Genotyping Mycoplasma hyorhinis by multi-locus sequence typing and multiple-locus variable-number tandem-repeat analysis

Mycoplasma hyorhinis is a swine pathogen bacterium, which causes significant economic losses. The infection spreads through direct contact between the animals. Powerful genotyping methods like PCR based multi-locus sequence typing (MLST) and multiple-locus variable-number tandem-repeat analysis (MLVA) are necessary to monitor the infections and to conduct epidemiological investigations; hence supporting the control of the disease. The aims of the present study were to examine M. hyorhinis isolates originating mainly from Hungary with MLST and MLVA developed in the study, and to compare the results of the two typing methods. To characterize 39 M. hyorhinis isolates and the type strain (NCTC 10,130), six house-keeping genes were selected for MLST and six tandem-repeat regions were chosen for MLVA. We were able to differentiate 31 sequence types and 37 genotypes within the 40 analyzed isolates by the MLST and the MLVA, respectively. With the combination of the two newly developed assays all examined isolates were distinguished with the exception of the ones originating from the same animal. The developed MLST assay provided a robust and high resolution phylogenetic tree, while the MLVA system is suitable for the differentiation of closely related isolates from the same farm, hence the assay is appropriate for epidemiologic studies

Anaplasmataceae closely related to Ehrlichia chaffeensis and Neorickettsia helminthoeca from birds in Central Europe, Hungary

Increasing amount of data attest that (in the context of vector-borne infections) birds are not only important as hosts of blood-sucking arthropod vectors, but also as reservoirs of vector-borne pathogens. From 2015 to 2019 cadavers of 100 birds (from 45 species, nine orders) were collected in Hungary, and their organs were screened for DNA from a broad range of vector-borne bacteria with PCR and sequencing. Molecular analyses revealed the presence of Anaplasmataceae, and sequencing identified bacteria closely related to Neorickettsia helminthoeca and Ehrlichia chaffeensis in a Eurasian teal (Anas crecca)and a song thrush (Turdus philomelos), respectively. All samples were PCR negative for rickettsiae, borreliae, Francisella and Coxiella spp., as well as for piroplasms. To our knowledge, this is the first report of a Neorickettsia and an Ehrlichia sp., which belong to the phylogenetic groups of N. helminthoeca and E. chaffeensis, respectively, from Europe. The potential presence of these two vector-borne bacteria needs to be taken into account during future studies on the eco-epidemiology of Anaplasmataceae in Europe

Establishment of a Mycoplasma hyorhinis challenge model in 5-week-old piglets

IntroductionMycoplasma hyorhinis is an emerging swine pathogen with high prevalence worldwide. The main lesions caused are arthritis and polyserositis, and the clinical manifestation of the disease may result in significant economic losses due to decreased weight gain and enhanced medical costs. We aimed to compare two challenge routes to induce M. hyorhinis infection using the same clinical isolate.MethodsFive-week-old, Choice hybrid pigs were inoculated on 2 consecutive days by intravenous route (Group IV-IV) or by intravenous and intraperitoneal routes (Group IV-IP). Mock-infected animals were used as control (control group). After the challenge, the clinical signs were recorded for 28 days, after which the animals were euthanized. Gross pathological and histopathological examinations, PCR detection, isolation, and genotyping of the re-isolated Mycoplasma sp. and culture of bacteria other than Mycoplasma sp. were carried out. The ELISA test was used to detect anti-M. hyorhinis immunoglobulins in the sera of all animals.ResultsPericarditis and polyarthritis were observed in both challenge groups; however, the serositis was more severe in Group IV-IV. Statistically significant differences were detected between the challenged groups and the control group regarding the average daily weight gain, pathological scores, and ELISA titers. Additionally, histopathological scores in Group IV-IV differed significantly from the scores in the control group. All re-isolated strains were the same or a close genetic variant of the original challenge strain.DiscussionOur results indicate that both challenge routes are suitable for modeling the disease. However, due to the evoked more severe pathological lesions and the application being similar to the hypothesized natural route of infection in Group IV-IV, the two-dose intravenous challenge is recommended by the authors to induce serositis and arthritis associated with M. hyorhinis infection

HLA-DQ2 homozygosis increases tTGA levels at diagnosis but does not influence the clinical phenotype of celiac disease: a multicenter study

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Sertések Mycoplasma hyorhinis okozta megbetegedése

A szerzők összefoglalják a Mycoplasma hyorhinis-ről és az általa okozott megbetegedésekről rendelkezésre álló legfontosabb ismereteket. Leírják, hogy a M. hyorhinis a savóshártyák és ízületek gyulladásával járó kórkép mellett, kötőhártyagyulladást és középfülgyulladást is okozhat. Megállapítják, hogy a megfelelő diagnózis felállításához a járványtani adatok, klinikai tünetek és kórbonctani elváltozások mellett laboratóriumi vizsgálatokra is szükség van, valamint, hogy az állományok célzott antibiotikumos kezelésével csökkenthető a M. hyorhinis fertőzés okozta gazdasági kár

Ivóvízben adagolt tilvalozin hatékonyságának vizsgálata Mycoplasma hyopneumoniae okozta légzőszervi betegség kezelésére hízósertésekben

