Morphological and molecular characterization of Sarcocystis wenzeli in chickens (Gallus gallus) in China

Abstract Background There has been considerable confusion concerning the number and classification of Sarcocystis spp. in chickens. Scarce nucleotide data of Sarcocystis spp. from chickens are provided in GenBank. The study aimed to investigate the morphological and molecular characteristics of Sarcocystis spp. found in chickens in China. Methods Tissues from 33 chickens were collected in 2019. Sarcocysts were observed using light (LM) and transmission electron microscopy (TEM). Individual sarcocysts from different chickens were selected for DNA extraction, and five loci, 18S rDNA, 28S rDNA, ITS1 region, the mitochondrial cox1 gene and the apicoplastic rpoB gene, were amplified from each sarcocyst, sequenced and analyzed. Results Only S. wenzeli was found in 14 of 33 (42.4%) chickens. Under LM, the sarcocysts were microscopic and exhibited palisade-like villar protrusions measuring 1.5–2.8 μm. Ultrastructurally, the sarcocyst wall contained numerous stubby hill-like villar protrusions. The protrusions included scattered microtubules, which extended from the tips of the protrusions into the ground substance. The five loci were successfully sequenced and the sequences deposited in GenBank. At 18S rDNA, ITS1 and cox1, the most similar sequences in GenBank were those of Sarcocystis sp. obtained from the brains of chickens, i.e. 99.9–100%, 98.1–98.5% and 99.3% identity, respectively. The five loci (18S rDNA, 28S rDNA, ITS1, cox1 and rpoB) showed different levels of interspecific sequence similarity with other closely related species of Sarcocystis (e.g. 99.8%, 99.0–99.2%, 89.3–89.7%, 98.5%, and 97.5%, respectively, with S. anasi). Phylogenetic analysis based on four of the loci (18S rDNA, cox1, rpoB and ITS1) revealed that S. wenzeli formed an independent clade with Sarcocystis spp. that utilize geese or ducks as intermediate hosts and canines as the known or presumed definitive host. Conclusions To our knowledge, the sequences of 28S rDNA and rpoB reported here constitute the first records of genetic markers of Sarcocystis spp. in chickens. Based on molecular analysis, S. wenzeli might be responsible for the neurological disease in chickens, and ITS1 and rpoB are more suitable for discriminating it from closely related Sarcocystis spp. Phylogenetic analysis revealed that S. wenzeli presents a close relationship with Sarcocystis spp. in geese or ducks.

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Morphological and molecular characterization of Sarcocystis wenzeli in chickens (Gallus gallus) in China

10.21203/rs.3.rs-49974/v4 ◽

2020 ◽

Author(s):

Jing Pan ◽

Chunli Ma ◽

Zhumei Huang ◽

Yulong Ye ◽

Hongxia Zeng ◽

...

Keyword(s):

