Molecular diagnosis of the tick-borne pathogen Anaplasma marginale in cattle blood samples from Nigeria using qPCR

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.

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A pilot study on molecular diagnosis of Hapalotrema mistroides (Digenea: Spirorchiidae) infection in blood samples of live loggerhead turtles Caretta caretta

BMC Veterinary Research ◽

10.1186/s12917-020-2232-y ◽

2020 ◽

Vol 16 (1) ◽

Cited By ~ 1

Author(s):

Erica Marchiori ◽

Giorgia Dotto ◽

Cinzia Tessarin ◽

Mario Santoro ◽

Andrea Affuso ◽

...

Keyword(s):

Pilot Study ◽

Molecular Diagnosis ◽

Caretta Caretta ◽

Blood Samples ◽

Loggerhead Turtles

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Polymorphism of ADIPOQ and EDG1 genes in Indonesian beef cattle

Journal of the Indonesian Tropical Animal Agriculture ◽

10.14710/jitaa.43.4.323-332 ◽

2018 ◽

Vol 43 (4) ◽

pp. 323

Author(s):

S. Sutikno ◽

R. Priyanto ◽

C. Sumantri ◽

J. Jakaria

Keyword(s):

Beef Cattle ◽

Promoter Region ◽

Marbling Score ◽

Blood Samples ◽

Adipoq Gene ◽

Pcr Rflp ◽

Hardy Weinberg Equilibrium ◽

Cattle Blood ◽

Bali Cattle ◽

Novel Snp

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.

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Influence of storage at different temperatures on inorganic phosphorus levels of cattle blood samples

Veterinary Record ◽

10.1136/vr.111.19.441 ◽

1982 ◽

Vol 111 (19) ◽

pp. 441-441 ◽

Cited By ~ 2

Author(s):

A. Hunter ◽

K. Matthews

Keyword(s):

Inorganic Phosphorus ◽

Blood Samples ◽

Different Temperatures ◽

Cattle Blood ◽

Phosphorus Levels

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Development of a Method to Detect Mycobacterium paratuberculosis in the Blood of Farmed Deer Using Actiphage® Rapid

Frontiers in Veterinary Science ◽

10.3389/fvets.2021.665697 ◽

2021 ◽

Vol 8 ◽

Author(s):

Anton Kubala ◽

Tania M. Perehinec ◽

Catherine Evans ◽

Andrea Pirovano ◽

Benjamin M. C. Swift ◽

...

Keyword(s):

Gradient Centrifugation ◽

Elisa Test ◽

Buffy Coat ◽

Test Results ◽

Blood Samples ◽

Diagnostic Potential ◽

Ficoll Gradient ◽

Blood Lysis ◽

Small Set ◽

Cattle Blood

Mycobacterium avium subsp paratuberculosis (MAP) is the causative agent of Johne's disease, which is an economically and clinically relevant pathogen for commercial deer production. The purpose of this study was to develop a method that could be used to rapidly detect MAP infection in deer using the Actiphage Rapid blood test. This test has previously been used to detect MAP in cattle blood following the purification of buffy coat using Ficoll gradients, however this method is quite laborious and costly. The purpose of this study was to develop a simpler method of blood preparation that was also compatible with deer blood and the Actiphage test. Initially differential lysis of RBCs using Ammonium Chloride-Potassium (ACK) blood lysis buffer was compared with the Ficoll gradient centrifugation method using cattle blood samples for compatibility with the Actiphage reagents, and it was found that the simpler ACK method did not have an impact on the Actiphage test reagents, producing an equivalent sensitivity for detection of low levels of MAP. When the two methods were compared using clinical blood samples from farmed deer, the ACK lysis method resulted in a cleaner sample. When a blinded test of 132 animals from 4 different production groups was carried out, the majority of the positive test results were found to be from animals in just one group, with a small number identified in a second group. The test results were found to be reproducible when a small set of positive animals were tested again 1 month after their initial testing. Finally a set of negative animals which had been previously screened using an ELISA test, all animals gave a negative Actiphage result. This study shows that this improved sample preparation method and Actiphage blood testing can be used to test blood samples from deer, and the full diagnostic potential of the method can now be evaluated.

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Bayesian estimation of ELISA and gamma interferon test accuracy for the detection of bovine tuberculosis in caudal fold test–negative dairy cattle in Kuwait

Journal of Veterinary Diagnostic Investigation ◽

10.1177/1040638718759574 ◽

2018 ◽

Vol 30 (3) ◽

pp. 468-470 ◽

Cited By ~ 2

Author(s):

Salwa Al-Mouqatea ◽

Mohammad Alkhamis ◽

Batool Akbar ◽

Abdulmohsen Ali ◽

Hameed Al-Aqeel ◽

...

