Effects of acute handling stress on cerebral monoaminergic neurotransmitters in juvenile Senegalese sole Solea senegalensis

Juvenile Senegalese sole Solea senegalensis were subjected for short periods to two different types of handling‐related stress: air exposure stress and net handling stress. The S. senegalensis were sacrificed 2 and 24 h after the stress events and the levels of serotonin (5‐HT), noradrenaline (NA), dopamine (DA) and their respective major metabolites, 5‐hydroxyindoleacetic acid (5‐HIAA), 3‐methoxy‐4‐hydroxyphenylglycol (MHPG) and 3,4‐dihydroxyphenylacetic acid (DOPAC), were measured in three brain regions (telencephalon, hypothalamus and optic tectum) and compared with those in control, non‐stressed S. senegalensis. Neither type of stress caused any significant alteration of serotoninergic activity (5‐HIAA:5‐HT ratio) or NA levels. Dopaminergic activity (DOPAC:DA ratio) was lower in stressed fish in all of the brain regions studied. For both air exposure stress and net handling stress, DA levels were significantly higher (P < 0·05) than in the control S. senegalensis. In addition, the higher DA levels after net handling stress were always significantly higher (P < 0·05) than those observed after acute air exposure stress, except in the telencephalon after 24 h. The significantly lower DOPAC:DA ratio (P < 0·05) in all of the brain regions studied was only observed in response to net handling stress.     

Effects of two parasites, Schistocephalus solidus (Cestoda) and Bunodera spp. (Trematoda), on the escape fast-start performance of three-spined sticklebacks

The fast‐start performance of three‐spined sticklebacks Gasterosteus aculeatus infected with Schistocephalus solidus and Bunodera spp. was determined and two distinct fast‐start responses (A and B) were observed. ‘A’ starts were of higher flexibility than B and three‐way ANOVA showed significant effects of A and B starts (P < 0·05), time (P < 0·05) and per cent standard body length, L_S (P < 0·05) on the orientation angle (angle of an individual segment of the fish with respect to the direction of travel). Schistocephalus solidus infection reduced maximum velocity (P < 0·05) and maximum acceleration (P < 0·05) of infected fish. Uninfected fish and fish infected with S. solidus up to a parasite index (parasite mass divided by the sum of fish and parasite mass) of 0·1 executed both types. Infected fish exclusively executed B starts for parasite index between 0·1 and 0·2. This was not due to a reduction in body flexibility associated with mechanical obstruction caused by S. solidus as no significant difference in the ratios of body width (P > 0·05) or depth (P > 0·05) to L_S were found between uninfected and infected fish. At a parasite index >0·2, infected fish were unable to perform escape fast‐starts increasing the likelihood of predation by their definitive hosts such as loons or belted kingfishers. Three‐spined sticklebacks infected with S. solidus with a parasite index of c. 0·2–0·3, however, were compromised by a suite of behavioural (e.g. increased foraging activity and amount of food consumed, increased risk associated with feeding and increased response latency to predatory stimuli), physiological (e.g. increased rate of oxygen consumption, slower growth, delayed sexual maturation and breeding success) and biomechanical (e.g. decreased fast‐start performance) factors. Bunodera spp. did not affect the escape fast‐start performance of three‐spined sticklebacks and no significant difference for maximum velocity (P > 0·05) and maximum acceleration (P > 0·05) was found.     

