Presence of Meloidogyne enterolobii Yang & Eisenback (=M. mayaguensis) in guava and acerola from Costa Rica

Nematology ◽  
2012 ◽  
Vol 14 (2) ◽  
pp. 199-207 ◽  
Author(s):  
Danny Antonio Humphreys ◽  
Valerie Moroz Williamson ◽  
Luis Salazar ◽  
Lorena Flores-Chaves ◽  
Luis Gómez-Alpizar

The presence of Meloidogyne enterolobii on two hosts and in several locations in Costa Rica is reported. Meloidogyne spp. females were extracted from root of acerola and of wild and introduced guava and their perineal patterns were examined. Females from acerola showed round or dorso-ventrally ovoid perineal patterns, in some cases with lateral lines or with a moderately high to high dorsal arch as is characteristic of M. incognita. In guava, perineal patterns were similar, but with a low to moderate dorsal arch. PCR amplification of the mitochondrial DNA region between COII and the 16S ribosomal RNA yielded a unique product of 705 bp for the five populations extracted from both crops. In addition, the amplified product spanning the 63 bp repeat region of the mitochondrial genome was a single fragment of 320 bp in contrast to other tropical species, which typically amplify multiple bands. Size of PCR products of both mitochondrial regions and the sequence of the 63 bp repeat region supported the identity of these isolates as M. enterolobii (=M. mayaguensis).

Parasitology ◽  
2002 ◽  
Vol 124 (2) ◽  
pp. 177-184 ◽  
Author(s):  
M. B. A. MENDONÇA ◽  
N. S. NEHME ◽  
S. S. SANTOS ◽  
E. CUPOLILLO ◽  
N. VARGAS ◽  
...  

Trypanosoma cruzi is currently classified into 2 major phylogenetic lineages, T. cruzi I and II, that correlate with the formerly described zymodeme 1 and 2, respectively. Another isoenzymic group (zymodeme 3–Z3) was also described. In this study, we analysed the genetic diversity among Z3 isolates of the Brazilian Amazon by restriction fragment length polymorphism of the intergenic transcribed spacers (ITSs) of the ribosomal RNA cistron and the size of the divergent domain D7 of the 24Sα rRNA gene. DNAs from 12 T. cruzi Z3 isolates obtained from humans (2), Panstrongylus geniculatus (1), and Rhodnius brethesi (9) were submitted to PCR amplification of the ITSs plus the 5·8S rDNA. The PCR products were digested with 4 distinct endonucleases and the profiles analysed by a numerical methodology. The phenetic dendrogram revealed a clear dichotomy in the Z3 group, defining 2 groups that were named Z3-A and Z3-B. Dimorphism was also found in the band sizes of the amplified D7 divergent domain of the 24Sα rDNA, which showed a perfect correlation with the ITSs clustering. The organization of the ribosomal cistron was investigated by Southern blotting and shown to be conserved in the genome of the 2 Z3 groups. This study shows that the rDNA cistron allows the definition of 2 distinct subclusters in Z3 isolates.


Parasitology ◽  
2000 ◽  
Vol 121 (5) ◽  
pp. 555-563 ◽  
Author(s):  
C. M. COLLINS ◽  
C. O. CUNNINGHAM

The intergenic spacer of the ribosomal RNA gene array from the monogenean Gyrodactylus salaris was isolated using PCR amplification. PCR products were cloned and sequenced. Three different fragments of 0·63, 1·0 and 2·62 kb, were consistently obtained. These showed homology at the 5′ and 3′ termini but differed in their overall size and intervening sequence. The 5′ end showed homology to various 28S ribosomal RNA gene sequences, suggesting that this represented the 3′ terminus of the G. salaris 28S ribosomal RNA gene. A number of features common to other eukaryotic intergenic spacers were found in the longest sequence, including A + T rich sequences, palindromic sequences and tandemly repeated elements. Two regions of 23 bp sequences arranged in non-identical tandem repeats were identified. There were 9 repeats in both regions, separated by 81 bp of non-repetitive sequence. The repeat units from the two regions shared some similarity at their 3′ ends. The G. salaris intergenic spacer sequence was examined for sequence motifs involved in the transcription of the ribosomal RNA genes in other species. Several regions with homology to transcription start sites were identified.


