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1.
Orv Hetil ; 165(33): 1303-1304, 2024 Aug 18.
Artículo en Húngaro | MEDLINE | ID: mdl-39154332
2.
Aquat Toxicol ; 273: 107020, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-39002427

RESUMEN

Concentrations of microplastics (MPs) were determined in three commonly used zebrafish housing systems to see if their levels could affect the final results of laboratory microplastic-related toxicology tests. MPs have received notable attention in the last few years, and their toxicology tests have also come to the fore. Zebrafish (Danio rerio), kept in fish housing systems, are widely used as models for MPs studies. Most of these systems contain a significant number of parts made of different polymers. As usage and amortization can erode these parts, MPs might appear in the keeping water or the fish body, which may represent a background load and possibly influence the results of microplastic-related toxicological tests. To take representative water samples from systems, two in-situ filtration techniques, a newly developed peristaltic pump-, and a jet pump-driven method were applied. The collected MP particles were analyzed with a Fourier-transform infrared microscope (detection limit 50 µm), and their possible origin was also investigated. The newly developed technique was more sufficient for sampling as it had a higher MPs recovery, especially in the smaller size range. Polyester, polyethylene and polypropylene were the most frequently detected polymers in the examined fish housing systems, the highest detected concentration was 0.31±0.12 particles/liter (0.22±0.16 µg/liter). These values are negligible compared to the literature data reporting enormously high applied MPs concentrations (104 - 2.21 × 108 particles/liter) during toxicology tests. The results also show that some detected MPs did not originate from the systems, their origin was presumed to be external.


Asunto(s)
Microplásticos , Pruebas de Toxicidad , Contaminantes Químicos del Agua , Pez Cebra , Pez Cebra/fisiología , Animales , Microplásticos/toxicidad , Microplásticos/análisis , Contaminantes Químicos del Agua/análisis , Contaminantes Químicos del Agua/toxicidad , Pruebas de Toxicidad/métodos , Vivienda para Animales , Monitoreo del Ambiente/métodos
3.
Artículo en Inglés | MEDLINE | ID: mdl-39009944

RESUMEN

Acute-phase inhibition of the pro-inflammatory alarmin S100A8/A9 improves cardiac function post-myocardial infarction (MI), but the mechanisms underlying the long-term benefits of this short-term treatment remain to be elucidated. Here, we assessed the effects of S100A8/A9 blockade with the small-molecule inhibitor ABR-238901 on myocardial neovascularization in mice with induced MI. The treatment significantly reduced S100A9 and increased neovascularization in the myocardium, assessed by CD31 staining. Proteomic analysis by mass-spectrometry showed strong myocardial upregulation of the pro-angiogenic proteins filamin A (~ 10-fold) and reticulon 4 (~ 5-fold), and downregulation of the anti-angiogenic proteins Ras homolog gene family member A (RhoA, ~ 4.7-fold), neutrophilic granule protein (Ngp, ~ 4.0-fold), and cathelicidin antimicrobial peptide (Camp, ~ 4.4-fold) versus controls. In-vitro, ABR-238901 protected against apoptosis induced by recombinant human S100A8/A9 in human umbilical vein endothelial cells (HUVECs). In conclusion, S100A8/A9 blockade promotes post-MI myocardial neovascularization by favorably modulating pro-angiogenic proteins in the myocardium and by inhibiting endothelial cell apoptosis.

4.
Magy Onkol ; 68(2): 171-176, 2024 Jul 16.
Artículo en Húngaro | MEDLINE | ID: mdl-39013091

RESUMEN

Previous twin studies show that genetic factors are responsible for 63% of the variability in breast density. We analyzed the mammographic images of 9 discordant twin pairs for breast cancer from the population-based Hungarian Twin Registry. We measured breast density using 3D Slicer software. Genetic variants predisposing to breast cancer were also examined. One of the examined twin pairs had a BRCA2 mutation in both members. There was no significant difference between the mean values of breast density in the tumor and non-tumor groups (p=0.323). In terms of parity and the presence of menopause, we found mostly no significant difference between the members of the twin pair. In our cohort of identical twins discordant for breast cancer, the average breast density showed no significant difference, which can be explained by the common genetic basis of breast cancer and breast density.


