Antiparasitic activity of the medicinal plant Dillenia suffruticosa against the marine leech Zeylanicobdella arugamensis (Hirudinea) and its phytochemical composition

Zeylanicobdella arugamensis (Annelida: Hirudinea), a marine parasitic leech, is currently affecting different species of cultured groupers, hybrid groupers, snappers and sea bass in Malaysia. Dillenia suffruticosa (Dilleniaceae), a medicinal plant found in Sabah, has been selected in our experiment to kill the leeches as a natural control method. The leech‐infested hybrid groupers were collected from aquaculture facilities, and the isolated leeches were challenged against methanol extract of D. suffruticosa leaves. The experiment was carried out using various concentrations of the extracts such as 25, 50 and 100 mg/ml. The methanol extract showed significant antiparasitic activity against Z. arugamensis with 100% mortality at a concentration of 100 mg/ml in 14.39 ± 3.75 min., followed by 50 and 25 mg/ml in 32.97 ± 9.29 and 41.77 ± 5.40 min., respectively. The phytochemical composition of the extract was determined using GC‐MS analysis to understand the nature of the principal compounds responsible for its antiparasitic properties. The leaves of D. suffruticosa demonstrated the presence of different bioactive compounds of various natures with varying percentages. Thus, it could be revealed that the methanol extract of D. suffruticosa mainly contains vital phytochemical compounds and showed an effective antiparasitic activity against the harmful leeches of hybrid groupers.     

Evaluation of limb autotomy as a promising strategy to improve production performances of mud crab (Scylla olivacea) in the soft‐shell farming system

Soft‐shell crab, a newly moulted crab having non‐calcified and hydrated soft exoskeleton, farming has been increasing tremendously in recent years throughout the world due to its high market price and consumer preferences. However, high aggressiveness and cannibalistic behaviour of mud crab limit its culture to an individual chamber system which ultimately makes the soft‐shell mud crab farming space and labour‐intensive. Therefore, the aim of this study was to improve the efficiency of soft‐shell crab production by autotomy of the claw and/or walking legs to reduce the moulting duration and intensify the culture system through double stocking in the existing individual chamber culture system. To achieve our objectives, a series of experiment were conducted at both indoor and on‐farm levels by removing the claw and/or walking legs of Scylla olivacea for two consecutive moulting cycles. Results showed that both claw ablation significantly (p < .01) shortened the moulting duration and increased the carapace width, weight gain, specific growth rate, feeding efficiency and yield parameter compared to the non‐ablated crabs more prominently at 1st moulting than the 2nd moulting in both indoor and on‐farm trials. Ablation of all walking legs while keeping the claw intact did not significantly influence (p > .05) the moulting duration and carapace width, but significantly (p < .01) improved the specific growth rate, weight gain, feeding efficiency and yield parameters at 1st moulting compared to the non‐ablation and unilateral three walking leg ablation groups. However, ablation of all appendages (both claw and walking legs) showed significant (p < .01) improvements of carapace width, weight gain, specific growth rate, feeding efficiency and yield parameters, and shortened the moulting duration up to 23 days from 34 days during 1st moulting and up to 27 days from 40 days at 2nd moulting. The survivals of different autotomized mud crabs were statistically non‐significant (p > .05) to the non‐autotomized crabs in various indoor and on‐farm trials. Besides improving the production performances, ablation of bilateral claw and all appendages allowed to double the stocking density in the existing individual chamber‐based soft‐shell crab farming that enhanced the production about threefold higher than the existing individual chamber‐based culture system. Finally, our results suggest that autotomy could be a promising technology for the commercial farming of soft‐shell mud crab in near future.     

