Molecular characterization of Brazilian avian pneumovirus isolates: comparison between immunochemiluminescent Southern blot and nested PCR

1999 ◽  
Vol 79 (2) ◽  
pp. 237-241 ◽  
Author(s):  
Maria Angela C Dani ◽  
Edison L Durigon ◽  
Clarice W Arns
Author(s):  
Gurupada Balol ◽  
C Channakeshava ◽  
M S Patil

Chickpea plants showing phytoplasma symptoms were observed in the research plots at University of Agricultural Sciences, Dharwad, Karnataka, India. The symptoms included phyllody, pale green leaves, bushy appearance and excessive axillary proliferation. The causal agent of the phyllody disease was identified based on symptoms, amplification of 16S rDNA of the phytoplasma by nested PCR with primers P1/P7 and R16F2n/R16R2 and 1,800 bp and 1,200 bp size products were amplified in first round PCR and nested-PCR respectively. The PCR product was sequenced and compared with the reference phytoplasma sequences collected from the database (NCBI). 16S rDNA sequences of Dharwad chickpea phytoplasma shared the highest nucleotide identity of (>98%) with Periwinkle phyllody16SrII-E (EU096500). This study indicated the association of ‘Candidatus Phytoplasma aurantifolia’ the 16SrII-E group infecting chickpea from Northern Karnataka.


2021 ◽  
Vol 41 (01) ◽  
pp. 160-162
Author(s):  
Aayesha Riaz

Bovine herpesvirus 1 (BHV-1) is a noteworthy reason for many Cattle/Buffalo diseases. Infectious bovine rhinotracheitis (IBR) is one of the diseases which are caused by the BHV-1. In the present study a cow which was suspected of IBR was examined. The animal was suffering from fever and respiratory distress along with rhinitis (red nose), in appetence, and dyspnea. The nasal mucosa and muzzle were distinctly inflamed, with nasal discharge. DNA from blood samples and nasal swabs were subjected to nested PCR using glycoprotein B gene (gB) Primers. The samples were found positive for BHV-1 gB gene. Sequencing and phylogenetic analysis revealed close similarities with other BHV-1 gB gene sequences. The accession numbers assigned to this pioneer sequences in GenBank are MT449510 (BHV-1-Pak 1) and MT449511 (BHV-1-Pak 2). In this study, we reported for the first time the detection of DNA of BHV-1 through nested PCR assay and molecular characterization of BHV-1 gB gene in Pakistan. This study will be useful in further diagnoses of BHV-1 in Pakistan and in development of BHV-1 vaccine to reduce economical losses due to IBR


2017 ◽  
Vol 45 (1) ◽  
pp. 6
Author(s):  
Mehmet Gultekin ◽  
Kerem Ural ◽  
Nuram Aysul ◽  
Adnam Ayan ◽  
Canberk Balikci ◽  
...  

Background: Giardia duodenalis (G. duodenalis) is an ubiquitous, flagellated intestinal protozoan with major public health significance worldwide. Limited data are available on the epidemiology of G. duodenalis in dairy cattle from Turkey. Determining the zoonotic potential of the Giardia infection requires molecular characterization. The aim of the present study was to investigate the prevalence and to molecularly characterize G. duodenalis in calves less than three months of age in Aydin, Aegean region of Turkey.Materials, Methods & Results: The study was conducted on different dairy farms in the south-western part of the Turkey, Aegean Region, Aydin. A total of 198 Holstein Friesian calves less than three months of age, of both sexes were enrolled into the study. Faecal samples from each calf were collected manually from the rectum using a disposable latex glove. The consistency of collected samples was recorded as diarrhoeic or non-diarrhoeic.  Diagnosis of G. duodenalis infection was made microscopically by detection of cysts in the faecal samples. One hundred and sixteen (58.5%) of the 198 faecal samples were diarrheic. Giardia cysts were found in 27 (23.28%) of the diarrheic samples and in 8 (9.76%) of nondiarrheic samples (P < 0.05). The overall prevalence of giardiosis in calves was determined as 17.67%. The prevalence of Giardia genotypes was identified by DNA sequence analysis of the beta-giardin gene for every PCR positive sample. The beta-giardin nested PCR assay was revealed assemblage A and sub-genotype A3 was detected in all of 35 samples (100%).Discussion: The highest prevalence of Giardia infection in calves is reported at the age between 1 and 6 months, and the prevalence shows decreased rate from the age of 6 months. The present study was conducted in Aydin, a province of south-western Turkey in the Aegean Region, and the overall prevalence from a total of 198 dairy calves was 17.67%. The prevalence rate in calves with diarrhoea was higher and reached up to 23.28%, whereas it was 9.76% in non-diarrhoeic calves. A prevalence study with molecular characterization of G. duodenalis isolates in cattle has not yet been reported from Turkey. Molecular studies have shown that mostly assemblage E predominates in cattle, but recent studies denoted that assemblage A is increasingly being detected and might be more widespread than expected before. In the present study, Giardia positive samples identified with a beta-giardin nested PCR assay. The sub-genotype A3 was identified in all samples. The same sub-genotype was identified in human and dog samples from different countries. Furthermore, sub-genotype A3 was found in humans and dogs from Turkey. In this context, results of the present study suggested an important role of calves as potential reservoirs of human infections in Turkey. In conclusion, epidemiological data revealed that G. duodenalis infection is frequent in calves with diarrhoea in Aydin, Turkey. The presence of the potentially zoonotic sub-genotype A3 and high prevalence of Giardia infection in diarrheic calves indicated the importance of treatment and necessary preventative measures. Further studies in human and animal populations living in this region are warranted regarding the zoonotic epidemiology of Giardia duodenalis.


