scholarly journals Optimisation of phagocytosis assay in rainbow trout (Oncorhynchus mykiss)

Author(s):  
H Minarova ◽  
P Ondrackova ◽  
M Palikova ◽  
J Mares ◽  
J Blahova ◽  
...  

A phagocytosis assay is one of the most commonly used functional immunological methods. There are many possible ways of assessing leucocytes and their ability to ingest different particles. The aim of this study was to optimise the phagocytosis assay in rainbow trout (Oncorhynchus mykiss) using labelled zymosan particles (Alexa Fluor 488 and Texas Red conjugate). Whole blood was incubated with the particles under different conditions and leucocytes were subsequently isolated by haemolysis in a hypotonic environment. The effect of the different incubation time, temperature, blood volume and dilution on the phagocytic activity was evaluated by flow cytometry. Our experiments showed that the incubation for at least 2 h at 15 °C provided optimal results, while the  blood volume and dilution had no significant effect. The optimised assay will be used for the examination of fish health and in further experimental studies.

2006 ◽  
Vol 291 (3) ◽  
pp. R711-R718 ◽  
Author(s):  
Erik Sandblom ◽  
Michael Axelsson

Central venous blood pressure (Pven) increases in response to hypoxia in rainbow trout ( Oncorhynchus mykiss), but details on the control mechanisms of the venous vasculature during hypoxia have not been studied in fish. Basic cardiovascular variables including Pven, dorsal aortic blood pressure, cardiac output, and heart rate were monitored in vivo during normoxia and moderate hypoxia (PWO2 = ∼9 kPa), where PWO2 is water oxygen partial pressure. Venous capacitance curves for normoxia and hypoxia were constructed at 80–100, 90–110, and 100–120% of total blood volume by transiently (8 s) occluding the ventral aorta and measure Pven during circulatory arrest to estimate the mean circulatory filling pressure (MCFP). This allowed for estimates of hypoxia-induced changes in unstressed blood volume (USBV) and venous compliance. MCFP increased due to a decreased USBV at all blood volumes during hypoxia. These venous responses were blocked by α-adrenoceptor blockade with prazosin (1 mg/kg body mass). MCFP still increased during hypoxia after pretreatment with the adrenergic nerve-blocking agent bretylium (10 mg/kg body mass), but the decrease in USBV only persisted at 80–100% blood volume, whereas vascular capacitance decreased significantly at 90–110% blood volume. In all treatments, hypoxia typically reduced heart rate while cardiac output was maintained through a compensatory increase in stroke volume. Despite the markedly reduced response in venous capacitance after adrenergic blockade, Pven always increased in response to hypoxia. This study reveals that venous capacitance in rainbow trout is actively modulated in response to hypoxia by an α-adrenergic mechanism with both humoral and neural components.


1997 ◽  
Vol 54 (12) ◽  
pp. 2901-2906 ◽  
Author(s):  
Kari Koponen ◽  
Ossi Ritola ◽  
Sirpa Huuskonen ◽  
Dan Linder ◽  
Katalin Monostory ◽  
...  

We assessed the intrastrain and interstrain variability of cytochrome P450 enzyme activities and conjugation reactions of different rainbow trout (Oncorhynchus mykiss) strains. The study protocol was divided into two parts: in two separate experiments, monooxygenase (7-ethoxyresorufin O-deethylase, EROD) and conjugation (glutathione S-transferase, GST; UDP-glucuronosyl transferase, UDPGT) enzyme activities were measured from liver and kidney samples of commonly used rainbow trout breeds. In the preliminary study, the differences in the basal biotransformation enzyme activities were detected in two separately cultivated rainbow trout strains. Results indicated significant intrastrain (UDPGT, GST) and interstrain (EROD) differences in biotransformation enzyme activities. In the experimental study, corresponding enzyme activities were measured from three different rainbow trout strains exposed to beta -naphthoflavone (BNF), a polycyclic aromatic hydrocarbon (PAH) type model compound.The results revealed significant interstrain differences, especially in hepatic EROD and GST\b activities. However, UDPGT activities in the liver, as well as the measured monooxygenase and conjugation activities in the kidney, showed no notable variance between strains.The present work addresses intrastrain differences between hatcheries and highlights the need for intercalibration of rainbow trout strains used in experimental studies in aquatic toxicology.


Planta Medica ◽  
2012 ◽  
Vol 78 (11) ◽  
Author(s):  
A Ghasemi Pirbalouti ◽  
E Pirali ◽  
G Pishkar ◽  
S Mohammadali Jalali ◽  
M Reyesi ◽  
...  

2014 ◽  
Author(s):  
Silvia Gonzalez-Rojo ◽  
Cristina Fernandez-Diez ◽  
Marta Lombo ◽  
Vanesa Robles Rodriguez ◽  
Herraez Maria Paz

Sign in / Sign up

Export Citation Format

Share Document