Plant Disease ◽  
2021 ◽  
Author(s):  
Aminat Korede Oyetunde ◽  
Yao Kolombia ◽  
Omowumi B Adewuyi ◽  
Steve Olaoluwa Afolami ◽  
Daniel Leigh Coyne

Meloidogyne enterolobii is a highly polyphagous tropical species of root knot nematode. It has been recorded to be causing major damage to a range of economically important crops and is increasingly recorded from new locations. The morphological similarity and overlap of characteristics with other commonly occurring species, especially M. incognita, has confused its diagnosis using morphometrics. Cassava (Manihot esculenta) is an important crop across the tropics, including Africa, where it is among the most important root and tuber crop for food security. Cassava can be heavily infected by root knot nematodes, which can incur heavy production losses. The main species known to affect cassava are M. incognita and M. javanica (Coyne and Affokpon, 2018). With the exception of one report of M. enterolobii morphologically identified from cassava roots during a survey in Brazil (Rosa et al., 2014), there is no record with molecular confirmation of it infecting the crop. In the absence of any molecular or isozyme confirmation, diagnosis of M. enterolobii is difficult to determine. In the current study, the species responsible for substantial galling damage (Fig. 1A) on several cassava roots growing in Ibadan, Nigeria (7°22′39″ N; 3°54′21″ E) were diagnosed. DNA isolated from juveniles recovered using a modified Baermann method (Hooper, 1986) from these roots was used for PCR amplification of the mitochondrial Nad5 using primer pair, NAD5F2 (5’-TATTTTTTGTTTGAGATATATTAG-3’) and NAD5R1 (5’-CGTGAATCTTGATTTTCCATTTTT-3’). The 515 bp PCR DNA product was sequenced on both strands (GenBank Accession No. MW965454) and found to be 100% identical to M. enterolobii with those in the DNA sequence database (KU372358, KU372359) (Janssen et al., 2016; Kolombia et al., 2017). In addition, M. incognita was also recovered from the galled roots and identified using the same primers (GenBank Accession No. MW965455) indicating a combined species infection (Fig. 2). Cultures of M. enterolobii, developed from single egg masses were maintained on tomato plants and used to assess infection on cassava in 10 L pots filled with steam sterilized loam soil in the screenhouse. Cassava cv. IITA-TMS-IBA070593 cuttings planted in June, 2018 and repeated in April, 2019 were inoculated with 1,000 juveniles per pot at three weeks after planting, and then maintained for four months before quantifying the nematode densities in both roots and soil. Nematode reproduction factor (RF), calculated from total nematode densities (n=8) from soil and roots was as high as 44.3, compared to uninoculated controls. Molecular diagnostics of M. enterolobii, as above, confirms unequivocally the host status of cassava to this nematode. This study reports for the first time the infection of cassava by M. enterolobii under field conditions in Africa and for the first time demonstrates the host suitability of cassava to this nematode (Fig. 1B). M. enterolobii is among the most commonly occurring root-knot nematode species in West Africa (dos Santos et al., 2019). It is therefore anticipated that M. enterolobii has long been infecting, especially in West Africa, but has been overlooked due to its morphological similarity with M. incognita. Given the high reproductive ability of M. enterolobii on cassava and its highly aggressive nature on a range of crops, it is likely that it is causing, or will result in, high levels of losses on cassava in Africa.


