Capoeta Damascina Research Articles

Ecological niche overlap from occurrence of two cyprinid sympatric species, Paracapoeta trutta and Capoeta damascina in the Sirvan River, Persian Gulf Basin

Ecological niche overlap from occurrence of two cyprinid sympatric species, Paracapoeta trutta and Capoeta damascina in the Sirvan River, Persian Gulf Basin

Antropojenik aktivitelerden etkilenen nehirden toplanan Cyprinidae familyasına ait balık türlerinin gonad sağlığı durumları

Many freshwater ecosystems are contaminated with heavy metals released by municipal wastewater, cultivation and factory wastewater. The aim of the study was to evaluate the negative impact of metal pollution on the fish reproductive system. It was performed using the gonadal histopathology, hepatosomatic index (HSI), and gonadosomatic index (GSI) of three fish species, Capoeta damascina, Squalius semae and Alburnus sellal inhabiting in Karasu River (Erzurum, Turkey) contaminated by various anthropogenic activities. The highest GSI values were determined for each sex according to the seasons, and lower GSI values were observed in polluted stations in all three species. It was observed that HSI values in fish varied significantly from station to station. In addition, an increase in ovarian and testicular lesions (degeneration in oocyte and spermatocytes, atresia, congestion, infiltration, edema, vascular hypertrophy, fibrosis) was detected in the polluted areas. The results clearly showed that the metal pollution in the river adversely affected the reproductive system of the fish species living in the river.

Hydrological alteration and biodiversity change along the river network caused by anthropogenic activities and climate variability

BackgroundPopulation growth and intensified human activities in conjunction with climate variability continue to affect the hydrologic cycle, aquatic and terrestrial flora and fauna. In this regard, understanding interactions among ecosystem functions, impacts of anthropogenic interventions and those of climate variability is vital for projecting future ecosystem responses to human activities and climate forcing. The objectives of this study are to determine the ecological flow state via eco-flow index based on discharge hydrograph, to model the ecological diversity through the Shannon diversity index, and to assess the degree of hydrologic alteration using indicators of hydrologic alteration and range of variability approach in six hydrometric stations along the Zayandeh-Rud River in central Iran. The river drains into Gavkhuni Marsh. Also, the streamflow-induced potential changes for Capoeta damascina (a cyprinid fish species of the genus Capoeta), Petroleuciscus esfahani (a small cyprinid fish) and Aphanius isfahanensis (a Cyprinodontid fish) are evaluated. The outcome is expected to assist managers with understanding the effects of anthropogenic activities and climate variability on Gavkhuni aquatic ecosystems so that management options that enhance species resilience and adaptability are outlined.ResultsHuman activities, a primary factor influencing the natural flow regime, caused a significant increase in the minimum flow, July to March streamflow, low pulse number, and the number of reversals in most studied stations. On the contrary, some other hydrologic indices declined in value. Reservoir impoundment, the most prominent factor among human interventions, resulted in an overall alteration degree of 74.8% in streamflow. Climate variability impacted the natural flow regime in the range of low degree hydrologic alteration (27.3%). In addition, the biodiversity of the study basin, as modeled by the Shannon diversity index, had strong relevance to the annual eco-surplus and was more sensitive to summer floods and autumn hydrological droughts than other factors.ConclusionsThis study corroborates the effectiveness of scenario-based hydrological modeling framework in evaluating the impacts of climate variability and human activities imposed on natural flow metrics. Additionally, the recently introduced eco-flow metrics based on discharge hydrographs and the Shannon diversity index based on indicators of hydrological alteration may be adopted in basins lacking ecological data. These two indices can effectively identify the most prominent factors in hydrological alteration and biodiversity change through a river network and may provide scientific decision-making support for water resource management in the study area.

