Development and evaluation of the polymerase chain reaction (PCR) method for diagnosis of spring viremia of carp virus (SVCV)

Aquaculture has been expanded rapidly to become a major commercial and food-producing sector worldwide in recent decade. In parallel, viral diseases rapidly spread among farms causing enormous economic losses. The accurate detection of pathogens at early stages of infection is a key point for disease control in aquaculture. Spring Viraemia of Carp Virus (SVCV) is a very severe pathogen of carp fishes in different parts of the world and is categorized as a reportable listed disease in the annual published list of World Organization for animal Health (OIE). The objective of this study was to develop and evaluate RT- PCR test for detecting SVC virus and also the sensitivity and specificity of this test. A semi nested RT- PCR was designed using combination of three primers: two external (SVCF , SVCR) and one internal (SVCS) primers which based on conserved region of G gen. The specificity of designed primers (only external ones) by examination on Viral Hemorrhagic Septicemia Virus (VHSV) and Infectious Hematopoietic Necrosis Virus (IHNV) was confirmed. For optimizing of the PCR test, primer concentration, primer annealing temperature, cycle number and Mgcl2 concentration were surveyed. Also for validity test, prevention of false negative and Assurance of its accuracy, a competitive internal control (mimic) designed and its suitable concentration was defined. Evaluation of the sensitivity of designed test were conducted first by comparing the different commercially available RNA isolation guidelines, two guidelines: isotiocyanate phenol���chloroform based protocols (RNX���Plus Iran, Iq2000 kit Taiwan ) and two column based protocols (Cinna pure RNA Iran , high pure viral RNA kit, Roche Germany ). The results indicated that the column based protocols (Roche method and Cinna pure), yield 36.77 ng/��l and 16/47 ng/��l RNA concentration respectively, which were significantly higher than other protocols(P<0.05). Then for evaluation of extracted RNA sensitivity, Serial dilution of SVCV strain 56.70 grown in EPC (1.9��105 TCID50/ml) was examined To compare sensitivity. Extracted RNA from serial dilution with stone's primers and commercial IQ-2000 kit were examined simultaneously. The result indicated that designed semi- nested RT- PCR was able to recognize SVC virus to 10-4 dilution and stone's primer recognize to 10-3 dilution whereas Iq-2000 commercial kit did not recognized in any dilution. In high virus titer in designed test two DNA band (462 bp and 266 bp) produced, and by decreasing virus titer 462 bp was omitted. In low virus titer or lack of virus, just DNA band (mimic) 729 bp can propagate. After designing and optimizing PCR test, a total of 400 suspected cultured Cyprinus carpio with high mortality from 4 aquaculture zone of Khuzestan province were collected and tested for SVCV during 2012- 2013 using developed PCR method and IQ- 2000. The results indicated that SVC virus was not observed in samples using both methods.

Rapid identification of European (Anguilla anguilla) and North American eel (Anguilla rostrata) by polymerase chain reaction.

A rapid and cost effective DNA test is described to identify European eel (Anguilla anguilla) and North American eel (Anguilla rostrata). By means of polymerase chain reaction (PCR) technique parts of the mitochondrial cytochrome b gene are amplified with species specific primers which are designed to produce PCR fragments of different characteristic sizes for European and American eel. The size differences can easily be made visible by agarose gel electrophoresis

A seven year study of the life cycle of the Mayfly Ephemera danica

Ephemera danica M��ller, 1764 (Ephemeroptera) is one of the largest mayflies found in the British Isles with some females reaching over 30 mm. It is a common and widespread species found in rivers, lakes and streams throughout Europe and is particularly abundant in many of the lowland rivers of the British Isles. The larvae are burrowers ��� mainly found where silt accumulates below macrophytes. This article gives a general overview of research work on the factors affecting the life cycle of Ephemera danica over a seven year period (1995���2002) on two rivers, the River Test at Leckford in Hampshire and the North Wey at Tilford in Surrey.

Life cycle of Pisidium obtusale Jenyns. [Translation from: Informatsionnyi Byulleten Biologiya Vnutrennikh Vod No.3 17-21, 1969.]

Pisidium obtusale in the neighbourhood of Borka occurs in deep pools on upland bogs. The majority of water-bodies where this mollusc lives are temporary. Their bottoms are covered with sedges, and sometimes mosses occur. Evidently there is a marked attraction of P. obtusale to places overgrown by willow. In pools remote from scrub or woody vegetation it does not appear. Temporary water-bodies fill up with melt water in the middle or the end of April and finally dry up at the end of July or the beginning of August. Observations on the life cycle of P. obtusale started on 21 April 1966, following on the filling-up of the water-body by melt water. The findings of the study are presented in this paper.