Asunto(s)
Densidad de la Mama , Neoplasias de la Mama , Mamografía , Humanos , Femenino , Neoplasias de la Mama/genética , Neoplasias de la Mama/patología , Neoplasias de la Mama/diagnóstico por imagen , Hungría , Persona de Mediana Edad , Gemelos Monocigóticos/genética , Adulto , Predisposición Genética a la Enfermedad , Sistema de Registros , Proteína BRCA2/genética , Anciano , Enfermedades en Gemelos/genética , Enfermedades en Gemelos/epidemiología , Mutación , Mama/diagnóstico por imagen , Mama/patología
5.
Children (Basel) ; 11(7)2024 Jun 28.
Artículo en Inglés | MEDLINE | ID: mdl-39062246

RESUMEN

INTRODUCTION: The corpus callosum is one of the five main cerebral commissures. It is key to combining sensory and motor functions. Its structure can be pathological (dysgenesis) or completely absent (agenesis). The corpus callosum dys- or agenesis is a rare disease (1:4000 live births), but it can have serious mental effects. METHODS: In our study, we processed the data of 64 pregnant women. They attended a prenatal diagnostic center and genetic counseling from 2005 to 2019 at the Department of Obstetrics and Gynecology at Semmelweis University. RESULTS: The pregnancies had the following outcomes: 52 ended in delivery, 1 in spontaneous abortion, and 11 in termination of pregnancy (TOP) cases (n = 64). The average time of detection with imaging tests was 25.24 gestational weeks. In 16 cases, prenatal magnetic resonance imaging (MRI) was performed. If the abnormality was detected before the 20th week, a genetic test was performed on an amniotic fluid sample obtained from a genetic amniocentesis. Karyotyping and cytogenetic tests were performed in 15 of the investigated cases. Karyotyping gave normal results in three cases (46,XX or XY). In one of these cases, postnatally chromosomal microarray (CMA) was later performed, which confirmed Aicardi syndrome (3q21.3-21.1 microdeletion). In one case, postnatally, the test found Wiedemann-Rautenstrauch syndrome. In other cases, it found X ring, Di George syndrome, 46,XY,del(13q)(q13q22) and 46,XX,del(5p)(p13) (Cri-du-chat syndrome). Edwards syndrome was diagnosed in six cases, and Patau syndrome in one case. CONCLUSIONS: We found that corpus callosum abnormalities are often linked to chromosomal problems. We recommend that a cytogenetic test be performed in all cases to rule out inherited diseases. Also, the long-term outcome does not just depend on the disease's severity and the associated other conditions, and hence proper follow-up and early development are also key. For this reason, close teamwork between neonatology, developmental neurology, and pediatric surgery is vital.

6.
Sensors (Basel) ; 24(11)2024 May 22.
Artículo en Inglés | MEDLINE | ID: mdl-38894089

RESUMEN

The measurement of electrical conductivity (EC) has long been a tool for understanding soil properties. Previous studies concluded that EC measurement is not an ion-selective method, but these papers did not address the measurement frequency. An experimental tool and method were developed for semi-factory conditions in a large-scale soil trough at the Institute of Technology of the Hungarian University of Agricultural and Life Sciences. A specially designed and built test apparatus mounted on the tractor's three-point hitch was used as a measuring device. The wear-resistant steel elements of the measuring device were also the sensors for measuring EC. This paper describes the conditions of the measurement series, the measurement results, and our conclusions from the experiments with the soil sensor. Different characteristics were measured in soil moistened with K and Ca solutions at different concentrations. The EC values show an increasing tendency with increasing salt concentration, and we also found that the rate of change of EC is different for different solution ratios. Based on our measurements, we found that the best method to isolate concentration differences is to use the test frequency range 20 Hz-250 kHz.