Muscle fibre growth dynamics in diploid and triploid rainbow trout

The effect of triploidy on muscle fibre growth was determined by comparing hyperplasia and hypertrophy of white muscle fibres in all-female, diploid and triploid rainbow trout Oncorhynchus mykiss (100–400 mm total length). Conventional morphometry and protein and DNA concentrations were used to assess muscle fibre hyperplasia and hypertrophy in white muscle samples derived from an anterio-dorsal location. Muscle fibre distributions were significantly different between triploids and diploids in trout <300 mm. The proportion of fibres <20 μm was higher in diploids than in triploids and the proportion of fibres in the 20–40 μm category was higher in triploids than in diploids. This indicates that the hyperplastic fibres of triploids are larger than those of diploids. Larger hyperplastic fibres in triploids are probably due to the combined effect of increased nuclear size in triploids and the relatively high nucleus: cell ratio observed in small muscle fibres. These larger fibres may be less favourable to cellular metabolic exchange because of their smaller surface area to volume ratios, and perhaps account for reduced viability and growth observed in triploids during early life stages. On the other hand, the lack of difference in the distribution of fibres <20 μm between diploids and triploids at larger body size ranges (301–400 mm) imply that triploid trout may have higher rates of new fibre recruitment and growth capacity at these sizes. There was no difference between diploid and triploid trout in the mean size of muscle fibres; however, the number of fibres per unit area was reduced by 10% in triploids. No differences were observed in protein or DNA concentrations in muscle tissues between the two genetic groups. Since triploid nuclei have 1·5 times more DNA than diploid nuclei, this deviation from the expected muscle DNA concentration (1·3–1·4 times more DNA in triploids when the 10% reduction in fibre density is considered) suggests that the number of nuclei per muscle fibre is reduced. In both diploids and triploids, mean fibre size increased with body length while fibre density decreased. Similarly, protein concentration in the muscle tissue increased and DNA concentration declined with increasing body length. Protein/DNA ratio was strongly and positively correlated with fibre size. These results demonstrate that changes in DNA and protein concentrations can be used to assess hyperplasia and hypertrophy in muscle tissues. However, the morphometric procedure provides better insight into muscle fibre growth as it enables the direct visualization and analysis of muscle fibre distribution patterns.     

Gross brain morphology of the armoured catfish Rineloricaria heteroptera,Isbrücker and Nijssen (1976), (Siluriformes: Loricariidae: Loricariinae): A descriptive and quantitative approach

The gross morphology of the brain of Rineloricaria heteroptera and its relation to the sensory/behavioural ecology of the species is described and discussed. The sexual and ontogenetic intraspecific variation in the whole brain length and mass, as well as within/between the eight different brain subdivisions volumes, is also examined and discussed. Negative allometry for the whole brain length/mass and relative growth of the telencephalon and optic tecta was observed. Positive allometry was observed for the relative growth of the olfactory bulbs and medulla oblongata . Univariate and multivariate statistical analyses did not reveal significant differences in the brain subdivision growth rates among sexes and/or developmental stages, except for the optic tectum and some portions of the medulla oblongata , with juveniles and males showing more developed optic tecta and medullary subdivisions, respectively. The growth rates for each brain subdivision were relatively constant, and the slopes of the growth equations were almost parallel, except for those of the olfactory bulbs and medulla oblongata subdivisions, suggesting some degree of tachyauxesis of subdivisions against the entire brain. The corpus cerebelli was the more voluminous brain subdivision in most specimens (principally adults), followed by the optic tectum (the more voluminous subdivision in juveniles), hypothalamus, and telencephalon, in that order. Differences in the number of lamellae and relative size of the olfactory organ were also detected among developmental stages, which were more numerous and larger in adults. Based on these results, it is possible to infer an ontogenetic shift in the habitat/resource use and behaviour of R. heteroptera . Vision, primarily routed through the optic tectum, could be fundamental in early stages, whereas in adults, olfaction and taste, primarily routed through the olfactory bulbs and medulla oblongata , play more important roles.     

Molecular detection and characterization of Theileria sp. from hedgehogs (Paraechinus aethiopicus) in Saudi Arabia

In this study, we conducted molecular detection and characterization of piroplasms that infect the Ethiopian or desert hedgehogs (Paraechinus aethiopicus) in Saudi Arabia. Blood samples from 112 (68 males and 44 females) desert hedgehogs from Unaizah, Central Saudi Arabia were screened for Theileria/Babesia DNA using the polymerase chain reaction (PCR) employing specific primers amplifying the partial 18S small subunit rRNA gene. Theileria DNA was detected in 51 samples (45·5%), giving a prevalence of 45·5%. Theileria DNA was found in 33 (48·5%) males and 18 (40·9%) females, and there was no significant difference (P > 0·05) in the prevalence between males and females. Similarly, there was no significant difference (P > 0·05) in the prevalence between juveniles (40%) and adults (46·7%). There was a significant difference in the prevalence of Theileria in hedgehogs collected from May to September and the period from October to April (P = 0·003). Four haplotypes of Theileria sp. in hedgehogs were detected and designated as H1–H4. H1 was the predominant haplotype and found in 80·8% of the positive individuals. Partial sequences of the 18S rRNA of Theileria sp. from hedgehogs grouped with Theileria spp. that are benign. This study is the first report of the occurrence of Theileria spp. in Saudi Arabian desert hedgehogs.     