Cytological aspects of gonadotropic cells and pre-opticoneurohypophysial neurosecretion during the ovarian cycle of Eudontomyzon danfordi. [Translation from: Studia Univ.Babes-Bolyai, Biologia 18 105-113, 1973.]

The connection between the activity of the gonadotropic cells of the hypophysis and the neurosecretion in the pre-optico neuro-hypophysial system in different groups of vertebrates has been examined by many authors. It has been established that in many species there exists some kind of synchronism between the sexual cycle and the cellular activity of neurosecretion, a fact that has led to the prevailing idea that gonadotropic activity in the hypophysis is regulated by the hypothalamus. This paper summarises the results of experiments made in this direction on Cyclostomata. The materials for research came from adult individuals of Eudontomyson danfordi Regan taken from the rivers Somes-Rece and Ivo (Harghita district) at different times of the year.

Reaction of fish to light (���Conclusions��� only). [Translation from: Voprosy Ikhtiologii 6 17-20, 1956]

Qualitative and quantitative characteristics of the reaction of certain fish to light were tested and reasons for the attraction of certain fish to light are discussed.

Individual growth and seasonal cycle of Daphnia middendorffiana in an Alpine lake [Translation from: Memorie dell'Istituto Italiano di Idrobiologia Dott.Marco de Marchi 26 41-83, 1970]

The problem of the peculiar reproductive biology of the cladoceran Daphnia middendorffiana is investigated from a cytological viewpoint, and by direct observation the meiotic phenomena of the eggs both subitaneous and resting is studied. and during maturation, the true mechanism of the succession of reproductive phases of different ecological significance. Samples were collected in the Italian Alpine Lake of Campo 4��.

The role of ciliates in the pelagic life cycle of the Mediterranean [Translation from: Rapports et Proces Verbaux des Reunions-Conseil International pour l'Exploration de la Mer 17, 511-512, 1963]

During hydrographic and plankton studies carried out since 1960 in the coastal zone between the Ebro and Castellon (western Mediterranean), data has been collected which confirms the importance of ciliates in the composition and activity of the plankton. The ciliates in 413 samples of 100 ml of water were counted, having been examined with the Utermohl microscope after sedimentation. The samples studied were distributed according to the density of their population. subject for study. The author concludes that recognition of the role of ciliates as an important link in the food chain of the sea would simplify the interpretation of certain problems posed by the nutrition of certain groups of planktonic animals.

Physiological aspects of the life cycle of the river lamprey, Lampetra fluviatilis L

The life cycle of the river lamprey, L. fluviatilis, is reviewed. The larval lamprey, or ammocoete, is a blind, filter-feeding animal, which normally lies concealed in the silt deposits of streams and rivers. After a period of 3-5 years in fresh water the ammocoete undergoes a metamorphosis in the summer months into a sexually immature, non-feeding stage known as the macrophthalia, which is active. This stage migrates downstream in late winter. It adopts a parasitic existence, in intertidal areas. After 18 months it returns to spawn in fresh water, after a final freshwater stage lasting up to 9 months. The river lamprey dies within a few days after the spawning period of 3-4 weeks, and none survive to spawn the following year.

Alterations of the global water cycle and their effects on river structure, function and services

River structure and functioning are governed naturally by geography and climate but are vulnerable to natural and human-related disturbances, ranging from channel engineering to pollution and biological invasions. Biological communities in river ecosystems are able to respond to disturbances faster than those in most other aquatic systems. However, some extremely strong or lasting disturbances constrain the responses of river organisms and jeopardise their extraordinary resilience. Among these, the artificial alteration of river drainage structure and the intense use of water resources by humans may irreversibly influence these systems. The increased canalisation and damming of river courses interferes with sediment transport, alters biogeochemical cycles and leads to a decrease in biodiversity, both at local and global scales. Furthermore, water abstraction can especially affect the functioning of arid and semi-arid rivers. In particular, interception and assimilation of inorganic nutrients can be detrimental under hydrologically abnormal conditions. Among other effects, abstraction and increased nutrient loading might cause a shift from heterotrophy to autotrophy, through direct effects on primary producers and indirect effects through food webs, even in low-light river systems. The simultaneous desires to conserve and to provide ecosystem services present several challenges, both in research and management.

