scholarly journals PCR-Based Detection ofBabesia ovisinRhipicephalus bursaand Small Ruminants

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
Bijan Esmaeilnejad ◽  
Mousa Tavassoli ◽  
Siamak Asri-Rezaei ◽  
Bahram Dalir-Naghadeh ◽  
Karim Mardani ◽  
...  

This study aimed to assess the prevalence ofBabesia ovisinfection in adultRhipicephalus bursaand small ruminants in West Azerbaijan province, Iran. Blood samples were collected from 280 sheep and 122 goats of forty randomly selected flocks. SpecificB. ovisfragment was detected in 67 animals (16.7%), of which 52 animals (18.6%) were sheep and 15 animals (12.2%) goats (P<0.05). Of the 848R. bursacollected from naturally infested small ruminants and farm dogs,Babesia oviswas detected by PCR in salivary glands of 94 adult ticks. The frequency ofB. ovisinfection was higher in flocks with tick in comparison with animals without tick (P<0.05). Positive amplification from blood of ruminants, ticks, oviposition ticks, eggs, and larvae was subjected to restriction digestion withHphI. One RFLP profile was produced. The PCR-RFLP results indicated that one strain ofB. ovisexists in this area. The results showed that the PCR was useful method to investigate the epidemiology of small ruminants’ babesiosis. Furthermore,R. Bursa, which can transovarially transmitB. ovisand as well as being widely distributed in West Azerbaijan province, Iran, might play an important role in the field as a natural vector ofB. ovis.

Animals ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 814
Author(s):  
Münir Aktaş ◽  
Sezayi Özübek ◽  
Mehmet Can Uluçeşme

Anaplasma phagocytophilum causes tick-borne fever in small ruminants. Recently, novel Anaplasma variants related to A. phagocytophilum have been reported in ruminants from Tunisia, Italy, South Korea, Japan, and China. Based on 16S rRNA and groEL genes and sequencing, we screened the frequency of A. phagocytophilum and related variants in 433 apparently healthy small ruminants in Turkey. Anaplasma spp. overall infection rates were 27.9% (121/433 analyzed samples). The frequency of A. phagocytophilum and A. phagocytophilum-like 1 infections was 1.4% and 26.5%, respectively. No A. phagocytophilum-like 2 was detected in the tested animals. The prevalence of Anaplasma spp. was comparable in species, and no significant difference was detected between sheep and goats, whereas the prevalence significantly increased with tick infestation. Sequencing confirmed PCR-RFLP data and showed the presence of A. phagocytophilum and A. phagocytophilum-like-1 variant in the sampled animals. Phylogeny-based on 16S rRNA gene revealed the A. phagocytophilum-like 1 in a separate clade together with the previous isolates detected in small ruminants and ticks. In this work, A. phagocytophilum-like 1 has been detected for the first time in sheep and goats from Turkey. This finding revealed that the variant should be considered in the diagnosis of caprine and ovine anaplasmosis.


1970 ◽  
Vol 1 (1) ◽  
pp. 14-21 ◽  
Author(s):  
Raunak Shrestha ◽  
Rubin Narayan Joshi ◽  
Kriti Joshi ◽  
Bal Hari Poudel ◽  
Bhupal Govinda Shrestha

Ser315Thr mutations in genes encoding the mycobacteria catalase-peroxidase (KatG) has been associated with the major resistance to isoniazid (INH) in Mycobacterium tuberculosis (MTB). Also G/C polymorphisms in INT4 region of the solute carrier family 11 member 1 gene (SLC11A1) and susceptibility towards tuberculosis (TB) has been demonstrated worldwide. 24 drug resistant MTB culture positive samples and 24 whole?blood samples were collected from different TB patients of Central Development Region of Nepal in 2009. A Polymerase Chain Reaction (PCR) - Restriction Fragment Length Polymorphism (RFLP) assay was carried out in order to investigate Ser315Thr KatG mutation and G/C polymorphism in INT4 region. 4 (16.67%) samples out of 24 MTB culture samples demonstrated the Ser315Thr KatG mutation whereas none of the 24 whole blood samples were found to contain G/C polymorphism in INT4. Though no significant correlation could be found between INT4 polymorphism and TB susceptibility, overall scenario of Nepal cannot be drawn from this data. Molecular diagnostic technique such as PCR-RFLP can be used in a robust scale to carry out base line studies in the TB population of Nepal. Key words: Multi?drug resistance; Tuberculosis; PCR; RFLP Nepal Journal of Biotechnology. Jan. 2011, Vol. 1, No. 1 : 14-21


2020 ◽  
Vol 23 (2) ◽  
pp. 228-235
Author(s):  
Adnan Ahmed ◽  
Jassim M Abdo

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.


