U radu su istraživane vrijednosti osnovnih parametara crvene loze po starosnim kategorijama kod jedinki potočne mrene (Barbus balcanicus). Hematološki status predstavlja pouzdano sredstvo za utvrđivanje fiziološkog stanja organizma, a posredno i stanja životne sredine, jer njegovi parametri reaguju na promjene uslova sredine. Hematološki parametri su obuhvatali: broj eritrocita, koncentraciju hemoglobina, hematokrit, MCV, MCH i MCHC. Praćeni parametri su analizirani u toku dvije godine i kod populacija iz dvije rijeke (Suturlija i Jakotinska rijeka). Kod jedinki iz rijeke Suturlije u prvoj godini istraživanja bile su zastupljene jedinke starosti 1+ do 5+, dok je tokom druge godine uzorak bio predstavljen jedinkama starosti 1+ do 6+. Istovremeno su tokom 2004. godine kod riba iz Jakotinske rijeke bile zastupljene jedinke starosti 1+ do 4+, a u drugoj godini jedinke starosti 1+ do 5+. Analiza po starosnim kategorijama pokazuje postojanje značajnih razlika u vrijednostima pojedinih parametara, a razlika je izraženija kod jedinki potočne mrene iz rijeke Suturlije, kod kojih su razlike utvrđene u obje godine. Međutim, kod potočne mrene iz Jakotinske rijeke statistički značajne razlike između vrijednosti konstatovanih kod različitih starosnih klasa bile su statistički značajne samo u drugoj godini istraživanja