2001 ◽  
Vol 39 (3) ◽  
pp. 1036-1041 ◽  
Author(s):  
M. J. Naylor ◽  
G. A. Harrison ◽  
R. P. Monckton ◽  
S. McOrist ◽  
P. R. Lehrbach ◽  
...  

2017 ◽  
Vol 7 (9) ◽  
pp. 548-552 ◽  
Author(s):  
Mohd Aiman Barudin ◽  
◽  
Muhammad Lokman Md Isa ◽  
Noratikah Othman ◽  
Afzan Mat Yusof ◽  
...  

Plant Disease ◽  
2012 ◽  
Vol 96 (9) ◽  
pp. 1372-1372 ◽  
Author(s):  
M. Cheng ◽  
J. Dong ◽  
L. Zhang ◽  
P. J. Laski ◽  
Z. Zhang ◽  
...  

Phytoplasmas have been reported from more than 70 plant species in China, most of which are from woody plants and very few are from potatoes (Solanum tuberosum). During the growing seasons of 2005 through 2011, potato disease surveys were conducted in seed and commercial fields in Yunnan Province and Inner Mongolia. Potato plants displayed symptoms of curled, yellowish and purplish leaves, shortened internodes, aerial tuber formation, and few small malformed underground tubers. Although the location of the fields surveyed each year varied, the disease seems to have become increasingly prevalent. In Yunnan, disease incidence was 5 to 24% in 2005 and 15 to 100% in 2010 and 2011. In Inner Mongolia, disease incidence in seed potato fields was 5 to 15% in 2006 and 25 to 50% in 2011. Total DNA was extracted from the leaves, stems, and roots of symptomatic and asymptomatic plants with a DNeasy Plant Mini Kit (Qiagen, Valencia, CA) according to the manufacturer's instruction. A nested PCR was performed by using primer pair P1/P7 followed by R16F2n/R16R2 to detect the presence of phytoplasmas (1,3). An approximate 1.25-kb PCR product was amplified from symptomatic plants but not from asymptomatic plants. Restriction fragment length polymorphism (RFLP) patterns were analyzed by digesting the 1.2-kb amplicon singly with restriction enzymes AluI, BfaI, HhaI, HpaI, KpnI, MseI, and TaqI. Comparing the RFLP patterns of samples with previously published phytoplasma strains, the phytoplasmas matched patterns of the stolbur group, subgroup E (16SrXII-E) (1). In addition, the PCR product from P1/P7, diluted 1:30, was amplified by using primer pair P1A/P7A (2). The nested PCR product was cloned into pCR8/GW/TOPO vector (Invitrogen, Carlsbad, CA) and sequenced by the Core Lab of the University of Alaska Fairbanks and GENEWIZ (South Plainfield, NJ). Nucleotide sequences (GenBank Accession No. EU293841) were analyzed by iPhyClassifier software (4), confirming the relationship of this phytoplasma to ‘Candidatus Phytoplasma fragariae’ with RFLP patterns identical to group 16SrXII-E. To our knowledge, this is the first molecular characterization of the stolbur group phytoplasmas associated with potato disease in China. The potato is becoming increasingly important in China. The impacts of stolbur on potato yield losses, disease distributions, and insect vectors are currently under investigation. References: (1) I.-M. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1998. (2) I.-M. Lee et al. Int. J. Syst. Evol. Microbiol 54:337, 2004. (3) C. D. Smart et al. Appl. Environ. Microbiol. 62:2988, 1996. (4) Y. Zhao et al. Int. J. Syst. Evol. Microbiol. 59:2582, 2009.


2015 ◽  
Vol 5 (3) ◽  
pp. 202-207 ◽  
Author(s):  
AMAM Zonaed Siddiki ◽  
Sohana Akter Mina ◽  
Zinat Farzana ◽  
Bibi Ayesa ◽  
Rasel Das ◽  
...  

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