Physical training prevents oxidative stress in L‐NAME‐induced hypertension rats

The present study investigated the effects of a 6‐week swimming training on blood pressure, nitric oxide (NO) levels and oxidative stress parameters such as protein and lipid oxidation, antioxidant enzyme activity and endogenous non‐enzymatic antioxidant content in kidney and circulating fluids, as well as on serum biochemical parameters (cholesterol, triglycerides, urea and creatinine) from Nω‐nitro‐L‐arginine methyl ester hydrochloride (L‐NAME)‐induced hypertension treated rats. Animals were divided into four groups (n = 10): Control, Exercise, L‐NAME and Exercise L‐NAME. Results showed that exercise prevented a decrease in NO levels in hypertensive rats (P < 0·05). An increase in protein and lipid oxidation observed in the L‐NAME‐treated group was reverted by physical training in serum from the Exercise L‐NAME group (P < 0·05). A decrease in the catalase (CAT) and superoxide dismutase (SOD) activities in the L‐NAME group was observed when compared with normotensive groups (P < 0·05). In kidney, exercise significantly augmented the CAT and SOD activities in the Exercise L‐NAME group when compared with the L‐NAME group (P < 0·05). There was a decrease in the non‐protein thiols (NPSH) levels in the L‐NAME‐treated group when compared with the normotensive groups (P < 0·05). In the Exercise L‐NAME group, there was an increase in NPSH levels when compared with the L‐NAME group (P < 0·05). The elevation in serum cholesterol, triglycerides, urea and creatinine levels observed in the L‐NAME group were reverted to levels close to normal by exercise in the Exercise L‐NAME group (P < 0·05). Exercise training had hypotensive effect, reducing blood pressure in the Exercise L‐NAME group (P < 0·05). These findings suggest that physical training could have a protector effect against oxidative damage and renal injury caused by hypertension. Copyright © 2012 John Wiley & Sons, Ltd.     

Difference in blood and water diffusion distance in gill lamellae of diploid and triploid tench Tinca tinca (L.)

Image analysis of sagittal sections of gill lamellae of diploid and triploid tench Tinca tinca revealed the blood and water diffusion distance in diploids (2·07 μm) to be significantly higher than that of their triploid siblings (1·46 μm; P < 0·01). Lamellae of diploids compared to triploids were found to be significantly shorter (105·84 v. 132·11 μm) and thicker (18·47 v. 14·21 μm; all at P < 0·05) than those of their triploid siblings but with similar mean sectional areas (1965·44 v. 1910·86 μm²).     

Characterization of Schizothorax prenanti cgnrhII gene: fasting affects cgnrhII expression

In this study, the role of chicken gonadotropin‐releasing hormone II (cgnrhII) in feeding regulation was investigated in Schizothorax prenanti. First, the full‐length S. prenanti cgnrhII cDNA consisted of 693 bp with an open reading frame of 261 bp encoding a protein of 86 amino acids. Next, cgnrhII was widely expressed in the central and peripheral tissues. Last, there were significant changes in cgnrhII mRNA expression in the fasted group compared to the fed group in the S. prenanti hypothalamus during 24 h fasting (P < 0·05). Furthermore, the cgnrhII gene expression presented a significant decrease in the fasted group compared with the fed group (P < 0·05) on days 3, 5 and 7, after re‐feeding, there was no significant changes in cgnrhII mRNA expression level between refed and fed group on day 9 (P > 0·05). Thus, the results suggest that cGnRH II expression is influenced by fasting and the gene may be involved in feeding regulation in S. prenanti.     