Bakrac-Beciraj, Azra

Cetkovic, Danijela

Dekic, Radoslav

Ivanc, Aleksandar

Lolic, Svjetlana

Maletin, Stevan

Obradovic, Sasa

English

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210 V INTERNATIONAL CONFERENCE “AQUACULTURE & FISHERY” - CONFERENCE PROCEEDINGSHeMatoloGical status of Different  aGe classes of barbus balcanicusRADOSLAV DEKIĆ1, ALEKSANDAR IVANC2, SVJETLANA LOLIĆ1, STEVAN MALETIN3, AZRA BAKRAČ-BEĆIRAJ4, SAŠA OBRADOVIĆ5, DANIJELA ĆET-KOVIĆ11Faculty of Natural Sciences and Mathematics, University Banja Luka Mladena Stojanovića 2, 78 000 Banja Luka, 2 Faculty of Biofarming, Megatrend University Belgrade, Maršala Tita 39, 24 300 Bačka Topola, 3 Faculty of Agriculture, University Novi Sad, 4 Biotehnical Faculty, University Bihać, Kulina bana 2, 77 000 Bihać, 5 Fac-ulty of economics and engineering management, Novi SadHEMATOLOŠKI STATUS RAZLIČITIH UZRASNIH KLASA  BARBuS BAlcANIcuSApstraktU radu su istraživane vrijednosti osnovnih parametara crvene loze po starosnim kate-gorijama kod jedinki potočne mrene (Barbus balcanicus). Hematološki status predstav-lja pouzdano sredstvo za utvrđivanje fiziološkog stanja organizma, a posredno i stanja životne sredine, jer njegovi parametri reaguju na promjene uslova sredine. Hematološki parametri su obuhvatali: broj eritrocita, koncentraciju hemoglobina, hematokrit, MCV, MCH i MCHC. Praćeni parametri su analizirani u toku dvije godine i kod populacija iz dvije rijeke (Suturlija i Jakotinska rijeka). Kod jedinki iz rijeke Suturlije u prvoj godini istraživanja bile su zastupljene jedinke starosti 1+ do 5+, dok je tokom druge godine uzorak bio predstavljen jedinkama starosti 1+ do 6+. Istovremeno su tokom 2004. go-dine kod riba iz Jakotinske rijeke bile zastupljene jedinke starosti 1+ do 4+, a u drugoj godini jedinke starosti 1+ do 5+. Analiza po starosnim kategorijama pokazuje posto-janje značajnih razlika u vrijednostima pojedinih parametara, a razlika je izraženija kod jedinki potočne mrene iz rijeke Suturlije, kod kojih su razlike utvrđene u obje godine. Međutim, kod potočne mrene iz Jakotinske rijeke statistički značajne razlike između vrijednosti konstatovanih kod različitih starosnih klasa bile su statistički značajne samo u drugoj godini istraživanja.Ključne riječi: hematologija, potočna mrena, starosne klaseV MEĐUNARODNA KONFERENCIJA ”AKVAKULTURA I RIBARSTVO” - ZBORNIK PREDAVANJA   211INTRODUcTIONHematological status represents reliable means for determining the physiological state of an organism and indirectly the state of its environment, because hematological parameters respond to environmental changes (Ivanc and Miljanović, 2001). The quan-titative characters of red and white blood cells are used for detecting the haematological status. They provide insight in a number of processes in the organism. On this bases, analyses of different blood components give valuable information concerning changes that appear in certain systems influenced by external and internal factors. Determination of hematological parameters and blood plasma biochemistry is used to assess the health of wild and domestic animals. Values of these parameters are useful for interpretation of the results that are related to different diseases and environmental conditions (Seker et al., 2005). Prerequisite for such hematology application is satisfactory knowledge of reference range of values of fish blood parameters. Age-related values of hematological parameters are even less known and insufficient (Hrubec et al., 2001).MATERIAL AND METHODS Fish sampling was done by electro-shocker providing pulsed direct current and with ability to customise the output voltage. IG 600 brand,  with 1.2 KW power. After appear-ance of galvanotaxis and galvano-narcosis, fish were collected with  landing net. Fish sampling was made during two years in monthly intervals. Handbooks from Vuković and Ivanović (1971), Simonović (2001), Banaresku and Bogutavskaya (2003), Kottelat and Freyhof (2007) were used for taxonomic determination of collected fish. The fish were placed into containers with sufficient water of appropriate quality. After that were transferred in keepnets, placed in native stream and let to reanimate for one hour. River Suturlija, by its course and its basin is located in the area southwest of Banja Luka, and its mouth into Vrbas river, as a left tributary, is placed in Srpske Toplice (Gornji Šeher), at an altitude of 159 m. River Jakotina is a left tributary of river Vrbanja with length of 15 km, witch originates at an altitude of 670 m, with mouth on 260 m.Hematological analysisBlood for hematological analysis was collected by heart puncture using sterile nee-dle (1.0 to 1.2 mm). Native blood, with anticoagulant is used for further analyses. Eryth-rocyte and leukocyte count was performed in hemocytometer using diluent by  Kekić and Ivanc (1982). Hemoglobin concentration (Hb) was determined by hemiglobin cya-nide method using Drabkin reagent (Blaxhall and Daisly, 1973). Hematocrit (Hct) was determined by microhematocrit centrifuge. Hematological parameters were calculated using values of hematocrit, erythrocyte number and hemoglobin concentration. Mean corpuscular volume (MCV)  Mean corpuscular hemoglobin (MCH) Mean corpuscular hemoglobin concentration (MCHC)  212 V INTERNATIONAL CONFERENCE “AQUACULTURE & FISHERY” - CONFERENCE PROCEEDINGSAge determinationMethods of scales observation and counting annual growth marks were used for age determination. Scales for age determination were collected from the shoulder area, above lateral line, and then, were placed in paper bags. Before age determination, scales were