Phylogenetic Analysis ◽

18S Rdna ◽

Sequence Similarity ◽

Rpob Gene ◽

28S Rdna ◽

Ground Substance ◽

Molecular Characteristics ◽

Cox1 Gene ◽

Intermediate Hosts ◽

Sarcocystis Spp

Abstract Background: There has been considerable confusion concerning the number and classification of Sarcocystis spp. in chickens. Scarce nucleotide data of Sarcocystis spp. from chickens are provided in GenBank. The study aimed to investigate the morphological and molecular characteristics of Sarcocystis spp. found in chickens in China.Methods: Tissues from 33 chickens were collected in 2019. Sarcocysts were observed using light (LM) and transmission electron microscopy (TEM). Individual sarcocysts from different chickens were selected for DNA extraction, and five loci, 18S rDNA, 28S rDNA, ITS1 region, the mitochondrial cox1 gene and the apicoplastic rpoB gene, were amplified from each sarcocyst, sequenced and analyzed.Results: Only S. wenzeli was found in 14 of 33 (42.4%) chickens. Under LM, the sarcocysts were microscopic and exhibited palisade-like villar protrusions measuring 1.5–2.8 μm. Ultrastructurally, the sarcocyst wall contained numerous stubby hill-like villar protrusions. The protrusions included scattered microtubules, which extended from the tips of the protrusions into the ground substance. The five loci were successfully sequenced and the sequences deposited in GenBank. At 18S rDNA, ITS1 and cox1, the most similar sequences in GenBank were those of Sarcocystis sp. obtained from the brains of chickens, i.e. 99.9–100%, 98.1–98.5% and 99.3% identity, respectively. The five loci (18S rDNA, 28S rDNA, ITS1, cox1 and rpoB) showed different levels of interspecific sequence similarity with other closely related species of Sarcocystis (e.g. 99.8%, 99.0–99.2%, 89.3-89.7%, 98.5%, and 97.5%, respectively, with S. anasi). Phylogenetic analysis based on four of the loci (18S rDNA, cox1, rpoB and ITS1) revealed that S. wenzeli formed an independent clade with Sarcocystis spp. that utilize geese or ducks as intermediate hosts and canines as the known or presumed definitive host.Conclusions: To our knowledge, the sequences of 28S rDNA and rpoB reported here constitute the first records of genetic markers of Sarcocystis spp. in chickens. Based on molecular analysis, S. wenzeli might be responsible for the neurological disease in chickens, and ITS1 and rpoB are more suitable for discriminating it from closely related Sarcocystis spp. Phylogenetic analysis revealed that S. wenzeli presents a close relationship with Sarcocystis spp. in geese or ducks.

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Morphological and Molecular Characterization of Sarcocystis Wenzeli in Chickens (Gallus gallus) in China

10.21203/rs.3.rs-49974/v1 ◽

2020 ◽

Author(s):

Jing Pan ◽

Chunli Ma ◽

Zhumei Huang ◽

Yulong Ye ◽

Hongxia Zeng ◽

...

Keyword(s):

Phylogenetic Analysis ◽

Related Species ◽

18S Rdna ◽

Sequence Similarity ◽

28S Rdna ◽

Ground Substance ◽

Molecular Characteristics ◽

Cox1 Gene ◽

Intermediate Hosts ◽

Closely Related Species

Abstract Background: There has been considerable confusion concerning the number and classification of Sarcocystis spp. found in chickens. To date, no nucleotide sequences of Sarcocystis spp. from chickens have been provided in GenBank. The present study aimed to investigate the morphological and molecular characteristics of Sarcocystis spp. in chickens in China.Methods: Tissues from 33 chickens were collected in 2019. Sarcocysts were observed using light (LM) and transmission electron microscopy (TEM). Individual sarcocysts from different chickens were selected for DNA extraction, and five loci, 18S rDNA, 28S rDNA, the ITS-1 region (ITS-1), the mitochondrial cox1 gene (cox1), and the apicoplastic rpoB gene (rpoB), were amplified from each sarcocyst, cloned, sequenced and analyzed.Results: Only S. wenzeli was found in 14 of 33 (42.4%) chickens. Under LM, the sarcocysts were microscopic and exhibited palisade-like villar protrusions measuring 1.5–2.8 μm. The bradyzoites were lancet-shaped, measuring 9.2–12.6 × 1.5–3.5 μm. Ultrastructurally, the sarcocyst wall contained numerous stubby hill-like villar protrusions that were up to 1.2 μm long and 1.0 μm wide. The protrusions included scattered microtubules, which extended from the tips of the protrusions into the ground substance. The intraspecific similarity of the five loci (18S rDNA, 28S rDNA, ITS-1, cox1, and rpoB) among the newly obtained sequences ranged from 99.8–100%, 99.7–100%, 99.0–99.9%, 100% and 98.9–100%, respectively. The five loci showed different levels of interspecific sequence similarity with closely related species of Sarcocystis (e.g., 99.8%, 99.0–99.2%, 89.3-89.7%, 98.5%, and 97.5%, respectively, with S. anasi). Phylogenetic analysis based on three of the loci (18S rDNA, cox1, and rpoB) revealed that S. wenzeli formed an independent clade with Sarcocystis spp. that employ geese or ducks as intermediate hosts and canines as the known or presumed definitive host.Conclusions: The sequences of 18S rDNA, 28S rDNA, ITS-1, cox1 and rpoB from S. wenzeli reported here constitute the first records of genetic markers of Sarcocystis from chickens made available in GenBank. Based on sequence analysis, ITS-1 and rpoB are more suitable for discriminating S. wenzeli from closely related species of Sarcocystis. Phylogenetic analysis revealed that S. wenzeli presents a close relationship with Sarcocystis spp. from geese or ducks.