Author(s):  
O.D. Biriukova ◽  
T.M. Suprovych ◽  
M.P. Suprovych ◽  
S.V. Laiter-Moskaliuk ◽  
I.O. Chornyi

Molecular genetic markers can detect polymorphism at the DNA level. This feature determines the possibility of their widespread use in genetics and breeding. Alleles of the BoLA-DRB3 gene (exon 2) can act as such markers if a statically significant association between the disease and the allele is established. The presence of such DNA markers in the genotype of animals makes it possible to judge the likelihood of disease in postnatal ontogenesis immediately after the birth of a heifer, based on which we can conclude about the conditions of further use of the animal in the main herd. According to the results of studying the polymorphism of the BoLA-DRB3 gene in cows of the Ukrainian black and white dairy breed resistant and susceptible to necrobacteriosis, four "informative" alleles were revealed. Two of them *03 and *22 are associated with resistance, and the other two - *16 and *23 with susceptibility to necrobacteriosis. The presence of these alleles in the genotype of the animal is determined by testing performed by PCR-RFLP. The method is time consuming, labor intensive and costly. To simplify it, the following technique is proposed. Restriction fragments of alleles *03, *16, *22 and *23 for endocluases RsaI, XhoII and HaeIII have the following DNA patterns: bbb, jbd, mba and nba. Due to the peculiarity of the restriction fragments, which is that endonuclease XhoII reveals in these alleles only one pattern b with length of 284 bp, the process of determining informative alleles can be simplified. Isolation of DNA from blood samples and amplification of a fragment of the BoLA-DRB3.2 gene with a size of 284 bp is carried out according to the established technique. Next, the restriction of the fragment by endonuclease XhoII and sampling having a pattern b. Selected samples are treated with RsaI endonuclease and only those with patterns b, j, m and n remain. The next step is to restrict the selected samples with HaeIII endonuclease and select heifers with bbb (*03) and nba (*23) genotypes. After the first restriction, blood samples without pattern b are eliminated from the experimental sample; after the second – two alleles with patterns RsaI + XhoII jb (*16) and mb (*22) are unambiguously determined, after the third – genotypes bbb and nba, which correspond to alleles *03 and *23. In total, only 75% of blood samples are typed, which reduces the material consumption, time and cost of work to identify heifers genetically susceptible (resistant) to necrobacteriosis.


2019 ◽  
Vol 6 (2) ◽  
pp. 232
Author(s):  
Sutikno Sutikno ◽  
Rudy Priyanto ◽  
Cece Sumantri ◽  
Jakaria Jakaria

