A PCR survey of vector-borne pathogens in different dog populations from Turkey

2017 ◽  
Vol 62 (3) ◽  
Author(s):  
Huanping Guo ◽  
Ferda Sevinc ◽  
Onur Ceylan ◽  
Mutlu Sevinc ◽  
Ege Ince ◽  
...  
Keyword(s):  
Blood Samples ◽  
Pet Dogs ◽  
Specific Pcr ◽  
Vector Borne ◽  
Positive Results ◽  
Substantial Morbidity

AbstractIn the present study, a total of 192 blood samples were collected from pet dogs, kennel dogs and shepherd dogs in Konya district, Turkey, and tested by specific PCR for the presence of vector-borne pathogens. Several pathogens were identified, most of which can cause substantial morbidity in dogs. PCR results revealed that 54 (28.1%) dogs were infected with one or more pathogens. Positive results were obtained for

scholarly journals Short Communication: Molecular characterization and blood hematology profile of dogs infected by Ehrlichia canis in Yogyakarta, Indonesia

2020 ◽  
Vol 21 (7) ◽  
Author(s):  
Yustinus oswin primajuni Wuhan ◽  
Aris Haryanto ◽  
Ida Tjahajati
Keyword(s):  
Molecular Characterization ◽  
Microscopic Examination ◽  
Short Communication ◽  
Pcr Amplification ◽  
Ehrlichia Canis ◽  
Infected Blood ◽  
Blood Samples ◽  
Glta Gene ◽  
Vector Borne ◽  
Positive Results

Abstract. Wuhan YOP, Haryanto A, Tjahajati I. 2020. Short Communication: Molecular characterization and blood hematology profile of dogs infected by Ehrlichia canis in Yogyakarta, Indonesia. Biodiversitas 21: 3242-3248. Ehrlichia canis is Gram-negative intracellular obligate bacteria that cause ehrlichiosis, a companion vector-borne disease is a potentially fatal disease that attacks dogs. The purpose of this study was to molecular characterize and determine the features of Ehrlichia-infected blood based on the amplification of the gltA gene in Ehrlichia infected dogs from Yogyakarta, Indonesia. Blood samples were collected from 51 dog patients from the Prof. Dr. Soeparwi Animal Hospital, animal clinics, and pet shops based on the anamnesis, clinical sign, and physical examination, followed by microscopic examination, routine hematology, PCR amplification, and DNA sequencing were carried out on the blood samples. Based on positive PCR amplification and blood hematology profile examination ehrlichiosis-positive in dogs showed that thrombocytopenia case was 82.3%, anemia was 70.5%, eosinopenia was 70.5%, neutropenia was 29.4%, monocytopenia was 23%, leukopenia was 17% and lymphopenia was 11.7%. Morulae of Ehrlichia sp.was not found in microscopic examination. Molecularly, detected of E. canis using the gltA gene showed that 34% of samples were positive results. Then 5 of positive Ehrlichia samples were DNA sequenced, they showed a high homology of 100% with Hat Yai isolates (KU765199.1). There was no genetic diversity between E. canis samples in Yogyakarta.

scholarly journals VECTOR-BORNE PATHOGENS-TESTING IN A ROMANIAN CANINE BLOOD BANK

2018 ◽  
Vol 6 ◽  
pp. 904-911
Author(s):  
Teodor-Stefan Ionescu ◽  
Sinziana Radulescu ◽  
Ruxandra Florea ◽  
Stelian Baraitareanu ◽  
Doina Danes
Keyword(s):  
Blood Donors ◽  
Holistic Approach ◽  
Blood Bank ◽  
Focused Attention ◽  
Donor Pool ◽  
Blood Samples ◽  
Negative Results ◽  
Blood Banks ◽  
Vector Borne ◽  
Positive Results

