1,720,961 research outputs found
Loop-mediated isothermal amplification in the field: Reality beyond the potentiality
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Traumatic cutaneous myiasis by Wohlfahrtia magnifica in a stray dog in Italy
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Spotted fever group rickettsiae associated with ixodid ticks in wild environment in Southern Italy
Full text linkIxodidae ticks are vectors and reservoirs of several species of rickettsiae, and tick-borne rickettsioses are reported worldwide. This study was aimed to verify the distribution of spotted fever group rickettsiae associated with ticks in a wild environment, the National Park of Gargano, where there is proximity between wild and domestic animals, and which is within an endemic area for rickettsiosis. Ticks were collected from animals or vegetation, morphologically identified and tested by a PCR targeting the 17kDa gene, and by a loop-mediated isothermal amplification (LAMP) targeting ompB gene. Out of 34 tested tick pools, 2 from Dermacentor marginatus, 1 from Ixodes ricinus, and 1 from Rhipicephalus turanicus resulted positive. Nucleotide sequences of amplicons showed high similarity with sequences from Rickettsia slovaca, Rickettsia raoultii, Rickettsia helvetica, and Rickettsia felis. The overall calculated infection rate was 26.19 per 1,000, while it rose up to 107.77 when only D. marginatus was considered. The results highlight the association among Ri. slovaca, Ri. raoultii, D. marginatus and wild boars from which infected ticks were collected. Finally, the study shows the low efficacy of the previously described LAMP method for the detection of Rickettsia spp., when compared to PCR, making urgent the development of most effective LAMP protocols
Identification and characterization of orf viruses isolated from sheep and goats in Southern Italy
No full textOrf virus (ORFV; Family: Poxviridae) is the causative agent of contagious ecthyma, or Orf disease in sheep, goats and other domestic or wild ruminants with a worldwide distribution. The disease is endemic in Italy, but few data are available about its distribution and epidemiology. In the present study we analysed 32 clinical samples, obtained from crusted scab lesions of 5 goats and 27 sheep, from 19 suspected outbreaks of contagious ecthyma in Apulia and Basilicata regions between 2012 and 2014. Negative staining electron microscopy (EM) and polymerase chain reaction (PCR) targeting the late transcription factor gene (VLTF-1) were used to identify the virus. Isolation was also attempted on BHK-21 cell line. PCR was proved to be more sensitive than EM, as it detected the virus in 28 out of 32 samples, whereas the EM detected it only in 26 out of the 32 samples. The majority of isolated strains forms a monophyletic group; these isolates, according to the VLTF-1 sequencing, are high related to ORFV strains previously shown to circulate in Southern Italy.L'ORF virus (ORFV; Family: Poxviridae) è l'agente causale dell'ectima contagioso, malattia infettiva contagiosa degli ovicaprini e di altri ruminanti domestici o selvatici, con distribuzione mondiale. La malattia è endemica in Italia, ma in letteratura scientifica sono disponibili pochi dati sulla sua distribuzione ed epidemiologia. In questo studio sono stati analizzati 32 campioni clinici (croste cutanee) prelevati da 27 ovini e 5 caprini provenienti da 19 focolai sospetti di ectima contagioso occorsi in Puglia e Basilicata tra il 2012 e il 2014. Per identificare il virus sono state effettuate sia l'analisi morfologica mediante microscopia elettronica (ME), sia una PCR mirata al rilevamento del gene del fattore di trascrizione tardiva (VLTF-1). I ceppi virali sono stati isolati anche su linea cellulare BHK-21. La PCR è risultata essere la metodica più sensibile, evidenziando la presenza di ORF virus in 28 dei 32 campioni analizzati, mentre la microscopia elettronica ha permesso di rilevare il virus solo in 26 dei 32 campioni. La maggior parte degli isolati forma un gruppo monofiletico; essi, sulla base delle sequenze del gene VLTF-1, risultano essere altamente correlati a ceppi di ORF virus già circolanti in Sud Italia in passato
Development and Application of a Loop-Mediated Isothermal Amplification (LAMP) Approach for the Rapid Detection of Dirofilaria repens from Biological Samples
