Identification of Theileria spp. in sheep and goats from Jeddah, Saudi Arabia, using molecular techniques

Background Thileriosis is a tick -born disease caused by hemoprotozoan parasites which has global veterinary and economic implications. Methods Blood samples were collected from 216 sheep and 83 goats from Jeddah, Saudi Arabia, were analyzed to determine whether the animals were infected with Theileria spp. parasites. The parasites were detected using a polymerase chain reaction (PCR) targeting the gene of 18S rRNA followed by sequencing. Results According to obtained findings, Theileria spp. were detected in sheep (57.8%, 48/83) and goats (51.9%, 112/216). Phylogenetic analysis to sequence data showed that T. ovis identified in this study were found to be closely connected to an isolate from Turkey, with 84.4–99.8% pairwise identity and 52.35–99.79% coverage.

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Diagnostic Accuracy of a Direct Panfungal Polymerase Chain Reaction Assay Performed on Stained Cytology Slides

Veterinary Pathology ◽

10.1177/0300985821991562 ◽

2021 ◽

pp. 030098582199156

Author(s):

Alexandra N. Myers ◽

Unity Jeffery ◽

Zachary G. Seyler ◽

Sara D. Lawhon ◽

Aline Rodrigues Hoffmann

Keyword(s):

Polymerase Chain Reaction ◽

Diagnostic Accuracy ◽

Molecular Techniques ◽

Fungal Culture ◽

Chain Reaction ◽

Identification Of Fungi ◽

Fungal Dna ◽

Polymerase Chain ◽

Panfungal Pcr ◽

Pcr Targeting

Molecular techniques are increasingly being applied to stained cytology slides for the diagnosis of neoplastic and infectious diseases. Such techniques for the identification of fungi from stained cytology slides have not yet been evaluated. This study aimed to assess the diagnostic accuracy of direct (without nucleic acid isolation) panfungal polymerase chain reaction (PCR) followed by sequencing for identification of fungi and oomycetes on stained cytology slides from dogs, cats, horses, and other species. Thirty-six cases were identified with cytologically identifiable fungi/oomycetes and concurrent identification via fungal culture or immunoassay. Twenty-nine controls were identified with no cytologically or histologically visible organisms and a concurrent negative fungal culture. Direct PCR targeting the internal transcribed spacer region followed by sequencing was performed on one cytology slide from each case and control, and the sensitivity and specificity of the assay were calculated. The sensitivity of the panfungal PCR assay performed on stained cytology slides was 67% overall, 73% excluding cases with oomycetes, and 86% when considering only slides with abundant fungi. The specificity was 62%, which was attributed to amplification of fungal DNA from control slides with no visible fungus and negative culture results. Direct panfungal PCR is capable of providing genus- or species-level identification of fungi from stained cytology slides. Given the potential of panfungal PCR to amplify contaminant fungal DNA, this assay should be performed on slides with visible fungi and interpreted in conjunction with morphologic assessment by a clinical pathologist.

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Application of advanced molecular techniques to detect vector borne pathogens from stray dogs and cats in two different climatic zones of Saudi Arabia

10.21203/rs.3.rs-21131/v1 ◽

2020 ◽

Author(s):

Abdullah D Alanazi ◽

Jan Šlapeta ◽

Abulaziz Alouffi ◽

Nichola Calvani ◽

Mohamed Alyousif ◽

...