Keyword(s):

Dairy Cattle ◽

Mycobacterium Bovis ◽

Bayesian Approach ◽

Bovine Tuberculosis ◽

Dairy Farms ◽

Test Accuracy ◽

Infected Cattle ◽

Blood Samples ◽

Ancillary Test ◽

Cattle Blood

Bovine tuberculosis (TB) is endemic in Kuwait; cattle identified as TB-positive using the caudal fold test (CFT) are culled. We used a Bayesian approach to estimate the sensitivity (Se) and specificity (Sp) of the IFNγ assay and ELISA, which are not routinely used in Kuwait in CFT-negative dairy cattle. Blood samples from CFT-negative cattle ( n = 384) collected from 38 dairy farms were tested by IFNγ assay and ELISA. The Se and Sp (95% CI) of the IFNγ were 85.0% (67.6–95.3%) and 90.4% (86.7–95.3%), respectively, whereas estimates for the ELISA were 61.1% (33.1–84.6%) and 85.4% (81.7–88.8%). TB prevalence (95 CI%) in CFT-negative cattle was estimated as 2.6% (0.5–9.5%). The IFNγ assay may play a role as an ancillary test for the identification of Mycobacterium bovis–infected cattle that are undetected by CFT.

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Differences in the occurrence of selenium, copper and zinc deficiencies in dairy cows, calves, heifers and bulls

Veterinární Medicína ◽

10.17221/5638-vetmed ◽

2012 ◽

Vol 50 (No. 9) ◽

pp. 390-400 ◽

Cited By ~ 9

Author(s):

L. Pavlata ◽

A. Podhorsky ◽

A. Pechova ◽

P. Chomat

Keyword(s):

Blood Serum ◽

Dairy Cows ◽

Zn Deficiency ◽

Blood Samples ◽

Cu Deficiency ◽

Status Assessment ◽

Cattle Blood ◽

Zn Status ◽

Se Status

This study was conducted to evaluate the mineral status of various bovine categories reared on the same farm, and to monitor the quality of their mineral nutrition in this way. Blood samples were collected on 20 farms in various regions of the CzechRepublicto diagnose the selenium (Se), copper (Cu) and zinc (Zn) status of dairy cows, calves, heifers and bulls. Blood samples were collected from 5 dairy cows and their calves, 5 heifers and, whenever possible, also from 5 bullocks (10 farms). To assess the Cu and Zn status of the cattle, blood serum concentrations of the elements were determined by flame AAS. The Se status assessment was based on the glutathione peroxidase (GSH-Px) activity in whole blood. Dairy cows and calves showed the higher GSH-Px activity in comparison with heifers and bulls (P < 0.001). Overall mean GSH-Px activity in the blood of dairy cows, calves, heifers and bulls was 720.47 ± 174.47 µkat/l, 688.34 ± 204.12 µkat/l, 555.69 ± 318.36 µkat/l and 516.17 ± 214.70 µkat/l, respectively. Se deficiency was diagnosed in 23% of the examined dairy cows (20% herds), 31% calves (25% herds), 51% heifers (50% herds) and in 58% of bulls (50% herds). Dairy cows showed lower Zn and higher Cu concentrations in comparison with the other categories (P < 0.001). Mean Zn concentrations in blood serum of dairy cows, calves, heifers and bulls were 12.21 ± 3.19 µmol/l, 18.91 ± 5.78 µmol/l, 17.80 ± 2.76 µmol/l and 16.69 ± 3.08 µmol/l, respectively. Zn deficiency was diagnosed in 41% of the examined dairy cows (45% herds) and 13% of bulls (10% herds). None of the calf or heifer herds was classified as Zn deficient when Zn deficiency was found in only 9% of calves and 1% of the examined heifers. Mean Cu concentrations in blood serum of dairy cows, calves, heifers and bulls were 13.62 ± 2.62 µmol/l, 10.18 ± 3.22 µmol/l, 10.96 ± 2.52 µmol/l and 11.18 ± 2.40 µmol/l, respectively. Cu deficiency was diagnosed in 28% of the examined dairy cows (20% herds), 70% of calves (80% herds), 65% of heifers (75% herds) and 70% of bulls (60% herds). Deficiency of at least one of the microelements monitored was diagnosed on all investigated farms.

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