El��ments nutritifs et production primaire dans les lagunes du C��te d'Ivoire : cycle annuel

This paper collects most of the information gathered between October 1974 and July 1976 within the framework of a research program concerning the hydrobioclimate of ivorian lagoons and especially the Ebri�� lagoon. Monthly surveys concerning the latter were carried out during 1975. The following parameters - and their vertical distribution wherever it had a meaning - were systematically gathered in a system of fifty-five stations: Transparency (Secchi), Temperature, Salinity, Chlorophylla, Dissolved oxygen, phosphate, nitrate, nitrite. These data provide an outline of the annual cycle of nutrients and primary production of the Ivoirian lagoons

White spot syndrome virus (WSSV) transmission risk through infected cooked shrimp products assessed by polymerase chain reaction (PCR) and bio-inoculation studies

The aim of the study was to evaluate the resistance of white spot syndrome virus (WSSV) in shrimps (Penaeus monodon) to the process of cooking. The cooking was carried out at 1000C six different durations 5, 10, 15, 20, 25 and 30 min. The presence of WSSV was tested by single step and nested polymerase chain reaction (PCR). In the single step PCR, the primers 1s5 & 1a16 and IK1 & IK2 were used. While in the nested PCR, primers IK1 &IK2 ��� IK3 & IK4 were used for the detection of WSSV. WSSV was detected in the single step PCR with the primers 1s5 and 1a16 and the nested PCR with the primers IK1 and IK2 ��� IK3 & IK4 from the cooked shrimp samples. The cooked shrimps, which gave positive results for WSSV by PCR, were further confirmed for the viability of WSSV by conducting the bio-inoculation studies. Mortality (100%) was observed within 123 h of intra-muscular post injection (P.I) into the live healthy WSSV-free shrimps (P. monodon). These results show that the WSSV survive the cooking process and even infected cooked shrimp products may pose a transmission risk for WSSV to the native shrimp farming systems.

Cycle sexuel et f��condit�� du machoiron Chrysichthys nigrodigitatus en Lagune Ebri��, C��te d'Ivoire

Changes in the seasonal development of the gonads of female Chrysichthys nigrodigitatus, in Ebri�� lagoon (C��te d'Ivoire) are described over an annual reproductive cycle. Seven macroscopic stages of gonad maturity were identified. There is a major spawning period from July to November. The mature fish spawn only once during the breeding season. There was a slightly higher correlation between fecundity and fish length than between fecundity and fish weight or gonad weight. Fecundity estimates ranged from 5438 to 36257 eggs and from 4878 to 87724 eggs, respectively for the fish in captivity and those in the natural environment.

��tude histologique du cycle sexuel de l'albacore (Thunnus albacares) Bonnaterre 1788

The evolution of the general microscopic structure of the ovary of Thunnus albacares related to the gonad index, and the ovocyte maturation process were studied. Some stages of the males sexual maturation were characterized.

Changes in catch rates and length and age at maturity, but not growth, of an estuarine plotosid (Cnidoglanis macrocephalus) after heavy fishing

The hypothesis that heavy fishing pressure has led to changes in the biological characteristics of the estuary cobbler (Cnidoglanis macrocephalus) was tested in a large seasonally open estuary in southwestern Australia, where this species completes its life cycle and is the most valuable commercial fish species. Comparisons were made between seasonal data collected for this plotosid (eeltail catfish) in Wilson Inlet during 2005���08 and those recorded with the same fishery-independent sampling regime during 1987���89. These comparisons show that the proportions of larger and older individuals and the catch rates in the more recent period were far lower, i.e., they constituted reductions of 40% for fish ���430 mm total length, 62% for fish ���4 years of age, and 80% for catch rate. In addition, total mortality and fishing-induced mortality estimates increased by factors of ~2 and 2.5, respectively. The indications that the abundance and proportion of older C. macrocephalus declined between the two periods are consistent with the perception of long-term commercial fishermen and their shift toward using a smaller maximum gill net mesh to target this species. The sustained heavy fishing pressure on C. macrocephalus between 1987���89 and 2005���08 was accompanied by a marked reduction in length and age at maturity of this species. The shift in probabilistic maturation reaction norms toward smaller fish in 2005���08 and the lack of a conspicuous change in growth between the two periods indicate that the maturity changes were related to fishery-induced evolution rather than to compensatory responses to reduced fish densities.