scholarly journals Gut Symbiotic Microbial Communities in the IUCN Critically Endangered Pinna nobilis Suffering from Mass Mortalities, Revealed by 16S rRNA Amplicon NGS

Pathogens ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 1002
Author(s):  
Athanasios Lattos ◽  
Ioannis A. Giantsis ◽  
Dimitrios Karagiannis ◽  
John A. Theodorou ◽  
Basile Michaelidis
Keyword(s):  
16S Rrna ◽  
Synergistic Effects ◽  
Disease Outbreaks ◽  
Healthy Population ◽  
Emerging Pathogens ◽  
Pinna Nobilis ◽  
Next Generation Sequencing Ngs ◽  
First Time ◽  
Vibrio Spp ◽  
Of Greece

Mass mortality events due to disease outbreaks have recently affected almost every healthy population of fan mussel, Pinna nobilis in Mediterranean Sea. The devastating mortality of the species has turned the interest of the research towards the causes of these events. After the haplosporidan infestation and the infection by Mycobacterium sp., new emerging pathogens have arisen based on the latest research. In the present study, a metagenomic approach of 16S rRNA next generation sequencing (NGS) was applied in order to assess the bacterial diversity within the digestive gland of diseased individuals as well as to carry out geographical correlations among the biodiversity of microbiome in the endangered species Pinna nobilis. The specimens originated from the mortalities occurred in 2019 in the region of Greece. Together with other bacterial genera, the results confirmed the presence of Vibrio spp., assuming synergistic effects in the mortality events of the species. Alongside with the presence of Vibrio spp., numerous bacterial genera were detected as well, including Aliivibrio spp., Photobacterium spp., Pseudoalteromonas spp., Psychrilyobacter spp. and Mycoplasma spp. Bacteria of the genus Mycoplasma were in high abundance particularly in the sample originated from Limnos island representing the first time recorded in Pinna nobilis. In conclusion, apart from exclusively the Haplosporidan and the Mycobacterium parasites, the presence of potentially pathogenic bacterial taxa detected, such as Vibrio spp., Photobactrium spp. and Alivibrio spp. lead us to assume that mortality events in the endangered Fan mussel, Pinna nobilis, may be attributed to synergistic effects of more pathogens.

scholarly journals The synergistic interference effect of silica nanoparticles concentration and the wavelength of ELISA on the colorimetric assay of cell toxicity

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Fariba Abbasi ◽  
Hassan Hashemi ◽  
Mohammad Reza Samaei ◽  
Amir SavarDashtaki ◽  
Abooalfazl Azhdarpoor ◽  
...  
Keyword(s):  
Mtt Assay ◽  
Interference Effect ◽  
Polynomial Regression ◽  
Colorimetric Assay ◽  
Synergistic Effects ◽  
Cell Toxicity ◽  
Diphenyltetrazolium Bromide ◽  
Nanoparticles Concentration ◽  
Artificial Neural Network Ann ◽  
First Time

AbstractThe 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay is the most common method for the determination of cell toxicity, but some factors limit the sensitivity of this method, such as pH. Less attention had been paid to the interference effect of optical and plasmonic properties of SiO2 nanoparticles (NPs) in the wavelength range assigned to MTT. This study investigated the synergistic interference effect of SiO2 NPs and wavelength on MTT assay for the first time. The examined variables included the type of SiO2 NPs concentrations (1, 10, and 100 mM) and different wavelengths (470, 490, 520, and 570 nm). The results showed that optical density (OD) increased (p < 0.05) when wavelength and the concentration of crystalline SiO2 NPs increased. So, the maximum OD at 10 and 100 mM were attributed to crystalline SiO2 NPs (p < 0.05) due to the functional group, whereas it was related to amorphous at 1 mM (p > 0.05). According to polynomial regression modeling (PRM), the maximum interference effect was predicted at crystalline SiO2 NPs and wavelength > 550 nm. Besides, the synergistic effects of SiO2 NPs, wavelength, and concentration of NPs had been a good fitting with first-order PRM. Thus, the concentration of SiO2 NPs had a confounder factor in colorimetric for MTT assay. The best artificial neural network (ANN) structure was related to the 3:7:1 network (Rall = 0.936, MSE = 0.0006, MAPE = 0.063). The correlation between the actual and predicted data was 0.88. As SiO2 NPs presence is an interfering factor in MTT assay concerning wavelength, it is suggested wavelength use with minimum confounding effect for MTT assay.