Keyword(s):

Saudi Arabia ◽

Middle Eastern ◽

High Elevation ◽

Molecular Techniques ◽

Limited Information ◽

Chain Reaction ◽

Climatic Zones ◽

Vector Borne Diseases ◽

Vector Borne ◽

Polymerase Chain

Abstract Background: Vector-borne diseases have been increasing worldwide and reported in many animals including dogs and cats. Limited or no data are currently available regarding canine and feline vector-borne diseases in Saudi Arabia and limited information is available from other Middle Eastern countries. The aim of this study was to compare vector-borne disease prevalence between two bio-climatically distinct regions of Saudi Arabia, Riyadh province that is arid positioned at low elevation and Asir province that is humid at high elevation. Methods: Blood samples from 74d ogs from Riyadh province and 70 dogs and 44 cats from Asirprovince were collected and examined for the presence of genomic DNA of Babesias pp, Anaplasma spp., Ehrlichias pp., Bartonella spp., Mycoplasma spp., and Hepatozoon spp. by polymerase chain reaction (PCR), Multiplex-tandem PCR (MT-PCR) and Sanger sequencing.Results: Seventy four dogs were tested from Riyadh province and found be negative of any pathogen. Of the 70 dogs examined from Asir province 45(64.3%) were positive. Specifically, 40 (57.1%) dogs were positive for A.platys, 20 (28.5%) for B.vogeli, 11(15.7%) for My.Haemocanis, two (2.85%) for Candidatus Mycoplasma haematoparvum and one (1.4%) for Br.henselae. Fourteen out of 44 cats (31.8%) were positive for one of the detected vector-borne pathogens. Six cats (13.6%) were positive for Candidatus Mycoplasma haemominutum and My.haemofelis, respectively, four cats (9.2%) were positive for Br.Henselae, two (4.54%) for Candidatus Mycoplasma haematoparvum and one (2.27%) for A. platys. Conclusions: The results of this study report the occurrence of A. platys, B. vogeli, Br. henselae, and My. haemocanis in dogs and of A. platys, Br. henselae, My.haemofelis and Candidatus Mycoplasma haemominutum in cats from Asir province Further molecular investigations are strongly recommended in order to reduce the risk of dogs and cats acquiring vector-borne diseases in Saudi Arabia.

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Assessment of the Nutraceutical Effects of Oleuropein and the Cytotoxic Effects of Adriamycin, When Administered Alone and in Combination, in MG-63 Human Osteosarcoma Cells

Nutrients ◽

10.3390/nu13020354 ◽

2021 ◽

Vol 13 (2) ◽

pp. 354

Author(s):

Katerina Gioti ◽

Anastasia Papachristodoulou ◽

Dimitra Benaki ◽

Nektarios Aligiannis ◽

Alexios-Leandros Skaltsounis ◽

...

Keyword(s):

Chemotherapeutic Agent ◽

Molecular Techniques ◽

Osteosarcoma Cells ◽

Elisa Method ◽

Chain Reaction ◽

Cytotoxic Effects ◽

Human Osteosarcoma ◽

Human Osteosarcoma Cells ◽

Anti Cancer ◽

Polymerase Chain

Oleuropein (OLEU) is the most distinguished phenolic compound found in olive fruit and the leaves of Olea europaea L., with several pharmacological properties, including anti-cancer actions. Adriamycin (ADR) is an anthracycline widely used as a chemotherapeutic agent, although it presents significant side effects. The aim of the present study was to investigate the effect of oleuropein alone (20 μg/mL) and in co-treatment with ADR (50 nM), in MG-63 human osteosarcoma cells. Therefore, cellular and molecular techniques, such as MTT assay, flow cytometry, real-time Polymerase Chain Reaction (PCR), western blot and Elisa method, as well as Nuclear Magnetic Resonance (NMR) spectroscopy, were applied to unveil changes in the signal transduction pathways involved in osteosarcoma cells survival. The observed alterations in gene, protein and metabolite levels denote that OLEU not only inhibits MG-63 cells proliferation and potentiates ADR’s cytotoxicity, but also exerts its action, at least in part, through the induction of autophagy.