Influence of environmental parameters on growth pattern and population structure of Carassius auratus gibelio in Eastern Ukraine

Population structure and growth parameters of Prussian carp (Carassius auratus gibelio) were studied in 12 freshwater ecosystems of the Donbass region (Eastern Ukraine). These ecosystems differed significantly with respect to their surface area, water transparency and annual concentrations of phosphorus and nitrogen. Amongst the studied ecosystems, diploid males and females as well as a smaller percentage of triploid females were found. The population structure of C. a. gibelio varied significantly in terms of the percentage of triploids and sex ratio amongst diploids. A considerable proportion of triploid females (>10%) was found in four ecosystems with intermediate surface area (38–50 ha) and relatively high growth rate of specimens. The sex ratio amongst diploids was significantly female-biased in seven of 12 ecosystems, including those where triploids were present in considerable numbers. The growth parameters of Prussian carp were significantly correlated with ecosystem characteristics, but the growth patterns of diploids and triploids were not significantly different from each other. The restricted distribution of triploid specimens suggests that the triploid form might counter diverse environmental challenges, whereas the diploid form of Prussian carp seems to be efficiently adapted to a wide range of ecosystem conditions.     

Oxidative stress is induced in the rat brain following repeated inhalation exposure to manganese sulfate

Eight-week-old rats inhaled manganese (Mn) in the form of MnSO₄ at 0, 0.03, 0.3, or 3.0 mg Mn/m³ for 6 h/d for 7 d/wk (14 consecutive exposures). Brain manganese concentrations in these animals were reported by Dorman et al. in 2001, noting the following rank order: olfactory bulb>striatum>cerebellum. We assessed biochemical end points indicative of oxidative stress in these three brain regions, as well as the hypothalamus and hippocampus. Glutamine synthetase (GS) protein levels and total glutathione (GSH) levels were determined for all five regions. GS mRNA and metallothionein (MT) mRNA levels were also evaluated for the cerebellum, hypothalamus, and hippocampus. Statistically significant increases (p<0.05) in GS protein were observed in the olfactory bulb upon exposure to the medium and high manganese doses. In the hypothalamus, statistically significant (p<0.05) but more modest increases were also noted in the medium and high manganese dose. Total GSH levels significantly (p<0.05) decreased only in the hypothalamus (high manganese dose), and MT mRNA significantly increased in the hypothalamus (medium manganese dose). No significant changes were noted in any of the measured parameters in the striatum, although manganese concentrations in this region were also increased. These results demonstrate that the olfactory bulb and hypothalamus represent potentially sensitive areas to oxidative stress induced by exceedingly high levels of inhaled manganese sulfate and that other regions, and especially the striatum, are resistant to manganese-induced oxidative stress despite significant accumulation of this metal.     

Origin of Triploids of Elatostema suzukii (Urticaceae) on Okinawa Island, the Ryukyus

Elatostema suzukii , an endemic species occurring only along streams in the Yanbaru area in the north of Okinawa Island, the Ryukyus, is known to comprise di-, tri-, and tetraploids with x=13. Molecular analysis of chloroplast DNA sequences has shown that two haplotypes, AT- and NI-types, characterized by the insertion or deletion of four nucleotides “ATAT”, exist in E. suzukii. The AT-type was found in some diploids and triploids only at Genkagawa, whereas the NI-type is widespread, occurring in both diploids and triploids of the Genkagawa site as well as at other localities. The triploids share both types with the diploids, indicating that the triploids have at least two independent origins. Because of its restricted distribution, the AT-type triploid may have arisen within the Genkagawa area. Received 16 October 2000/ Accepted in revised form 18 June 2001     

The Effects of Dietary Selenium and Vitamin E and their Combination on the Fatty Acids of Erythrocytes,Bone Marrow and Spleen Tissue Lipids of Lambs

The object was to determine the influence of dietary vitamin E, selenium and their combination on the fatty acid con-tent of erythrocytes, bone marrow and spleen lipids of Akkaraman lambs. After supplementation for 15 days, the amount of all fatty acids was slightly higher (p < 0·05) in the vitamin E as compared to the control group, whereas the amount of longer fatty acids was significantly higher (p < 0·01, p < 0·001) in the selenium and combination groups. On the thirtieth day, the amount of all fatty acids was slightly high (p < 0·5) in all the supplemented groups in comparison with the control group. In the bone marrow lipids, the amount of longer fatty acids was decreased (p < 0·05, p < 0·01, p < 0·001) in the vitamin E and combination groups as compared to the control. Although the amount of some fatty acids was high (p < 0·05, p < 0·01) in the selenium group compared to the control, linoleic (18:2), linolenic (18:3) and the polyunsaturated fatty acids (PUFA) were lower (p < 0·05, p < 0·001). In the spleen lipids, the amount of longer fatty acids was slightly decreased (p < 0·05) in the vitamin E group as compared with the control; however the amount of longer fatty acids was significantly higher (p < 0·05, p < 0·01) in the selenium and combination groups in comparison to the control group. Thus dietary supplementation with selenium was more effective than dietary vitamin E supplementation in altering the fatty acid content of the erythrocyte, bone marrow and spleen lipids. © 1997 John Wiley & Sons, Ltd.     