scholarly journals Molecular Detection and Phylogeny of Anaplasmaphagocytophilum and Related Variants in Small Ruminants from Turkey

Animals ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 814
Author(s):  
Münir Aktaş ◽  
Sezayi Özübek ◽  
Mehmet Can Uluçeşme
Keyword(s):  
16S Rrna ◽  
Small Ruminants ◽  
Rrna Gene ◽  
Infection Rates ◽  
Sheep And Goats ◽  
Significant Difference ◽  
Pcr Rflp ◽  
First Time ◽  
Japan And China ◽  
Apparently Healthy

Anaplasma phagocytophilum causes tick-borne fever in small ruminants. Recently, novel Anaplasma variants related to A. phagocytophilum have been reported in ruminants from Tunisia, Italy, South Korea, Japan, and China. Based on 16S rRNA and groEL genes and sequencing, we screened the frequency of A. phagocytophilum and related variants in 433 apparently healthy small ruminants in Turkey. Anaplasma spp. overall infection rates were 27.9% (121/433 analyzed samples). The frequency of A. phagocytophilum and A. phagocytophilum-like 1 infections was 1.4% and 26.5%, respectively. No A. phagocytophilum-like 2 was detected in the tested animals. The prevalence of Anaplasma spp. was comparable in species, and no significant difference was detected between sheep and goats, whereas the prevalence significantly increased with tick infestation. Sequencing confirmed PCR-RFLP data and showed the presence of A. phagocytophilum and A. phagocytophilum-like-1 variant in the sampled animals. Phylogeny-based on 16S rRNA gene revealed the A. phagocytophilum-like 1 in a separate clade together with the previous isolates detected in small ruminants and ticks. In this work, A. phagocytophilum-like 1 has been detected for the first time in sheep and goats from Turkey. This finding revealed that the variant should be considered in the diagnosis of caprine and ovine anaplasmosis.

scholarly journals Genetic characterization at the species and symbiovar level of indigenous rhizobial isolates nodulating Phaseolus vulgaris in Greece

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Evdoxia Efstathiadou ◽  
Georgia Ntatsi ◽  
Dimitrios Savvas ◽  
Anastasia P. Tampakaki
Keyword(s):  
Phaseolus Vulgaris ◽  
Common Bean ◽  
Phylogenetic Affiliation ◽  
Rhizobial Inoculation ◽  
Rhizobial Species ◽  
The Common ◽  
First Time ◽  
Of Greece ◽  
Rhizobial Isolates

AbstractPhaseolus vulgaris (L.), commonly known as bean or common bean, is considered a promiscuous legume host since it forms nodules with diverse rhizobial species and symbiovars. Most of the common bean nodulating rhizobia are mainly affiliated to the genus Rhizobium, though strains belonging to Ensifer, Pararhizobium, Mesorhizobium, Bradyrhizobium, and Burkholderia have also been reported. This is the first report on the characterization of bean-nodulating rhizobia at the species and symbiovar level in Greece. The goals of this research were to isolate and characterize rhizobia nodulating local common bean genotypes grown in five different edaphoclimatic regions of Greece with no rhizobial inoculation history. The genetic diversity of the rhizobial isolates was assessed by BOX-PCR and the phylogenetic affiliation was assessed by multilocus sequence analysis (MLSA) of housekeeping and symbiosis-related genes. A total of fifty fast-growing rhizobial strains were isolated and representative isolates with distinct BOX-PCR fingerpriniting patterns were subjected to phylogenetic analysis. The strains were closely related to R. anhuiense, R. azibense, R. hidalgonense, R. sophoriradicis, and to a putative new genospecies which is provisionally named as Rhizobium sp. I. Most strains belonged to symbiovar phaseoli carrying the α-, γ-a and γ-b alleles of nodC gene, while some of them belonged to symbiovar gallicum. To the best of our knowledge, it is the first time that strains assigned to R. sophoriradicis and harbored the γ-b allele were found in European soils. All strains were able to re-nodulate their original host, indicating that they are true microsymbionts of common bean.