INTRODUCTION: Canine blood banking in veterinary medicine is an expanding market. Once the demand for blood products increased all over the world, canine blood banks have focused attention on the risk of spreading diseases through blood transfused products. The need to preserve a healthy donor-pool, free of blood-borne infectious diseases, mainly in endemic areas, led to the implementation of appropriate protocols for screening canine blood donors using specific tests.OBJECTIVES: The aim of this study was to evaluate the presence of Anaplasma phagocytophilum/Anaplasmaplatys, Echrlichiacanis/Echrlichiaewingii, Dirofilariaimmitis and Borrelia burgdorferi using the enzyme immunoassay technology (EIA) among the donors of a Romanian canine blood bank, from January 2015 to December 2016.METHODS: Blood samples from 575 donors were collected and 1253 tests were performed with SNAP 4DX Plus® (IDEXX Laboratories, Fremont, CA) to reveal the presence of D. immitis antigens and the antibodies toward A. phagocytophilum and/or A. platys, E. canis and/or E. ewingii and B. burgdorferi.RESULTS: The results of this holistic approach show that all blood samples provided negative results for B. burgdorferi and E.canis/E. ewingii (0/1253), while 0.87% (11/1253) samples provided positive results for A. phagocytophilum/A. platys and 6.94% (87/1253) for D. immitis.CONCLUSION: The next studied topic would be to compare the results provided by the EIA technology with results of real time PCR and qPCR regarding these vector-borne pathogens.

Wolbachia and Cytoplasmic Incompatibility in the CaliforniaCulex pipiensMosquito Species Complex: Parameter Estimates and Infection Dynamics in Natural Populations

Genetics ◽  
2003 ◽  
Vol 165 (4) ◽  
pp. 2029-2038 ◽  
Author(s):  
Jason L Rasgon ◽  
Thomas W Scott
Keyword(s):  
Species Complex ◽  
Cytoplasmic Incompatibility ◽  
Natural Populations ◽  
Wolbachia Infection ◽  
Field Conditions ◽  
Parameter Estimates ◽  
Infection Prevalence ◽  
Infection Frequency ◽  
Specific Pcr ◽  
Vector Borne

AbstractBefore maternally inherited bacterial symbionts like Wolbachia, which cause cytoplasmic incompatibility (CI; reduced hatch rate) when infected males mate with uninfected females, can be used in a program to control vector-borne diseases it is essential to understand their dynamics of infection in natural arthropod vector populations. Our study had four goals: (1) quantify the number of Wolbachia strains circulating in the California Culex pipiens species complex, (2) investigate Wolbachia infection frequencies and distribution in natural California populations, (3) estimate the parameters that govern Wolbachia spread among Cx. pipiens under laboratory and field conditions, and (4) use these values to estimate equilibrium levels and compare predicted infection prevalence levels to those observed in nature. Strain-specific PCR, wsp gene sequencing, and crossing experiments indicated that a single Wolbachia strain infects Californian Cx. pipiens. Infection frequency was near or at fixation in all populations sampled for 2 years along a >1000-km north-south transect. The combined statewide infection frequency was 99.4%. Incompatible crosses were 100% sterile under laboratory and field conditions. Sterility decreased negligibly with male age in the laboratory. Infection had no significant effect on female fecundity under laboratory or field conditions. Vertical transmission was >99% in the laboratory and ∼98.6% in the field. Using field data, models predicted that Wolbachia will spread to fixation if infection exceeds an unstable equilibrium point above 1.4%. Our estimates accurately predicted infection frequencies in natural populations. If certain technical hurdles can be overcome, our data indicate that Wolbachia can invade vector populations as part of an applied transgenic strategy for vector-borne disease reduction.

scholarly journals Seroprevalence of Hepatitis E Virus Infection among Blood Donors in Bulgaria

Viruses ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 492
Author(s):  
Magdalena Baymakova ◽  
Krasimira Terzieva ◽  
Rumen Popov ◽  
Elisaveta Grancharova ◽  
Todor Kundurzhiev ◽  
...  
Keyword(s):  
Virus Infection ◽  
Hepatitis E Virus ◽  
Blood Donors ◽  
Hepatitis E ◽  
Elisa Test ◽  
Blood Samples ◽  
Wild Boars ◽  
Domestic Pigs ◽  
Specific Questionnaire ◽  
Positive Results