Recent update on the prevalence of Vibrio species among cultured grouper in Peninsular Malaysia

Vibrio infections are common among marine fish and lead to serious problems in the aquaculture sector. This study reports a recent occurrence of Vibrio species (spp.) isolated from cultured groupers in Peninsular Malaysia using the gyrB and pyrH genes. A total of 147 Vibrio strains were successfully isolated from 77 (64%) groupers using culture method and subjected to gyrB and pyrH sequencing for species identification and confirmation. Results showed that 89% of Vibrio strains were identified and clustered to six groups of Vibrio spp., while 11% were not clustered to any Vibrio spp. using the gyrB sequences. Meanwhile, by analysis of the pyrH sequences all the 147 Vibrio strains (100%) were successfully identified and clustered into 11 groups of Vibrio spp., including the gyrB non‐identified strains. The pyrH gene provides a better resolution for identification of Vibrio spp. compared with the gyrB gene. Thus, the pyrH gene was more suitable for a rapid determination of Vibrio spp. distribution in Peninsular Malaysia. Using the pyrH gene, our study found higher prevalence of Vibrio vulnificus (33%), V. alginolyticus (24%) and V. parahaemolyticus (22%), followed by V. rotiferianus (5%), V. harveyi (3%), V. tubiashii (2%), V. campbellii (2%), V. ponticus (1%), V. diabolicus (1%), V. owensii (1%) and others Vibrio sp. (7%). Thus, the results of this study revealed that the occurrence of pathogenic vibrios among grouper fish is still high in Malaysian aquaculture. In addition, the pyrH gene was proved as a suitable marker for rapid identification of Vibrio species compared with the gyrB gene.     

Sperm quality in male <Emphasis Type="Italic">Barbus barbus</Emphasis> L. fed different diets during the spawning season

Sperm quality of Barbus barbus L. was compared among the three following dietary regimes: Group A, fed 100% commercial diet (Karpico™ containing 33% crude protein and 6% fat), Group B, fed 78% commercial diet and 22% frozen chironomid (Chironomus plumosus) larvae, and Group C, fed 56% commercial diet and 44% frozen chironomid larvae. Concentrations of polyunsaturated fatty acids (PUFAs) in Group A, B, and C were 39.1, 42.0, and 44.6, respectively, as a percentage of total fatty acids. Sperm morphology, volume, concentration and motility, total number of spermatozoa, and osmolality of the seminal plasma were compared during the spawning season. Dietary regime did not influence sperm volume, concentration, or total number of spermatozoa, osmolality of seminal plasma, or the percentage of motile sperm, but significantly affected sperm morphology (except for anterior and posterior parts of the midpiece) and sperm velocity (P < 0.05). Groups B and C showed similar sperm characteristics during the spawning season compared to Group A. Almost all parameters changed either among or within groups during the spawning season, suggesting differences in terms of the optimal time for sperm collection. The best time for sperm collection was March for Group A, but April for Groups B and C, when the osmolality of the seminal plasma measured 289 mOsmol kg⁻¹ and sperm motility was maximal. Spermatogenesis, hydration, and cell decomposition were confirmed as the three major parameters controlling sperm characteristics during the spawning season. The possible correlation between sperm morphology and motility requires further study.     

Calcitriol Treatment Attenuates Uric Acid-Induced Kidney Injury via Super Oxide Dismutase-1 (SOD-1) Upregulation and Fibrosis Reduction

Background:Hyperuricemia induces nephropathy through the mediation of oxidative stress, tubular injury, inflammation, and fibrosis. The high uric acid level is associated with the reduction of vitamin D levels. However, the reno-protective effects of this vitamin in hyperuricemia condition remain unknown. This study aimed to elucidate calcitriol treatment in a uric acid-induced hyperuricemia mice model. Methods:Uric acid (125 mg/kg BW) was administered intraperitoneally for 7 (UA7) and 14 (UA14) days. Calcitriol (0.5 g/kg BW) was intraperitoneally injected for the following seven days, after 14 days of uric acid induction (UA14VD7 group). The control group received NaCl 0.9%, by the same route. Serum creatinine was measured using calorimetric method, and uric acid levels were assessed using enzymatic calorimetric assay. Tubular injury and fibrosis were assessed using PAS and Sirius red staining. RT-PCR and qRT-PCR were carried out for the analyses of SOD-1, Collagen-1, and TGF-1 mRNA expression in the kidney. Immunostaining of SOD-1 was performed to detect its expression in the kidney. Results:Uric acid and creatinine levels markedly increased in UA14 groups, followed by an exacerbation of tubular injury. RT-PCR revealed the upregulation of Collagen-1 and TGF-1, along with the downregulation of SOD-1. Calcitriol treatment attenuated the injury with reducing uric acid and creatinine levels, as well as tubular injury. This was associated with lower Collagen-1 and TGF-1 mRNA expression compared to the UA7 and UA14 groups. SOD-1 was upregulated in epithelial cells in the UA14VD7 group. Conclusion:Calcitriol treatment after uric acid induction may attenuate kidney injury through upregulation of SOD-1 and downregulation of Collagen-1 and TGF-1 gene expression. Key Words: Fibrosis, Hyperuricemia, Kidney injury, Superoxide dismutase-1, Vitamin D     

