The in vitro hemolytic activity of erythrocytes from free-living mute swan (Cygnus olor) spread out in Gdynia and Sopot Coasts

• Autorzy: Tkachenko H. , Kurhaluk N. , Hetmanski T.

Warianty tytułu

PL

In vitro aktywność hemolityczna erytrocytów samców i samic ładędzia niemego (Cygnus olor) z wybrzeża Gdyni i Sopotu

• Języki publikacji: EN

Abstrakty

EN

The erythrocytes could be considered as a valuable model in studying the cytotoxic effect of environmental xenobiotics owing to its particular structure and the important components of their plasma membrane especially lipids and proteins. The aim of our study was to investigate the erythrocyte hemolytic activity of mute swan (Cygnus olor) from Baltic coastal region in two points, i.e. Sopot and Gdynia. These comparisons will show whether mute swans on the Baltic coastal regions have elevated levels of hemolysis plus gender variation in erythrocyte damage. Forty adult mute swans were captured from Sopot and Gdynia in February 2013. Collected birds were measured and weighted, and the blood was sampled. The birds were divided into 4 groups (after the 2nd/3rd year of life): males from Sopot (n = 10), females birds from Sopot (n = 10), males from Gdynia (n = 10), and females from Gdynia (n = 10). Blood samples were collected from the medial metatarsal vein in sodium citrate for erythrocyte sedimentation rate. Blood was stored in tubes and held on the ice until centrifugation. The plasma was removed. The erythrocyte suspension (one volume) was washed with five volumes of saline solution three times. The acid resistance of erythrocytes was induced by 0.1M HCl, while osmotic resistance was induced by incubation erythrocyte suspension with the different solution of urea and NaCl, as well as in solutions of different NaCl concentration (0.1-0.9%). The erythrocyte hemolysis was measured spectrophotometrically. Our study revealed that the highest level of the hemolysis percentage was observed among females from Gdynia, males from Sopot, then females from Sopot, and males from Gdynia. The significant differences between the hemolysis percentage among females from Gdynia and Sopot from 13.5 to 18.0 min after HCl-induced hemolysis was observed. The results also indicated significant differences between the hemolysis percentage among males from Gdynia and Sopot from 14.0 to 19.0 min. Hemolysis percentage showed to be increased not significantly among males from Gdynia with time from hemolysis initiation to 10.0 min but close to linearity with a value of hemolysis percentage among males from Sopot after 25 min of hemolysis initiation. The maximal level of hemolysis percentage was significantly higher among females from Gdynia (at 15.0 min) compared to females from Sopot (at 14.0 min). The maximal level of hemolysis percentage among males from Gdynia and Sopot was noted at 0.5 min after hemolysis initiation. The maximal level of hemolysis percentage was 4.40% and 4.11%, respectively. The highest percentage of hemolyzed erythrocytes after incubation with solutions of various urea concentration was observed both among males and females from Gdynia. The lowest percentage of hemolyzed erythrocytes was exhibited by the erythrocyte suspension of males from Sopot. Comparing the percentage of hemolyzed erythrocytes both among males and females from Sopot, the higher values of hemolyzed erythrocytes was noted among females. The highest percentage of hemolyzed erythrocytes after incubation with solutions of various NaCl concentration was observed among males and females from Sopot and further among males and females from Gdynia after incubation with 0.5-0.9% NaCl solution. Comparing the percentage of hemolyzed erythrocytes after incubition with 0.1-0.4% NaCl solution, the highest percentage of hemolyzed erythrocytes was observed among males and females from Gdynia and then among males and females from Sopot. Our study showed more individual variation, probably related to the impact of the environmental habitat and consumption of ingested prey items by mute swans. Of those males and females, females from Gdynia showed the highest level of hemolyzed erythrocytes.

