The role of Ca2+ and Na+ membrane transport in brook trout (Salvelinus fontinalis) spermatozoa motility

The role of environmental ion composition and osmolality in Ca²⁺ signaled activation was assessed in spermatozoa of brook trout Salvelinus fontinalis. Milt from ten mature males was obtained by abdominal massage. Spermatozoa motility was evaluated in 0, 100, and 300 mOsm/kg NaCl or sucrose solutions, buffered by 10 mM Tris–HCl pH 8.5. For investigation of spermatozoa reaction to external Ca²⁺ concentration, 2 mM ethylene glycol tetraacetic acid (EGTA) was added to the activation media as a calcium ions chelator. For investigation of the effect of external Na⁺ concentration in conditions of low external Ca²⁺, 100 µM amiloride was added to the EGTA-containing solutions as a Na⁺ transport blocker. Low motility was observed in sucrose (Na⁺ free) solutions containing 2 mM EGTA but not in Na⁺ solutions containing 2 mM EGTA. Addition of amiloride led to significantly increased motility (P < 0.05) compared with sucrose (Na⁺ free) solutions containing 2 mM EGTA. We conclude that Na⁺ transport in Ca²⁺-free solutions plays a regulatory role in brook trout spermatozoa activation. The influence of competitive Na⁺ and Ca²⁺ transport on the control of spermatozoa activation requires further study with respect to its application for improvement of artificial activation and storage media.     

Changes of sperm morphology,volume, density,and motility parameters in northern pike during the spawning period

Sexually mature males (BW?=?1600?±?150 g and TL?=?235?±?30 mm) of northern pike (Esox lucius L.) were randomly selected from a pond to record changes in their sperm quality parameters (spermatozoa morphology, sperm volume, density, and motility parameters) during the spawning season. The morphological and motility parameters changed significantly during the reproductive season with following trends. Only, head width was not changed during the spawning season. The longest spermatozoa and its flagellar length were found at the middle of spawning period (TL?=?38.24?±?0.37 μm and 35.14?±?0.26 μm) and shortest at the beginning of spawning period (TL?=?34.81?±?0.29 μm and 32.53?±?0.18 μm). Other morphological characters were always the lowest at the beginning of spawning period. Sperm volume was changed from 0.33?±?0.3 ml in February, 0.43?±?0.2 ml in March to 0.24?±?0.1 ml in April, and density from 16.2?±?0.2?×?109 spermatozoa ml^?1 in February, 19.4?±?0.2?×?109 spermatozoa ml^?1 in March to 4.8?±?0.2?×?109 spermatozoa ml^?1 in April. Same sperm velocity was observed in all spawning terms at 10 and 20 s after activation. Higher velocity was found at 30 and 40 s after activation in sperm collected at the middle and the end of spawning period. Significantly, higher percentage of motile sperm was observed at 20, 30, and 40 s after activation in sperm sampled at the end of spawning period. This study supports the hypothesis that longer spermatozoa swim faster.     

Effectiveness of dietary vitamin supplementation to the performance of common carp (Cyprinus carpio L.) larvae in intensive rearing condition

Study of enrichment of different vitamins (C, B₁, B₆ and E) in common carp larvae through 4 weeks feeding in recirculation system was carried out. For this purpose, procedure for enrichment of Artemia nauplii with different vitamins was developed and investigated the effectiveness in larvae nutrition. The habituation of larvae to dry feeds was done using supplemented dry diets with the same vitamins. To study the effect of vitamins in young fish, the most common stress situation was planned for carp nursed fries, which occurs during transportation. Levels of vitamins and parameters of immune response in mucus were monitored before and after stress situation. At the end of the feeding trials using enriched Artemia nauplii, level of vitamin B₁, B₆ and C were increased in the body of fish, but remained on similar level or decreased after dry feeding period. Therefore, we concluded that Artemia is a good delivery vector of these vitamins. While handling stress, vitamin C and E concentration was decreased and sparing effect of antioxidant vitamins was confirmed in groups supplemented with both vitamins. Vitamin B₁ was consumed intensively during the stress, vigorously in groups without supplementation. Contrary to this, vitamin B₆ level increased in stress condition in group without supplementations. The levels of immunoglobulins in mucus were increased in all treated groups independently of treatment. Our results confirm that carp larvae reared in tank condition needs sufficient amount of vitamins C, B₁ and B₆ supplements to protect them from the mild handling stress.     

Livelihood implications of in situ-on farm conservation strategies of fruit species in Uzbekistan

The aim of this paper is to analyze the impact of a set of interventions related to on-farm/in situ conservation and use of fruit species (cultivated and wild) on farmers’ livelihoods and species diversity in Central Asia. Specifically, a difference-in-differences propensity score matching is used to evaluate the outcome of a development research program in Uzbekistan between 2005 and 2010. Species crop diversity maintained by farmers before and after the project increased as a result of the interventions, showing the efficacy of the interventions promoted by the projects in terms of conservation. Furthermore, innovations provided by the program increased both household propensity of marketing and self-consumption of target fruit. However, the program’s interventions did not seem to impact significantly any of the indicators related to household livelihoods. The short time elapsed between the end of the project and the impact assessment may be too brief to capture any observable impact on livelihoods.     