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Molecular Identification of Trypanosoma evansi Isolated from Arabian Camels (Camelus dromedarius) in Riyadh and Al-Qassim, Saudi Arabia

Animals ◽

10.3390/ani11041149 ◽

2021 ◽

Vol 11 (4) ◽

pp. 1149

Author(s):

Dina M. Metwally ◽

Isra M. Al-Turaiki ◽

Najwa Altwaijry ◽

Samia Q. Alghamdi ◽

Abdullah D. Alanazi

Keyword(s):

Saudi Arabia ◽

Infection Rate ◽

Ribosomal Rna ◽

Phylogenetic Analyses ◽

Trypanosoma Evansi ◽

Camelus Dromedarius ◽

18S Ribosomal Rna ◽

18S Ribosomal Rna Gene ◽

Polymerase Chain ◽

Pcr Targeting

We analyzed the blood from 400 one-humped camels, Camelus dromedarius (C. dromedarius), in Riyadh and Al-Qassim, Saudi Arabia to determine if they were infected with the parasite Trypanosoma spp. Polymerase chain reaction (PCR) targeting the internal transcribed spacer 1 (ITS1) gene was used to detect the prevalence of Trypanosoma spp. in the camels. Trypanosoma evansi (T. evansi) was detected in 79 of 200 camels in Riyadh, an infection rate of 39.5%, and in 92 of 200 camels in Al-Qassim, an infection rate of 46%. Sequence and phylogenetic analyses revealed that the isolated T. evansi was closely related to the T. evansi that was detected in C. dromedarius in Egypt and the T. evansi strain B15.1 18S ribosomal RNA gene identified from buffalo in Thailand. A BLAST search revealed that the sequences are also similar to those of T. evansi from beef cattle in Thailand and to T. brucei B8/18 18S ribosomal RNA from pigs in Nigeria.

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Myostatin Gene Expression and Its Application on Goat Breeding Programme

Indonesian Bulletin of Animal and Veterinary Sciences ◽

10.14334/wartazoa.v27i2.1537 ◽

2018 ◽

Vol 27 (2) ◽

pp. 89 ◽

Cited By ~ 1

Author(s):

Aron Batubara

Keyword(s):

Muscle Hypertrophy ◽

Body Weight Gain ◽

Muscle Growth ◽

Single Strand Conformation Polymorphism ◽

Molecular Techniques ◽

Chain Reaction ◽

Myostatin Gene ◽

Production Parameters ◽

Polymerase Chain ◽

Conformation Polymorphism

Characteristics of double muscled growth in animals are influenced by myostatin gene (MSTN). Myostatin gene is known as a member of the growth gene's superfamily (TGF-β) which works to suppress the muscle growth. However, the presence of six mutations on MSTN cause the gene inactive, and trigger the occurrence of muscle hypertrophy. Identification of myostatin gene was conducted by molecular techniques, and the most common method is polymerase chain reaction followed by single-strand conformation polymorphism (PCR-SSCP). Research on sheep and goat in several countries showed that there had been several variations occurred in myostatin gene but further studies are required to correlate these variations to body weight gain and other important production parameters. For goat production in Indonesia, myostatin mutations cause double muscling that can be utilised for genetic improvement in goat breeding plan to produce a new breed with high quality meat.

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Molecular diagnosis and biochemical studies of tick-borne diseases (anaplasmosis and babesiosis) in Aberdeen Angus Cattle in New Valley, Egypt

Veterinary World ◽

10.14202/vetworld.2020.1884-1891 ◽

2020 ◽

Vol 13 (9) ◽

pp. 1884-1891

Author(s):

Nani Nasreldin ◽

Rania M. Ewida ◽

Hatem Hamdon ◽

Yasser F. Elnaker

Keyword(s):

