Lytic bacteriophages specific to Flavobacterium columnare rescue catfish, Clarias batrachus (Linn.) from columnaris disease

This investigation was aimed to find out appropriate strategy against antibiotic resistant bacterial fish pathogen, F. columnare. This pathogen was found persistently associated with fishes causing columnaris disease and ensuing mass mortality in hatchery and culture system of Sub - Himalayan region. Nine lytic F. columnare phages (FCP1 - FCP9) specific to its fifteen isolates were isolated from the water and bottom sediments of various geo-climatic regions of North India. The F. columnare phage FCP1 (made of hexagonal head and non contractile long tail belonging to family Podovariedae, a member of DNA virus) exhibited broader host range to lyse 9 out of 15 isolates of F. columnare. Therapeutic ability of FCP1 phage was assessed in C. batrachus inoculated intramuscularly (im) with virulent bacterial isolate FC8 and post inoculated (PI) with FCP1 phage (@ 10(8) : 10(6):: cfu : pfu) through intramuscular (im), immersion (bath) and oral (phage impregnated feed) treatment. Significant (p < 0.001) reduction (less than 10(-3) cfu ml(-1)) in host bacterium in the sera, gill, liver and kidney of challenged fishes was noted after 6 hr of phage treatment. Quantum of phage played a significant role in bringing down bacterial population as in the sera of dose 1 (@ 4.55 x 10(6) pfu ml(-1)) and dose 2 (@ 9.15 x 10(6) pfu ml(-1)) treated fishes mean log10 cfu value reduced by 3 logs (58.39%) and 5 logs (73.77%) at 96 hr, respectively. Phage treatment led to disappearance of gross symptoms, negative bacteriological test, detectable phage and 100% survival in experimentally infected C. batrachus. Result of this study provides evidence of profound lytic impact of FCP1 phage and represents its interesting therapeutic importance against antibiotic resistant F. columnare.     

Distribution and abundance of fish eggs and larvae in Arasalar estuary, Karaikkal, south-east coast of India

The distribution and abundance of fish eggs and larvae have been studied at two Stations of Arasalar estuary. Eggs of Stolephorus sp., Mugil cephalus, Sardinella sp., Cynoglossus sp and the larvae of Stolephorus indicus, Ambassis commersoni, Terapon jarbua, Chanos chanos, Trichiurus sp Anguilla sp and Thryssa sp have been collected and identified. Environmental parameters such as temperature, salinity, pH and dissolved oxygen were also recorded. The present study clearly indicate the higher occurrence and abundant of fish eggs and larvae during March - July. The fair numbers of eggs and larvae indicate the existence of breeding ground within the estuary. Study on the physico-chemical parameters found that the abundance and distribution of fish eggs and larvae were influenced by the salinity of this estuary.     

Physico-chemical and microbial characteristics of the coral reef environment of the Gulf of Mannar marine biosphere reserve, India

Investigation on physico-chemical parameters and bacteial characteristics of the coral reef environs of the Gulf of Mannar biosphere reserve was studied. The study found the influence of different physico-chemical parameters on one another and also on the distribution of the total heterotrophic bacteria (THB) in the coral reef areas. Nutrients exhibited considerable seasonal and spatial variations with influence on the bacterial population. Coral reef areas recorded higher bacterial population density both in water (3.5 to 18 x 10(5) CFU ml(-1)) and sediment (1 to 14 x 10(7) CFU g(-1)) samples than the non coral reef areas (3.4 to 10.5 x 10(4) CFU ml(-1) in water and 0.9 to 7 x 10(6) CFU g(-1)). The study also found the dominance of gram negative groups at all the three stations (64.73, 63,5 and 72.59%) with Pseudomonas contributing maximum number of strains in all the samples. In addition Vibrio, Aeromonas, Flavobacterium, Cytophaga, Enterobacter and Alcaligenes were also recorded. The gram positive group was represented by Bacillus, Micrococcus, Arthrobacter and Corynebacterium. The genetic composition of THB isolated from the coral mucus revealed the presence of Vibrio and Micrococcus in all the coral mucus.     