Swimming performance and associated ionic disturbance of juvenile pink salmon Oncorhynchus gorbuscha determined using different acceleration profiles

Swimming performance was assessed in juvenile pink salmon Oncorhynchus gorbuscha (body mass <5·0 g) using five different protocols: four constant acceleration tests each with a different acceleration profile (rates of 0·005, 0·011, 0·021 and 0·053 cm s^?2) and a repeated ramped‐critical swimming speed test. Regardless of the swim protocol, the final swimming speeds did not differ significantly (P > 0·05) among swim tests and ranged from 4·54 to 5·20 body lengths s^?1. This result supports the hypothesis that at an early life stage, O. gorbuscha display the same fatigue speeds independent of the swimming test utilized. Whole body and plasma [Na⁺] and [Cl^?] measured at the conclusion of these tests were significantly elevated when compared with control values (P < 0·05) and appear to be predominantly associated with dehydration rather than net ion gain. Given this finding for a small salmonid, estimates of swim performance can be accurately measured with acceleration tests lasting <10 min, allowing a more rapid processing than is possible with a longer critical swim speed test.     

Reduction of Salmonella on alfalfa seeds using peroxyacetic acid and a commercial seed washer is as effective as treatment with 20 000 ppm of Ca(OCl)2

Aims: The efficacy of a commercial seed washer and 1 and 3% peroxyacetic acid or 20 000 ppm calcium hypochlorite for reducing Salmonella on alfalfa seeds was investigated. Methods and Results: Alfalfa seeds were inoculated with Salmonella Stanley to achieve c. 5 log CFU g^?1. Seeds were then treated with 1 or 3% peroxyacetic acid or 20 000 ppm calcium hypochlorite for 15 min in a commercial seed washer that uses air to enhance contact of the sanitizer with the seed. Experiments were also conducted using industry and laboratory methods. An c. 1‐log reduction in number of Salm. Stanley was demonstrated regardless of the chemical treatment or method of treatment. Although this 1‐log reduction was significant (P < 0·05), differences among the treatments were not significant. Treating the seed with 1 and 3% peroxyacetic acid resulted in similar Salm. Stanley reductions of 1·77 and 1·34 log, respectively, not being statistically significant (P > 0·05). Conclusions: These results suggest that under conditions tested, 1 or 3% peroxyacetic acid solutions are equally effective as 20 000 ppm of Ca(OCl)₂ in the reduction of Salm. Stanley on alfalfa seed when used in conjunction with a commercial seed washer. Significance and Impact of the Study: A 1% peroxyacetic acid solution could potentially be used in place of 20 000 ppm of Ca(OCl)₂ for treatment of seeds used for sprouting_. The commercial seed washer did not enhance removal of Salm. Stanley from alfalfa seeds, but did facilitate removal of excess soil from seeds.     

POSTNATAL CHANGES IN FREE AMINO ACID,DNA, RNA AND PROTEIN CONCENTRATIONS OF MINIATURE SWINE BRAIN1

Postnatal developmental characteristics of miniature swine brain were evaluated through the first 9 weeks of age. Differential growth rates of cerebrum, cerebellum and brain stem were defined in terms of DNA, RNA, protein and free amino acid concentrations at ages 5, 21, 35 and 63 days. Within the experimental conditions provided, hyperplasia ceased just prior to ages 21, 35 and 63 days for cerebellum, brain stem and cerebrum, respectively. An additional cerebral growth spurt, observed between weaning at age 35 days and sacrifice at age 63 days, may be indicative of impaired brain development due to inadequate nutrition provided by the dam's milk. Developmental changes in mean concentrations of brain free amino acids varied with anatomical area and differed somewhat from those of other species previously reported. For example, mean cerebral concentrations of aspartic acid, γ-aminobutyric acid and asparagine + glutamine decreased significantly (P < 0·05) with age and mean glutamic acid concentration was 5 times that of taurine.     