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Morphological and molecular characterization of Sarcocystis wenzeli in chickens (Gallus gallus) in China

10.21203/rs.3.rs-49974/v2 ◽

2020 ◽

Author(s):

Jing Pan ◽

Chunli Ma ◽

Zhumei Huang ◽

Yulong Ye ◽

Hongxia Zeng ◽

...

Keyword(s):

Phylogenetic Analysis ◽

18S Rdna ◽

Sequence Similarity ◽

28S Rdna ◽

Ground Substance ◽

Molecular Characteristics ◽

Cox1 Gene ◽

Intermediate Hosts ◽

Rdna Its ◽

Sarcocystis Spp

Abstract Background: There has been considerable confusion concerning the number and classification of Sarcocystis spp. in chickens. Scarce nucleotide data of Sarcocystis spp. from chickens are provided in GenBank. The study aimed to investigate the morphological and molecular characteristics of Sarcocystis spp. found in chickens in China. Methods: Tissues from 33 chickens were collected in 2019. Sarcocysts were observed using light (LM) and transmission electron microscopy (TEM). Individual sarcocysts from different chickens were selected for DNA extraction, and five loci, 18S rDNA, 28S rDNA, the ITS-1 region ( ITS-1 ), the mitochondrial cox1 gene ( cox1 ), and the apicoplastic rpoB gene ( rpoB ), were amplified from each sarcocyst, sequenced and analyzed. Results: Only S . wenzeli was found in 14 of 33 (42.4%) chickens. Under LM, the sarcocysts were microscopic and exhibited palisade-like villar protrusions measuring 1.5–2.8 μm. Ultrastructurally, the sarcocyst wall contained numerous stubby hill-like villar protrusions. The protrusions included scattered microtubules, which extended from the tips of the protrusions into the ground substance. The five loci were successfully sequenced and deposited in GenBank. At 18S rDNA, ITS-1 and cox1 , the most similar sequences in GenBank were those of Sarcocystis sp. obtained from the brains of chickens, i.e., 99.9–100%, 98.1–98.5%, and 99.3% identity, respectively. The five loci (18S rDNA, 28S rDNA, ITS-1 , cox1 and rpoB ) showed different levels of interspecific sequence similarity with other closely related species of Sarcocystis (e.g., 99.8%, 99.0–99.2%, 89.3-89.7%, 98.5%, and 97.5%, respectively, with S . anasi ). Phylogenetic analysis based on three of the loci (18S rDNA, cox1 , and rpoB ) revealed that S . wenzeli formed an independent clade with Sarcocystis spp. that employ geese or ducks as intermediate hosts and canines as the known or presumed definitive host. Conclusions: The sequences of 28S rDNA and rpoB reported here constitute the first records of genetic markers of Sarcocystis spp. in chickens. Based on molecular analysis, S . wenzeli might be responsible for the neurological disease in chickens, and ITS-1 and rpoB are more suitable for discriminating it from closely related Sarcocystis spp. Phylogenetic analysis revealed that S . wenzeli presents a close relationship with Sarcocystis spp. in geese or ducks.

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Morphological and molecular characterization of Sarcocystis wenzeli in chickens (Gallus gallus) in China

10.21203/rs.3.rs-49974/v3 ◽

2020 ◽

Author(s):

Jing Pan ◽

Chunli Ma ◽

Zhumei Huang ◽

Yulong Ye ◽

Hongxia Zeng ◽

...

