scholarly journals Prevalence of Ticks and detection of Rickettsial Agents from blood of Tick-Infested animals in Lower Sindh: A MOLECULAR APPROACH

2021 ◽  
Vol 3 (3) ◽  
pp. 70-78
Author(s):  
Iram Shaikh
Keyword(s):  
Blood Smear ◽  
Survey Study ◽  
Molecular Approach ◽  
Chain Reaction ◽  
Rickettsial Infection ◽  
Blood Samples ◽  
Animal Pathogens ◽  
Polymerase Chain ◽  
Wise Pattern ◽  
Molecular Parasitology

Ticks are important vectors of human and animal pathogens. They are considered as main vectors for transmission of rickettsial agents affecting animal and human health. The study was designed to investigate district wise pattern and detection of rickettsial agents by using molecular and conventional techniques in blood samples of infected cattles, buffalos, sheeps and goats. A survey study was carried out in lower Sindh (Tharparkar, Badin, Hyderabad, Karachi, Tando Muhammad khan, Thatta and Mirpurkhas). Blood samples were collected randomly from infected Cattles, buffalos, sheeps and goats and transported to the Molecular Parasitology laboratory, Sindh Agriculture University, Tandojam, followed by examinations under stereomicroscope and Polymerase Chain Reaction (PCR). The study showed that overall infection of Rickettsial agents among infected animals was recorded follwoing Microscopy/ Blood smear test in cattles, buffalos, sheeps and goats was 41.79, 49.09, 46 and 41.66% respectively, whereas overall infection through PCR in cattle, buffalo, sheep and goat was 39.55, 43.55, 46 and 55.55% respectively. Whereas animal-wise data through PCR indicates that in case of Goats (55.55%) were more susceptible to rickettsial infection as compared to sheep (46%), buffaloes (43.55%) and cattle (39.55%). The highest rate of rickettsial agents was found in district Tharparkar and lowest rate was found in district Karachi. Microscopy/Blood smear method indicates that Buffaloes were more susceptible for infection. Whereas PCR indicates Goats were more susceptible for infection.

scholarly journals Evaluation on the presence of Anaplasma, Ehrlichia, and Babesia spp. in goats (Capra hircus) in Cebu, the Philippines

2019 ◽  
Vol 12 (6) ◽  
pp. 774-777 ◽  
Author(s):  
Adrian P. Ybañez ◽  
Orgil V. Arrabis ◽  
Dennis Justin M. Alvarez ◽  
Eloiza May S. Galon ◽  
Rhea Mae P. Jayag ◽  
...  
Keyword(s):  
Blood Smear ◽  
Peripheral Blood Smear ◽  
The Philippines ◽  
Capra Hircus ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Blood Samples ◽  
Blood Smear Examination ◽  
Polymerase Chain ◽  
Babesia Spp

Background: Tick-borne diseases are caused by a wide variety of viruses, pathogens, and diseases. Anaplasma, Ehrlichia, and Babesia spp. are among the most known tick-borne pathogens in Asia. In the Philippines, these pathogens were already reportedly present in dogs and large ruminants, but no study has been reported yet evaluating their presence in goats. Aim: The present study aimed to evaluate the presence of Anaplasma, Ehrlichia, and Babesia spp. in goats in Cebu, the Philippines. Materials and Methods: A total of 100 blood samples from goats were collected in Cebu, the Philippines. Profile of sampled goats including age, body score, and sex was obtained. Peripheral blood smear examination and DNA extraction were performed. Nested polymerase chain reaction assay was used to evaluate the presence of Anaplasma, Ehrlichia, and Babesia spp. Results: None of the samples were found positive with Anaplasma, Ehrlichia, and Babesia spp. infection. Conclusion: Tested goats were negative with Anaplasma, Ehrlichia, and Babesia spp. and calls for continuous surveillance of these pathogens due to the reported detection of these pathogens in other livestock animals in the area.