一株菲降解菌的特性及相关降解基因的克隆

从石油污染土壤中分离到一株菲降解菌2F5-2.根据该菌株生理生化特征和16S rDNA序列相似性分析,将其初步鉴定为鞘氨醇杆菌属(Sphingobium sp.).该菌株在10 h内对100 mg/L的菲的降解率为100%.降解菲的最适温度为30℃,最适pH为7.对降解途径的初步研究显示,该菌株通过水杨酸途径降解菲.克隆了编码芳香烃双加氧酶α亚基的基因phdA,它与菌株Sphingomonas sp.P2、Sphingobium yanoikuyae B1、Sphingomonas sp.ZP1中phdA的同源性分别为97.9%、98%和100%,表明该基因具有保守性.图6参16     

Zooplankton diversity and physico-chemical conditions in three perennial ponds of Virudhunagar district, Tamilnadu

Plankton diversity and physico-chemical parameters are an important criterion for evaluating the suitability of water for irrigation and drinking purposes. In this study we tried to assess the zooplankton species richness, diversity and evenness and to predict the state of three perennial ponds according to physico-chemical parameters. A total of 47 taxa were recorded: 24 rotifers, 9 copepods, 8 cladocerans, 4 ostracods and 2 protozoans. More number of zooplankton species were recorded in Chinnapperkovil pond (47 species) followed by Nallanchettipatti (39 species) and Kadabamkulam pond (24 species). Among the rotifers, Branchionus sp. is abundant. Diaphanosoma sp. predominant among the cladocerans. Among copepods, numerical superiority was found in the case of Mesocyclopes sp. Cypris sp. repeated abundance among ostracoda. Present study revealed that zooplankton species richness (R1 and R2) was comparatively higher (R1: 4.39; R2: 2.13) in Chinnapperkovil pond. The species diversity was higher in the Chinnapperkovil pond (H': 2.53; N1: 15.05; N2: 15.75) as compared to other ponds. The water samples were analyzed for temperature, pH, electrical conductivity alkalinity salinity, phosphate, hardness, dissolved oxygen and biological oxygen demand. Higher value of physico-chemical parameters and zooplankton diversity were recorded in Chinnapperkovil pond as compared to other ponds. The zooplankton population shows positive significant correlation with physico-chemical parameters like, temperature, alkalinity phosphate, hardness and biological oxygen demand, whereas negatively correlated with rainfall and salinity. The study revealed that the presence of certain species like, Monostyla sp., Keratella sp., Lapadella sp., Leydigia sp., Moinodaphnia sp., Diaptomus sp., Diaphanosoma sp., Mesocyclopes sp., Cypris sp. and Brachionus sp. is considered to be biological indicator for eutrophication.     

Studies on influence of natural biowastes on cellulase production by Aspergillus niger

The objective of this study was to determine the influence of natural biowaste substrates such as banana peel powder and coir powder at varying environmental parameters of pH (4-9) and temperature (20-50 degrees C) on the cellulase enzyme production by Aspergillus niger. The cellulase enzyme production was analyzed by measuring the amount of glucose liberated in IU ml(-1) by using the dinitrosalicylic acid assay method. The substrates were pretreated with 1% NaOH (alkaline treatment) and autoclaved. The maximum activity of the enzyme was assayed at varying pH with temperatures being constant and varying temperatures with pH being constant. The highest activity of the enzyme at varying pH was recorded at pH 6 for banana peel powder (0.068 +/- 0.002 IU ml) and coir powder (0.049 +/- 0.002 IU ml(-1)) and the maximum activity of the enzyme at varying temperature was recorded at 35 degrees C for both banana peel powder (0.072 +/- 0.001 IU ml(-1)) and coir powder (0.046 +/- 0.003 IU ml(-1)). At varying temperatures and pH the high level of enzyme production was obtained at 35 degrees C and pH 6 by using both the substrates, respectively. However among the two substrates used for the production of cellulases by Aspergillus niger banana peel powder showed maximum enzymatic activity than coir powder as substrate.     

Effects of cadmium on metallothionein and histology in gills of tilapia (Oreochromis niloticus) at different salinities

The objectives of this study were to evaluate the effects of sublethal cadmium concentrations on the levels of cadmium, metallothionein (MT) and histological changes in gills of East Java strain tilapia (Oreochromis niloticus) at different salinity levels. The levels of cadmium in control gills were not significantly different at 0, 5 and 10 practical salinity unit (PSU). The cadmium concentrations in gills of cadmium-exposed fish were significantly higher at 0 PSU than at 5, 10 and 15 PSU. The MT concentrations in control gills were not significantly different at 0, 5, and 10 PSU. The MT concentrations of cadmium-exposed fish were significantly higher than those in respective control groups at 0, 5 and 10 PSU. Significant gill damage occurred in fish exposed to cadmium at lower salinity. The epithelial lifting was noted at gills of fish exposed to 2.5 mg/L of cadmium at 0 PSU, and telangiectasis was observed at gills exposed to 5 mg/L of cadmium at 0 PSU. The level of gill damage decreased with increasing salinity of media. The increased MT and histological changes in gills of our findings could be a protective response of animals to toxic effect of cadmium.     

