scholarly journals Biochemical detection of E-ADA on Neospora caninum tachyzoites and the effects of a specific enzymatic inhibitor

2015 ◽  
pp. 4455-4460
Author(s):  
Alexandre A. Tonin ◽  
Aleksandro S. Da Silva ◽  
Victor C. Pimentel ◽  
Daniele Zanini ◽  
Maria Rosa C. Schetinger ◽  
...  
Keyword(s):  
Cell Culture ◽  
High Activity ◽  
Adenosine Deaminase ◽  
Protein Concentration ◽  
Negative Impact ◽  
Neospora Caninum ◽  
Free Form ◽  
Biochemical Detection ◽  
Caninum Tachyzoites ◽  
Enzymatic Inhibitor

Objective. This study aimed to investigate the presence and activity of the ecto adenosine deaminase (E-ADA) enzyme in tachyzoites of Neospora caninum (Nc-1 strain), as well as to assess the activity of a well-known E-ADA inhibitor, the deoxycoformycin. Materials and methods. The parasites were grown in cell culture, being subsequently separated in a pellet of tachyzoites, on which the E-ADA activity was tested using the concentrations 0 (control), 0.2, 0.4 and 0.8 mg mL-1. Results. The E-ADA showed high activity, progressively increasing its activity according to the enhancement of the protein concentration. The test was carried out with different concentrations of deoxycoformycin, showing that it was able to inhibit the E-ADA present on the free form of the parasite. Conclusions. Based on these results we conclude that the E-ADA is present on tachyzoites of N. caninum, and deoxycoformycin is able to inhibit this enzyme. In this sense, knowing the negative impact of N. caninum on reproductive issue in cattle (mainly abortion), might it is an alternative in order to deal with this parasitic infection.Key words: adenosine deaminase, deoxycoformycin, neosporosis (Source: CAB, MeSH).

scholarly journals PHYTOHAEMAGGLUTININ’S EFFECT ON NEOSPORA CANINUM ANTIGEN PRODUCTION

2013 ◽  
Vol 18 (1) ◽  
Author(s):  
Mariana Kikuti ◽  
Virgínia Bodelão Richini-Pereira ◽  
Rodrigo Costa Da Silva ◽  
Helio Langoni
Keyword(s):  
Cell Culture ◽  
Vero Cell ◽  
Real Time Pcr ◽  
Gold Standard ◽  
Peritoneal Fluid ◽  
Negative Impact ◽  
Neospora Caninum ◽  
Meriones Unguiculatus ◽  
Antigen Production

The parasite Neospora caninum affects mainly cattle and dogs. This study aimed to evaluate the effect of phytohaemagglutinin (PHE) in antigen production of N. caninum NC-1 strain in gerbils (Meriones unguiculatus) and in vitro. 20 gerbils were used, 10 inoculated intraperitoneally with 1 x 106 tachyzoites and 10 with 1 x 106 tachyzoites plus 300 µL/mL of PHE. 16 bottles of Vero cell culture were inoculated, 8 with 1.5 x 105 tachyzoites and 8 with 1.5 x 105 tachyzoites plus 30 µL/mL of PHE. Serology of gerbils was performed on day 0 and before euthanasia. Tachyzoites present in peritoneal fluid and cell culture bottles were quantified by Neubauer chamber and by real-time PCR (qPCR). PHE has not interfered in the production of tachyzoites of N. caninum in intraperitoneal inoculated gerbils and the effect of PHE in cell culture had a negative impact, considering the qPCR technique as the gold standard.

scholarly journals Entrapment of the Fastest Known Carbonic Anhydrase with Biomimetic Silica and Its Application for CO2 Sequestration

Polymers ◽  
2021 ◽  
Vol 13 (15) ◽  
pp. 2452
Author(s):  
Chia-Jung Hsieh ◽  
Ju-Chuan Cheng ◽  
Chia-Jung Hu ◽  
Chi-Yang Yu
Keyword(s):  
Carbonic Anhydrase ◽  
High Activity ◽  
Co2 Sequestration ◽  
Residual Activity ◽  
Entrapment Efficiency ◽  
Free Form ◽  
Uncatalyzed Reaction ◽  
The Stability ◽  
Time Required

Capturing and storing CO2 is of prime importance. The rate of CO2 sequestration is often limited by the hydration of CO2, which can be greatly accelerated by using carbonic anhydrase (CA, EC 4.2.1.1) as a catalyst. In order to improve the stability and reusability of CA, a silica-condensing peptide (R5) was fused with the fastest known CA from Sulfurihydrogenibium azorense (SazCA) to form R5-SazCA; the fusion protein successfully performed in vitro silicification. The entrapment efficiency reached 100% and the silicified form (R5-SazCA-SP) showed a high activity recovery of 91%. The residual activity of R5-SazCA-SP was two-fold higher than that of the free form when stored at 25 °C for 35 days; R5-SazCA-SP still retained 86% of its activity after 10 cycles of reuse. Comparing with an uncatalyzed reaction, the time required for the onset of CaCO3 formation was shortened by 43% and 33% with the addition of R5-SazCA and R5-SazCA-SP, respectively. R5-SazCA-SP shows great potential as a robust and efficient biocatalyst for CO2 sequestration because of its high activity, high stability, and reusability.