A Study on Molecular Diagnosis of Theileria Species Infection by PCR Amplification in Sheep and Goats in Multan, Pakistan

2017 ◽  
Vol 60 (1) ◽  
pp. 36-45
Author(s):  
Muhammad Riaz ◽  
Zahida Tasawar
Keyword(s):  
Polymerase Chain Reaction ◽  
Mixed Infection ◽  
Blood Smear ◽  
Pcr Amplification ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Blood Samples ◽  
Sheep And Goats ◽  
Theileria Lestoquardi ◽  
Polymerase Chain

In this present study polymerase chain reaction (PCR) assay was used for identification anddifferentiation of Theileria species infection in Multan, Pakistan. Out of 220 blood samples collected fromsheep and goats, 31.2% (70/220) were found positive for Theileria species by PCR amplification comparedto only 9.1% (20/220) on blood smear. Theileria infection was observed in 39.3% (57/145) of sheep and18.6% (13/75) of goats sampled. The prevalence of Theileria ovis and Theileria lestoquardi in the 70positive samples was found to be 57% (40/70) and 30% (21/70), respectively with only 12.3% (9/70) ofblood samples having a mixed infection of both T. ovis and T. lestoquardi. Overall the prevalence ofT. ovis infection was higher than T. lestoquardi in both sheep and goats. Herds with sheep only hadsignificantly higher parasitic prevalence. The results confirm that PCR is direct, specific and sensitive toolfor diagnosis of ovine theileriosis.

Comparison of Droplet Digital Polymerase Chain Reaction (ddPCR) and Real-Time Quantitative Polymerase Chain Reaction (qPCR) in Detecting Neonatal Invasive Fungal Infections

2021 ◽  
Vol 11 (3) ◽  
pp. 373-379
Author(s):  
Huitao Li ◽  
Xueyu Chen ◽  
Xiaomei Qiu ◽  
Weimin Huang ◽  
Chuanzhong Yang
Keyword(s):  
Polymerase Chain Reaction ◽  
Limit Of Detection ◽  
Quantitative Polymerase Chain Reaction ◽  
Diagnostic Methods ◽  
Chain Reaction ◽  
Fungal Isolation ◽  
Blood Samples ◽  
Fungal Strains ◽  
Polymerase Chain ◽  
Digital Polymerase Chain Reaction

Invasive fungal infection (IFI) is the leading cause of death in neonatal patients, yet the diagnosis of IFI remains a major challenge. At present, most IFI laboratory diagnostic methods are based on classical, but limited, methods such as fungal isolation and culture and histopathological examination. Recently, quantitative polymerase chain reaction (qPCR) and droplet digital polymerase chain reaction (ddPCR) technology have been adopted to quantify nucleic-acid identification. In this study, we established qPCR and ddPCR assays for IFI diagnosis and quantification. qPCR and ddPCR were carried out using identical primers and probe for the amplification of 18S rRNA. Assay results for three fungal strains were positive, whereas ten non-fungal strains had negative results, indicating 100% specificity for both ddPCR and qPCR methods. Genomic DNA of Candida albicans was tested after a serial dilution to compare the sensitivity of the two PCR methods. The limit of detection of ddPCR was 3.2 copies/L, which was a ten-fold increase compared with that of the qPCR method (32 copies/L). Blood samples from 127 patients with high-risk factors and clinical symptoms for IFI were collected from a NICU in Shenzhen, China, and analyzed using qPCR and ddPCR. Thirty-four blood samples from neonates had a proven or probable diagnosis of IFI, and 25 of these were positive by qPCR, whereas 30 were positive by ddPCR. Among the 93 blood samples from neonates who had a possible IFI or no IFI, 24 were positive using qPCR, and 7 were positive using ddPCR. In conclusion, ddPCR is a rapid and accurate pan-fungal detection method and provides a promising prospect for IFI clinical screening.

scholarly journals Bartonella spp. in human and animal populations in Gauteng, South Africa, from 2007 to 2009