Keyword(s):

Phylogenetic Analysis ◽

18S Rdna ◽

Sequence Similarity ◽

Rpob Gene ◽

28S Rdna ◽

Ground Substance ◽

Molecular Characteristics ◽

Cox1 Gene ◽

Intermediate Hosts ◽

Rdna Its

Abstract Background: There has been considerable confusion concerning the number and classification of Sarcocystis spp. in chickens. Scarce nucleotide data of Sarcocystis spp. from chickens are provided in GenBank. The study aimed to investigate the morphological and molecular characteristics of Sarcocystis spp. found in chickens in China.Methods: Tissues from 33 chickens were collected in 2019. Sarcocysts were observed using light (LM) and transmission electron microscopy (TEM). Individual sarcocysts from different chickens were selected for DNA extraction, and five loci, 18S rDNA, 28S rDNA, ITS-1 region (ITS-1), the mitochondrial cox1 gene (cox1), and the apicoplastic rpoB gene (rpoB) , were amplified from each sarcocyst, sequenced and analyzed.Results: Only S. wenzeli was found in 14 of 33 (42.4%) chickens. Under LM, the sarcocysts were microscopic and exhibited palisade-like villar protrusions measuring 1.5–2.8 μm. Ultrastructurally, the sarcocyst wall contained numerous stubby hill-like villar protrusions. The protrusions included scattered microtubules, which extended from the tips of the protrusions into the ground substance. The five loci were successfully sequenced and deposited in GenBank. At 18S rDNA, ITS-1 and cox1, the most similar sequences in GenBank were those of Sarcocystis sp. obtained from the brains of chickens, i.e., 99.9–100%, 98.1–98.5% and 99.3% identity, respectively. The five loci (18S rDNA, 28S rDNA, ITS-1, cox1and rpoB) showed different levels of interspecific sequence similarity with other closely related species of Sarcocystis (e.g., 99.8%, 99.0–99.2%, 89.3-89.7%, 98.5%, and 97.5%, respectively, with S. anasi). Phylogenetic analysis based on four of the loci (18S rDNA, cox1, rpoB and ITS-1) revealed that S. wenzeli formed an independent clade with Sarcocystis spp. that employ geese or ducks as intermediate hosts and canines as the known or presumed definitive host.Conclusions: The sequences of 28S rDNA and rpoB reported here constitute the first records of genetic markers of Sarcocystis spp. in chickens. Based on molecular analysis, S. wenzeli might be responsible for the neurological disease in chickens, and ITS-1 and rpoB are more suitable for discriminating it from closely related Sarcocystis spp. Phylogenetic analysis revealed that S. wenzeli presents a close relationship with Sarcocystis spp. in geese or ducks.

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Morphological and Ultrastructural Characterization of Hemocytes in an Insect Model, the Hematophagous Dipetalogaster maxima (Hemiptera: Reduviidae)

Insects ◽

10.3390/insects12070640 ◽

2021 ◽

Vol 12 (7) ◽

pp. 640

Author(s):

Natalia R. Moyetta ◽

Fabián O. Ramos ◽

Jimena Leyria ◽

Lilián E. Canavoso ◽

Leonardo L. Fruttero

Keyword(s):

Cell Biology ◽

Cell Types ◽

Transmission Electron ◽

Ultrastructural Characterization ◽

Current Classification ◽

Contrast Microscopy ◽

First Time ◽

Controversial Aspect

Hemocytes, the cells present in the hemolymph of insects and other invertebrates, perform several physiological functions, including innate immunity. The current classification of hemocyte types is based mostly on morphological features; however, divergences have emerged among specialists in triatomines, the insect vectors of Chagas’ disease (Hemiptera: Reduviidae). Here, we have combined technical approaches in order to characterize the hemocytes from fifth instar nymphs of the triatomine Dipetalogaster maxima. Moreover, in this work we describe, for the first time, the ultrastructural features of D. maxima hemocytes. Using phase contrast microscopy of fresh preparations, five hemocyte populations were identified and further characterized by immunofluorescence, flow cytometry and transmission electron microscopy. The plasmatocytes and the granulocytes were the most abundant cell types, although prohemocytes, adipohemocytes and oenocytes were also found. This work sheds light on a controversial aspect of triatomine cell biology and physiology setting the basis for future in-depth studies directed to address hemocyte classification using non-microscopy-based markers.