7.
Animals (Basel) ; 14(11)2024 May 23.
Artículo en Inglés | MEDLINE | ID: mdl-38891584

RESUMEN

Porcine Reproductive and Respiratory Syndrome (PRRS) significantly impacts the pig farming industry globally, leading to economic losses due to reduced productivity. This study focuses on assessing the presence and impact of PRRS within Hungarian wild boar populations amidst efforts to eradicate the virus from domestic pig herds. We used a combination of serological and virological tests on samples collected from wild boars across Hungary to evaluate the prevalence of PRRS virus and its potential transmission risks to domestic pigs. Our findings reveal a low seropositivity rate in wild boars, suggesting a minimal role of wild boars in the transmission of PRRS to domestic pig populations. Moreover, no relationship was found between domestic pig and wild boar densities, emphasizing the limited interaction and consequent risk of disease spread between these populations. We confirm the effectiveness of Hungary's PRRS eradication measures among domestic herds and highlight the negligible risk posed by wild boars in re-introducing the PRRS virus.

8.
Diagnostics (Basel) ; 14(10)2024 May 08.
Artículo en Inglés | MEDLINE | ID: mdl-38786277

RESUMEN

Background: Exercise-induced modifications in ECG parameters among individuals with an early repolarization pattern (ERP) have not been evaluated in detail. We aimed to assess this phenomenon, with potential associations with arrhythmogenesis. Methods: Twenty-three young, healthy males with ERP (ERP+) participated in this study, alongside a control group, which consisted of nineteen healthy males without ERP (ERP-). ECGs at baseline, at peak exercise (Bruce protocol), and during the recovery phase were analyzed and compared between the two groups. Results: The treadmill test demonstrated strong cardiovascular fitness, with similar chronotropic and pressor responses in both groups. In the baseline ECGs, the QRS complex and the QT interval were shorter in the ERP+ group. During exercise, the P-wave duration was significantly longer and the QRS was narrower in the ERP+ group. In the recovery phase, there was a longer P wave and a narrower QRS in the ERP+ group. During the treadmill test, the J wave disappeared or did not meet the criteria required for ERP diagnosis. Conclusions: The slowed intra-atrial conduction found during exercise could be predictive of atrial arrhythmogenesis in the setting of ERP. The disappearing of J waves during exercise, due to increased sympathetic activity, has potential clinical significance.

9.
Microbiol Resour Announc ; 13(7): e0105623, 2024 Jul 18.
Artículo en Inglés | MEDLINE | ID: mdl-38809039

RESUMEN

A subgroup of Salmonella (S.) enterica subsp. enterica serovar Paratyphi B is significantly associated with invasive infections in humans. We report the complete genome sequence of a potentially invasive. S. Paratyphi B isolated from a mute swan (Cygnus olor) found dead at an urban recreation park in Berlin, Germany.

10.
Polymers (Basel) ; 16(9)2024 Apr 24.
Artículo en Inglés | MEDLINE | ID: mdl-38732662

RESUMEN

The goal of the study was to compare the surface characteristics of typical implant materials used in orthopedic surgery and traumatology, as these determine their successful biointegration. The morphological and chemical structure of Vortex plate anodized titanium from commercially pure (CP) Grade 2 Titanium (Ti2) is generally used in the following; non-cemented total hip replacement (THR) stem and cup Ti alloy (Ti6Al4V) with titanium plasma spray (TPS) coating; cemented THR stem Stainless steel (SS); total knee replacement (TKR) femoral component CoCrMo alloy (CoCr); cemented acetabular component from highly cross-linked ultrahigh molecular weight polyethylene (HXL); and cementless acetabular liner from ultrahigh molecular weight polyethylene (UHMWPE) (Sanatmetal, Ltd., Eger, Hungary) discs, all of which were examined. Visualization and elemental analysis were carried out by scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS) and X-ray photoelectron spectroscopy (XPS). Surface roughness was determined by atomic force microscopy (AFM) and profilometry. TPS Ti presented the highest Ra value (25 ± 2 µm), followed by CoCr (535 ± 19 nm), Ti2 (227 ± 15 nm) and SS (170 ± 11 nm). The roughness measured in the HXL and UHMWPE surfaces was in the same range, 147 ± 13 nm and 144 ± 15 nm, respectively. EDS confirmed typical elements regarding the investigated prosthesis materials. XPS results supported the EDS results and revealed a high % of Ti4+ on Ti2 and TPS surfaces. The results indicate that the surfaces of prosthesis materials have significantly different features, and a detailed characterization is needed to successfully apply them in orthopedic surgery and traumatology.