2012 ◽  
Vol 79 (2) ◽  
Author(s):  
Anastasia N. Trataris ◽  
Jennifer Rossouw ◽  
Lorraine Arntzen ◽  
Allan Karstaedt ◽  
John Frean
Keyword(s):  
Health Care Professionals ◽  
Deoxyribonucleic Acid ◽  
Bartonella Henselae ◽  
Hiv Positive ◽  
Host Immune System ◽  
Wild Rodent ◽  
Chain Reaction ◽  
Blood Samples ◽  
Animal Populations ◽  
Polymerase Chain

Bartonellae are highly adaptive organisms that have the ability to evade the host immune system and cause persistent bacteraemia by occupying the host’s erythrocytes. Bartonella spp. is under-studied and health care professionals often misdiagnose Bartonella-related infections. The aim of this study was to investigate the carriage of Bartonella spp. circulating in human and animal populations in Gauteng using culturing and polymerase chain reaction (PCR) detection. A total of 424 human, 98 cat, 179 dog, and 124 wild rodent blood samples were plated onto specialised media and incubated for 7–21 days at 37 ºC in CO2. Culture isolates morphologically similar to Bartonella control strains were confirmed by PCR and sequenced to determine species. Deoxyribonucleic acid (DNA) was extracted from all blood samples and tested by nested PCR. Bartonella could only be cultured from the cat and rodent specimens. Cat isolates were > 99% similar to Bartonella henselae URBHLIE 9, previously isolated from an endocarditis patient, and rat isolates were > 98% similar to either RN24BJ (candidus ‘Bartonella thailandensis’) or RN28BJ, previously isolated from rodents in China. The PCR prevalences were 22.5% in HIV-positive patients, 9.5% in clinically healthy volunteers, 23.5% in cats, 9% in dogs and 25% in rodents. Findings of this study have important implications for HIV-positive patients.

scholarly journals PCR - based diagnosis to evaluate the performance of malaria reference centers

2004 ◽  
Vol 46 (4) ◽  
pp. 183-187 ◽  
Author(s):  
Silvia Maria Di Santi ◽  
Karin Kirchgatter ◽  
Karen Cristina Sant'Anna Brunialti ◽  
Alessandra Mota Oliveira ◽  
Sergio Roberto Santos Ferreira ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Gold Standard ◽  
Blood Smear ◽  
Plasmodium Species ◽  
Thick Blood Smear ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Positive Results ◽  
Ssrrna Gene

Although the Giemsa-stained thick blood smear (GTS) remains the gold standard for the diagnosis of malaria, molecular methods are more sensitive and specific to detect parasites and can be used at reference centers to evaluate the performance of microscopy. The description of the Plasmodium falciparum, P. vivax, P. malariae and P. ovale ssrRNA gene sequences allowed the development of a polymerase chain reaction (PCR) that had been used to differentiate the four species. The objective of this study was to determine Plasmodium species through PCR in 190 positive smears from patients in order to verify the quality of diagnosis at SUCEN's Malaria Laboratory. Considering only the 131 positive results in both techniques, GTS detected 4.6% of mixed and 3.1% of P. malariae infections whereas PCR identified 19.1% and 13.8%, respectively.

scholarly journals AMPLIFIKASI GEN COI DARI SAMPEL DARAH ULAR DENGAN MENGGUNAKAN BEBERAPA PASANGAN PRIMER UNIVERSAL

ZOOTEC ◽  
2019 ◽  
Vol 39 (2) ◽  
pp. 314
Author(s):  
Dylan R. Pahlevi ◽  
Edwin De Queljoe ◽  
Beivy J. Kolondam
Keyword(s):  
Coi Gene ◽  
Polymerase Chain Reaction Reaction ◽  
Cytochrome Oxidase Subunit ◽  
Chain Reaction ◽  
Dna Ladder ◽  
Blood Samples ◽  
Total Dna ◽  
Polymerase Chain ◽  
Snake Blood ◽  
I Gene

