scholarly journals Epidemiological and Molecular Characterization of Echinococcus granulosus Isolated from Small Ruminants in Kashmir Valley, India

2021 ◽  
Author(s):  
Akeel Beigh ◽  
Mohmommad Darzi ◽  
Samina Bashir ◽  
Parvaiz Dar ◽  
Nazir Ganai ◽  
...  
Keyword(s):  
Small Ruminants ◽  
Restriction Enzymes ◽  
Economic Problem ◽  
Kashmir Valley ◽  
Predominant Genotype ◽  
Pcr Product ◽  
Relative Prevalence ◽  
Female Sheep ◽  
Sheep Strain

Background: Cystic Echinococcosis (CE) is an emergent or re-emergent zoonosis and remains a public health and economic problem all over the world. Methods:  The present study was carried on the prevalence and genotypes of Echinococcus present in small ruminants in Kashmir valley. A total of 2100, sheep (2052) and goats (48), slaughtered or spontaneously dead, from various areas of Kashmir valley were screened for the presence of hydatidosis. In case of goat none of the cases were found positive for hydatidosis, whereas, all the positive cases (85) were recorded in sheep only. The overall prevalence of hydatidosis was 4.04%. The prevalence was higher in female sheep (5.46%) compared to males (2.83%). Season-wise highest prevalence was in summer (4.55%), followed by autumn (4.1%), spring (3.89%) and winter (2.5%).The liver was observed to be the most frequently infected organ with relative prevalence of 61.17% followed by lungs (38.82%).The rDNA-ITS1 fragment of positive samples was amplified with BD1 / 4S primers. Results: The length of amplified fragment for all isolated samples was 1000bps. The products obtained on PCR were digested with four restriction enzymes (Rsa 1, Alu 1, Msp 1 and Taq1). Rsa 1, Alu 1, Msp 1 yielded identical fragments, 300 and 700 bp in sheep. TaqI restriction enzyme had no effect on PCR product and after digestion; intact 1000bps fragment was seen. Conclusion: Phylogenetic analysis of ITS1 gene revealed that the common sheep strain (G1) is the predominant genotype in sheep in Kashmir valley.

scholarly journals Molecular Typing ofNeisseria gonorrhoeaeIsolates by Opa-Typing and Ribotyping in New Delhi, India

2009 ◽  
Vol 2009 ◽  
pp. 1-6 ◽  
Author(s):  
Pejvak Khaki ◽  
Preena Bhalla ◽  
Ahmad Mir Fayaz ◽  
Sohiela Moradi Bidhendi ◽  
Majid Esmailzadeh ◽  
...  
Keyword(s):  
Molecular Typing ◽  
Preventive Measures ◽  
Restriction Enzymes ◽  
New Delhi ◽  
Pcr Product ◽  
Sexual Contacts ◽  
High Level ◽  
Epidemiological Characteristics ◽  
Epidemiological Characterization

Control and preventive measures for gonococcal infections are based on precise epidemiological characteristics ofN. gonorrhoeaeisolates. In the present study the potential utility of opa-typing and ribotyping for molecular epidemiological study of consecutive gonococcal strains was determined. Sixty gonococcal isolates were subjected to ribotyping with two restriction enzymes,AvaII andHincII, and opa-typing withTaqI andHpaII for epidemiological characterization of gonococcal population. Ribotyping withAvaII yielded 6 ribotype patterns while twelve RFLP patterns were observed withHincII. Opa-typing of the 60 isolates revealed a total 54 opa-types, which 48 were unique and 6 formed clusters. Fifty-two opa-types were observed withTaqI-digested PCR product while opa-typing withHpaII demonstrated 54 opa-types. The opa-types from isolates that were epidemiologically unrelated were distinct, whereas those from the sexual contacts were identical. The results showed that opa-typing is highly useful for characterizing gonococcal strains from sexual contacts and has more discriminatory than ribotyping that could differentiate between gonococci of the same ribotype. The technique even with a single restriction enzyme has a high level of discrimination (99.9%) between epidemiologically unrelated isolates. In conclusion, the molecular methods such as opa-typing and ribotyping can be used for epidemiological characterization of gonococcal strains.

scholarly journals Molecular Characterization of Stolbur Group Subgroup E (16SrXII-E) Phytoplasma Associated with Potatoes in China

Plant Disease ◽  
2012 ◽  
Vol 96 (9) ◽  
pp. 1372-1372 ◽  
Author(s):  
M. Cheng ◽  
J. Dong ◽  
L. Zhang ◽  
P. J. Laski ◽  
Z. Zhang ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Nested Pcr ◽  
Inner Mongolia ◽  
Disease Incidence ◽  
Restriction Enzymes ◽  
Potato Disease ◽  
Growing Seasons ◽  
Pcr Product ◽  
Disease Surveys

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.