ABSTRAK Gen FTO berfungsi sebagai regulasi homeostasis, deposisi lemak dan pengaturan obesitas. Penelitian ini bertujuan untuk mengidentifikasi polimorfisme SNP g.125550A>T di ekson 3 gen FTO pada bangsa sapi potong Indonesia. Sampel darah diperoleh dari 209 ekor sapi, terdiri atas sapi bali (44), madura (20), Pesisir (20), katingan (20), Peranakan ongole (PO) (22), Pasundan (20), Sumba Ongole (SO) (11), brahman (20), simental (15), dan limousin (18). Polimorfisme gen FTO dianalisis menggunakan metode PCR-RFLP (HpyCH4III) dan direct sequencing. Hasil genotiping SNP g.125550A>T adalah polimorfik (genotipe AA, AT, dan TT) pada sapi madura, pesisir, katingan, PO, pasundan, SO, brahman, simental, dan limousin. Frekuensi alel A dan T masing-masing adalah 0,70, 0,68, 0,84, 0,89, 0,70, 0,86, 0,90, 0,73, 0,69 dan 0,30, 0,33, 0,16, 0,11, 0,30, 0,14, 0,10, 0,27, 0,31. Nilai Ho dan He masing-masing adalah 0,60-0,14 dan 0,44-0,18 serta dalam keseimbangan Hardy-Weinberg (P>0.05). Sementara pada sapi bali bersifat monomorfik hanya bergenotipe AA. Hasil sekuensing SNP g.125550A>T ditemukan mutasi tranvesi A menjadi T pada posisi nukleotida  g.125550. Berdasarkan hasil penelitian ini, dapat disimpulkan bahwa SNP 125550A>T gen FTO beragam dan berpotensi dijadikan marka genetik untuk kualitas daging pada bangsa sapi potong Indonesia.Kata Kunci: gen FTO, PCR-RFLP, Sapi, SNP g.125550A>TABSTRACTThe FTO gene functions as regulation of homeostasis, fat deposition and regulation of obesity. This study aimed to identify the polymorphism of SNP g.125550A>T in exon 3 of FTO gene in Indonesian beef cattle. Blood samples were collected from 209 cattle, including bali (44), madura (20), pesisir (20), katingan (20), PO (22), pasundan (20), SO (11), brahman (20), simental (15), and limousin (18). Polymorphism of the FTO gene was analyzed using PCR-RFLP (HpyCH4III) and direct sequencing methods. The results of genotyping SNP g.125550A>T was polymorphic (AA, AT and TT genotypes) in madura, pesisir, katingan, PO, pasundan, SO, brahman, simental, and limousin cattle. The frequency of A and T alleles were 0,70, 0,68, 0,84, 0,89, 0,70, 0,86, 0,90, 0,73, 0,69 and 0,30, 0,33, 0,16, 0,11, 0,30, 0,14, 0,10, 0,27, 0,31 respectively. The values of Ho and He were 0,60-0,14 and 0,44-0,18 respectively and in Hardy-Weinberg equilibrium (P>0,05). While in Bali cattle was monomorphic (AA genotype). Results of sequencing SNP g.125550A>T of the FTO gene found a transverse mutation A to T at the nucleotide position g.125550. As a result of this study, it can be concluded that SNP 125550A>T of the FTO gene was diverse and potentially used as genetic markers for meat quality in Indonesian beef cattle.Keywords: cattle, FTO gene, PCR-RFLP, SNP g.125550A>T.


2021 ◽  
Vol 6 (1) ◽  
pp. 60-68
Author(s):  
O. V. Suntsova ◽  
V. A. Rar ◽  
O. V. Lisak ◽  
I. V. Meltsov ◽  
E. K. Doroschenko ◽  
...  

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.


Author(s):  
T. I. Trukhina ◽  
I. A. Solovieva ◽  
G. A. Bonadrenko

The authors focus on the fight against the most common helminthic diseases of agricultural animals as they see it as an important part of the veterinary science. Bunostomosis is one of such diseases; it is a helminthiasis of ruminants caused by vipostomum trigonocephalum nematodes (in sheep and goats) and vipostomum phlebotomum (mainly in the cattle), which are parasitic in the small intestines. The disease is characterized by diarrhea, exhaustion and death of ruminants. The disease is widespread and causes great economic damage. The authors explored the prevalence of tuberculosis and age-related infection with tuberculosis by ovoscopy and levoscopy in the Amur region. They examined 2655 animals, including 1415 small ruminants and 1240 cattle from five districts of the Amur region. The researchers examined 4 calves and 4 lambs aged 4 inorder to study the period of bunostom development in the organism of animals, which were infected with percutaneously and orally. Brown larvae received from faeces of spontaneously infested animals is considered to be the material for infection. The development and survival of larvae in the environment were investigated in 2016-2017. The authors found out that in the Amur region larvae develop at the temperatures above 40С. The period of development at the temperature from 40С tо 9 0С was 23 days. When the temperature was increasing, the development period was reduced to 5-7 days. In October larvae stop their development. The underdeveloped eggs and larvae left die in winter. At the farms of the Amur region, the infection rate of cattle and sheep is 31.9 and 49.6%. The bunostome infection of the cattle aged 1-11 months is 59,8%; 1-2 years - 26,0; 3 years and more - 19,9; sheep aged 4-11 months infection rate is 52,9; 1 year - 60,9, 3 years and more - 38,7%. The authors observed bunostomes development during 89-102 days in the ruminant organism before the sexually mature stage in the conditions of the Amur region.


Biomolecules ◽  
2020 ◽  
Vol 10 (5) ◽  
pp. 773
Author(s):  
Edgar Jesús Delgado-Núñez ◽  
Alejandro Zamilpa ◽  
Manasés González-Cortazar ◽  
Agustín Olmedo-Juárez ◽  
Alexandre Cardoso-Taketa ◽  
...  