12.
Nutrients ; 16(6)2024 Mar 16.
Artículo en Inglés | MEDLINE | ID: mdl-38542774

RESUMEN

Dermal photoaging refers to the skin's response to prolonged and excessive ultraviolet (UV) exposure, resulting in inflammation, changes to the tissue, redness, swelling, and discomfort. Betanin is the primary betacyanin in red beetroot (Beta vulgaris) and has excellent antioxidant properties. Yet, the specific molecular mechanisms of betanin in HaCaT cells have not been fully clarified. The objective of this study was to investigate the activity of betanin and the underlying mechanisms in HaCaT cells; furthermore, in this study, we explored the protective effect of various concentrations of betanin against UVB irradiation on HaCaT cells. Additionally, we assessed its influence on the transcription of various epigenetic effectors, including members of the DNA methyltransferase (DNMT) and histone deacetylase (HDAC) families. Our findings demonstrate a notable downregulation of genes in HaCaT cells, exhibiting diverse patterns upon betanin intake. We considered the involvement of DNMT and HDAC genes in distinct stages of carcinogenesis and the limited exploration of the effects of daily exposure dosages. Our results indicate that betanin may protect the skin from damage caused by UV exposure. Further investigation is essential to explore these potential associations.


Asunto(s)
Betacianinas , Neoplasias Cutáneas , Humanos , Betacianinas/farmacología , Fragmentación del ADN , Células HaCaT , Neoplasias Cutáneas/genética , Neoplasias Cutáneas/prevención & control , Epigénesis Genética , Quimioprevención , Rayos Ultravioleta/efectos adversos
13.
Animals (Basel) ; 14(1)2024 Jan 04.
Artículo en Inglés | MEDLINE | ID: mdl-38200906

RESUMEN

Porcine reproductive and respiratory syndrome (PRRS) is the cause of the most severe economic losses in the pig industry worldwide. PRRSV is extremely diverse in Europe, which poses a significant challenge to disease control within a country or any region. With the combination of phylogenetic reconstruction and network analysis, we aimed to uncover the major routes of the dispersal of PRRSV clades within Hungary. In brief, by analyzing >2600 ORF5 sequences, we identified at least 12 clades (including 6 clades within lineage 1 and 3 clades within lineage 3) common in parts of Western Europe (including Denmark, Germany and the Netherlands) and identified 2 novel clades (designated X1 and X2). Of interest, some genetic clades unique to other central European countries, such as the Czech Republic and Poland, were not identified. The pattern of PRRSV clade distribution is consistent with the route of the pig trade among countries, showing that most of the identified clades were introduced from Western Europe when fatteners were transported to Hungary. As a result of rigorous implementation of the national eradication program, the swine population was declared officially free from PRRSV. This map of viral diversity and clade distribution will serve as valuable baseline information for the maintenance of PRRSV-free status in the post-eradication era.