Genetic variation for shoot elongation before winter and its correlation with vernalization requirement in winter oilseed rape (<Emphasis Type="Italic">Brassica napus</Emphasis> L.)

Changing climatic conditions in north-western Europe are accompanied by occasional extreme weather conditions. This requires breeding of winter oilseed rape cultivars which are resilient to diverse abiotic stress factors, e.g. frost, drought and heat. The degree of vernalization requirement of winter oilseed rape has been found to be related to frost tolerance and winter hardiness. Shoot elongation before winter in particular has been identified as one decisive factor for frost tolerance in winter oilseed rape. However, the relationship between vernalization requirement and shoot elongation before winter is not known. In the present study the genetic variation for shoot elongation before winter and vernalization requirement of 19 genetic diverse breeding lines and cultivars were analyzed. Autumn and spring sown field experiments in multiple environments were performed to determine shoot elongation before winter and vernalization requirement, respectively. In spring sown field experiments, genotypes with a low vernalization requirement were characterized by the occurrence of long bolting plants with flower buds. Large and significant genotypic variation was found for shoot length in the autumn sown and spring sown environment. Broad sense heritability was quite high for shoot length in the spring sown environment (h²?=?97%), whereas it was only of medium size for shoot length before winter (h²?=?62%). Although the correlation between shoot length before winter and shoot length in the spring sown environment was positive (Spearman’s rank r_S?=?0.48*), a number of genotypes with reduced shoot elongation before winter and low vernalization requirement were identified. Results indicate that genotypes with a reduced shoot elongation before winter independent of their vernalization requirement can be selected in breeding programs.     

Identification of superior parents and hybrids for improving canola production under optimum and late sowing conditions

Major advancement in canola breeding depends on heterotic hybrids that require high general combining ability (GCA) and specific combining ability (SCA) inbred lines. In order to estimate heritability, gene action type, GCA, SCA and heterosis and to identify superior hybrids with wider adaptation to cold, one hundred canola hybrids were produced by crossing 10 lines and 10 testers in a Line?×?Tester mating design. The F₁ and F₂ generations were sown in α-lattice design in 2012 and 2013 growing seasons under optimum (early October) and late sowing (early November) conditions to be evaluated for days to flowering, days to physiological maturity, number of pods per plant, number of seeds per pod, thousand seed weight, seed yield and leaf electrical conductivity. The combined analysis indicated sufficient genetic diversity in the population and significant difference between two sowing date. The Line?×?Tester analysis presented significant GCA and SCA effects for all studied traits across optimum and late sowing conditions. The main gene action type was found to be non-additive, especially incomplete dominance and over-dominance in both conditions. Narrow-sense heritability ranged from low to moderate whereas broad-sense heritability was recorded more than 60% for all of the studied traits in both generations and conditions. The average heterosis in F₂ population for all studied traits was lower than that in F₁ representing this fact that heterosis is generally related to the heterozygosity at the population level and poorly correlated with heterozygosity at the individual level.     