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Sarcocystis Attenuati N. Sp. (Apicomplexa: Sarcocystidae) Infecting the Asian Gray Shrew, Crocidura Attenuata (Insectivora: Soricidae), in China

10.21203/rs.3.rs-907878/v1 ◽

2021 ◽

Author(s):

junjie hu ◽

Jun Sun ◽

Yanmei Guo ◽

Hongxia Zeng ◽

Yunzhi Zhang ◽

...

Keyword(s):

Host Specificity ◽

Definitive Host ◽

18S Rdna ◽

Common Species ◽

Molecular Characteristics ◽

Intermediate Hosts ◽

Parasite Life Cycle ◽

Rdna Sequences ◽

Transmission Electron ◽

18S Rdna Sequences

Abstract Background: There are limited data on Sarcocystis in insectivores. The Asian gray shrew, Crocidura attenuata, is one of the most common species of insectivores in the family Soricidae distributed in South Asia and Southeast Asia. To date, Sarcocystis has never been recorded in this host.Methods: Tissues from 42 Asian gray shrews were collected in China in 2017 and 2018. Sarcocysts were observed using light (LM) and transmission electron microscopy (TEM). To complete the parasite life cycle, muscle tissues of the host infected with sarcocysts were force-fed to two beauty rat snakes, Elaphe taeniura. Individual sarcocysts from different Asian gray shrews and oocysts/sporocysts isolated from the small intestines and feces of the experimental snakes were selected for DNA extraction, and seven genetic markers, including two nuclear loci (18S rDNA and ITS1), three mitochondrial genes (cox1, cox3 and cytb), and two apicoplastic genes (rpoB and clpC), were amplified, sequenced and analyzed.Results: Sarcocysts were found in 17 of 42 (40.5%) Asian gray shrews. Under LM, the microscopic sarcocysts were exhibited saw-tooth-like protrusions measuring 3.3–4.5 μm. Ultrastructurally, the sarcocyst wall contained numerous lancet- or leaf-like villous protrusions, similar to type 9h. The experimental beauty rat snakes shed oocysts/sporcysts measuring 11.9–16.7 × 9.2–10.6 μm with a prepatent period of 10 to 11 days. Comparing these sequences with those previously deposited in GenBank revealed that the 18S rDNA sequences and cox1 sequences shared the highest similarity with those of S. scandentiborneensis recorded in tree shrews, Tuaia minor and T. tana (i.e., 97.6–98.3% and 100% identity, respectively). Phylogenetic analysis based on 18S rDNA, ITS1 or cox1 sequences revealed that this parasite formed an independent clade with Sarcocystis spp. that utilize small animals as intermediate hosts and snakes as the known or presumed definitive host. On the basis of morphological and molecular characteristics and host specificity, the parasite was proposed as a new species, named S. attenuati.Conclusions: Sarcocysts were recorded in Asian gray shrews for the first time. The sarcocysts were characterized morphologically and molecularly. The 18S rDNA and cox1 sequences of S. attenuati, named in the present study, shared the highest identities with those of S. scandentiborneensis. However, the sarcocysts of the two species of Sarcocystis were quite different under LM and TEM. Based on experimental infection, beauty rat snakes have been proven to be a definitive host of S. attenuati. As more species of Sarcocystis from insectivores and other small mammals are properly morphologically and molecularly characterized, we may gain a better understanding of the biodiversity, host specificity and evolution of Sarcocystis in the future.

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Molecular characterization of isolated infectious bronchitis viruses from affected vaccinated broiler flocks in Syria

BMC Veterinary Research ◽

10.1186/s12917-020-02672-1 ◽

2020 ◽

Vol 16 (1) ◽

Author(s):

Tamara Al-Jallad ◽

Morshed Kassouha ◽

Mohamad Salhab ◽

Anouar Alomar ◽

Mouhamad AL-Masalma ◽

...