Studies on seed germination and growth in weed species of rice field under salinity stress

An investigation was made to see the salt tolerance of 10 weed species of rice. Properly dried and treated seeds of weed species were placed on 9 cm diameter petridishes lined with Whatman No. 1 filter paper under 6 salinity regimes, viz. 0 (control), 4, 8, 16, 24 and 32 dS m(-1). The petri dishes were then kept in germinator at 25 +/- 1.0 degrees C and 12 hr light. The number of germinated seeds were recorded daily. The final germination percentage, germination index (GI), seedling vigour index, mean germination time and time for 50% germination were estimated. Root and shoot lengths of the weed seedlings were measured at 20 days after salt application and relative growth values were calculated. Results revealed that salinity decreased final germination percentage, seed of germination as measured by GI, and shoot and root length in all the species. Germination of most of the weed seeds was completely arrested (0) at 32 dS m(-1) salinity except in E. colona (12%) and C. iria (13.9%). The species C. iria, E. colona, J. linifolia and E. crusgalli showed better germination (above 30%) upto 24 dS m(-1) salinity level and were regarded as salt-tolerant weed species. J. linifolia, F. miliacea, L. chinensis and O. sativa L. (weedy rice) were graded as moderately tolerant and S. zeylanica, S. grosus and C. difformis were regarded as least tolerant weed species.     

Effect of phosphogypsum amendment on soil physico-chemical properties, microbial load and enzyme activities

Phosphogypsum (PG) is produced as a solid waste from phosphatic fertilizer plants. The waste slurry is disposed off in settling ponds or in heaps. This solid waste is now increasingly being used as a calcium supplement in agriculture. This study reports the effectof PG amendmenton soil physico chemical properties, bacterial and fungal count and activities of soil enzymes such as invertase, cellulase and amylase over an incubation period of 28 days. The highest mean percent carbon loss (55.98%) was recorded in 15% PG amended soil followed by (55.28%) in 10% PG amended soil and the minimum (1.68%) in control soil. The highest number of bacterial colonies (47.4 CFU g(-1) soil), fungal count (17.8 CFU g(-1) soil), highest amylase activity (38.4 microg g(-1) soil hr(-1)) and cellulase activity (38.37 microg g(-1) soil hr(-1)) were recorded in 10% amended soil. Statistically significant difference (p<0.05) has been recorded in the activities of amylase and cellulase over the period of incubation irrespective of amendments. Considering the bacterial and fungal growth and the activities of the three soil enzymes in the control and amended sets, it appears that 10% PG amendment is optimal for microbial growth and soil enzyme activities.     

Effects of salinity and dietary DHA (22:6n - 3) content on lipid composition and performance of Penaeus kerathurus postlarvae

A two-way ANOVA experiment was designed to study the effects of salinity and dietary docosahexaenoic acid (DHA; 22:6n-3) on lipid composition and performance of postlarvae from the marine shrimp Penaeus kerathurus (Forskål, 1775). Shrimp were reared from 1- to 8-d-old postlarvae at 35 and 25‰S with Kelko-enriched Artemia sp. [20.0 μg (n-3)?HUFA mg^?1 dry weight; 9.1 μg DHA mg^?1 dry weight] and nonenriched Artemia sp. [14.2 μg (n-3)?HUFA mg^?1 dry weight; 0.3 μg DHA mg^?1 dry weight]. Dietary DHA content did not affect either total length or survival but influenced the nutritional status represented by condition indices (triacylglycerol/total polar lipid and triacylglycerol/free cholesterol) of 8-d-old postlarvae at the end of the experiment. Culture salinity affected final total length and condition indices but did not show any effect on survival in the different experimental treatments. The interaction of dietary DHA and culture salinity was not significant for total length and survival but was significant for both condition indices used. P. kerathurus 8-d-old postlarvae showed better growth, survival and nutritional condition when reared at 35‰S and when fed on Kelko-enriched Artemia sp., but the differences with postlarvae from other treatments were very poorly marked. The results demonstrate that 8-d-old postlarvae may have sufficiently developed osmoregulatory capabilities to resist 25‰S under good conditions, although (n-3) HUFA-enriched diets may also enhance osmotic stress resistance, general performance and disease resistance.     