Hepatitis E virus (HEV) infection is widespread among domestic pigs, industrial swine, and wild boars in Bulgaria. The aim of the current research was to present the HEV seroprevalence among blood donors in Bulgaria. In the present study, 555 blood donors (479 males and 76 females) were enrolled from five districts in the country (Shumen, Pleven, Stara Zagora, Plovdiv, and Sofia districts). All blood samples were tested for anti-HEV IgG using the recomWell HEV IgG ELISA test (Mikrogen GmbH, Neuried, Germany). Each participating donor completed a short, structured, and specific questionnaire to document data on the current study. Anti-HEV IgG positive results were detected in 144 (25.9%) blood donors, including 129 (26.9%) males and 15 (19.7%) females. The established HEV seropositivity was 28.8% (23/80) in Shumen district, 23.2% (22/95) in Pleven district, 27.1% (38/140) in Stara Zagora district, 27.5% (44/160) in Plovdiv district, and 21.3% (17/80) in Sofia district. A high HEV seroprevalence was found for persons who declared that they were general hunters (48.7%; 19/39; p = 0.001) and hunters of wild boars (51.6%; 16/31; p = 0.001). We present the first seroprevalence rates of HEV infection in blood donors from Bulgaria. The results of our research showed high HEV seropositivity among blood donors.

scholarly journals Molecular detection of vector borne pathogens in anemic and thrombocytopenic dogs in southern Brazil

2018 ◽  
Vol 27 (4) ◽  
pp. 505-513 ◽  
Author(s):  
Anna Cláudia Baumel Mongruel ◽  
Priscila Ikeda ◽  
Keyla Carstens Marques de Sousa ◽  
Jyan Lucas Benevenute ◽  
Margarete Kimie Falbo ◽  
...  
Keyword(s):  
18S Rrna ◽  
Molecular Detection ◽  
Phylogenetic Analyses ◽  
Southern Brazil ◽  
Conventional Pcr ◽  
Pathogenic Potential ◽  
Vector Borne ◽  
Pcr Assays ◽  
Etiological Agents ◽  
Positive Results

Abstract Arthropod-borne pathogens are medically important because of their ability to cause diseases in their hosts. The purpose of this study was to detect the occurrence of Ehrlichia spp., piroplasmids and Hepatozoon spp. in dogs with anemia and thrombocytopenia in southern Brazil. EDTA-whole blood was collected from 75 domestic dogs presenting anemia or/and thrombocytopenia from Guarapuava, state of Paraná, Brazil. DNA samples were subjected to conventional PCR assays for Ehrlichia spp. (dsb), piroplasmids (18S rRNA) and Hepatozoon spp. (18S rRNA), followed by sequencing and phylogenetic analyses. Among the 75 dogs, one (1.33%) was positive for Hepatozoon sp. and six (8%) were positive for piroplasmids in 18S rRNA cPCR assays. None of the dogs showed positive results in Ehrlichia spp.-cPCR targeting dsb gene. The phylogenetic analyses revealed that three piroplasm sequences were clustered with Rangellia vitalii, while one sequence was grouped with B. vogeli. The only sequence obtained from Hepatozoon spp.-PCR protocol was pooled with H. canis. Therefore, there is urgent need for differential molecular diagnosis of the two piroplasm species cited as etiological agents in clinical cases of canine hemoparasitic diseases, given the higher pathogenic potential of R. vitalii than of B. vogeli.

scholarly journals Genotypic detection of some virulence factors among Aeromonas hydrophila Isolated from Diarrhea Cases (Iraq)

2018 ◽  
Vol 9 (SPL1) ◽  
Author(s):  
Ilham A Bunyan ◽  
Israa K Obais
Keyword(s):  
Virulence Factors ◽  
Aeromonas Hydrophila ◽  
Molecular Detection ◽  
Virulence Gene ◽  
Molecular Study ◽  
Protease Gene ◽  
Allelic Ladder ◽  
Specific Pcr ◽  
Molecular Length ◽  
Positive Results