A widespread pathway for substitution of adenine by diaminopurine in phage genomes

Science ◽  
2021 ◽  
Vol 372 (6541) ◽  
pp. 512-516
Author(s):  
Yan Zhou ◽  
Xuexia Xu ◽  
Yifeng Wei ◽  
Yu Cheng ◽  
Yu Guo ◽  
...  
Keyword(s):  
Large Scale ◽  
Restriction Enzymes ◽  
Diverse Range ◽  
Host Restriction ◽  
Form And Function ◽  
Multienzyme System ◽  
Dna Modifications ◽  
Dna Production ◽  
And Function

DNA modifications vary in form and function but generally do not alter Watson-Crick base pairing. Diaminopurine (Z) is an exception because it completely replaces adenine and forms three hydrogen bonds with thymine in cyanophage S-2L genomic DNA. However, the biosynthesis, prevalence, and importance of Z genomes remain unexplored. Here, we report a multienzyme system that supports Z-genome synthesis. We identified dozens of globally widespread phages harboring such enzymes, and we further verified the Z genome in one of these phages, Acinetobacter phage SH-Ab 15497, by using liquid chromatography with ultraviolet and mass spectrometry. The Z genome endows phages with evolutionary advantages for evading the attack of host restriction enzymes, and the characterization of its biosynthetic pathway enables Z-DNA production on a large scale for a diverse range of applications.

scholarly journals Molecular and Pathogenicity Characterization of Sphaceloma manihoticola Isolates from South-Central Brazil

Plant Disease ◽  
2003 ◽  
Vol 87 (11) ◽  
pp. 1322-1328 ◽  
Author(s):  
Elizabeth Alvarez ◽  
Juan Fernando Mejia ◽  
Teresa L. Valle
Keyword(s):  
Morphological Characteristics ◽  
Restriction Pattern ◽  
Poly Merase Chain Reaction ◽  
Highly Pathogenic ◽  
Central Brazil ◽  
Host Interaction ◽  
South Central ◽  
Pcr Product ◽  
Pathogenic Variation

Isolates of Sphaceloma manihoticola, the asexual stage of Elsinoe brasiliensis, were collected from several regions of south-central Brazil. The isolates were obtained from samples of leaves, stems, and petioles of cassava (Manihot esculenta) and the weedy Euphorbia heterophylla (“amendoim bravo”) by directly plating infected tissue onto acidified potato dextrose agar. For pathogenicity studies, 19 isolates were inoculated onto each of two cassava cultivars, MBRA 703 as a susceptible cultivar and MBRA 12 as a resistant cultivar to S. manihoticola. MBRA 703, with the greatest pathogenicity to 58% (11) of the isolates, showed an intermediate pathogenic reaction to 16% (3) of the isolates, and was less pathogenic to 26% (5) of the isolates. MBRA 12, with a less pathogenic reaction to 63% (12) of the isolates, showed an intermediate pathogenic reaction to 16% (3) of the isolates, and was highly pathogenic to 21% (4) of the isolates. The isolates were verified as belonging to the genus Sphaceloma based on their morphological characteristics, including conidia and hyphae of monoconidial isolate. Conidia of isolates were small, thin-walled, ellipsoid to (rarely) globose, commonly with one or two gut-tules. Conidiophores were phialides, hyaline to slightly pigmented 0-to-1 septate; conidiophores from the weedy specie were phialides, hyaline to brown 0-to-2 septate producing hyaline conidia. The isolates also were verified as belonging to the genus Sphaceloma by using a poly-merase chain reaction (PCR) assay, which detected a 645-bp band in all isolates except two (1 and 6) for which the PCR product had 600 bp. Digestion of the amplified product with the enzymes MspI and CfoI allowed differences to be detected in restriction patterns among isolates. A homogeneous banding pattern was obtained for 17 of the isolates but a different restriction pattern was obtained for isolates 1 and 6 of E. heterophylla. This suggests the possibility of another species within this group of isolates. The results indicate the presence of pathogenic variation among isolates of the fungus and an isolate-host interaction, because statistically significant differences were observed between the two cassava cultivars in response to inoculation with the isolates of S. manihoticola.