Capoeta svanetica (Teleostei, Cyprinidae), a New Species from the Luchunis River (Rioni River Drainage) in Georgia

Recent Georgia’s fish fauna includes four Capoeta species: C. capoeta (Kura River basin), C. kaput (Araxes River and its left tributaries, the Akhuryan and Mezamor rivers), C. sieboldii (East and South–East Black Sea river basins), and C. banarescuii (Chorokh River basin). Based on morphological data analysis new species of the genus Capoeta — C. svanetica sp. n. is described. It is more similar to the Capoeta damascina complex group (Anatolian–Iranian group) which also includes C. banarescui, C. baliki, and C. sieboldii spread at East and South-East Black Sea coastal rivers. The Anatolian–Iranian group, also known as small-scales species group well distinguished from Aral-Caspian species (C. capoeta and C. kaput in Georgia) by highest scales number in lateral line. C. svanetica sp. n. is distinguished from other Capoeta species of Black Sea basin rivers in Georgia and adjacent waters by combination of characters: two pairs of barbels are present (C. sieboldii and C. oguzelii have one barbels pair); no spots on the body (C. oguzelii has small black spots); lower lip has keratinised edge without fringe (C. sieboldii and C. oguzelii are characterized by fringed lips); scales small, 70–74 total lateral line scales (in C. sieboldii only 52–60 scales); 10–12 scale rows above lateral line (C. banarescui, C. baliki and C. oguzelii have more than 12 scale rows) and 7–8 scale rows below lateral line (C. baliki and C. oguzelii have more than 10 scale rows); 12–15 gill rakers on the first gill arch (C. sieboldii and C. baliki have more then 16); last unbranched fin ray soft, serrae number on it is 7–9 in adult and 0 in juvenile samples (all other Capoeta species of Black Sea basin rivers have well ossified last unbranched ray with high serrae number); anal fin base length is only 7.4–9.4 % of SL (more shorter than C. banarescui, C. baliki and also C. tinca); as well as mouth width (25.4–29.4 % of HL); eye diameter (19.0–28.7 % of HL) is biggest than same in C. banarescui, C. baliki and also C. tinca.

Kuzgun Baraj Gölü ve Drenajlarının Balık Faunası (Erzurum, Türkiye)

This study was carried out to determine the fish fauna of Kuzgun Dam Lake dranaige. Eight (Barbus lacerta, Capoeta damascina, Alburnus sellal, Alburnoides velioglui, Squalius semae, Oxynoemacheilus araxensis, O. bergianus and Salmo euphrataeus) fish species belonging to 4 families (Cyprinidae, Leuciscidae, Nemachelidae and Salmonidae) were determined.

A comparison of the metals and metalloid levels in wild and cultured Capoeta damascina fish and assessment of its potential health risks to humans in Iran

This research aimed to assess metals/metalloids (Fe, Cr, Cu, Mn, Mo, Se, Zn, Ni, Sr, Ba, Co, Hg, Pb, As, and Cd) in liver and muscle tissue of wild and culture Capoeta damascina fish using (ICP/OES). The healthy assessment revealed that the accumulation of Hg in adults and children consuming 7 meals/week might harm their health. However, the high Se/Hg ratio in the fish alleviates the adverse effects of Hg. Furthermore, the carcinogenic risk of As and Ni accumulation should be considered for adults having 7 meals/week or more. It is recommended to continually monitor these metals and metalloids in the fish and cultured fish feed.

Three new species of Neoechinorhynchus (Acanthocephala: Neoechinorhynchidae) from fresh-water fish collected from Tigris River, Iraq.

Three new species of Neoechinorhynchus are described from freshwater fish of Tigris River, at Salah Al-Din province, mid Iraq, namely Neoechinorhynchus tigrisensis n. sp. from Shilik Leuciscus vorax (Heckel) and Brond-snout Chondrostoma regium (Heckel), N. planilizai n. sp. from Abu mullet Planiliza abu (Heckel) and Tigris catfish Silurus triostegus (Heckel) and N. barbi n. sp. from Mesopotamian barb Capoeta damascina (Valenciennes). The first two new species shared the para-receptacle structure (PRS) with seven nominal valid species of the genus Neoechinorhynchus but they differed in other taxonomic traits, while the third new species shared the vaginal vestibular muscles (para vaginal patch) at the end of female trunk with N. spiramuscularis Amin, Heckmann & Ha, 2014 and N. zabensis Amin, Abdullah & Mhaisen, 2003 but differed for further morphological features. Proboscis hooks of middle and posterior circles in N. barbi n. sp. are remarkably bigger than in the other two new species (N. tigrisensis and N. planilizai) while the hooks in first circle is almost similar in the three species. Furthermore, N. tigrisensis n. sp. and N. planilizai n. sp. differ from some marine species by having smallest hooks size at anterior circle, with different shape and size of hooks. The three new species reported in the present study are compared with other eight species of Neoechinorhynchus reported so far from fish in Iraq. This work contributes to increase the knowledge on the biodiversity of fish parasites all over the world, especially for acanthocephalans.