Preparation and epitope characterization of monoclonal antibodies suitable for detection of <Emphasis Type="Italic">Tomato yellow leaf curl virus</Emphasis>

Tomato yellow leaf curl virus (TYLCV) is a begomovirus that seriously threatens tomato crops worldwide. Current immunodiagnostic methods for this pathogen employ commercially produced mAbs raised against TYLCV. However, despite the existence of these mAbs, little information regarding their characterization or strategy of production has been published. In addition, research on TYLCV would certainly benefit were more mAbs available, thus allowing efficient examination of the virus life cycle, modes of pathogenesis and possibly the development of resistant cultivars. The coat protein (CP) of TYLCV is the only known building block of the viral capsid. Therefore, in this study we used CP as an immunogen for the production of novel mAbs. We employed a strategy in which the CP was truncated at its C-terminus to avoid intra- and inter-protein interactions that could impair epitope exposure. For the same reason, we used a denaturated antigen to expose linear epitopes during the immunization. This effort yielded three mAbs: they were characterized biochemically and immunologically, and their epitopes were mapped. Possible applications of these mAbs are discussed.     

Effect of restricted suckling on milk yield,composition and flow,udder health,and postpartum anoestrus in grazing Holstein cows

The effect of restricted suckling on milk yield and composition, udder health, and postpartum anoestrus in dairy cows in pasture-based systems, was studied in 32 Holstein multiparous cows and their calves. At calving, each cow–calf pair was randomly assigned to one of two treatments: restricted suckling (RS) of the cows by her own or another calf, twice daily for 30 min or artificial rearing (AR) of the calves with milk obtained from the bulk tank, offered twice a day in buckets. Treatments were applied until week 8 after calving. The diet of the cows consisted of direct grazing in improved pastures, corn silage and a commercial concentrate which was offered at milking. Milk production and composition, udder health, body condition score of the cows, body weight and milk intake of the calves were measured weekly, and the first postpartum ovulation was determined three times a week by ovarian ultrasonography. Cows with RS management had a lower machine-milked milk yield (17.9 vs. 24.8 kg/d), a lower fat percentage (3.21 vs. 4.11%) and 4% fat-corrected milk yield (16.2 vs. 25.7 kg/d), and also a lower average milk flow (1.35 vs. 1.76 kg/min) than cows in the AR treatment. There was no effect of treatment on milk protein percentage or udder health as measured by milk electrical conductivity. The interval from calving to first postpartum ovulation was shorter in the AR cows than in the RS cows (18.5 vs. 21.8 days). The RS calves consumed more milk (7.2 vs. 5.4 kg/d), gained more body weight (0.813 vs. 0.656 kg/d), and had a higher body weight at weaning (84.3 vs. 73.3 kg) than AR calves. Restricted suckling of grazing dairy cows had a negative effect on machine-milked milk yield, fat percentage and 4% fat-corrected milk yield, but had no effect on udder health or on improved weight gain and body weight at weaning of the calves.     

香菇多糖对鲤鱼（cyprinus carpio）离体培养免疫细胞的活性调节作用*

采用Percoll密度离心等技术,对鲤鱼的头肾巨噬细胞和外周血白细胞进行分离纯化,并离体培养。体外暴露不同浓度的黄芪多糖（APS）和香菇多糖（LNT）后,分别采用 MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide)法测定它们对鲤鱼外周血白细胞增殖的影响;NBT(nitroblue tetrazolium)还原法和Griess试剂显色法测定对头肾巨噬细胞的呼吸爆发的影响;实时定量PCR法测定头肾巨噬细胞细胞因子IL-1β诱导表达的影响。结果显示,香菇多糖能显著诱导巨噬细胞的氧爆发活性;其低浓度时对细胞的氮呼吸爆发活性无显著影响,随着作用浓度的增加表现为高浓度抑制作用;香菇多糖可以显著促进鲤鱼外周血白细胞的增殖,且能显著增强IL-1β的体外诱导表达。结果表明香菇多糖对离体培养鲤鱼免疫细胞有明显活性作用,对鲤鱼非特异性免疫和特异性免疫具有促进作用,是一种有潜力的鱼用免疫增强剂。     

Development of a PCR test to diagnose Haemophilus parasuis infections.

A polymerase chain reaction (PCR) test was developed in order to improve the accuracy and speed of diagnosis of Haemophilus parasuis, an economically important respiratory pathogen that affects swine. The gene sequence of the 16S small subunit ribosomal RNA of H. parasuis (GenBank M75065) was compared with 56 16S sequences of related bacteria, including those frequently isolated from pig tissues. Two species-specific primers were designed: HPS forward and HPS reverse. The predicted size of the amplified PCR product was 821 bp. The PCR test could detect a minimum of 102 bacteria and 0.69 pg of DNA. Thirty-one H. parasuis isolates, including 12 different serovars and 19 field isolates, were positive using the PCR test. No amplification was observed when the test was run using DNA from 15 other bacterial species commonly isolated from swine tissues. A weak band was observed when the PCR test was performed using Actinobacillus indolicus DNA as template. Clinical samples tested by PCR included tissues and swabs from 5 animals naturally infected with H. parasuis and 1 experimentally infected animal. The PCR was positive in 26 of 30 clinical samples. Four samples showed weak bands, and these results were not considered positive. Haemophilus parasuis was isolated from 18 of 30 of these samples. Tissues from specific pathogen-free (SPF) pigs and from unrelated species were negative for H. parasuis isolation and PCR. The developed PCR was successfully used in the diagnosis of H. parasuis infection, especially when compared with traditional microbiology techniques.