scholarly journals Histopathological and molecular characterization of encephalitic listeriosis in small ruminants from northern Paraná, Brazil

2013 ◽  
Vol 44 (3) ◽  
pp. 889-896 ◽  
Author(s):  
Selwyn Arlington Headley ◽  
Lívia Bodnar ◽  
Juliana T.T. Fritzen ◽  
Dalton Evert Bronkhorst ◽  
Alice Fernandes Alfieri ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Small Ruminants

scholarly journals Impact of a Novel W2027L Mutation and Non-Target Site Resistance on Acetyl-CoA Carboxylase-Inhibiting Herbicides in a French Lolium multiflorum Population

Genes ◽  
2021 ◽  
Vol 12 (11) ◽  
pp. 1838
Author(s):  
Shiv Shankhar Kaundun ◽  
Joe Downes ◽  
Lucy Victoria Jackson ◽  
Sarah-Jane Hutchings ◽  
Eddie Mcindoe
Keyword(s):  
Lolium Multiflorum ◽  
Restriction Enzymes ◽  
Target Site ◽  
Cereal Crops ◽  
Acetyl Coa Carboxylase ◽  
Wild Type ◽  
Resistance Mutations ◽  
Acetyl Coa ◽  
Pcr Product ◽  
Target Site Resistance

Herbicides that inhibit acetyl-CoA carboxylase (ACCase) are among the few remaining options for the post-emergence control of Lolium species in small grain cereal crops. Here, we determined the mechanism of resistance to ACCase herbicides in a Lolium multiflorum population (HGR) from France. A combined biological and molecular approach detected a novel W2027L ACCase mutation that affects aryloxyphenoxypropionate (FOP) but not cyclohexanedione (DIM) or phenylpyraxoline (DEN) subclasses of ACCase herbicides. Both the wild-type tryptophan and mutant leucine 2027-ACCase alleles could be positively detected in a single DNA-based-derived polymorphic amplified cleaved sequence (dPACS) assay that contained the targeted PCR product and a cocktail of two discriminating restriction enzymes. Additionally, we identified three well-characterised I1781L, I2041T, and D2078G ACCase target site resistance mutations as well as non-target site resistance in HGR. The non-target site component endowed high levels of resistance to FOP herbicides whilst partially impacting on the efficacy of pinoxaden and cycloxydim. This study adequately assessed the contribution of the W2027L mutation and non-target site mechanism in conferring resistance to ACCase herbicides in HGR. It also highlights the versatility and robustness of the dPACS method to simultaneously identify different resistance-causing alleles at a single ACCase codon.

scholarly journals Identification and Characterization of Spontaneous Auxotrophic Mutants in Fusarium langsethiae

2017 ◽  
Author(s):  
Olga Gavrilova ◽  
Anna Skritnika ◽  
Tatiana Gagkaeva
Keyword(s):  
Intraspecific Diversity ◽  
Northern Europe ◽  
Growth Defect ◽  
Organic Media ◽  
Auxotrophic Mutants ◽  
Pcr Product ◽  
Subgroup Ii ◽  
Free Media ◽  
Identification And Characterization

Analysis of 49 strains of F. langsethiae originating from northern Europe (Russia, Finland, Sweden, UK, Norway, and Latvia) revealed the presence of spontaneous auxotrophic mutants that reflect natural intraspecific diversity. Our investigations detected that 49.0% of F. langsethiae strains were auxotrophic mutants for biotin, and 8.2% of the strains required thiamine as a growth factor. They failed to grow on vitamin-free media. For both prototrophic and auxotrophic strains, no growth defect was observed in rich organic media. Without essential vitamins, a significant reduction in the growth of the auxotrophic strains results in a decrease of the formation of T-2 toxin and diacetoxyscirpenol. In addition, all analysed F. langsethiae strains were distinguished into two subgroups based on PCR product sizes. According to our results, 26 and 23 strains of F. langsethiae belong to subgroups I and II respectively. We determined that the deletion in the IGS region of the rDNA of F. langsethiae belonging to subgroup II is linked with temperature sensitivity and causes a decrease in strain growth at 30 °C. Four thiamine auxotrophic strains were found in subgroup I, while 21 biotin auxotrophic strains were detected in subgroups II. To the best of our knowledge, the spontaneous mutations in F. langsethiae observed in the present work have not been previously reported.