AMPLIFICATION OF COI GENE FROM SNAKE BLOOD SAMPLES USING TWO UNIVERSALPRIMER PAIRS. This study aims to amplify COI (Cytochrome Oxidase Subunit I) gene fragments from snake blood. Four samples were obtained from four different snake individuals that were captured in Tapahan Telu Waterfall, Kali Village, Minahasa Regency. Total DNA from the sample was isolated and then the COI gene was amplified through the PCR (Polymerase Chain Reaction) reaction using two primer pairs, LCO1490-HCO2198 and FF2d-FR1d. These four samples were successfully amplified using different primers, i.e. DRP1 and DRP3 by FF2d-FR1d and DRP2 and DRP4 by LCO1490-HCO2198 primers. The success of amplification marked by the presence of 710 bp (LCO1490-HCO2198) and 707 bp (FF2d-FR1d) bands, which were indicated by those bands were located close to the standard 750 bp DNA ladder. Key words: Snake, Amplification, COI gene, Primers, PCR

scholarly journals Molecular detection and occurrence of equine theileriosis in Arabian horses in Al-Najaf province/Iraq

2020 ◽  
Vol 57 (3) ◽  
pp. e166996
Author(s):  
Hayder Mohammad Al-Rammahi ◽  
Abdulameer Abed Hatem ◽  
Asaad Chasib Al-Atabi
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Extraction ◽  
Molecular Detection ◽  
Molecular Technique ◽  
Chain Reaction ◽  
Blood Samples ◽  
Equine Piroplasmosis ◽  
Polymerase Chain

This study was designed to detect equine piroplasmosis using the molecular technique in Al-Najaf province during the season that showed an increment in tick activities. Blood samples were collected from 110 horses with more than two signs of piroplasmosis. After DNA extraction, the product was examined by a polymerase chain reaction to amplify 18SrRNA. The results showed that the overall percentage of equine theileriosis was 38.18%. According to gender, the percentage of infection was 43.48% and 29.27% in females and males, respectively. Significant variations appeared between infected horses according to age, and the percentage of infection was 50% and 35.22% in less than 2 years and more than 2 years age, respectively. Moreover, the percentage of infection was 62.5% and 19.35% in animals with and without acariasis, respectively. Significant variations were also seen in equine theileriosis according to geographical areas, and the higher percentage was reported in Hera district (60.87%), while the lowest percentage was in the center of Al-Najaf (21.43%). This difference may be due to different distribution of vector of disease (tick), which may be the availability of the suitable weather that helped in the multiplication of the intermediate vectors. In conclusion, this study proved the variations in the occurrences of equine piroplasmosis according to gender, age, and geographical areas.

scholarly journals Molecular identification by multiplex polymerase chain reaction of Pasteurella multocida in cattle and buffaloes in Baghdad

2014 ◽  
Vol 38 (1) ◽  
pp. 99-106
Author(s):  
Ihab G. M. AL-Shemmari
Keyword(s):  
Polymerase Chain Reaction ◽  
Multiplex Pcr ◽  
Multiplex Polymerase Chain Reaction ◽  
Pasteurella Multocida ◽  
Type B ◽  
Chain Reaction ◽  
Blood Samples ◽  
Nasal Swabs ◽  
Polymerase Chain ◽  
B 31

The aim of this study was to identify pasteurella multocida and their types by PCR in cattle’s and buffaloesi bagdad from March to August 2012 on 204 animals , including 102 cattle and 102 buffaloes at slaughter houses from Baghdad .Blood samples and nasal swaps were collected , before slaughtering and lung tissues of slaughtered animal , and from 54 clinically suspected cases of pasteurellosis , including 27 bovines ,and 27 buffaloes the samples taken included blood and nasal swabs . Pasteurellamultocida were isolated from 94 animals include 49 cattle 45 buffaloes. The typing of the isolates by multiplex PCR for genotyping Pasteuerllamultocida revealed 93 isolates of type B , 31 from cattle and 62 from buffaloes ,and 81 isolates of type A , 55 from cattle and 26 from buffaloes .

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