Neospora caninum (Protozoa: Apicomplexa) infections in rats

1990 ◽  
Vol 68 (7) ◽  
pp. 1595-1599 ◽  
Author(s):  
David S. Lindsay ◽  
J. P. Dubey
Keyword(s):  
Rattus Norvegicus ◽  
Neospora Caninum ◽  
Clinical Signs ◽  
Hepatic Necrosis ◽  
Female Rats ◽  
Methylprednisolone Acetate ◽  
Laboratory Rats ◽  
Experimental Inoculation ◽  
Caninum Tachyzoites ◽  
Subcutaneous Inoculation

The susceptibility of laboratory rats (Rattus norvegicus) to experimental inoculation with tachyzoites of Neospora caninum was examined. Groups of female rats were intramuscularly injected with 0, 2, or 4 mg of methylprednisolone acetate (MPA) 7 days prior to, and on the day of, subcutaneous inoculation with 0 or 1.5 × 105 tachyzoites. Clinical signs of disease or deaths did not occur in rats given nothing or only N. caninum tachyzoites. Rats given only 4 mg MPA failed to grow as well as rats given nothing or only N. caninum tachyzoites but were otherwise healthy. All of 20 rats given 4 mg MPA and tachyzoites died of hepatitis and pneumonia within 12 days postinoculation. Hepatic necrosis was the most striking lesion seen in these rats, and other milder lesions consisted of pneumonia, encephalitis, and myositis. The response of rats given 2 mg MPA and tachyzoites was less severe. Three of 20 rats died with encephalitis, myositis, hepatitis, and pancreatitis. Mild lesions, but no N. caninum tachyzoites, were seen in 3 of 14 rats inoculated only with tachyzoites. Rats given the 4 mg MPA treatment and inoculated with N. caninum tachyzoites appear to be suitable subjects for examining acute neosporosis and could be used in studies designed to examine treatment of acute disease.

scholarly journals Редокс-реакція насіння кукурудзи Оржиця 237 МВ на дію гербіцидів

2015 ◽  
pp. 50-53
Author(s):  
Ю. В. Лихолат ◽  
Г. С. Россихіна-Галича
Keyword(s):  
Superoxide Dismutase ◽  
Lipid Oxidation ◽  
High Activity ◽  
Negative Impact ◽  
Lipid Hydroperoxides ◽  
Antioxidant Protection ◽  
Hybrid Corn ◽  
Ripening Process ◽  
Protection Systems

Висвітлено вплив гербіцидних препаратів на нако-пичення ТБК-активних продуктів, гідропероксидівліпідів і активність супероксиддисмутази (СОД) тапероксидази (ПО) у насінні кукурудзи гібриду Оржиця237МВ. У достигаючому насінні кукурудзи, обробле-ної гербіцидами, виявлено інтенсифікацію перебігуреакцій окиснення ліпідів різної сили та функціону-вання компонентів антиоксидантного захисту (СОД,ПО). Уповільнення накопичення продуктів перокси-дації, відзначене у стиглому насінні, пов’язано з висо-кою активністю супероксиддисмутази, стабільнимфункціонуванням каталази та пероксидази. Наведеневище дає змогу дійти висновку, що за дії гербіцидів унасінні рослин кукурудзи гібриду Оржиця 237МВ упроцесі достигання підвищена здатність до належ-ної активації досліджених ферментних захисних сис-тем та їх функціонування на рівні, який забезпечив бивідновлення і підтримку гомеостазу. Встановлено,що найменша негативна дія спричинена гербіцидами«Харнес», «Майстер» і «Аденго». The paper deals with the effect of herbicidal agents on accumulation of TBA-active products, lipid hydroperoxides and activity of superoxide dismutase (SOD) and peroxidase (PO) in seeds of Orzhytsa 237MV hybrid corn. In the ripening corn seeds treated with herbicides, intensification of the course of reactions of lipid oxidation of the varying degree and functioning of antioxidant protection components (SOD, PО) has been revealed. Slowed accumulation of peroxidation products recorded in ripe seeds is connected with high activity of superoxide dismutase, and stable functioning of catalase and peroxidase. The above leads to the conclusion that the action of herbicides on seeds of plants of Orzhytsa 237MV hybrid corn in their ripening process promotes the ability to proper activation of studied enzymatic protection systems and functioning thereof at the level which would ensure the restoration and maintenance of homeostasis. It is found that Harness, Meister and Adengo herbicides have the least negative impact.

scholarly journals Two-dimensional gel electrophoresis and immunoblot analysis of Neospora caninum tachyzoites

2004 ◽  
Vol 5 (2) ◽  
pp. 139 ◽  
Author(s):  
Eung Goo Lee ◽  
Jae Hoon Kim ◽  
Yong Seung Shin ◽  
Gee Wook Shin ◽  
Yong Hwan Kim ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Neospora Caninum ◽  
Immunoblot Analysis ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Caninum Tachyzoites

scholarly journals Comparison of an ELISA assay for the detection of adhesive/invasive Neospora caninum tachyzoites