A szerzők hízósertéseken, nagyüzemi körülmények között vizsgálták 10 mg/ttkg, ivóvízben, 5 napon át adagolt tilvalozin (TVN) hatékonyságát Mycoplasma hyopneumoniae okozta légzőszervi betegség gyógy- és metafilaktikus kezelésére. A gyógyulási arány 91,7% volt a TVN, 86,7% a referens kontrollcsoportban, ami szignifikáns egyenértékűséget jelent. Az új megbetegedés aránya szignifikánsan kisebb volt (0,9% szemben a 4,5%-kal), valamint a légzőszervi tünetek enyhébb formában jelentek meg a TVN (átlag pontszám: 0,09, ill. 0,26), mint a kezeletlen kontrollcsoportban. Az eredmények alátámasztják a tilvalozin hatékonyságát a sertések Mycoplasma-pneumoniájának gyógy- és metafilaktikus kezelésére. | The objective of this field trial was to investigate the efficacy of tilvalozin (TVN) administered via drinking water at a dose of 10 mg/kg bw for 5 days for the treatment and metaphylaxis of respiratory disease caused by Mycoplasma hyopneumoniae (hereafter: M. hyopneumoniae) for fattening pigs under farm conditions. The primary parameters were the cure rate of clinically ill pigs and the new incidence rate among in contact pen mates. The two groups consisted of mixed sex fattening pigs of the same age (16 weeks) housed in the same fattening barn in 11–11 pens (average 22 pigs per pen). In addition to M. hyopneumoniae, Pasteurella multocida, Glässerella parasuis and Mycoplasma hyorhinis were detected in bronchoalveolar fluid taken before the first treatment and Actinobacillus pleuropneumoniae was isolated from the lung of a dead pig. The TVN group received Aivlosin® 625 mg granules for use in drinking water for pigs (ECO Animal Health), while the clinically ill animals in the control group were treated with tylosin injection (Pharmasin 200 mg/ml inj. Huvepharma NV) at a dose of 10 mg/kg bw for 3 days. Clinically healthy pigs in the control group were not treated. Thus, the study was a positive (reference) controlled (non-inferiority) from curative treatment and a negative (untreated) controlled from metaphylaxis point of view. The cure rate was 91.7% in the TVN group vs 86.7% in the positive control group, which was statistically significant equivalence. The incidence of new cases as well as the severity of the respiratory signs proved to be significantly lower in the TVN (0.9%, mean respiratory score 0.09) than in the negative (untreated) control group (4.5% and mean score of 0.26). The results support the efficacy of tylvalosin in a dose of 10 mg/kg bw via drinking water for the treatment and metaphylaxis of respiratory disease associated with M. hyopneumoniae under field conditions

HLA-DQ2 homozygosis increases tTGA levels at diagnosis but does not influence the clinical phenotype of coeliac disease : A multicentre study

Magnitude of gluten-specific T-cell responses in coeliac disease (CD) might be dependent on HLA-DQ2 gene dose. We aimed to investigate the effects of HLA-DQB1*02 allele dose on clinical outcomes.We reviewed the charts of all coeliac patients attending to three Hungarian university clinics after 1997 and included those patients, who (a) were diagnosed with CD, (b) underwent high-resolution HLA typing and (c) were ≥18 years at the time of data collection. HLA typing was performed to determine DQB1*02 allele dose. Patients were divided into risk groups by DQB1*02 allele dose, as follows: high-, intermediate- and low-risk groups corresponded to a double, single and zero doses, respectively. We used ANOVA and Pearson's chi-squared test to explore association between HLA risk and clinical variables.A total of 727 coeliac patients attended the clinics but only 105 (14.4%) patients were eligible for inclusion. High, intermediate and low HLA risk patients comprised 35.3%, 52.3% and 12.3% of the study population, respectively. Double dose of HLA-DQB1*02 was more frequent in patient with high tTGA level (>10 times the upper limit of normal; p = 0.045). Gene dose was not associated with younger age at diagnosis (p = 0.549), gender (p = 0.739), more severe diagnostic histology (p = 0.318), more frequent classical presentation (p = 0.846), anaemia (p = 0.611), metabolic bone disease (p = 0.374), dermatitis herpetiformis (p = 0.381) and autoimmune diseases (p = 0.837).Our study shows a significant gene dose effect in terms of tTGA level at diagnosis, but no significant association between HLA-DQB1*02 allele dose and the clinical outcomes in CD

Preventing Plasmon Coupling between Gold Nanorods Improves the Sensitivity of Photoacoustic Detection of Labeled Stem Cells <i>in Vivo</i>

Gold nanorods are excellent contrast agents for imaging technologies which rely on near-infrared absorption such as photoacoustic imaging. For cell tracking applications, the cells of interest are labeled with the contrast agent prior to injection. However, after uptake into cells by endocytosis, the confinement and high concentration in endosomes leads to plasmon band broadening and reduced absorbance. This would limit the potential of multispectral optoacoustic tomography in terms of spectral processing and, consequently, sensitivity. Here, we show that steric hindrance provided by silica coating of the nanorods leads to the preservation of their spectral properties and improved photoacoustic sensitivity. This strategy allowed the detection and monitoring of as few as 2 × 10⁴ mesenchymal stem cells in mice over a period of 15 days with a high spatial resolution. Importantly, the silica-coated nanorods did not affect the viability or differentiation potential of the transplanted mesenchymal stem cells