14.
Surg Endosc ; 38(2): 529-539, 2024 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-38062181

RESUMEN

BACKGROUND: Endometriosis is a chronic condition affecting 6-10% of women of reproductive age, with endometriosis-related pain and infertility being the leading symptoms. Currently, the gold standard treatment approach to surgery is conventional laparoscopy (CL); however, the increasing availability of robot-assisted surgery is projected as a competitor of CL. This study aimed to compare the perioperative outcomes of robot-assisted laparoscopy (RAL) and CL in endometriosis surgery. OBJECTIVES: We aimed to compare the effectiveness and safety of these two procedures. METHODS: A systematic search was conducted in three medical databases. Studies investigating different perioperative outcomes of endometriosis-related surgeries were included. Results are presented as odds ratios (OR) or mean differences (MD) with 95% confidence intervals (CI). RESULTS: Our search yielded 2,014 records, of which 13 were eligible for data extraction. No significant differences were detected between the CL and RAL groups in terms of intraoperative complications (OR = 1.07, CI 0.43-2.63), postoperative complications (OR = 1.3, CI 0.73-2.32), number of conversions to open surgery (OR = 1.34, CI 0.76-2.37), length of hospital stays (MD = 0.12, CI 0.33-0.57), blood loss (MD = 16.73, CI 4.18-37.63) or number of rehospitalizations (OR = 0.95, CI 0.13-6.75). In terms of operative times (MD = 28.09 min, CI 11.59-44.59) and operating room times (MD = 51.39 min, CI 15.07-87.72;), the RAL technique remained inferior. CONCLUSION: RAL does not have statistically demonstrable advantages over CL in terms of perioperative outcomes for endometriosis-related surgery.


Asunto(s)
Endometriosis , Laparoscopía , Procedimientos Quirúrgicos Robotizados , Robótica , Femenino , Humanos , Endometriosis/cirugía , Procedimientos Quirúrgicos Robotizados/métodos , Laparoscopía/métodos , Complicaciones Intraoperatorias/cirugía
15.
Environ Pollut ; 342: 123113, 2024 Feb 01.
Artículo en Inglés | MEDLINE | ID: mdl-38072021

RESUMEN

In this study, the disrupting effects of glyphosate (GLY), aminomethylphosphonic acid (AMPA), and three glyphosate-based herbicides (GBHs) on vitellogenesis in a non-concentration-dependent manner are reported for the first time in 120 h of acute exposure of zebrafish at environmentally relevant concentrations. GBHs are commonly used worldwide in weed control management. Due to their extensive application, they frequently occur in aquatic ecosystems and may affect various organisms. The active substance GLY and its major by-product, AMPA, are the most thoroughly studied chemicals; however, the adverse effects of the complex formulas of GBHs with diverse and unknown content of co-formulants are still not sufficiently researched. This study focused on the embryotoxicity, sublethal malformations, and estrogenic potency of GLY, AMPA, and four commonly used GBHs on zebrafish embryos using a wild type and an estrogen-sensitive, transgenic zebrafish line (Tg(vtg1:mCherry)). After 120 h of exposition, AMPA did not cause acute toxicity, while the LC50 of GLY was 160 mg/L. The GBHs were more toxic with LC50 values ranging from 31 to 111 GLY active equivalent (a.e.) mg/L. Exposure to 0.35-2.8 mg/L GBHs led to sublethal abnormalities: typical symptoms were structural deformation of the lower jaw and anomalies in the olfactory region. Deformity rates were 10-30% in the treated groups. In vivo, fluorescently expressed vtg1 mCherry protein in embryonic liver was detected by a non-invasive microscopic method indicating estrogenic action through vitellogenin production by GLY, AMPA, and GBHs. To confirm the in vivo findings, RT-qPCR method was performed to determine the levels of the estrogenicity-related vtg1 mRNA. After 120 h of exposure to GLY, AMPA, and three GBHs at a concentration of 0.35 mg/L, the expression of vtg1 gene was significantly up-regulated. Our results highlight the risk that short-term GLY and GBH exposure can cause developmental malformations and disrupt the hormonal balance in zebrafish embryos.