The present study included the detection ofsome virulence factorsof Aeromonas hydrophila under molecular level to clinical isolates were taken from patients suffering from diarrhea during the period from July (2017) to October (2017). Molecular detection of Hemolysin gene (ahh) was done for all isolates. The results showed that all isolates (100%) gave positive results for this virulence gene. the positive results were detected by the presence of (130) bp bands when compared with allelic ladder. The genomic DNA of the samples was extracted and bands were observed by performing agarose gel electrophoresis. When PCR was performed,results clearly indicate that all isolated organisms contained serine protease gene and all the amplified products produced a band at the level of (900 bp) when compared with the allelic ladder. Molecular detection of this gene was carried out by using a specific PCR primer were done by comparison with allelic ladder which gave a (309bp) It was found that (Aerolysin) gene present in (12) (75%) of the positive samples. Lip gene was also detected in A. hydrophila samples and found that all 16 samples (100%) gave positive results to this gene which gave molecular length (382) bp. Molecular study was carried out to show the sequence identity of cytotonic enterotoxins gene in Aeromonas spp. to that in A. hydrophila. Analysis of the A. hydrophila genome revealed a number of a putative virulence factors,including a gene that heat-labile cytotonic enterotoxin (alt). Our study showed that all (16) isolates (100%) gave positive results to this gene,which gave molecular length (442)bp. Molecular detection of cytotonic enterotoxins gene (ast) was done for (16) A. hydrophila isolates and the results showed that all isolates have this gene (100%). The positive results for (ast) virulence were detected by the presence of (331) bp band compared with allelic ladder.

scholarly journals A Cost-Effective Method for Identifying Enterobacterales with OXA-181

2019 ◽  
Vol 57 (11) ◽  
Author(s):  
Gisele Peirano ◽  
Yasufumi Matsumura ◽  
Diego Nobrega ◽  
Johann D. D. Pitout
Keyword(s):  
Indian Subcontinent ◽  
Cost Effective ◽  
Accurate Information ◽  
Surveillance Program ◽  
Molecular Surveillance ◽  
Specific Pcr ◽  
Cost Effective Method ◽  
Molecular Studies ◽  
Surveillance Programs ◽  
Positive Results

ABSTRACT OXA-181 is the second most common global OXA-48-like carbapenemase and is endemic in the Indian subcontinent. Molecular studies have shown that Enterobacterales with OXA-181 are often introduced into regions of nonendemicity. Distinguishing OXA-181 from other OXA-48-like enzymes often requires sequencing, which is rather expensive and time-consuming. A specific PCR (i.e., OXA181PCR) for the detection of blaOXA-181 was validated using a global collection (n = 315) of bacteria with well-characterized carbapenemases and showed 100% sensitivity and specificity (95% confidence interval [CI], 94.1 to 100 and 98.6 to 100, respectively) for detecting bacteria with OXA-181. The OXA181PCR subsequently gave positive results on 58/160 (36%) Enterobacterales with OXA-48-like carbapenemases from the 2015 INFORM surveillance program. The blaOXA-181-positive Enterobacterales were present in 9 countries spanning 5 continents, illustrating the global distribution of OXA-181. This methodology can easily be incorporated into molecular surveillance programs to provide accurate information about the prevalence of OXA-181. A loop-mediated isothermal amplification (LAMP)-OXA48 assay overall performed well for detecting OXA-48-like enzymes but showed poor specificity due to false-positive results with non-OXA carbapenemases.

scholarly journals Methylation analysis of MIR200 family in Mexican patients with colorectal cancer

2019 ◽  
Vol 68 (3) ◽  
pp. 782-785
Author(s):  
Carlos Rogelio Alvizo-Rodriguez ◽  
Maria de la Luz Ayala-Madrigal ◽  
Jesus Arturo Hernandez-Sandoval ◽  
Helen Haydee Fernanda Ramirez-Plascencia ◽  
Christian Octavio Gonzalez-Villaseñor ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Peripheral Blood ◽  
Reference Group ◽  
Polyacrylamide Gel Electrophoresis ◽  
Methylation Pattern ◽  
Methylation Analysis ◽  
Blood Samples ◽  
Normal Tissues ◽  
Negative Results ◽  
Specific Pcr