2004 ◽  
Vol 29 (4) ◽  
pp. 441-446 ◽  
Author(s):  
Laura Otero ◽  
Daniel Ducasse ◽  
Robert N. G. Miller

Using PCR-based assays with specific primers for amplification of the ribosomal DNA intergenic spacer region (IGS) and a portion of the mitochondrial DNA small subunit ribosomal RNA gene (mtDNA SSU rRNA), the genetic variability among Verticillium dahliae isolates from olive (Olea europaea) and other host species from Argentina and Brazil was estimated. The derived UPGMA-generated phenograms based upon the restriction fingerprinting data of rDNA IGS products revealed genetic differences, correlating with the host of origin. Isolates infecting olive genetically distinct from those from cocoa (Theobroma cacao) and sunflower (Helianthus annuus). Digestion of mitochondrial DNA SSU rRNA PCR products revealed less variability, distinguishing only one isolate from sunflower. Ribosomal DNA ITS restriction patterns were identical for all isolates of V. dahliae, irrespective of host of origin. These preliminary results may have relevance for Verticillium wilt control practices, possibly reflecting a different evolutionary origin, or reproductive isolation of the pathogen in olive, distinct from populations of other hosts.


Genome ◽  
2000 ◽  
Vol 43 (3) ◽  
pp. 584-587 ◽  
Author(s):  
Pawel Brzuzan

Length variation of the mitochondrial DNA control region was observed with PCR amplification of a sample of 138 whitefish (Coregonus lavaretus). Nucleotide sequences of representative PCR products showed that the variation was due to the presence of an approximately 100-bp motif tandemly repeated two, three, or five times in the region between the conserved sequence block-3 (CSB-3) and the gene for phenylalanine tRNA. This is the first report on the tandem array composed of long repeat units in mitochondrial DNA of salmonids.Key words: Coregonus lavaretus, D-loop mitochondrial DNA, tandemly repeated sequences.


Gut Pathogens ◽  
2021 ◽  
Vol 13 (1) ◽  
Author(s):  
Sohyun Lee ◽  
Nanjoo Park ◽  
Sujung Yun ◽  
Eunseon Hur ◽  
Jiwon Song ◽  
...  

AbstractNon-typhoidal salmonellosis remains a pressing public health problem worldwide. Quinolones, particularly fluoroquinolones, are widely used to treat various infections, including non-typhoidal salmonellosis, which can be a serious illness. The emergence of fluoroquinolone-resistant Salmonella has resulted in treatment failure and high mortality rates. In this study, we estimated the presence of plasmid-mediated quinolone resistance (PMQR) genes in Salmonella enterica isolated from human salmonellosis patients in South Korea from 2016 to 2019. We evaluated the association of these genes with fluoroquinolone susceptibility. Antimicrobial susceptibility tests for Salmonella isolates were performed using the Vitek II system, and the minimum inhibitory concentrations (MIC) of ciprofloxacin and levofloxacin were determined using the E-test method. Plasmid-mediated quinolone resistance (PMQR) genes were detected by PCR amplification and quinolone resistance-determining regions (QRDRs) of the gyrA and parC genes were analyzed following Sanger sequencing of the PCR products. Thirty-four Salmonella strains with reduced susceptibility to fluoroquinolones (ciprofloxacin MIC ≥ 0.125 µg/mL and levofloxacin MIC ≥ 0.25 µg/mL) were selected from 208 human clinical Salmonella isolates. Among them, 22 Salmonella strains harbored one PMQR gene (qnrA, qnrB, or qnrS), and three Salmonella strains carried two PMQR genes (qnrS and aac(6′)-Ib-cr or qnrA and qnrB). qnrS was the most common PMQR gene. Serotyping revealed that Salmonella 4,[5]12:i:- (32.4%, 11/34) and Salmonella Typhimurium (29.4%, 10/34) were the two most predominant serovars, and Multi-locus sequence typing (MLST) showed that ST19 and ST34 were the most frequent sequence types. In conclusion, qnr gene-positive Salmonella 4,[5],12:i:- and Salmonella Typhimurium were the main serovars responsible for reduced susceptibility to fluoroquinolones. Therefore, our findings suggest that PMQR-positive Salmonella strains, which can be isolated from various samples including human, food, and the environment, should be carefully monitored.