Pollution and Ecological Risk Assessment of Heavy Metals in Farmland Soils in Yanqi County,Xinjiang, Northwest China

A total of 50 farmland soil samples were collected from the Yanqi County, Xinjiang, China, and the concentrations of eight heavy metal elements (As, Cd, Cr, Cu, Mn, Ni, Pb and Zn) were determined by standard methods. The spatial distribution, pollution level and ecological risk status of heavy metals were analyzed based on GIS technology, the Geo-accumulation Index (Igeo), the Pollution Load Index (PLI) and the Potential Ecological Risk Index (RI). Results indicated that: (1) The average contents of Cd, Cr, Ni, Pb, and Zn of farmland soils exceeded the background values of irrigation soils in Xinjiang by 1.5, 1.40, 1.33, 2.63, and 4.92 times, respectively. Cd showed a no-pollution level, Zn showed a partially moderate pollution level, Pb showed a slight pollution level, and Cr, Cu, As, Mn, and Ni showed no-pollution level, compared to the classification standard. The PLI values of heavy metal elements of farmland soils varied from 0.83 to 1.89, with an average value of 1.29, at the moderate pollution level. (2) The Individual Potential Ecological Risk Index for heavy metals in the study area was ranked in the order of: As > Ni > Cu > Cd > Pb > Cr> Zn. The RI values of heavy metals of farmland soils varied from 3.45 to 11.34, with an average value of 6.13, at the low ecological risk level. (3) Cu and Mn of farmland soils were mainly originated from the soil parent material and topography of the study area. As, Cd, Ni and Pb were mainly originated from human activities, and Cr and Zn may originated from both natural and anthropogenic factors in the study area.     

Economic feasibility of goldfish (Carassius auratus Linn.) recirculating aquaculture system

Economic analysis of a goldfish (Carassius auratus Linn.) recirculating aquaculture system (RAS) with rearing tank capacity of 5 m³ (RT5) has been evaluated based on two culture period in 2009 and 2010. The stocking density of goldfish was kept at 100 numbers per m³ of rearing water volume. Furthermore, based on the data obtained from RT5, four more hypothetical RASs of rearing tank capacities of 10 m³ (RT10), 20 m³ (RT20), 50 m³ (RT50) and 100 m³ (RT100) were conceptualized for economic analysis that assess and compare the effects of economies of scale on profitability. The payback period, accounting rate of return (ARR), net present value (NPV), profitability index (PI) and modified internal rate of return (MIRR) were evaluated for the different water volumes (up to 100 m³) assuming project life to be 10 years. The economic analysis revealed that a goldfish RAS with rearing tank capacity 20 m³ onwards can be feasible with a payback period less than 4 years, accounting rate of return 54%, net present value of Rs 241820 and modified internal rate of return of 23%. Sensitivity analyses revealed that market price of fish was most sensitive followed by survival rate, labour, maintenance, FCR, initial investment, discount rate, electricity cost and feed cost respectively. Interestingly, feed cost was found to be the least sensitive one.     

Evaluating and modeling the spatiotemporal pattern of regional-scale salinized land expansion in highly sensitive shoreline landscape of southeastern Iran

Taking an area of about 2.3×10⁴ km² of southeastern Iran, this study aims to detect and predict regional-scale salt-affected lands. Three sets of Landsat images, each set containing 4 images for 1986, 2000, and 2015 were acquired as the main source of data. Radiometric, atmospheric and cutline blending methods were used to improve the quality of images and help better classify salinized land areas under the support vector machine method. A set of landscape metrics was also employed to detect the spatial pattern of salinized land expansion from 1986 to 2015. Four factors including distance to sea, distance to sea water channels, slope, and elevation were identified as the main contributing factors to land salinization. These factors were then integrated using the multi-criteria evaluation (MCE) procedure to generate land sensitivity map to salinization and also to calibrate the cellular-automata (CA) Markov chain (CA-Markov) model for simulation of salt-affected lands up to 2030, 2040 and 2050. The results of this study showed a dramatic dispersive expansion of salinized land from 7.7 % to 12.7% of the total study area from 1986 to 2015. The majority of areas prone to salinization and the highest sensitivity of land to salinization was found to be in the southeastern parts of the region. The result of the MCE-informed CA-Markov model revealed that 20.3% of the study area is likely to be converted to salinized lands by 2050. The findings of this research provided a view of the magnitude and direction of salinized land expansion in a past-to-future time period which should be considered in future land development strategies.