Asunto(s)
Glifosato , Herbicidas , Organofosfonatos , Animales , Pez Cebra , Ácido alfa-Amino-3-hidroxi-5-metil-4-isoxazol Propiónico , Glicina/toxicidad , Ecosistema , Herbicidas/toxicidad , Animales Modificados Genéticamente , Estrona
16.
Antibiotics (Basel) ; 12(12)2023 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-38136774

RESUMEN

(1) Background: Microinjection of zebrafish (Danio rerio) embryos offers a promising model for studying the virulence and potential environmental risks associated with Pseudomonas aeruginosa. (2) Methods: This work aimed to develop a P. aeruginosa infection model using two parallel exposition pathways on zebrafish larvae with microinjection into the yolk and the perivitelline space to simultaneously detect the invasive and cytotoxic features of the examined strains. The microinjection infection model was validated with 15 environmental and clinical strains of P. aeruginosa of various origins, antibiotic resistance profiles, genotypes and phenotypes: both exposition pathways were optimized with a series of bacterial dilutions, different drop sizes (injection volumes) and incubation periods. Besides mortality, sublethal symptoms of the treated embryos were detected and analyzed. (3) Results: According to the statistical evaluation of our results, the optimal parameters (dilution, drop size and incubation period) were determined. (4) Conclusions: The tested zebrafish embryo microinjection infection model is now ready for use to determine the in vivo virulence and ecological risk of environmental P. aeruginosa.

17.
Animals (Basel) ; 13(24)2023 Dec 05.
Artículo en Inglés | MEDLINE | ID: mdl-38136786

RESUMEN

Porcine reproductive and respiratory syndrome (PRRS) is a widespread infectious disease that is currently a major cause of economic losses in pig production. In Hungary, a National PRRS Eradication Program has been introduced to attain a more efficient, economic, and competitive international market position. The program has been also approved by the EU, but the resulting legal obligations have imposed a burden on Hungarian producers to comply with EU competition rules. The implementation of the program has been carried out by the veterinary authorities with the consent of, continuous support from and monitoring conducted by organisations within the pig sector as well as a scientific committee. The PRRS eradication program in Hungary was based on a regional territorial principle and was compulsory for all pig holdings within the regions. In Hungary, large fattening farms operate as all-in/all-out or continuous flow systems. Large-scale breeding herds are predominantly farrow-to-finish types. Although its significance has decreased in recent decades, 20% of the Hungarian pig population is still kept on small (backyard) farms (<100 animals). All PRRSV-infected large-scale farms had to develop a unit-adapted eradication plan, including external and internal biosecurity measures, vaccinations, etc. It was crucial to render each fattening unit free of the disease, as fattening units play a significant role in spreading the virus within the country. The eradication efforts mainly implemented were depopulation-repopulation methods, but on some farms a testing and removal method has been used. As the eradication progressed over the years, the introduction of infected fattening pigs was restricted. Thanks to these measures, Hungarian large-scale fattening farms became PRRSV-free by the end of 2018. The PRRSV-free status of small-scale herds was achieved by the end of 2015 and was maintained between 2016 and 2021. By 31 December 2021, all breeding pigs in large-scale farms in Hungary were free of wild-type PRRS virus. By 31 March 2022, the total pig population of the country, including all backyard farms and fattening units, achieved PRRSV-free status. The future goal is to ensure and maintain the PRRSV-free status of Hungary via strict import regulations of live animals combined with the continuous and thorough screening of incoming and resident herds for the presence of the virus.