Haemonchus contortus affect small ruminants all over the world. Anthelmintics cause resistance, contamination, and a risk of public health. Prosopis laevigata is a plant used as a home remedy against many diseases in Mexico. This study arose from a preliminary study where a P. laevigata hydroalcoholic extract (Pl-hae) showed anthelmintic activity (aa) against H. contortus. Searching for bioactive compounds (bac) with high aa, the Pl-hae was fractioned obtaining an aqueous (Aq-F) and an ethyl acetate fraction (EtAc-F), and a flavonoid with aa identified as isorhamnetin was obtained from EtAc-F. Both fractions were in vitro assessed by the egg hatch test (eht) and larval mortality (lm) assays. The bac obtained from EtAc-F were characterised by NMR analysis. The highest aa were recorded with EtAc-F, resulting in 100% eht and 80.45% lm at 0.75 and 30 mg/mL, respectively. Alterations in eggs and larvae attributed to isorhamnetin were recorded by environmental scanning electron microscopy, confocal laser scanning and by high-resolution digital-coupled camera. This flavonoid caused 100% eht at 0.07 mg/mL after 48 h and 100% lm at 7.5 mg/mL after 72 h exposure. Isorhamnetin has promising potential as an anthelmintic against sheep haemonchosis.


2018 ◽  
Vol 43 (4) ◽  
pp. 323
Author(s):  
S. Sutikno ◽  
R. Priyanto ◽  
C. Sumantri ◽  
J. Jakaria

The ADIPOQ and EDG1 genes were responsible in intramuscular fat deposition and marbling scores. This study was aimed to identify polymorphism of indel g.81966364D>I in promoter region of ADIPOQ gene and SNP c.-312A>G in 5' UTR of EDG1 gene in Indonesian beef cattle. Blood samples were collected from 211 cattle, including Bali (44), Madura (20), Pesisir (18), Katingan (20), PO (22), Pasundan (20), SO (12), Brahman (20), Simmental (15) and Limousin (18). Polymorphism of ADIPOQ gene was analyzed using PCR and direct sequencingmethods, whereas EDG1 gene was analyzed using PCR-RFLP (MscI enzyme) and direct sequencingmethods. Results of genotyping indel g.81966364D>I was monomorphic (DD genotype). The SNP c.-312A>G was polymorphic (AA and AG genotype) in Madura, Pesisir, Pasundan, Brahman, and Limousine. The Frequencies of allele A and G were 0.95, 0.92, 0.98, 0.95, 0.94 and 0.05, 0.08, 0.02, 0.05, 0.06 respectively. The values of Ho and He were 0.05-0.17 and 0.05-0.15 respectively and in Hardy-Weinberg equilibrium (P>0.05). In Bali, Katingan, PO, SO and Simmental were monomorphic (GG genotype). In Bali cattle, two novel SNP candidates were found in position of c.-399C>T and c.-273C>G which were potential to be used as genetic markers of marbling score for Bali cattle. As result this study, it can be concluded that ADIPOQ gene was similar while EDG1 gene was different in Indonesian beef cattle. in addition, found two candidates potential SNP in Bali cattle.


2014 ◽  
Vol 63 (5) ◽  
pp. 659-666 ◽  
Author(s):  
Kazumasa Kamei ◽  
Masahiro Asakura ◽  
Srinuan Somroop ◽  
Noritoshi Hatanaka ◽  
Atsushi Hinenoya ◽  
...  

Although Campylobacter jejuni and Campylobacter coli are the most common bacterial causes of human gastrointestinal diseases, other Campylobacter species are also involved in human and animal infections. In this study, we developed a cytolethal distending toxin (cdt) gene-based PCR-RFLP assay for the detection and differentiation of C. jejuni, C. coli, C. fetus, C. hyointestinalis, C. lari, C. helveticus and C. upsaliensis. Previously designed common primers, which can amplify the cdtB gene of C. jejuni, C. coli and C. fetus, were used for detecting seven Campylobacter species and differentiating between them by restriction digestion. The PCR-RFLP assay was validated with 277 strains, including 35 C. jejuni, 19 C. coli, 20 C. fetus, 24 C. hyointestinalis, 13 C. lari, 2 C. helveticus, 22 C. upsaliensis, 3 other Campylobacter spp. and 17 other species associated with human diseases. Sensitivity and specificity of the PCR-RFLP assay were 100 % except for C. hyointestinalis (88 % sensitivity). Furthermore, the PCR-RFLP assay successfully detected and differentiated C. jejuni, C. coli and C. fetus in clinical and animal samples. The results indicate that the PCR-RFLP assay is useful for the detection and differentiation of seven Campylobacter species important for human and animal diseases.


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