cleaned with NaCl and KOH solutions and water, and then were observed using binocular magnifier.RESULTS AND DIScUSSIONValues of monitored hematological parameters were correlated with age groups. Mean values of erythrocyte parameters for different age classes, rivers and years of researches are shown in table 1. Blood values of Large-spot barbel (Barbus balcanicus) from River Suturlija in 2004. showed significant age-related differences. Erythrocyte number was the highest in age class 5+, and the lowest in class 2+. Significant differ-ence was recorded between values of age classes 2+ and 4+ (0.021), 2+ and  5+ (0.020), as well as between 1+ and 5+ (0.029). The highest value of hemoglobin concentration was recorded in age class 4+, and the lowest in 1+ class.Table 1. Mean values of erythrocyte parameters of Barbus balcanicus from Rivers Su-turlija and Jakotina by age classes and years of researchRiverYearAge classMean valueErythrocyte numberx 1012/lHb(g/l)Hct(l/l)MCV(fl)MCH(pg)MCHC(g/l eryt.)Suturlija20041+x1.254 69.02 0.398 318.750 55.241 175.2722+ 1.246 72.13 0.422 341.563 58.393 172.8253+ 1.294 71.64 0.431 333.855 55.609 167.7684+ 1.310 73.61 0.411 315.757 56.490 180.7975+ 1.390 69.44 0.416 301.609 50.139 169.56220051+x1.270 67.64 0.389 308.489 53.603 175.5282+ 1.300 70.44 0.417 323.889 54.652 170.7583+ 1.297 74.26 0.442 342.574 57.791 172.1044+ 1.366 74.49 0.445 329.548 55.159 169.0745+ 1.343 75.34 0.438 329.086 56.846 173.6606+ 1.430 80.56 0.440 322.938 59.423 187.237Jakotina20041+x1.225 73.25 0.443 362.869 60.329 168.1502+ 1.207 74.75 0.451 374.768 62.316 168.4433+ 1.250 76.30 0.451 376.408 63.896 170.7514+ 1.355 77.77 0.457 345.420 58.155 171.69520051+x1.258 72.49 0.437 349.536 58.081 168.9892+ 1.240 75.10 0.453 375.276 61.671 168.1313+ 1.273 75.87 0.466 371.632 60.594 165.2234+ 1.282 73.56 0.451 360.319 58.366 164.9545+ 1.269 71.91 0.450 358.376 57.126 161.5496+ 1.245 70.37 0.317 256.317 56.585 223.562Significant differences were established comparing values of class 1+ with values of age classes 2+ (0.014) and  4+ (0.013). Hematocrit values also were related with fish age. Significant differences in hematocrit values existed between class 1+ and 2+ (0.005) and 1+ and 3+ class (0.004). The highest value was in age class 3+, and the low-est in 1+. Values of mean corpuscular volume (MCV) ranged from 301.608 fl (age class V MEĐUNARODNA KONFERENCIJA ”AKVAKULTURA I RIBARSTVO” - ZBORNIK PREDAVANJA   2135+) to 341.563 fl (class 2+). Significant differences were between age class 2+ and 1+ (0.003) and 2+ and 4+(0.014). Values of MCH showed significant differences by com-paring class 2+ with class 1+ (0.007) and 5+ (0.022). The highest value of this parameter was in age class 2+, and the lowest in 5+. In the same time, significant difference was recorded in MCHC values between age classes 3+ and 4+ (0.039).In 2005 in research of Barbus balcanicus from River Suturlija, the individuals from age 1+ to 6+ were found. Analysis by age classes showed presence of significant dif-ference in most of monitored parameters. The values of erythrocyte number ranged from 1.270 x 1012/l (in age class 1+) to 1.430 x 1012/l (class 6+). Relation of this param-eter with fish age showed significant difference between age class 1+ on one side, and classes 4+ (0.000), 5+ (0.035) and 6+ (0.016) on the other side. Comparing values of class 2+ with values of 4+ (0.010) and 6+ (0.050). Differences were significant between values from class 3+  and 4+ (0.006) and 3+ and 6+ (0.044). Hemoglobin concentration was constantly increasing from classes 1+ to 6+. Significant differences were recorded comparing values from age class 1+ with classes 3+ (0.000), 4+ (0.000), 5+ (0.001) and 6+ (0.001). Values of  class 2+ were significantly different from those in classes 3+ (0.012), 4+ (0.013), 5+ (0.024) and 6+ (0.017). The lowest hematocrit value was in age class 1+, and the highest in class 4+. Values from age class 1+ were significantly different from those in classes 2+ (0.015), 3+ (0.000), 4+ (0.000) and 5+ (0.003). Also, statistically significant difference was recorded comparing values from age class 2+ with values from classes 3+(0.030) and 4+ (0.021). Age class 3+ had signifaicantly higher values than class 1+ (0.002). This is also true rom MCH (0.013). MCHC values did not show significant difference by age classes. Correlation between hematological parameters and age was less expressed in barbel from River Jakotina.In 2004, significant differences between age classes were  not recorded. However, it should be pointed out that the highest values of erythrocyte number, hemoglobin concentration, hematocrit and MCHC were recorded in age class 4+, while the highest values of MCV and MCH were recorded in age class 3+. Lack of significant differen-cess in 2004 could be explained by different sample sizes. So, more than 50% of total sample belonged to age class 1+, followed by age class 2+, while the rest of age classes were presented by fewer individuals.  