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The effects exerted by human recombinant interleukin-1β (hrIL-1β) and the prostaglandin inhibitor indomethacin on the course of Cryptosporidium baileyi infection in chickens were studied. Daily oocyst shedding was monitored by a quantitative method throughout the experiment. Humoral immune response to C. baileyi was assessed by ELISA at 3 weeks of age while the level of cellular immune response to phytohaemagglutinin-P (PHA-P) by a skin test at 23 days of age. Parenteral application of hrIL-1b decreased oocyst shedding to 62%, but the infection ran a similar course in treated and control birds. The PHA-P skin test demonstrated increased cellular immune reaction in chickens receiving IL-1b, but there was no significant difference in the humoral responses of the two groups as detected by ELISA. On the other hand, indomethacin mixed to the feed lessened oocyst shedding to 13.7% and also shortened its duration. Immunological parameters as reflected by PHA-P skin test and ELISA results indicated enhanced cellular but unaltered humoral immune response. These data suggest that the sys- temic application of interleukin-1 can induce partial protection against C. baileyi in chickens and that prolonged, abundant oocyst shedding is due to an indometha- cin-sensitive immunodepression via the prostaglandin pathway.

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The vaccine-induced maternal immunity against classical swine fever (CSF) was investigated in this study. Eight sows were vaccinated with the Chinese strain of classical swine fever virus (CSFV). The length of time between vaccination and farrowing was 167-217 days. Milk samples from the front, middle and back udder sections and blood samples were taken from the sows on days 3 and 14 after farrowing. Blood samples were obtained from the piglets at the age of 3, 6 and 10 weeks. The antibody level of the milk was examined by ELISA, and that of blood samples by the virus neutralization (VN) test as well. In all 3-week-old piglets and in 80% of the 6-week-old animals the neutralizing antibody level reached the titre of 1:40. In none of the 10-week-old piglets did the titre exceed the value of 1:20, but in about 25% of the piglets it reached 1:20; the half of these piglets came from two litters. In none of the piglets did the antibody level reach the negative threshold in the ELISA test during the study. No significant differences were found between the udder sections in milk antibody level by ELISA.

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Acta Veterinaria Hungarica
Authors:
Š. Vilček
,
Jana Mojžišová
,
Viera Bajová
,
Š. Paulík
,
L. Strojný
,
B. Ďurkovič
, and
Vlasta Hipíková

A serological survey for bovine viral diarrhoea virus (BVDV) antibodies on a collection of 1295 serum samples obtained from 6-12 months old cattle originating from 45 farms in Slovakia was carried out. On 13 farms more than 90% of the examined animals were seropositive, on 14 farms 71-90% seroprevalence was observed, on 13 farms only 50-70% animals were found to be positive for BVDV antibodies, while the remaining 5 farms showed fewer than 50% seropositive animals. The average incidence of BVDV antibodies (around 70%) was similar as determined 30 years ago. Of 84 serum samples from seronegative animals originating from 14 farms in which 70-98% seropositivity was observed, six were positive in Ag-BVDV ELISA indicating persistently infected (PI) cattle. On a farm to which animals were imported from abroad, a BVD outbreak was observed. Of 110 animals tested, four were positive in Ag-ELISA indicating the presence of PI cattle on this farm. Genetic typing of two isolates from imported animals performed by RT-PCR (324/326 primers from 5´-UTR), sequencing of PCR products and computer-assisted phylogenetic analysis revealed that they belong to BVDV-1h group.

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Simple real-time PCR assay with one set of primer and probe for rapid, sensitive qualitative and quantitative detection of Entamoeba histolytica has been used. Consensus sequences were used to amplify a species-specific region of the 16S rRNA gene, and fluorescence resonance energy transfer hybridization probes were used for detection in a LightCycler platform (Roche). The anchor probe sequence was designed to be a perfect match for the 16S rRNA gene of Entamoeba species, while the acceptor probe sequence was designed for Entamoeba histolytica, which allowed differentiation. The performed characteristics of the real-time PCR assay were compared with ELISA antigen and microscopical detection from 77 samples of individuals with suspected clinical diagnosis of imported E. histolytica infection. Stool and liver abscess pus samples were examined with analytical sensitivity of 5 parasites per PCR reaction. The melting curve means Tms (standard deviation) in clinical isolates were 54°C. The real-time assay was 100% sensitive and specific for differentiation of Entamoeba histolytica, compared with conventional ELISA or microscopy. This real-time PCR assay with melting curve analysis is rapid, and specific for the detection and differentiation of Entamoeba histolytica. The suitability for routine use of this assay in clinical diagnostic laboratories is discussed.