PL

Erytrocyty można uznać za cenny model do badania cytotoksycznego działania środowiskowych ksenobiotyków ze względu na ich szczególną strukturę i ważne składniki ich błony komórkowej, zwłaszcza lipidy i białka. Celem naszych badań była ocena aktywności hemolitycznej erytrocytów łabędzia niemego (Cygnus olor) z regionu morza Bałtyckiego (plaże w Sopocie i Gdyni). Porównania te miały pokazać, czy populacja łabędzia niemego w nadbałtyckich regionach przybrzeżnych ma podwyższoną hemolizę erytrocytową oraz czy te zmiany zależą od płci ptaków. Czterdzieści dorosłych łabędzi niemych przebadano w Sopocie i Gdyni w lutym 2013 r. Ptaki podzielono na 4 grupy (po II/III roku życia): samce z Sopotu (n = 10), samice z Sopotu (n = 10), samce z Gdyni (n = 10) i samice z Gdyni (n = 10). Próbki krwi pobierano z przyśrodkowej żyły skoku w probówki z cytrynianem sodu. Krew przechowywano w probówkach i trzymano na lodzie aż do odwirowania. Osocze zostało usunięte. Zawiesinę erytrocytów (jedna objętość) przemyto pięcioma objętościami roztworu soli fizjologicznej trzykrotnie. Kwasową odporność erytrocytów indukowano 0,1M HCl, podczas gdy odporność osmotyczną indukowano przez inkubację zawiesiny erytrocytów w różnych roztworach mocznika i NaCl, a także w roztworach o różnym stężeniu NaCl (0,1-0,9%). Hemolizę erytrocytów mierzono spektrofotometrycznie. Najwyższy poziom hemolizy zanotowano u samic z Gdyni, samców z Sopotu, następnie samic z Sopotu i samców z Gdyni. Stwierdzono istotne różnice między odsetkiem hemolizy u samic z Gdyni i Sopotu z 13,5 do 18,0 min po hemolizie indukowanej HCl. Wyniki wskazują również na istotne różnice pomiędzy odsetkiem hemolizy wśród samców z Gdyni i Sopotu od 14,0 do 19,0 min indukowanej hemolizy. Procent hemolizy zwiększył się nieznacznie u samców z Gdyni w czasie od rozpoczęcia hemolizy do 10 min. Maksymalny odsetek hemolizy był istotnie wyższy wśród samic z Gdyni (po 15,0 min) w porównaniu z samicami z Sopotu (14,0 min). Maksymalny poziom hemolizy wśród samców z Gdyni i Sopotu zanotowano w 0,5 minuty po inicjacji hemolizy. Maksymalny poziom hemolizy wynosił odpowiednio 4,40% i 4,11%. Najwyższy odsetek hemolizowanych erytrocytów po inkubacji z roztworami o różnym stężeniu mocznika obserwowano zarówno u samców, jak i u samic z Gdyni. Najmniejszy odsetek hemolizowanych erytrocytów wykazywała zawiesina erytrocytów samców z Sopotu. Porównując odsetek hemolizowanych erytrocytów zarówno u samców, jak i samic z Sopotu, wyższe wartości hemolizowanych erytrocytów odnotowano wśród samic. Najwyższy odsetek hemolizowanych erytrocytów po inkubacji z roztworami o różnym stężeniu NaCl zaobserwowano u samców i samic z Sopotu, a następnie u samców i samic z Gdyni po inkubacji z 0,5-0,9% roztworem NaCl. Porównując odsetek hemolizowanych erytrocytów po inkubacji z 0,1-0,4% roztworem NaCl, najwyższy odsetek hemolizowanych erytrocytów zaobserwowano u samców i samic z Gdyni, a następnie u samców i samic z Sopotu. Nasze badanie wykazuje większą zmienność osobniczą, prawdopodobnie związaną z oddziaływaniem środowiska naturalnego i spożyciem połkniętych przedmiotów przez nieme łabędzie na plażach Trójmiasta. Spośród samic i samców łabędzi niemych samice z Gdyni wykazały najwyższy poziom hemolitycznych erytrocytów.

Słowa kluczowe

EN

Gdynia town; Sopot town; mute swan; Cygnus olor; erythrocyte; hemolysis; hemolytic activity

• Wydawca: -
• Czasopismo: Baltic Coastal Zone. Journal of Ecology and Protection of the Coastline
• Rocznik: 2018
• Tom: 22
• Opis fizyczny: p.73-90,fig.,ref.

Twórcy

autor

Tkachenko H.

• Institute of Biology and Environmental Protection, Pomeranian University in Slupsk, ul. Arciszewskiego 22B, 76-200 Slupsk, Poland

autor

Kurhaluk N.

• Institute of Biology and Environmental Protection, Pomeranian University in Slupsk, ul. Arciszewskiego 22B, 76-200 Slupsk, Poland

autor

Hetmanski T.

• Institute of Biology and Environmental Protection, Pomeranian University in Slupsk, ul. Arciszewskiego 22B, 76-200 Slupsk, Poland

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