Oxidative Stress ◽

Polymerase Chain Reaction ◽

Molecular Techniques ◽

Babesia Bovis ◽

Blood Parasites ◽

Chain Reaction ◽

Infected Female ◽

Angus Cattle ◽

Polymerase Chain ◽

Female Cattle

Background and Aim: Anaplasmosis and babesiosis are tick-borne diseases that threaten livestock production with subsequent considerable economic losses. This study was conducted to diagnose Anaplasma and Babesia infection using molecular techniques in imported Aberdeen Angus cattle imported from Uruguay to El-Kharga Oasis in New Valley, Egypt, and to investigate the effects of disease on some serum biochemical and oxidative stress parameters. Materials and Methods: Blood samples were collected from 31 cattle, 21 diseased and ten apparently normal, of varying ages and sex. The blood was used for the preparation of blood smears, polymerase chain reaction assay, and separation of serum for biochemical investigation. The experimental production farm at the Faculty of Agriculture, New Valley University, was infested with ticks and variable clinical manifestations during the period from December 2017 to March 2018. One calf died of a suspected blood parasite infection. Results: The blood film examination revealed infection by blood parasites in 21 samples. Anaplasma marginale and Babesia bovis were identified in 12 and 14 samples, respectively. A total of 14 samples were examined by polymerase chain reaction (PCR) to make these identifications. Biochemical parameters showed significantly elevated serum alanine aminotransferase, aspartate aminotransferase, total bilirubin (T. Bil), and urea in blood from parasite-infected female cattle and male calves compared with controls. Increased serum total protein, globulin, and creatinine were recorded only in infected female cattle. The blood glucose level was significantly decreased in infected female cattle and male calves compared with controls. Furthermore, albumin and albumin/globulin ratio was significantly reduced in the infected female cattle. Oxidative stress profiles of infected animals showed a significant increase in serum nitric oxide and malondialdehyde, and both total antioxidant capacity and reduced glutathione (GSH) were significantly reduced in comparison with control animals. Conclusion: The incidence of A. marginale and B. bovis infection is high in imported Aberdeen Angus cattle in New Valley Province. PCR methods provide a short-term assessment of disease. An extensive epidemiological survey, employing serology together with molecular genetic methods, monitoring of abundance and distribution of tick vectors, availability of vaccination programs, and tracking of animal transport is also needed for control of blood parasites.

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High Resolution Microsatellite Marker Analysis of Some Rice Landraces Using Metaphor Agarose Gel Electrophoresis

Indonesian Journal of Biotechnology ◽

10.22146/ijbiotech.15269 ◽

2016 ◽

Vol 20 (1) ◽

pp. 54 ◽

Cited By ~ 1

Author(s):

K. Kristamtini ◽

T. Taryono ◽

Panjisakti Basunanda ◽

Rudi Hari Murti

Keyword(s):

Microsatellite Markers ◽

Gel Electrophoresis ◽

Polyacrylamide Gel Electrophoresis ◽

Molecular Techniques ◽

Agarose Gel ◽

Chain Reaction ◽

Rice Landraces ◽

Polymorphic Pattern ◽

Polymerase Chain ◽

Microsatellite Marker Analysis

Microsatellite markers or simple sequences repeats are DNA - based molecular techniques that areused to see the different among accessions and inbred lines. There are three methods to analysis the results ofthe polymerase chain reaction of microsatellite markers namely polyacrylamide gel electrophoresis (PAGE),capillary electroforesis, and Metaphor Agarose Gel Electroforesis (MAGE), and the Use of MAGE assessedmore easily and economically the polymorphic pattern of DNA markers. This study aimed to obtain fast,effective and efficient in term of easy and cheap technique to identify microsatellite markers of some blackrice cultivars and F2 populations from crosses between black with white rice. The results showed that MAGEsuccessfully separated clearly SSRs alleles with different sizes of less than 25 bp .

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Sensitive SYBR Green—Real Time PCR for the Detection and Quantitation of Avian Rotavirus A

Veterinary Sciences ◽

10.3390/vetsci6010002 ◽

2018 ◽

Vol 6 (1) ◽

pp. 2 ◽

Cited By ~ 2

Author(s):

David De la Torre ◽

Claudete Astolfi-Ferreira ◽

Ruy Chacon ◽

Antonio Piantino Ferreira

Keyword(s):