Survey of Fish Fauna from Five Stations in Duhok Province, Kurdistan Region, Iraq

A survey of fish species was carried out from five stations in Duhok province during the period from August 2016 to September 2017. A total of 517 freshwater fishes, belong to 19 species, namely: Acanthobrama marmid, Arabibarbus grypus, Capoeta damascina, C. trutta, Carasobarbus luteus, Carassius carassius, Chondrostoma regium, Cyprinion kais, C. macrostomum, Cyprinus carpio, Garra rufa, Leuciscus vorax, Luciobarbus barbulus, L. esocinus, L. xanthopterus, Mastacembelus mastacembelus, Mesopotamichthys sharpeyi, Planiliza abu and Silurus triostegus were recorded. The survey showed that the abundant fish at all five stations was C. carassius which is one of the nuisance fish species while the little abundance was recorded for C. luteus.

Niğde İli (Türkiye) İhtiyofaunası

Bu çalışma Mayıs 2016-Temmuz 2017 tarihleri arasında Niğde İli tatlı su balık faunasını belirlemek amacıyla yapılmıştır. Niğde il sınırları içerisinde yapılan arazi çalışmaları sonucunda iki takımdan beş familyaya mensup olmak üzere 12 tür tespit edilmiştir; Capoeta pestai, Capoeta baliki, Capoeta damascina, Cyprinus carpio, Gobio gymnostethus, Pseudophoxinus battalgilae, Salmo labecula, Squalius cappadocicus, Oncorhynchus mykiss, Oxynoemacheilus ciceki, Oxynoemacheilus cilicicus ve Oxynoemacheilus eregliensis. Bu türlerden dokuz tanesi bulundukları havzalara endemik türlerdir. C. baliki, C. pestai, O. ciceki ve O. cilicicus türleri ise il için yeni kayıt niteliği taşımaktadır.

A comparison between sedation using propofol and clove oil in Levantine scraper (Capoeta damascina)

Anesthetic agents are used in fish to decrease stress and mortality during sorting, surgery, diagnosis and artificial fertilization procedure. This study was done to evaluate and compare the efficacy of propofol and clove oil for anesthetizing Capoeta damascina. For this purpose, 40 fish were caught from Arvand River and divided into two groups. After a pilot study, propofol and clove oil were examined in 12.5 mg/L and 125 mg/L concentrations, respectively. The onset of anesthesia, duration period of anesthesia and recovery time were recorded separately for each group and compared. Results showed that the anesthesia onset of specimens in propofol treatment, was significantly shorter than that of clove oil treatment (p <0.05). Also, recovery time in propofol treatment was longer (p <0.05) than that of clove oil. Based on the results, it was recommended to use propofol instead of clove oil in performing anesthesia for this species due to rapid induction, longer duration and more prolonged effect in immersion method.

Efficiency and Suitability of the Fish Passages of River Ceyhan, Turkey

The efficiencies of the two fish passages of River Ceyhan (Turkey) were investigated by using trap catches, external tags (T-bar anchor tags), PIT telemetry and radio telemetry. According to the results of the study, the pool and weir type fish passage were not effective; no fish entered the trap in this fish passage. Further, it was observed that none of the 50 external tagged fishes, 44 PIT-tagged fishes, and 47 radio-tagged fishes passed through this fish passage. A total of 900 Capoeta damascina and 520 Alburnus adanensis entered the trap installed in the vertical slot fish passage. A relationship was found between the gender ratio of the migratory C. damascina and their migration times. Fish passage performance was different according to the applications. These differences in fish passage success among the applications originated from the sizes of the tagged fish. Because the sizes of the radio-tagged fish were much larger than that of the external tagged fish and PIT-tagged fish so the success of the fish passage according to the radio telemetry was estimated higher than that of the other applications. The further passing ratio of the fish over 20 cm in length was higher than that of the small individuals.