The larvae of three Greek species of Hydropsyche (Trichoptera: Hydropsychidae) and key for larvae of known Aegean Hydropsyche species

Zootaxa ◽  
2018 ◽  
Vol 4382 (2) ◽  
pp. 381
Author(s):  
IOANNIS KARAOUZAS
Keyword(s):  
Balkan Peninsula ◽  
Diagnostic Features ◽  
Aegean Islands ◽  
First Time ◽  
Of Greece

The larvae of Hydropsyche cetibeli Malicky & Sipahiler 1993 from the Aegean, Hydropsyche mostarensis Klapálek 1898, endemic of the Balkan Peninsula, and Hydropsyche pygmalion Malicky 2001 endemic of Serifos Island (Cyclades, Greece) are described for the first time. The diagnostic features of the species are described and illustrated, and some information regarding their ecology is included. In addition, a tabular key for larvae of the known Hydropsyche species of Greece and the Aegean Islands is included. 

scholarly journals First record of Ornithodoros faccinii (Acari: Argasidae) on toads of genus Rhinella (Anura: Bufonidae) in Brazil

2018 ◽  
Author(s):  
Hermes Ribeiro Luz ◽  
Bruna Barboza Bezerra ◽  
Walter Flausino ◽  
Arlei Marcili ◽  
Sebastián Muñoz-Leal ◽  
...  
Keyword(s):  
16S Rrna ◽  
Tick Species ◽  
Rio De Janeiro ◽  
Bird Species ◽  
First Record ◽  
Molecular Approach ◽  
Phylogenetic Relatedness ◽  
Mitochondrial Sequence ◽  
First Time

Abstract Although a group of soft ticks (Argasidae) associated with amphibians was recently discovered in Brazilian rainforests, parasitism by these ticks on cold-blooded animals remains less common than on mammal and bird species. In this study, we identified ticks that were collected from toads that had been caught in December 2016 and January 2017, at Itinguçú waterfall (22°54’05” S; 43°53’30” W) in the municipality of Itaguaí, state of Rio de Janeiro. Tick specimens were identified using a morphological and molecular approach. In total, twelve larvae of Ornithodoros ticks were collected from three individuals of Rhinella ornata and were identified as Ornithodoros faccinii. Our results include a longer 16S rRNA mitochondrial sequence for O. faccinii that supports its phylogenetic relatedness to Ornithodoros saraivai, and we report this tick species parasitizing Rhinella toads for the first time in Brazil.

scholarly journals Α revision of tenuipalpid mites of Greece (Acari: Tenuipalpidae)

2017 ◽  
Vol 5 ◽  
pp. 47 ◽  
Author(s):  
Ε.Ν. Hatzinikolis
Keyword(s):  
New Species ◽  
The Family ◽  
The World ◽  
Two New Species ◽  
First Time ◽  
Of Greece

The family Tenuipalpidae from Greece is revised and keys to the Greek species of the gene­ra Aegyptobia, Brevipalpus, Cenopalpus, Pentamerismus and Tenuipalpus are provided. A key to the Greek tenuipalpid genera is also given. The species: Aegyptobia leiahensis, Phytoptipaipusparadoxus, Brevipalpus recki, Pentamerismus coronatus, P. juniperi, P. o­regonensis, Pseudoleptus zelihae, Dolichotetranychus floridanus, Raoiella macfarlanei and Obdulia tamaricis are recorded for the first time. Two new species Aegyptobia karys­tensis and Aegyptobia aliartensis are described and illustrated. Hosts, distributional data and relation to hosts are presented for each species. A revaluation of the world genera and subgenera of the Tenuipalpidae is presented.

Evidence for direct transmission of the cat lungworm Troglostrongylus brevior (Strongylida: Crenosomatidae)

Parasitology ◽  
2013 ◽  
Vol 140 (7) ◽  
pp. 821-824 ◽  
Author(s):  
EMANUELE BRIANTI ◽  
GABRIELLA GAGLIO ◽  
ETTORE NAPOLI ◽  
LUIGI FALSONE ◽  
SALVATORE GIANNETTO ◽  
...  
Keyword(s):  
Pulmonary Disease ◽  
Scientific Information ◽  
Observational Evidence ◽  
Direct Transmission ◽  
Emerging Pathogens ◽  
Troglostrongylus Brevior ◽  
New Knowledge ◽  
Paratenic Hosts ◽  
First Time

SUMMARYMetastrongyloids of cats are emerging pathogens that may cause fatal broncho-pulmonary disease. Infestation of definitive hosts occurs after ingestion of intermediate or paratenic hosts. Among metastrongyloids of cats, Troglostrongylus brevior and Troglostrongylus subcrenatus (Strongylida: Crenosomatidae) have recently been described as agents of severe broncho-pulmonary disease. Here, we provide, for the first time, observational evidence suggesting the direct transmission of T. brevior from queen cat to suckling kittens. This new knowledge will have a significant impact on current scientific information of this parasite and shed new light into the biology and epidemiology of metastrongyloid nematodes.