Polymerase Chain Reaction ◽

Real Time ◽

Sybr Green ◽

Molecular Techniques ◽

Economic Losses ◽

Rt Pcr ◽

Chain Reaction ◽

Rotavirus A ◽

Avian Nephritis Virus ◽

Polymerase Chain

Avian rotavirus A (ARtV-A) is a virus that affects young birds, causing acute diarrhea and economic losses in the poultry industry worldwide. The techniques used for the diagnosis of ARtV-A include electron microscopy, isolation in cell culture, and serology, as well as molecular techniques, such as the reverse transcription-polymerase chain reaction (RT-PCR). The objective of this work was to standardize a real-time RT-polymerase chain reaction (RT-qPCR) using SYBR Green chemistry for the rapid detection and quantification of ARtV-A from bird tissues and materials fixed on FTA cards on the basis of the nucleotide sequence of segment 6 (S6), which codes for the structural VP6 protein of ARtV-A. The results show the efficient amplification of the proposed target, with a limit of detection (LoD) of one copy gene (CG) per microliter of cDNA and a limit of quantification (LoQ) of 10 CGs per microliter. The efficiency of the primers was determined to be 95.66% using a standard curve, with an R2 value of 0.999 and a slope of −3.43. The specificity was determined using samples coinfected with ARtV-A, the chicken parvovirus, the chicken astrovirus, and the avian nephritis virus as positive controls and commercially available vaccines of the infectious bronchitis virus, infectious bursa disease virus, avian reovirus and healthy organs as negative controls. This technique, which lacks nonspecific PCR products and dimers, demonstrated greater sensitivity and specificity than conventional RT-PCR, and it reduced the analysis time by more than 50%.

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Extraction of microsporidial DNA from modified trichrome-stained clinical slides and subsequent species identification using PCR sequencing

Parasitology ◽

10.1017/s0031182008004411 ◽

2008 ◽

Vol 135 (6) ◽

pp. 701-703 ◽

Cited By ~ 16

Author(s):

K. S. CHAN ◽

T. H. KOH

Keyword(s):

Polymerase Chain Reaction ◽

Species Identification ◽

Environmental Samples ◽

Species Level ◽

Molecular Techniques ◽

Chain Reaction ◽

Clinical Specimens ◽

Polymerase Chain ◽

Pcr Conditions ◽

Hiv Negative

SUMMARYMolecular techniques involving polymerase chain reaction (PCR) and sequencing provide a relatively simple and objective means of identifying microsporidia to species level. We modified previously described methods of DNA extraction and PCR conditions for identification of microsporidia from museum slides, clinical specimens and environmental samples and successfully identifiedVittaforma corneaein 11 out of 13 cases of microsporidial infection from used trichrome-stained slides of corneal scrapings from HIV-negative patients with keratoconjunctivitis.

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Portable sequencer in the fight against infectious disease

Journal of Human Genetics ◽

10.1038/s10038-019-0675-4 ◽

2019 ◽

Vol 65 (1) ◽

pp. 35-40 ◽

Cited By ~ 7

Author(s):

Arthur Elia Mongan ◽

Josef Sem Berth Tuda ◽

Lucky Ronald Runtuwene

Keyword(s):

Infectious Disease ◽

Polymerase Chain Reaction ◽

Nucleic Acid ◽

Disease Management ◽

Acid Composition ◽

Molecular Techniques ◽

Chain Reaction ◽

Major Threat ◽

The World ◽

Polymerase Chain

Abstract Infectious disease is still a major threat in the world today. Five decades ago, it was considered soon to be eradicated, but the adaptation of pathogens to environmental pressure, such as antimicrobials, encouraged the emergence and reemergence of infectious disease. The fight with infectious disease starts with prevention, diagnosis, and treatment. Diagnosis can be upheld by observing the cause of disease under the microscope or detecting the presence of nucleic acid and proteins of the pathogens. The molecular techniques span from classical polymerase chain reaction (PCR) to sequencing the nucleic acid composition. Here, we are reviewing the works have been undertaken to utilize a portable sequencer, MinION, in various aspects of infectious disease management.

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