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The aims of the present study were to determine (a) the effectiveness of an attenuated live Cryptobia salmositica vaccine; (b) the effects of food deprivation on the immune response and its duration in rainbow trout ( Oncorhynchus mykiss ) immunised with a live C. salmositica vaccine or with a killed Aeromonas salmonicida vaccine. The fish were divided into three groups (I, II and III; 14 fish per group), those in Groups I and II were under food deprivation (0.40% of body weight), while Group III fish were fed to satiety. The study showed that the attenuated strain of C. salmositica did not cause anaemia and disease, and the fish were protected from clinical disease when they were challenged with virulent parasites. Parasitaemia in all fish vaccinated and challenged with virulent C. salmositica fluctuated and was relatively low; however, fish in Group III had higher parasitaemia than those in Groups I and II between weeks 8 and 14. The numbers of activated neutrophils increased [nitroblue tetrazolium (NBT) assay] after immunisation with both Cryptobia and Aeromonas vaccines and they remained high throughout the experiment. Antibody production (ELISA values) increased after vaccination and were slightly higher in Group III. ELISA titres against A. salmonicida increased after vaccination and decreased after 5 weeks. The titres increased again after the vaccinated fish were given booster, and they were higher than those in the first vaccinated fish.

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Orvosi Hetilap
Authors:
Melinda Vanya
,
Károly Szili
,
Szilvia Zámolyi
,
Hajnalka Magyar
,
Gábor Kőrösi
,
Árpád Benkő
,
Anna Hornyák
,
Zsolt Hegedűs
, and
Lajos György

Absztrakt

A szerzők egy 46 éves fogva tartott férfi beteg esetét mutatják be, akinél Coxiella burnetii által okozott atípusos pneumoniát állapítottak meg. A mellkasröntgen-felvétel interstitialis pneumoniát mutatott ki. Western-blot és ELISA-tesztek Coxiella burnetii-antitestre pozitívak voltak. A beteg klinikai állapota 10 napos doxycyclin- és amoxicillin-, valamint B6-vitamin-kezelést követően javult. A szerzők kiemelik, hogy a Q-láz diagnózisának megállapítása specifikus antitestvizsgálaton alapul. Atípusos pneumonia esetén a hasonló tüneteket okozó egyéb baktérium, illetve vírusantitest-vizsgálatnak is fontos szerepe van. Orv. Hetil., 2015, 156(18), 741–743.

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Herpes simplex virus type 2 infection is a quite common but frequently asymptomatic, therefore undiagnosed condition. Genital HSV-2 infection may cause neonatal herpes, enhances HIV transmission and may play a role in infertility. To evaluate the prevalence of HSV-2 in Hungary we tested 2500 serum samples for the presence of anti-HSV-2 IgG by ELISA method. According to our results Hungary belongs to the low-infected countries, the HSV-2 seroprevalence grows with age and is significantly higher among women than in men. We also examined the serostatus of 512 pregnant women and 539 women attending infertility clinics. Results show that the HSV-2 prevalence is significantly higher among women attending infertility clinics and the seropositivity of pregnant women is similar to that of the general Hungarian women population with the same age.

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Abstract

Background: Amyotrophic lateral sclerosis (ALS) is a degenerative disease characterized by progressive depletion of motor neurons in brain, brain stem, and spinal cord, whereas Hirayama disease (HD) results from anterior horn cell loss in the lower cervical cord. On the other hand, matrix metalloproteinases (MMPs) are known to digest components of the extracellular matrix. Of these, MMP-9 is considered as a marker of neuroinflammation. Materials and methods: We have measured MMP-9 levels in the serum of 30 patients with ALS, 10 patients of HD, and 25 healthy controls using ELISA method. Results: MMP-9 levels were significantly elevated in the patients suffering from ALS (P < 0.01) and HD (P < 0.05). Furthermore, MMP-9 levels in ALS have a significant positive correlation (R 2 = 0.9) with the duration of the disease. Conclusions: These results suggest that neuroinflammation is an underlying component in ALS pathology and further opens up the search for suitable biomarkers.

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Magyarországra behurcolt trópusi arbovírusfertőzések 2016 és 2020 között

Imported tropical arbovirus infections in Hungary between 2016 and 2020

Orvosi Hetilap
Authors:
Orsolya Nagy
,
Anna Nagy
,
Szilvia Tóth
,
Anita Koroknai
, and
Mária Takács