18.
Front Microbiol ; 14: 1284929, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-38033583

RESUMEN

Salmonella enterica subsp. enterica serovar Agona has a history of causing food-borne outbreaks and any emergence of multidrug-resistant (MDR) isolates in novel food products is of concern. Particularly, in food products frequently consumed without sufficient heating prior to consumption. Here, we report about the MDR isolate, 18-SA00377, which had been isolated from a dietary supplement in Germany in 2018 and submitted to the German National Reference Laboratory for Salmonella. WGS-based comparative genetic analyses were conducted to find a potential reservoir of the isolate itself or mobile genetic elements associated with MDR. As a phylogenetic analysis did not yield any closely related S. Agona isolates, either globally or from Germany, a detailed analysis of the largest plasmid (295,499 bp) was performed as it is the main carrier of resistances. A combined approach of long-read and short-read sequencing enabled the assembly of the isolate's chromosome and its four plasmids. Their characterization revealed the presence of 23 different antibiotic resistance genes (ARGs), conferring resistance to 12 different antibiotic drug classes, as well as genes conferring resistance to six different heavy metals. The largest plasmid, pSE18-SA00377-1, belongs to the IncHI2 plasmid family and carries 16 ARGs, that are organized as two distinct clusters, with each ARG associated with putative composite transposons. Through a two-pronged approach, highly similar plasmids to pSE18-SA00377-1 were identified in the NCBI database and a search for Salmonella isolates with a highly similar ARG resistance profile was conducted. Mapping and structural comparisons between pSE18-SA00377-1 and these plasmids and Salmonella isolates showed that both the plasmid backbone and identical or similar ARG clusters can be found not only in Salmonella isolates, originating mostly from a wide variety of livestock, but also in a diverse range of bacterial genera of varying geographical origins and isolation sources. Thus, it can be speculated that the host range of pSE18-SA00377-1 is not restricted to Salmonella and its spread already occurred in different bacterial populations. Overall, this hints at a complex history for pSE18-SA00377-1 and highlights the importance of surveilling multidrug-resistant S. enterica isolates, especially in novel food items that are not yet heavily regulated.

19.
Animals (Basel) ; 13(19)2023 Oct 02.
Artículo en Inglés | MEDLINE | ID: mdl-37835686

RESUMEN

This study reports on the molecular epidemiology of Ingelvac-PRRS-MLV-associated cases in Hungary for the period 2020-2021. Field epidemiology investigations led the experts to conclude that imported pigs, which were shipped through transit stations in Denmark, introduced the vaccine virus. The movement of fatteners and the neglect of disease control measures contributed to the spread of the virus to PRRS-free pig holdings in the vicinity. Deep sequencing was performed to genetically characterize the genes coding for the virion antigens (i.e., ORF2 through ORF7). The study isolates exhibited a range of 0.1 to 1.8% nucleotide sequence divergence from the Ingelvac PRRS MLV and identified numerous polymorphic sites (up to 57 sites) along the amplified 3.2 kilo base pair genomic region. Our findings confirm that some PRRSV-2 vaccine strains can accumulate very high number of point mutations within a short period in immunologically naive pig herds.

20.
Animals (Basel) ; 13(20)2023 Oct 15.
Artículo en Inglés | MEDLINE | ID: mdl-37893946

RESUMEN

Porcine reproductive and respiratory syndrome virus (PRRSV) is a major concern worldwide. Control of PRRSV is a challenging task due to various factors, including the viral diversity and variability. In this study, we evaluated an amplicon library preparation protocol targeting the ORF7 region of both PRRSV species, Betaarterivirus suid 1 and Betaarterivirus suid 2. We designed tailed primers for a two-step PCR procedure that generates ORF7-specific amplicon libraries suitable for use on Illumina sequencers. We tested the method with serum samples containing common laboratory strains and with pooled serum samples (n = 15) collected from different pig farms during 2019-2021 in Hungary. Testing spiked serum samples showed that the newly designed method is highly sensitive and detects the viral RNA even at low copy numbers (corresponding to approx. Ct 35). The ORF7 sequences were easily assembled even from clinical samples. Two different sequence variants were identified in five samples, and the Porcilis MLV vaccine strain was identified as the minor variant in four samples. An in-depth analysis of the deep sequencing results revealed numerous polymorphic sites along the ORF7 gene in a total of eight samples, and some sites (positions 12, 165, 219, 225, 315, 345, and 351) were found to be common in several clinical specimens. We conclude that amplicon deep sequencing of a highly conserved region of the PRRSV genome could support both laboratory diagnosis and epidemiologic surveillance of the disease.

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