Osteological Characters to Define Six Species of the Capoeta damascina Species Complex (Cypriniformes: Cyprinidae)

This paper presents the results of a comparative study of eight osteological characters: the counts, dimension and regionalization of the vertebral column, the pattern of interdigitation of the dorsal- and anal-fin pterygiophores with the neural and haemal spines of the vertebrae, the distribution of the dorsal- and ventral-procurrent caudal-fin rays, the distribution of the principal caudal-fin rays, and the morphology of the caudal-fin skeleton of six species of the Capoeta damascina complex (Cypriniformes: Cyprinidae). Potential taxonomically valuable osteological characters are described and used to separate the six species of Capoeta studied. Formulae for the structure of the vertebral column, the dorsal- and anal-fin pterygiophores’ interdigitation with the neural and haemal spines of the vertebrae, the distribution of the dorsal and ventral procurrent caudal-fin rays, and the distribution of the principal caudal-fin rays are developed. These morphological descriptive parameters express a morphotype that may be linked with the cypriniform mode of swimming of the Capoeta species investigated. The morphological study of the vertebral column of six species of the Capoeta damascina complex enables the division of this bony structure into five or six morphologically distinct regions, depending on the species, and the drawing of characteristic vertebral profiles. This regionalization is more complicated than the classical division in abdominal and caudal parts only.

Embryogenesis and early larval development in wild-caught Levantine scraper, Capoeta damascina (Valenciennes, 1842).

Levantine scraper, Capoeta damascina is a candidate species for future stock assessments, conservation studies, and hatchery efforts. Herein, we documented embryonic and early larval development, from egg activation to the exogenous feeding period, using morphological and histological landmarks. Embryos were obtained by in vitro fertilization from hormonally induced wild-caught broodstock, and subsequent development was monitored at temperatures coinciding with native conditions. Embryonic development from fertilization to hatch lasted ~105-110 hr. Larvae emerged with unpigmented eyes and body morphology, as well as an undifferentiated digestive tract. The mouth was closed at hatch by the oropharyngeal membrane and opened by the early endogenous feeding period. Trabeculae cartilage, quadrate bone, and Meckel's cartilage of the endoskeleton were present during the endogenous feeding period. During this period, the larvae underwent considerable changes in craniofacial morphology, locomotion, and organogenesis of the digestive tract. The cartilaginous floor of the neurocranium developed and the first four ceratobranchials appeared simultaneously at the end of endogenous feeding period. The digestive tract was differentiated into buccopharynx, esophagus, and small intestine during the endogenous feeding period. The intestinal valve and numerous longitudinal folds at the posterior region of the intestine formed together by the endo-exogenous feeding period. Major developmental events in retinogenesis occurred during the endogenous feeding period. When larvae entered exogenous feeding the mouth was fully-functional. Additionally, liver size and eye diameter increased. Our analysis of embryonic and early larval development in Levantine scraper aligned with other freshwater fishes.

Spawning performance, serum sex steroids, and ovarian histology in wild-caught Levantine scraper, Capoeta damascina (Valenciennes, 1842) treated with various doses of sGnRHa + domperidone.

As global aquaculture continues to expand, increasing efforts are focusing on assisted reproductive technologies. This study sought to test whether salmon GnRH [D-Arg6, Pro9NEt]) analogue (sGnRHa) + domperidone injection at 0.25 mL/body weight (BW; 5-μg sGnRHa + 2.5-mg domperidone), 0.5 mL/kg BW (10-μg sGnRHa + 5-mg domperidone), or 1.0 mL/kg BW (20-μg sGnRHa + 10-mg domperidone) affects spawning performance and gamete quality in wild-caught Levantine scraper, Capoeta damascina. The ability of these treatments to elicit a response was further examined by in vivo stimulation of estradiol (E2) and 17α,20β-dihydroxy-4-pregnen-3-one (DHP) and by its in vivo potency to induce oocyte maturation (OM). Females that received saline injection (control) did not spawn, whereas sGnRHa + domperidone induced ovulation and spawning across the hormonal gradient. Spawning success was highest with the 0.5 mL/kg dosage (80%) and female latency period decreased with increasing dosage. Females treated with 0.5 mL/kg had a significantly higher fecundity than those injected with 0.25 or 1.0 mL/kg. Mean oocyte diameter significantly increased in females treated with 0.5 or 1.0 mL/kg. Fertilization success, hatching rate, larvae morphology, and survival were not affected by hormonal treatment. At 12 h postinjection, E2 levels significantly declined in females treated with 0.5 or 1.0 mL/kg, whereas DHP levels significantly increased across the hormonal gradient. This steroidogenic shift is supported by histological analyses, where OM was accelerated by administration of sGnRHa + domperidone in a dose-dependent manner. In conclusion, the 0.5 mL/kg dosage of sGnRHa + domperidone is recommended for assisted reproduction of Levantine scraper.