The present study aimed to analyze the methylation pattern of the MIR200 family in the colorectal tissues and peripheral blood of colorectal cancer (CRC) patients. Previous informed consent, 102 samples of colorectal tissues (tumor and adjacent normal tissues) and 40 peripheral blood samples were collected from CRC patients. Additionally, we included a reference group of 40 blood samples. DNA extraction was done for colorectal tissues and peripheral blood. For methylation-specific PCR, we used bisulfite-treated DNA and controls for methylated and unmethylated DNA were included to each assay. PCR fragments were separated by 6% polyacrylamide gel electrophoresis. Methylation-positive and methylation-negative results were confirmed by bisulfite genomic sequencing technique. We analyzed 102 colorectal tissues and 40 blood samples from 51 CRC patients. MIR200B/MIR200A/MIR429 methylation analysis discloses no differences among tissues (p>0.05). However, MIR200C/MIR141 methylation showed differences between colorectal tissues and peripheral blood of CRC patients (p<0.0001) and mainly methylated alleles were observed in peripheral blood. These findings suggest a tissue-specific methylation pattern for the MIR200C/MIR141 promoter.

scholarly journals Pyrosequencing versus methylation-specific PCR for assessment of MGMT methylation in tumor and blood samples of glioblastoma patients

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Anna Estival ◽  
Carolina Sanz ◽  
Jose-Luis Ramirez ◽  
Jose Maria Velarde ◽  
Marta Domenech ◽  
...  
Keyword(s):  
Mgmt Methylation ◽  
Blood Samples ◽  
Methylation Specific Pcr ◽  
Specific Pcr

Incidence of the JAK2V617F Mutation In Mexican Patients with Myeloproliferative Neoplasms (MPNs)

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 5180-5180
Author(s):  
Gregorio Ignacio ◽  
Rosa María Arana-Trejo ◽  
Verónica Gónzalez ◽  
Maria Paula Hérnandez ◽  
Yolanda Lugo ◽  
...  
Keyword(s):  
Conflicts Of Interest ◽  
Myeloproliferative Neoplasms ◽  
Direct Sequencing ◽  
Jak2v617f Mutation ◽  
Normal Blood ◽  
Base Substitution ◽  
Blood Samples ◽  
Specific Pcr ◽  
Dna And Rna ◽  
Mexican Patients

Abstract Abstract 5180 Introduction: The V617F mutation in JAK2 gene has been described in approximately 50–90% of patients with ET, MF AND PV [essential trombocythaemia, idiopathic myelofibrosis and policithemia vera]; but has also been reported, albeit at a lower frequency, in patients with other myeloid malignancies such as atypical CML, CMML, AML, MDS, JMML and CNL. A single G>T base substitution in exon 12 results in the conversion of valine to a phenylalanine aminoacid at position 617 of the JAK2 gene. The identification in this study were using techniques such as allele-specific PCR, RFLP-PCR and direct sequencing; for to determine the incidence in Mexican patients with MPNs. Patients and Methods: The JAK2V617F mutation was determined in 88 patients and 5 normal blood samples for healthy individuals as controls. About the patients, 60 were cataloged like MPNs and 28 patients with features suggestive of MPNs vs CML. Samples for bone marrow or peripherical blood were taken either at time of diagnosis of MPNs or during treatment with cytoreductive or anti-thrombotic agents. DNA and RNA were extracted using the QIAamp DNA and RNeasy mini kit (Qiagen) and amplified by the three techniques mentioned for JAK2V617F and by nested RT-PCR for BCR/ABL. Results: The five normal blood samples for controls were negative for JAK2V617F mutation and to BCR/ABL. Patients had median age 65 years (47–85 years old), 46% male and 54% female. In de overall patients: 60 patients with MPNs all were BCR/ABL negative and 20 (33%) had JAK2V617F. In the 28 patients with likely MPNs vs CML, 23 were BCR/ABL positive/JAK2 negative, two had the coexistence of both genetics defects [BCR/ABL+ and JAK2V617F+] and 3 BCR/ABL and JAK2 negative. Finally the patiens with JAK2V617F+, were 12 ET, PV 1, MF 2, CML 2, and 5 continued like MPNs. Discussion: The incidence of the JAK2V617F in this study for MPNs patiens were 33% and the incidence varied between MPNs subtype. Less than ten cases of BCR/ABL+ CML with JAK2V617F have been published; we report two patients with the coexistence and we agree with previous reports that screening for JAK2V617F mutation should be considered in any BCR/ABL+ CML patients and the clinical outcome will be define in long period. Disclosures: No relevant conflicts of interest to declare.

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