Keyword(s):

Sequence Similarity ◽

Hypervariable Region ◽

Effective Vaccine ◽

Molecular Characteristics ◽

Poultry Industry ◽

S1 Gene ◽

Infectious Bronchitis ◽

Epidemiological Monitoring ◽

Immunological Protection

Abstract Background Avian Infectious Bronchitis Virus (IBV) is a highly contagious disease that imposes a huge economic burden on the global poultry industry. IBV contains numerous serotypes and variants with incomplete tenuous cross immunological protection. The failure of currently used vaccines to protect against diverse, circulating IBV strains that are specific to a given region poses a major problem for the poultry industry. Thus, there is an urgent need to conduct studies aimed at genotyping field IB viruses. In this study, we have determined the molecular characteristics of circulating IBV by sequencing the S1 gene of viral isolates from affected previously vaccinated broiler flocks suffering from the disease. Results Ten isolates propagated in embryonated eggs showed an ability to induce typical IBV lesions after three successive viral passages. We performed a nested RT–PCR assay that targeted the hypervariable region 3 (HVR-3) of the S1 gene, and identified the isolates as IBV through sequence analysis. The IBV isolates showed sequence similarity between the Syrian isolates that vary from 96.20 to 100%, and those being closer to the Variant-2 strain IS/1494/06 (EU780077.2) with 97.5 to 99.4% similarities. However, less nucleotide identity was found with sequences belonging to the used vaccine strains such as H120, Mass5, and 4/91. Conclusions This study showed the presence of the Variant-2 strain circulating in Syrian broiler flocks showing signs of IBV disease. Currently, there is no commercial effective vaccine which protects birds against the Variant-2 strain. Continuous monitoring procedures should be taken to control and limit the spread of the IBV Variant-2 strain. This research emphasizes both the importance of epidemiological monitoring in intensive poultry farming for novel pathogens and the use of local isolates as future vaccine targets.

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Molecular characterization and phylogenetic analysis of Taenia multiceps from China

Acta Parasitologica ◽

10.1515/ap-2018-0085 ◽

2018 ◽

Vol 63 (4) ◽

pp. 721-727 ◽

Cited By ~ 1

Author(s):

L. Tan ◽

A.B. Wang ◽

S.Q. Zheng ◽

X.L. Zhang ◽

C.J. Huang ◽

...

Keyword(s):

Phylogenetic Analysis ◽

Dna Sequences ◽

Hunan Province ◽

Biological Study ◽

Economic Losses ◽

Molecular Characteristics ◽

12S Rrna ◽

Intermediate Hosts ◽

Taenia Multiceps ◽

Coenurus Cerebralis

Abstract Taenia multiceps, one of the most widely distributed zoonotic tapeworm parasites, is able to parasitize the small intestine of canids. The metacestode of T.multiceps is fatal to ruminants and causes important economic losses in livestock. However, molecular characteristics of T.multiceps and coenurus in China are still unclear. In this study, 36 goat isolates of the coenurus stage and 18 dog isolates of the adult stage of T.multiceps were obtained from three geographical areas in China and the isolated parasite above were analyzed by amplifying the partial of cytochrome coxidase subunit 1(pcox1), 12S ribosomal RNA (12S rRNA) from mitochondrial DNA (mtDNA) regions and an internal transcribed spacer (ITS) of ribosomal DNA (rDNA). These DNA sequences obtained from T.multiceps and coenurus were employed to evaluate the nucleotide diversity and confirm the relationship between T.multiceps and coenurus. Sequences variation were 0–1.4%, 0–1.5%, 0–4.2% for pcox1, 12S rRNA and ITS, respectively, among T.multiceps and coenurus isolates obtained in this study. In Sichuan province, sequence variations for Coenurus cerebralis isolated from Yaan city were 0–1.4% for pcox1, 0–1.0% for 12S rRNA and 0–2.1% for ITS. In Hunan province, variations were 0–1.0%, 0–1.5% and 0–3.3% for corresponding genes for non-coenurus cerebralis isolated from Changsha city, while variations of T.multiceps isolates from Xiangxi autonomous prefecture were 0–1.0%, 0–1.1% and 0–3.4% for pcox1, 12S rRNA and ITS, respectively. Phylogenetic analysis based on pcox1 sequences indicated that all cerebral and noncerebral metacestodes belong to T.multiceps. These results provide reference values for future molecular epidemiological and biological study on T.multiceps in dogs and intermediate hosts.