Effect of hCG and Ovaprim™ on reproductive characteristics of male Levantine scraper, Capoeta damascina (Valenciennes, 1842)

Species richness and abundance within the genus Capoeta has been depleted. As such, there is great need for developing assisted reproductive technologies for controlling reproduction in captivity. Here, we conducted in vivo studies with single administrations of human chorionic gonadotropin (hCG) and Ovaprim™ [(D-Arg6, Pro9NEt)-sGnRH + domperidone] in wild-caught Levantine scraper, Capoeta damascina and then evaluated milt characteristics, fertilization success, serum sex steroids, and spermatogenesis via histological testicular development. Spermiation responses were significantly stronger for Ovaprim injected fish than those injected with hCG or saline. hCG had a negative effect on milt quality by reducing the percentage of motile sperm and fertilization success at 12–48 h post injection (hpi), which was not observed after treatment with Ovaprim or the saline injection. Hormonal therapy resulted in higher sperm densities and spermatocrit, although sperm longevity was not impacted. Sex steroids were not impacted by hCG or saline injection, but Ovaprim effectively induced androgen and progestin release, as evident by higher serum levels of testosterone, and 17α,20β-dihydroxy-4-pregnen-3-one. Consequently, their levels peaked at 12 hpi, which coincided with maximal milt production. Histological analysis of the testes and quantification of germ cell types revealed that Ovaprim significantly stimulated spermiogenesis, as a higher number of accumulated spermatozoa were observed at 12 h and 24 hpi. Testes from saline and hCG-injected fish remained unchanged through the experiment, and contained all stages of germ cells, predominantly spermatocytes with few spermatozoa. In conclusion, Ovaprim treatment successfully induced steroidogenesis and maturation of spermatogenic germ cells, leading to spermiation and milt production without having any negative impacts on sperm quality and fertility in wild-caught C. damascina.

Occurrence and description of Clinostomum complanatum (Rudolphi, 1819) metacercariae in freshwater fishes from Gheshlagh basin, West of Iran

Clinostomum spp. have a long uncertain taxonomic history which also have attracted great attentions. This could be due to their zoonotic potential and the presence of yellow grubs in the fish as a second intermediate host. In the current study, a total of 3oo freshwater fish belonging to the nine species were collected from two stations in the Gheshlagh basin, Kurdistan Province. Four species including Alburnus mossulensis, Capoeta damascina, Garra rufa and Squalius cephalus were found to be infected with the metacercariae. The highest prevalence (4.1%) and mean abundance (0.31±0.37) were observed in C. damascina. The metacercariae were identified using molecular (Internal Transcribed Spacer (ITS)), SEM and morphological analysis as Clinostomum complanatum. The phylogenetic analysis of four sequences of ITS gene were conducted. The specimens were placed within a lineage of C. complanatum and formed a clade with other Clinostomum species in the Palearctic region. The current study revealed the C. damascina, G. rufa and A. mossulensis as new hosts for C. complanatum and first report of this metacercariae in the region. Furthermore, the present study demonstrate the first molecular and morphological data on C. complanatum of the Iranian freshwater fish.

Mitochondrial phylogeny and taxonomic status of the Capoeta damascina species group (Actinopterygii: Cyprinidae) in Iran with description of a new species

The members of the genus Capoeta show an abstruse taxonomic status, a complex evolutionary history with the highest diversification in the Middle East and are closely related to the genus Luciobarbus . Our molecular and morphological results confirm the existence of eight species within the Capoeta damascina group in Iran. In this paper all eight recognized species are reviewed, and diagnoses are presented for all species. Capoeta birunii is considered as new species from Zayandehrud drainage basins. This species is distinguished from other small scaled Capoeta in the C. damascina group by a combination of morphological and molecular characters.