Összefoglaló. Bevezetés: A Dengue-, Zika- és Chikungunya-vírus-fertőzések a trópusokról importált leggyakoribb arbovírusfertőzések. Földrajzi elterjedésük átfedő, közös vektoraik és hasonló tüneteik miatt szerológiai és molekuláris módszerek együttes alkalmazásán alapuló mikrobiológiai vizsgálatokkal különíthetők el megbízhatóan. Célkitűzés: Munkánk célja a 2016 és 2020 között endémiás területen járt, tünetes és tünetmentes utazók vizsgálata volt, minden esetben mindhárom vírusfertőzés irányában. A diagnosztikus tesztek során az alvadásgátolt teljes vér és vizelet bevonásával vizsgáltuk a vírus-RNS kimutathatóságának esélyét a különböző mintatípusokból. Módszer: Savópárminták szerológiai analízise során a Dengue-, Zika- és Chikungunya-vírus-specifikus ellenanyagválasz alakulását vizsgáltuk ELISA-módszerrel. Reaktív eredmények esetében a szerológiai keresztreakciók kizárására immunfluoreszcens és ELISA-technikán alapuló további vizsgálatokat végeztünk a hazai és az utazás során érintett területeken előforduló flavi- és alphavirusok irányában. Vérsavó-, alvadásgátolt teljes vér és vizeletmintákból reverztranszkripciót követő valós idejű polimeráz-láncreakcióval vírus-RNS-kimutatást végeztünk. Eredmények: Az 1037 vizsgált utazó közül 133 esetben kaptunk reaktív szerológiai és/vagy molekuláris eredményt. Az alvadásgátolt teljes vér mintából sikerült a legnagyobb arányban vírusnukleinsavat kimutatni mind a Dengue- és Zika-, mind a Chikungunya-vírus esetében. Megbeszélés: Endémiás területről hazatért utazók vizsgálatát a tünetek hasonlósága miatt mindhárom vírusfertőzés irányában együttesen indokolt elvégezni. A flavi- és alphavirusokra jellemző nagyfokú szerológiai keresztreaktivitás miatt a nukleinsav-kimutatás javíthatja a mikrobiológiai diagnosztika pontosságát. Következtetés: A három vírus mikrobiológiai diagnosztikáját segíti a korai mintavétel és a molekuláris vizsgálatok kiterjesztése további mintatípusokra: alvadásgátolt teljes vér és vizelet. A behurcolt vírusfertőzések azonosítása fokozott jelentőségű, mert az Európában is jelen lévő vektorszúnyogfajok felvetik az autochton átvitel lehetőségét. Orv Hetil. 2021; 162(50): 2000–2009.

Summary. Introduction: Dengue-, Zika- and Chikungunya infections are among the most frequently imported tropical arbovirus infections. Due to their shared endemic regions, vectors and similar clinical symptoms, differential diagnosis is based on serological and molecular analysis. Objective: The aim of our study was to identify the imported arbovirus infections of travellers between 2016 and 2020. Furthermore, to improve the diagnostic sensitivity, anticoagulated whole blood and urine samples were involved in molecular diagnosis. Method: Virus-specific antibody kinetics was tested in paired sera of patients by ELISA method. In case of reactive results, further serological analysis was performed using immunofluorescence assays and/or ELISA tests to exclude serological cross-reactions caused by other members of the flavi- and alphaviruses. Detection of viral RNA was attempted from serum, anticoagulated whole blood and urine specimens using reverse transcription and real-time polymerase chain reaction. Results: Out of the tested 1037 travellers, reactive serological and/or molecular results were obtained in 133 cases. Anticoagulated whole blood proved to be the most suitable specimen for viral RNA detection of the three viruses. Discussion: Parallel testing of Dengue-, Zika- and Chikungunya infections is recommended, as symptom-based differential diagnosis is challenging. Due to the characteristic serological cross-reactivity of flavi- and alphaviruses, microbiological diagnosis relies on both serological and molecular tests. Conclusion: Involving anticoagulated whole blood and urine samples into molecular analysis and early sample collection improve the sensitivity of microbiological diagnostics. Identification of imported tropical arbovirus infections is of high importance as the presence of vector mosquitos in Europe raises the possibility of autochthon transmission. Orv Hetil. 2021; 162(50): 2000–2009.

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In six healthy mares and 24 mares showing reproductive disorders swab samples were taken from the fossa clitoridis to isolate Taylorella equigenitalis, and from the uterus to isolate mycoplasmas, ureaplasmas and other aerobic bacteria. Swab samples were also taken from the uterus for Chlamydiaantigen ELISA and ChlamydiaPCR studies. The uterus of 27 mares was examined cytologically, and biopsy samples were taken from the endometrium for histological examinations and for immunohistochemical examinations aimed at the detection of chlamydiae. T. equigenitalis, mycoplasmas, ureaplasmas and chlamydiae could not be detected from any of the mares examined. Aerobic facultative pathogenic bacteria were isolated from mares with endometritis in four cases. In 18 out of 22 mares with endometritis (82%) no infective agents could be demonstrated. Further studies are needed to elucidate the relative importance of non-infectious causes of endometritis and of anaerobic bacteria often detectable in the uterus in the aetiology of the reproductive disorders observed.

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