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Characterization of the Xanthomonas translucens Complex Using Draft Genomes, Comparative Genomics, Phylogenetic Analysis, and Diagnostic LAMP Assays

Phytopathology ◽

10.1094/phyto-08-16-0286-r ◽

2017 ◽

Vol 107 (5) ◽

pp. 519-527 ◽

Cited By ~ 25

Author(s):

Paul A. Langlois ◽

Jacob Snelling ◽

John P. Hamilton ◽

Claude Bragard ◽

Ralf Koebnik ◽

...

Keyword(s):

Phylogenetic Analysis ◽

Diagnostic Tools ◽

Cereal Crops ◽

Genome Sequences ◽

Germplasm Exchange ◽

Geographic Spread ◽

A Genome ◽

Xanthomonas Translucens

Prevalence of Xanthomonas translucens, which causes cereal leaf streak (CLS) in cereal crops and bacterial wilt in forage and turfgrass species, has increased in many regions in recent years. Because the pathogen is seedborne in economically important cereals, it is a concern for international and interstate germplasm exchange and, thus, reliable and robust protocols for its detection in seed are needed. However, historical confusion surrounding the taxonomy within the species has complicated the development of accurate and reliable diagnostic tools for X. translucens. Therefore, we sequenced genomes of 15 X. translucens strains representing six different pathovars and compared them with additional publicly available X. translucens genome sequences to obtain a genome-based phylogeny for robust classification of this species. Our results reveal three main clusters: one consisting of pv. cerealis, one consisting of pvs. undulosa and translucens, and a third consisting of pvs. arrhenatheri, graminis, phlei, and poae. Based on genomic differences, diagnostic loop-mediated isothermal amplification (LAMP) primers were developed that clearly distinguish strains that cause disease on cereals, such as pvs. undulosa, translucens, hordei, and secalis, from strains that cause disease on noncereal hosts, such as pvs. arrhenatheri, cerealis, graminis, phlei, and poae. Additional LAMP assays were developed that selectively amplify strains belonging to pvs. cerealis and poae, distinguishing them from other pathovars. These primers will be instrumental in diagnostics when implementing quarantine regulations to limit further geographic spread of X. translucens pathovars.

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Contribution to the Chromatic Characterization of Unifloral Honeys from Galicia (NW Spain)

Foods ◽

10.3390/foods8070233 ◽

2019 ◽

Vol 8 (7) ◽

pp. 233 ◽

Cited By ~ 5

Author(s):

Olga Escuredo ◽

María Shantal Rodríguez-Flores ◽

Sergio Rojo-Martínez ◽

María Carmen Seijo

Keyword(s):

Physicochemical Properties ◽

Principal Component ◽

Total Content ◽

Physicochemical Characteristics ◽

Dependent Variables ◽

Close Relationship ◽

Forward Stepwise ◽

Unifloral Honey

Honey color and other physicochemical characteristics depend mainly on the botanical and geographical origin. The study of these properties could make easier a correct classification of unifloral honey. This work determined the palynological characteristics and some physicochemical properties such as pH, electrical conductivity, and color (Pfund scale and the CIELa*b* coordinates), as well as the total content of the bioactive compounds phenols and flavonoids of ninety-three honey samples. Samples were classified as chestnut, blackberry, heather, eucalyptus, and honeydew honey. The study showed a close relationship between the physicochemical variables and the botanical origin. The five types of honey presented different physicochemical properties among them. A principal component analysis showed that Hue, lightness, b*, and Chroma variables were important for the honey types classification, followed by Erica pollen, pH, Cytisus, and Castanea variables. A forward stepwise regression analysis was performed introducing as dependent variables the color (mm Pfund) and the Chroma and the Hue variables. The regression models obtained explained 86%, 74%, and 86% of the variance of the data, respectively. The combination of the chromatic and physicochemical and pollen variables through the use of multivariable methods was optimal to characterize and group the honey samples studied.

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