ticks (n = 54) tested. Rickettsia (R.) massiliae was the most common pathogen found (42.6% of ticks) followed by Theileria spp. (33.3%), Anaplasma (A.) ovis and R. slovaca (25.9% each). Anaplasma centrale, A. marginale, Ehrlichia spp., R. aeschlimannii, R. conorii and endosymbionts (Francisella- and Coxiella-like) were detected at much lower rates (1.9–22.2%) in ticks. Ticks from goats (83.9%) carried significantly higher microorganisms than those from sheep (56.5%). This study demonstrates that ticks of small ruminants from the FATA are carrying multiple microorganisms of veterinary and medical health significance and provides the basis for future investigations of ticks and tick-borne diseases of animals and humans in this and neighboring regions.

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