2014 ◽  
Vol 23 (1) ◽  
pp. 36-43 ◽  
Author(s):  
Luiz Miguel Pereira ◽  
Ana Patrícia Yatsuda
Keyword(s):  
Cell Culture ◽  
Neospora Caninum ◽  
Culture Cell ◽  
Quantitative Detection ◽  
Lacz Gene ◽  
Galactosidase Activity ◽  
Apicomplexan Parasites ◽  
Effects Of Temperature ◽  
Colorimetric Reaction

Neospora caninum belongs to the phylum Apicomplexa, the causative agent of neosporosis, which leads to economic impacts on cattle production. A common feature among apicomplexan parasites is the invasive process driven mostly by the parasite. As a first evaluation of candidate molecules that play a possible role by interfering in this invasive process, the in vitro invasion assay is a fast and direct way to screen future agonists or antagonists. This work involved the development of a new cell culture ELISA and transient β-galactosidase activity applied to the semi-quantitative detection of N. caninum in Vero cell culture. Cell culture ELISA is based on histochemistry and immunology, resulting in a colorimetric reaction. The β-galactosidase activity was obtained by the transient transfection of the lacZ gene under control of RPS13 promoter of N. caninum. These methods were used to evaluate the effects of temperature (37°C and 85°C) on the invasion and adhesion of tachyzoites. The three tested methods (real time PCR, β-galactosidase activity and ELISA) showed a similar pattern, indicating that different methods may be complementary.

Vaccination with gamma-Irradiated Neospora caninum Tachyzoites Protects Mice Against Acute Challenge with N. caninum

2006 ◽  
Vol 53 (2) ◽  
pp. 151-156 ◽  
Author(s):  
S. RAMAMOORTHY ◽  
D. S. LINDSAY ◽  
G. G. SCHURIG ◽  
S. M. BOYLE ◽  
R. B. DUNCAN ◽  
...  
Keyword(s):  
Neospora Caninum ◽  
Gamma Irradiated ◽  
Caninum Tachyzoites ◽  
Acute Challenge

scholarly journals Experimental Neosporosis in Pregnant Ewes and Their Offspring

1996 ◽  
Vol 33 (6) ◽  
pp. 647-655 ◽  
Author(s):  
M. M. McAllister ◽  
A. M. McGuire ◽  
W. R. Jolley ◽  
D. S. Lindsay ◽  
A. J. Trees ◽  
...  
Keyword(s):  
Neospora Caninum ◽  
Pathological Features ◽  
Clinically Normal ◽  
Naturally Occurring ◽  
Inoculum Dose ◽  
Bovine Neosporosis ◽  
Cerebral Necrosis ◽  
The Brain ◽  
Caninum Tachyzoites ◽  
Clinical And Pathological Features

Six groups of six pregnant ewes each were inoculated with 170,000 or 1,700,000 tachyzoites of Neospora caninum on gestation day 65, 90, or 120. All ewes seroconverted, and none showed signs of illness other than abortion. Regardless of the inoculum dose, all ewes inoculated on gestation day 65 aborted: ewes inoculated on gestation day 90 aborted, gave birth to weak lambs, or gave birth to clinically normal lambs; and all ewes inoculated on gestation day 120 gave birth to clinically normal lambs. Using an immunohistological procedure that stains bradyzoites, we observed protozoal cysts in brains of 11 of 29 (38%) aborted fetuses, in one of four (25%) weak lambs, and in seven of 18 (39%) clinically normal lambs. Cysts were not observed in extraneural tissues from two clinically normal lambs that had cysts in the brain. No evidence of infection was observed in tissues of five ewes examined using an immunohistological procedure that stains N. caninum tachyzoites and bradyzoites. Multifocal nonsuppurative encephalitis was observed in 46 of 51 (90%) aborted, weak, or clinically normal lambs. Cerebral necrosis, dystrophic mineralization, and meningitis were also commonly identified in live and aborted lambs (even when severely autolyzed). Nonsuppurative, necrotizing placentitis was observed in 15 of 17 (88%) placentas. Nonsuppurative myositis was common in fetuses but not in live lambs. Inflammation occurred less frequently in liver and lung. Clinical and pathological features of neosporosis in sheep closely resemble those of bovine neosporosis and ovine toxoplasmosis. Although abortion caused by naturally occurring neosporosis in sheep has not been reported, diagnosticians should carefully distinguish between neosporosis and toxoplasmosis in cases of ovine protozoal abortion unless future investigations exclude the likelihood of naturally acquired neosporosis in sheep.

Detection of surface-associated and intracellular glycoconjugates and glycoproteins in Neospora caninum tachyzoites

1999 ◽  
Vol 29 (10) ◽  
pp. 1597-1611 ◽  
Author(s):  
N Fuchs ◽  
K Ingold ◽  
S Sonda ◽  
P Bütikofer ◽  
A Hemphill
Keyword(s):  
Neospora Caninum ◽  
Caninum Tachyzoites
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