Water ◽  
2021 ◽  
Vol 13 (15) ◽  
pp. 2086
Author(s):  
Maciej Karpowicz ◽  
Sabina Smolska ◽  
Magdalena Świsłocka ◽  
Joanna Moroz

Our results are the first insight into groundwater copepods of the Polish Lowland. The sampling was conducted in 28 wells in north-eastern Poland, and Copepoda were present in 16 wells. We have identified six Copepoda species and one Cladocera. We have classified four species as stygophiles—Eucyclops serrulatus, Diacyclops bisetosus, Diacyclops crassicaudis, and Cyclops furcifer. These species were frequently found in studied wells of different regions of north-eastern Poland, often in high numbers, and females with egg sacs were observed. We present a detailed morphological description of these species, together with molecular characteristics based on mitochondrial DNA markers (COI gene) for E. serrulatus, D. bisetosus, and D. crassicaudis, and 12S ribosomal RNA for C. furcifer. We also present the development of abnormal structures in one specimen of D. crassicaudis, where the upper part of furcal rami was fused to form a single plate.


1999 ◽  
Vol 122 (2) ◽  
pp. 323-328 ◽  
Author(s):  
M. T. E. P. ALLSOPP ◽  
C. M. HATTINGH ◽  
S. W. VOGEL ◽  
B. A. ALLSOPP

A panel of 16S ribosomal RNA gene probes has been developed for the study of the epidemiology of heartwater; five of these detect different cowdria genotypes, one detects five distinct genotypes; one detects any Group III Ehrlichia species other than Cowdria and one detects any Group II Ehrlichia species. These probes have been used on PCR-amplified rickettsial 16S rRNA genes from over 200 Amblyomma hebraeum ticks. Control ticks were laboratory-reared and either uninfected or fed on sheep experimentally infected with different cowdria isolates, field ticks were collected from animals in heartwater-endemic areas. All tick-derived DNA samples were also examined by PCR amplification and probing for two other cowdria genes (map1 and pCS20) which have previously been used for heartwater epidemiology. This paper describes the first direct comparison of all currently available DNA probes for heartwater-associated organisms.


2019 ◽  
Vol 14 (1) ◽  
Author(s):  
Xiaoxiao Zhu ◽  
Hoi-Yan Wu ◽  
Pang-Chui Shaw ◽  
Wei Peng ◽  
Weiwei Su

Abstract Background Pheretima is a minister drug in Naoxintong capsule (NXTC), a well-known traditional Chinese medicine (TCM) formula for the treatment of cardiovascular and cerebrovascular diseases. Owing to the loss of morphological and microscopic characteristics and the lack of recognized chemical marker, it is difficult to identify Pheretima in NXTC. This study aims to evaluate the feasibility of using DNA techniques to authenticate Pheretima, especially when it is processed into NXTC. Methods DNA was extracted from crude drugs of the genuine and adulterant species, as well as nine batches of NXTCs. Based on mitochondrial cytochrome c oxidase subunit I (COI) gene, specific primers were designed for two genera of genuine species, Metaphire and Amynthas, respectively. PCR amplification was performed with the designed primers on crude drugs of Pheretima and NXTCs. The purified PCR products were sequenced and the obtained sequences were identified to species level with top hit of similarity with BLAST against GenBank nucleotide database. Results Primers MF2R2 and AF3R1 could amplify specific DNA fragments with sizes around 230–250 bp, both in crude drugs and NXTC. With sequencing and the BLAST search, identities of the tested samples were found. Conclusion This study indicated that the molecular approach is effective for identifying Pheretima in NXTC. Therefore, DNA identification may contribute to the quality control and assurance of NXTC.


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