First Complete Genome of Lake Sinai Virus Lineage 3 and Genetic Diversity of Lake Sinai Virus Strains From Honey Bees and Bumble Bees

2020 ◽  
Vol 113 (3) ◽  
pp. 1055-1061 ◽  
Author(s):  
Laura Šimenc ◽  
Urška Kuhar ◽  
Urška Jamnikar-Ciglenečki ◽  
Ivan Toplak
Keyword(s):  
Honey Bee ◽  
Honey Bees ◽  
Complete Genome ◽  
Bumble Bees ◽  
Bumble Bee ◽  
Rt Pcr ◽  
Genome Region ◽  
Pcr Method ◽  
Next Generation Sequencing Ngs ◽  
First Time

Abstract The complete genome of Lake Sinai virus 3 (LSV3) was sequenced by the Ion Torrent next-generation sequencing (NGS) technology from an archive sample of honey bees collected in 2010. This strain M92/2010 is the first complete genome sequence of LSV lineage 3. From October 2016 to December 2017, 56 honey bee samples from 32 different locations and 41 bumble bee samples from five different locations were collected. These samples were tested using a specific reverse transcriptase-polymerase chain reaction (RT-PCR) method; 75.92% of honey bee samples and 17.07% of bumble bee samples were LSV-positive with the RT-PCR method. Phylogenetic comparison of 557-base pair-long RNA-dependent RNA polymerase (RdRp) genome region of selected 23 positive samples of honey bees and three positive bumble bee samples identified three different LSV lineages: LSV1, LSV2, and LSV3. The LSV3 lineage was confirmed for the first time in Slovenia in 2010, and the same strain was later detected in several locations within the country. The LSV strains detected in bumble bees are from 98.6 to 99.4% identical to LSV strains detected among honey bees in the same territory.

scholarly journals Prospective assessment of NGS-detectable mutations in CML patients with nonoptimal response: the NEXT-in-CML study

Blood ◽  
2020 ◽  
Vol 135 (8) ◽  
pp. 534-541 ◽  
Author(s):  
Simona Soverini ◽  
Luana Bavaro ◽  
Caterina De Benedittis ◽  
Margherita Martelli ◽  
Alessandra Iurlo ◽  
...  
Keyword(s):  
Clinical Relevance ◽  
Kinase Inhibitors ◽  
Clinical Decision Making ◽  
Mutation Screening ◽  
Clinical Decision ◽  
Kinase Domain ◽  
Low Level ◽  
Prospective Assessment ◽  
Next Generation Sequencing Ngs ◽  
First Time

Abstract In chronic myeloid leukemia (CML) patients, tyrosine kinase inhibitors (TKIs) may select for drug-resistant BCR-ABL1 kinase domain (KD) mutants. Although Sanger sequencing (SS) is considered the gold standard for BCR-ABL1 KD mutation screening, next-generation sequencing (NGS) has recently been assessed in retrospective studies. We conducted a prospective, multicenter study (NEXT-in-CML) to assess the frequency and clinical relevance of low-level mutations and the feasibility, cost, and turnaround times of NGS-based BCR-ABL1 mutation screening in a routine setting. A series of 236 consecutive CML patients with failure (n = 124) or warning (n = 112) response to TKI therapy were analyzed in parallel by SS and NGS in 1 of 4 reference laboratories. Fifty-one patients (22 failure, 29 warning) who were negative for mutations by SS had low-level mutations detectable by NGS. Moreover, 29 (27 failure, 2 warning) of 60 patients who were positive for mutations by SS showed additional low-level mutations. Thus, mutations undetectable by SS were identified in 80 out of 236 patients (34%), of whom 42 (18% of the total) had low-level mutations somehow relevant for clinical decision making. Prospective monitoring of mutation kinetics demonstrated that TKI-resistant low-level mutations are invariably selected if the patients are not switched to another TKI or if they are switched to a inappropriate TKI or TKI dose. The NEXT-in-CML study provides for the first time robust demonstration of the clinical relevance of low-level mutations, supporting the incorporation of NGS-based BCR-ABL1 KD mutation screening results in the clinical decision algorithms.

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