Full text linkDirofilariasis by Dirofilaria repens is an important mosquito vector borne parasitosis, and the dog represents the natural host and reservoir of the parasite. This filarial nematode can also induce disease in humans, and in the last decades an increasing number of cases have been being reported. The present study describes the first loop mediated isothermal amplification (LAMP) assay to detect D. repens DNA in blood and mosquitoes. Two versions of the technique have been developed and described: in the first, the amplification is followed point by point through a real time PCR instrument (ReT-LAMP); in the second, the amplification is visualized by checking UV fluorescence of the reaction mixture after addition of propidium iodide (PI-LAMP). The two variants use the same set of 4 primers targeting the D. repens cytochrome oxidase subunit I (COI) gene. To assess the specificity of the method, reactions were carried out by using DNA from the major zoonotic parasites of the family of Onchocercidae, and no amplification was observed. The lower limit of detection of the ReT-LAMP assay was 0.15 fg/μl (corresponding to about 50 copy of COI gene per μl). Results suggest that the described assay is specific, and its sensitivity is higher than the conventional PCR based on the same gene. It is also provide a rapid and cost-effective molecular detection of D. repens, mainly when PI-LAMP is applied, and it should be performed in areas where this emerging parasitosis is endemic
First report of Coxiella burnetii and Borrelia burgdorferi sensu lato in poultry red mites, Dermanyssus gallinae (Mesostigmata, Acari), related to urban outbreaks of dermatitis in Italy
Full text linkThe poultry red mite (PRM), Dermanyssus gallinae, is a nonburrowing haematophagous nest-dwelling ectoparasite of birds; occasionally it bites humans, inducing dermatitis. The possibility that this parasite may also be involved in transmission of pathogens is an additional concern. We investigated the presence of zoonotic agents in PRMs from bird nests and pets, and related them to urban outbreaks of dermatitis. A total of 98 PRMs from 12 outbreaks of PRM dermatitis that occurred in Italian cities from 2001 to 2017 were molecularly investigated for detection of Coxiella spp. (16S rRNA), Chlamydophila spp. (16S rRNA), Rickettsia spp. (17 kDa protein-encoding gene), Borrelia burgdorferi sensu lato (groEL gene) and Bartonella spp. (16S–23S rRNA intergenic spacer). Of the 12 tested mite pools, one was positive for Coxiella burnetii (100% identity) and two for B. burgdorferi sensu lato (99% with Borrelia afzelii). For the first time, the presence of B. burgdorferi sensu lato and C. burnetii is reported in PRMs from urban areas. Birds, mainly pigeons, can harbour both pathogens. Therefore, birds and their nest-dwelling PRMs may play a role in the epidemiology of these infections
Coxiella-like endosymbiont associated to the "Anatolian brown tick" Rhipicephalus bursa in Southern Italy
No full textSeveral different ticks have been reported to harbor microbes related to Coxiella burnetii, the agent of the Q fever. Rhipicephalus bursa is an important vector of tick-borne diseases in livestock in Mediterranean area; it is also abundant in ovi-caprine farms with C. burnetii infection, in Southern Italy. 60 females of Rh. bursa (15 pools) and 40 their eggs (2 pools) were screened for C. burnetii by a conventional PCR targeting the insertion sequence IS1111 and by Loop mediated isothermal amplification assay (LAMP) targeting com1 gene. One of 15 tick pools (1/15) and both egg pools (2/2) were found positive by LAMP assay and negative by PCR targeting IS1111 gene. 16S rRNA gene was amplified by PCR from the LAMP-positive pools, amplicons were sequenced and found 95% similar to the corresponding sequences from C. burnetii. This let us to hypothesize the presence of a new Coxiella-like endosymbiont associated with Rh. bursa which could be vertically transmitted, described here for the first time. The lack of detection of IS1111 in Coxiella endosymbiont of Rh. bursa could be related to the possible absence of the Pathogenicity island of C. burnetii, to which IS1111s are associated
Case report: Molecular detection of dirofilaria repens in an italian patient after a stay in tanzania
Full text linkA 35-year-old man was admitted to a hospital in the south of Italy because of a periocular nodule and subpalpebral edema. The patient reported having been stayed in Tanzania five months before. Hematologic parameters were within the normality range, the Acanthocheilonema viteae ELISA did not detect significant levels of antifilarial IgG, and no further symptoms were described. The surgical inspection of the nodule led to the isolation of two filarioid parasites, identified as Dirofilaria repens by scanning electron microscope (SEM), and then by molecular assays. Knott's test did not reveal microfilaremia, whereas loop-mediated isothermal amplification andPCRdetected D. repens DNA. The patient was treated with doxycycline, and he was found no more positive at the follow-up
Going Beyond Counting First Authors in Author Co-citation Analysis
Full text linkThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
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