Catch composition, length frequency and biomass of commercial carps in Zayandehrud dam, Isfahan Province, Iran

Abbasi MR, Paighambari SY, Pouladi M, Ghorbani R. 2017. Catch composition, length frequency and biomass of commercial carps in Zayandehrud dam, Isfahan Province, Iran. Biodiversitas 18: 939-944. The reservoir of Zayandehrud with an area of 4800 hectares due to its economic role is one of the important lakes in Isfahan Province. This study was performed from September 2015 to March 2016, to evaluate catch composition, length frequency and biomass of carps for sustainable utilization of fish resources. Sampling operations were performed using gill nets with mesh sizes including 40, 50, 60, 70, 80, 90, 120, and 150 mm (stretched) with a length of 1 to 3 km seasonally. A total of 617 fish with a total weight of 1109.7 kg were caught. The important species which were caught by fishing operations were including common carp (Cyprinus carpio), silver carp (Hypophthalmichthys molitrix), bighead carp (Hypophthalmichthys nobilis), grass carp (Ctenopharyngodon idella) and mesopotamian barb (Capoeta damascina), respectively. Due to natural reproduction season (spring, in July), the fishing season usually starts from August to the end of winter. Common carp with 292 pieces (47.32%), silver carp with 87 pieces (14%) and mesopotamian barb with 221 pieces (35.81%) had the most abundance; whereas crucian carp with 4 pieces (0.64%) and grass carp with 1 piece (0.16%) showed the lowest abundance. Also as the main species of study, highest length ranges of caught common carps were in the range of 38-45 cm and 46-50 cm in autumn and winter, whereas highest length ranges of caught silver carps were in the range of 56-86 cm and 48-54 cm in autumn and winter. Length - weight relationships of carp showed an isometric growth model that indicates homogeneous growth in fish.

Age and growth of the Mesopotamian barb, Capoeta damascina, in central Iran

Age and growth of Mesopotamian barb, Capoeta damascina, were studied in the Zayandehrud River, in central Iran, from March 2007 to April 2008. Among the 689 specimens studied, age groups in males ranged from 1+ to 9+ and that in females ranged from 1+ to 10+. The sex ratio (1M:1.6F) was not significantly different from 1:1 ratio. Maximum fork length and weight was 39 cm and 1,115 g for males and 54.2 cm and 2,340 g for females. In all age classes, females were larger than males. The most frequent age groups were 3+ in males and 4+ in females. The highest value for condition factor was observed in June. The growth of C. damascina was described by von Bertalanffy growth curve as k=0.01, L∞=56.2 cm, to=-0.63 for males and k = 0.05, L∞=117.1 cm and to =-0.43 years for females. The length-weigh relationship was described as W=0.0169L2.95 (r2=0.98) for males and as W=0.0155L2.99 (r2=0.99) for females, indicating an isometric growth pattern in both sexes. Growth performance index φ` was estimated as 5.73 for males and 6.53 for females, indicating a faster growth rate in females.

Reproductive characteristics of Siahmahi, Capoeta damascina (family: Cyprinidae) in Beheshtabad river, Tigris basin

A total of 426 specimens of Mesopotamian barb (Siahmahi) Capoeta damascina (Cyprinidae) were caught by gillnets (1–6 cm mesh sizes) from Beheshtabad River, Chaharmahal and Bakhtiari Province from May 2013 to June 2014. Fish specimens were anesthetized in 1% clove oil solution, fixed in 10% buffered formalin and transported to the laboratory for further analyses. Fork lengths of the sampled fish ranged from 8.94 cm to 42.45 (23.32±6.53SD) cm and total weight from 10.3 to 1255.5 (242.5±213.4) g. Based on scale readings, the maximum age was found to be 7+ years for males and 8+ for females. Six reproductive characteristics, viz: sex ratio, Gonadosomatic Index (GSI), Hepatosomatic Index (HSI), relative fecundity, absolute fecundity and ova diameter were determined. The smallest mature male and female were 11.2 and 18.5 cm in fork length respectively. Sex ratio was 1M:0.7F. The GSI showed that spawning occurred from March to June. Oocyte diameters ranged from 0.57 to 2.48 (1.30±0.41) mm. Absolute fecundity ranged between 2260 and 51770 (15360±12030) eggs. Absolute fecundity and fish size (fork length and total weight) were highly correlated. Relative fecundity varied from 11 to 65 (33±12) eggs per gram of total body weight. The average hepatosomatic index was 2.25 ± 0.05 and the highest value was observed in March (3.05 ± 0.31).