Slightly different correlation between hemato-logical parameters from barbel inhabiting River Suturlija was recorded in 2005. During that year, significant differences of hemoglobin concentration values were determined between age class 1+ on one side, and age classes 2+ (0.044) and 3+ (0.044) on the other side. The highest value of this parameter was in age class 3+, and the lowest in class 6+. The lowest hematocrit value was recorded in age class 6+, and it was significantly different from age classes 1+ (0.007), 2+ (0.002), 3+ (0.001), 4+ (0.003) and 5+ (0.005). Statistically significant difference was also recorded between age classes 1+ and 3+ (0.037).Erythrocyte number did not differ significantly with age, while MCV values were different between age class 6+ (in which were recorded the lowest values) and classes 2+ (0.020), 3+ (0.027) and 4+ (0.046). Also, significant difference was recorded be-tween age classes 1+ and 2+ (0.036). MCH differed significantly between classes 1+ and 2+ (0.030). MCHC values had  highest values in class 6+ which was significantly different from classes 1+ (0.002), 2+ (0.002), 3+ (0.001), 4+ (0.001) and 5+ (0.001).The other researches also showed presence of significant differences in values of hematological parameters between different age classes. Age-related analysis of blood parameters of hybrid Morone chrysops and Morone saxatilis showed presence of differ-214 V INTERNATIONAL CONFERENCE “AQUACULTURE & FISHERY” - CONFERENCE PROCEEDINGSence. In the research, the individuals from 4 to 19 month of age were presented, divided into age classes: 4 months, 6 months, 9 months, 15 months and 19 months. Values of hematocrit and MCV were significantly lower in the youngest individuals in compari-son with other age classes. Individuals aged 6 months had significantly lower values of hemoglobin concentration, MCH and MCHC in comparison with other age groups. The values of erythrocyte number were the highest in individuals aged 9 and 15 months (Hrubec et al., 2001). Also, research regarding to correlation between hematological parameters and fish biology in Silurus triostegus showed that hemoglobin concentra-tion and hematocrit values were growing with age, until the age of four, and then were decreasing. Research in correlation between hemoglobin concentration and hematocrit value showed high correlation coefficient (Abood, 1992). According to Chanchal et al., (Chanchal et al., 1979), young fishes usually show high physical activity and intense feeding during growth period which explains high recorded values of hemoglobin concentration and hematocrit. However, the older fishes show slow metabolic activity after certain age (Joshi and Tandon, 1977). Research of erythrocyte parameters in species Rhamdia quelen showed that hematological param-eters were also correlated with body length. Individuals of this species were divided in two groups, based on difference in standard body length. Standard body length values of fishes from the first group were ranged from 20.5 to 30.0 cm, and the second group from 30.5 to 40.0 cm. Analysis of hematological parameters showed significantly high-er erythrocyte number in the first group, while the values of hemoglobin concentration, hematocrit and MCHC were approximately the same in the both groups. Higher values of MCV and MCH were recorded in fishes from the second group (Camargo and Pouey, 2004).Results of haematological parameters in Capoeta capoeta umbla showed increased values of erythrocyte number, hemoglobin concentration and hematocrit with age (Örün and Erdemli, 2003).Based on all presented hematological parameters it can be noted that they are differ-ent in different age classes, but specific for every fish species.cONcLUSIONSErythrocyte parameters of Large-spot barbel (Barbus balcanicus) from researched rivers show significant difference with age.Major differences of monitored parameters correlated with age groups were recorded in individuals from River Suturlija.Differences were not recorded between barbel individuals from River Jakotina in the first year of research, while they were recorded in the second year.It can be noted that all presented haematological parameters are different for differ-ent age classes, but specific for every fish species.REFERENcESAbood, A.Y.A.L. (1992): Studies on the relationship between some haematological parameters and the biology of the fish, Silurus triostegus. Acta Ichthyologica et Pisca-toria. XXII, 2: 197- 201.Banaresku, P. (1964): Pisces-Osteichtyes Fauna R. P. Romane, 23. Edit Acad., Bu-curesti, 962 pp. u The Freshwater Fishes of Europe, Vol. 5/II, Cyprinidae 2, Part II: V MEĐUNARODNA KONFERENCIJA ”AKVAKULTURA I RIBARSTVO” - ZBORNIK PREDAVANJA   215Barbus, 2003.Blaxhall, P. C. and Daisly, K. W. (1973): Routine hematological methods for use With fish blond. J. Fish. biol., 5: 771-781.Camargo, S. O. and Pouey, J. F. (2004): Hematological parameters of jundia (Rham-dia quelen) originating from the natural environment. Sixth International Congress on the Biology of  Fish, Manaus Brasil, 2004.Hrubec, T.C., Smith, S.A. and Robjertson, J.L. (2001): Age-Related Changes in He-matology and Plasma Chemistry Values of Hybrid Striped Bass (Morone chrysops x Morone saxatilis). Veterinary Clinical Pathology, 30, 1: 8-15.Kekić, H. and Ivanc, A. (1982): A new direct method for counting fish blood cells. Ichtyologia, 14, 1 : 55.Kottelat, M. J. Freyhof (2007): Handbook of European freshwater fishes. Kottelat, Cornol.Switzerland and Freyhof, Berlin, GermanyÖrün, I. and A. Erdemli, Ü (2003): A Study on Blood Parameters of Capoeta capoeta umbla (Heckel, 1843) Captured From Karakaya Dam LakeSeker, Y., Karatas, M. and Sezer, M. (2005): Determination of Biochemical Para-meter Values of Chub (Leuciscus cephalus) Population in Almus Dam Lake, Turkey. Journal of Animal Andveterinary Advanses 4, 11: 927 – 929. Simonović, P. (2001): Ribe Srbije, Beograd.Vuković,T., Ivanović, B. (1971): Slatkovodne ribe Jugoslavije, Zemaljski muzej BiH, Sarajevo.

Hematološki status različitih uzrasnih klasa barbus balcanicus

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Hematološki status različitih uzrasnih klasa barbus balcanicus

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