The effects of selenium,vitamin E and their combination on the composition of fatty acids and proteins in Saccharomyces cerevisiae

The aim of our studies was to test the effect and role of vitamin E and selenium supplements on yeast cell. In this study, the effects of selenium (Se), vitamin E (Vit. E), and their combination (Se plus Vit. E) on the composition of fatty acids and proteins were examined in Saccharomyces cerevisiae strains WET136 and 522. S. cerevisiae cells were grown up in YEPD medium supplemented with Se, Vit. E or their combination. It was found that the level of stearic acid was increased in all supplemented groups (p<0·05; p<0·001). The content of saturated and unsaturated fatty acids was decreased (p<0·05; p<0·01; p<0·001) in Vit. E and Vit. E plus Se supplemented S. cerevisiae. On the other hand, Se alone caused an increase (p<0·001) in the saturated fatty acids but a decrease (p<0·05; p<0·001) in the unsaturated fatty acids. Total proteins in S. cerevisiae were significantly increased (p<0·001) by Vit. E supplement. There was no significant change observed in S. cerevisiae supplemented with Se. These findings indicate that membrane composition of S. cerevisiae is affected by both Vit. E and Se supplements. © 1997 John Wiley & Sons, Ltd.     

Stimulation of natural killer cells by homoeopathic complexes: An in vitro and in vivo pilot study in advanced cancer patients

The present study was designed in order to evaluate the effects of five homoeopathic complex preparations on functional activity natural killer cells (NKCs) in advanced cancer patients. We examined the effects of Coenzyme Compositum®, Ubichinon Compositum®, Glyoxal Compositum®, Katalysatoren® and Traumeel® on the functional activity of NKCs. Experimental procedures included in vitro and in vivo trials. The in vitro trials were performed in NKCs isolated from 12 healthy volunteers (aged 44 ± 4 years) and incubated with the five homoeopathic complex preparations. The in vivo trials were performed in 15 advanced cancer patients (aged 55 ± 12 years) supplemented for 3 months with the homoeopathic preparations. All five homoeopathic preparations significantly increased the cytotoxic activity of the NKCs at the lowest NKCs/target cell ratio 12:1 (p < 0·05). The order of activity was: Ubichinon Compositum® > Glyoxal Compositum® > Katalysatoren® > Traumeel® > Coenzyme Compositum®. In the advanced cancer patients, the homoeopathic preparation significantly increased NKCs cytotoxic activity (p < 0·05). The homoeopathic complex preparations tested in this study can be used as an adjuvant immunotherapy in advanced cancer patients. Copyright © 2013 John Wiley & Sons, Ltd.     

Spermatozoa in triploids of the rosy bitterling Rhodeus ocellatus ocellatus

Artificially induced triploid male Rhodeus ocellatus ocellatus showed typical nuptial colorations, irrespective of spermiation. In milt from triploids, abnormal spermatozoa (malformation of the head and mitochondrion, excessive formation of the head, mitochondrion and flagellum, and no flagellum) occurred at 78°4% frequency. Spermatozoa with multiflagella were most common, often with a saccate-like organ. Many triploid spermatozoa moved actively as long as those of diploids (10·92±0·91 min=mean±S.D., P >0·05), but did not advance like diploids, spinning around until movement ceased. The sperm density in triploids was < 2% of that from diploids. In triploid testes, deformed and variously sized spermatids were often observed, and normal spermatids and spermatozoa were seldom recognized. The DNA content of triploid spermatozoa varied greatly, compared with that of diploids. Peak of sperm DNA content differed slightly between two triploid samples with two peaks at 1·5 n and 1·9 n ( P <0·0001 in both), respectively. Triploids had the greatest average sperm head diameter of 2·25±0·67 μm (mean±S.D.), while that of diploids was 1·83±0·15 μm ( P =0·002). In the fertilization test using the eggs of diploids ( n =1500, 30 trials), only one egg developed. The embryo chromosome number was 60 (2·5 n) and the ploidy of spermatozoa contributing to fertilization appears to be 1·5 n. The extremely low fertility of triploid R. o. ocellatus spermatozoa seems to be caused by the reduced motility and large head size of spermatozoa, and the low sperm density of the milt. The ploidy of spermatozoa that are successful in fertilization is likely to be related to the distribution pattern in the DNA content of cells.