scholarly journals Restriction endonuclease characterization of resistant plasmids in Enterobacteriaceae isolated from children in the Sudan

1989 ◽  
Vol 103 (3) ◽  
pp. 487-496 ◽  
Author(s):  
P. Shears ◽  
G. Suliman ◽  
C. A. Hart
Keyword(s):  
Antimicrobial Resistance ◽  
Restriction Endonuclease ◽  
Restriction Enzymes ◽  
Surveillance Systems ◽  
High Prevalence ◽  
Resistance Patterns ◽  
Reproducible Method ◽  
Restriction Endonuclease Digest

SUMMARYThe investigation of plasmid similarity is an important component in the surveillance of antimicrobial resistance and in the detection of epidemic plasmids. The use of restriction endonucleases in the classification of transferable, multiply-resistant plasmids from faecal Enterobacteriaceae isolated at the Children's Emergency Hospital, Khartoum was investigated. Twenty-four transconjugant plasmids, coding for 11 different resistance patterns, each of molecular weight 62 MDa. were studied using four restriction enzymes;PstI,EcoR I,HindIII andAraII. Fifteen different digest profiles were obtained. Restriction profiles discriminated between plasmids with differing resistance patterns and demonstrated homology of plasmids with common resistance patterns. Restriction endonuclease digest patterns provide a potentially rapid and reproducible method of plasmid classification, that could contribute towards surveillance systems in tropical countries with a high prevalence of antimicrobial resistance.

Characterization of cry Genes in a Mexican Bacillus thuringiensis Strain Collection

1998 ◽  
Vol 64 (12) ◽  
pp. 4965-4972 ◽  
Author(s):  
Alejandra Bravo ◽  
Sergio Sarabia ◽  
Lorena Lopez ◽  
Hernesto Ontiveros ◽  
Carolina Abarca ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Specific Primers ◽  
Cry Proteins ◽  
Pcr Product ◽  
Strain Collection ◽  
Lepidopteran Insects ◽  
Ecological Patterns ◽  
Biogeographical Regions ◽  
Selection Of

ABSTRACT Mexico is located in a transition zone between the Nearctic and Neotropical biogeographical regions and contains a rich and unique biodiversity. A total of 496 Bacillus thuringiensis strains were isolated from 503 soil samples collected from the five macroregions of the country. The characterization of the strain collection provided useful information on the ecological patterns of distribution of B. thuringiensis and opportunities for the selection of strains to develop novel bioinsecticidal products. The analysis of the strains was based on multiplex PCR with novel general and specific primers that could detect the cry1,cry3, cry5, cry7, cry8,cry9, cry11, cry12,cry13, cry14, cry21, andcyt genes. The proteins belonging to the Cry1 and Cry9 groups are toxic for lepidopteran insects. The Cry3, Cry7, and Cry8 proteins are active against coleopteran insects. The Cry5, Cry12, Cry13, and Cry14 proteins are nematocidal. The Cry11, Cry21, and Cyt proteins are toxic for dipteran insects. Six pairs of general primers are used in this method. Strains for which unique PCR product profiles were obtained with the general primers were further characterized by additional PCRs with specific primers. Strains containingcry1 genes were the most abundant in our collection (49.5%). Thirty-three different cry1-type profiles were identified. B. thuringiensis strains harboringcry3 genes represented 21.5% of the strains, and 7.9% of the strains contained cry11 and cyt genes.cry7, cry8, and cry9 genes were found in 0.6, 2.4, and 2.6% of the strains, respectively. No strains carrying cry5, cry12, cry13,cry14, or cry21 genes were found. Finally, 14% of the strains did not give any PCR product and did not react with any polyclonal antisera. Our results indicate the presence of strains that may harbor potentially novel Cry proteins as well as strains with combinations of less frequently observed cry genes.

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