Herring (Clupea harengus) filter-feeding in the dark

The behaviour of juvenile herring (Clupea harengus L.) feeding on Artemia sp. nauplii in both light and dark was recorded using an infra-red-sensitive television-recording technique. In the light, two modes of feeding were observed, particle biting and filtering, but in the dark only filtering was observed. Marked differences in swimming behaviour were seen between light and dark. In the light, the fish continued to school while feeding in both modes; in the dark, the school dispersed, the fish swam slower in tight circular paths and fed only by filtering. In the dark, filtering fish swam faster (0.11 m s^-1) than non-filtering fish (0.07 m s^-1). In the light, no difference in speed was measured between filtering and non-filtering fish (0.34 m s^-1). Owing to the lower filtering speeds in the dark, the removal rate of nauplii from the water was much lower than in light, except in the highest prey concentrations. This suggests that if night-time filter-feeding takes place in the sea, it will be of importance only when exploiting dense patches of food.     

Foraging behavior in early Atlantic cod larvae (Gadus morhua) feeding on a protozoan (Balanionsp.) and a copepod nauplius (Pseudodiaptomussp.)

Food limitation is likely to be a source of mortality for fish larvae in the first few weeks after hatching. In the laboratory, we analyzed all aspects of foraging in cod larvae (Gadus morhua Linnaeus) from 5 to 20 d post-hatching using protozoa (Balanion sp.) and copepod nauplii (Pseudodiaptomus sp.) as prey. A camera acquisition system with two orthogonal cameras and a digital image analysis program was used to observe patterns of foraging. Digitization provided three-dimensional speeds, distances, and angles for each foraging event, and determined prey and fish larval head and tail positions. Larval cod swimming speeds, perception distances, angles, and volumes increased with larval fish size. Larval cod swam in a series of short intense bursts interspersed with slower gliding sequences. In 94% of all foraging events prey items were perceived during glides. Larval cod foraging has three possible outcomes: unsuccessful attacks, aborted attacks, and successful attacks. The percentage of successful attacks increased with fish size. In all larval fish size classes, successful attacks had smaller attack distances and faster attack speeds than unsuccessful attacks. Among prey items slowly swimming protozoans were the preferred food of first-feeding cod larvae; larger larvae had higher swimming speeds and captured larger, faster copepod nauplii. Protozoans may be an important prey item for first-feeding larvae providing essential resources for growth to a size at which copepod nauplii are captured. Received: 20 April 1999 / Accepted: 12 January 2000     

Combined effects of temperature and salinity on the complete development ofEurytemora velox (Crustacea: Calanoidea)

The calanoid copepodEurytemora velox was collected from rock pools at Castletown, Isle of Man, UK. Its optimum environmental requirements, particularly temperature and salinity, were determined, with a view to its possible future use as living food in intensive fish and shellfish farming. The species was cultured in 21 different temperature and salinity combinations. Investigations covered a period of two years from December 1983 to December 1985. Complete development from hatching to adult stage was followed in 21 temperature and salinity combinations. Nauplii suffered relatively high mortalities, indicating the sensitivity of this development stage to variations in temperature and salinity. Highest nauplii survival was observed in the combinations 15°C with 25 and 20 S and 20°C with 20 S, the highest copepodite survival at 10°C and 20 S. Lower salinities were tolerated better at higher temperatures and higher salinities at lower temperatures. Development time varied with the temperature and salinity combinations. Lower salinities at the lower temperatures of 10° and 15°C and both lower and higher salinities at 20°C prolonged development, particularly of the naupliar stage. Highest Q₅ values (i.e., rate of change of development with a 5 C° increase in temperature) were recorded for the naupliar stage. Statistical analysis indicated that salinity influences the survival of both nauplii and copepodites; however, this effect is not linear.     

Degradation characteristics and metabolic pathway of 4-nitrophenol by a halotolerant bacterium Arthrobacter sp. CN2

Arthrobacter sp. strain CN2, capable of degrading 4-nitrophenol, was isolated from activated sludge. Degradation of 4-nitrophenol was optimized at pH 7.7, 30?°C, and 0.53% of glucose. Salt tolerance of 4-nitrophenol degradation was as high as 6% (w/v). Several biodegradation intermediates were identified and quantified by high-performance liquid chromatography and mass spectrometry. 4-Nitrocatechol is involved in the degradation of 4-nitrophenol by CN2. Scale-up of 4-nitrophenol degradation was conducted in a bioreactor with different salinity. When the salinity was below 7%, the degradation rate of 4-nitrophenol was above 90% (100 mg L^?1, 3 L).     

Combined effects of heavy metal (Hg) concentration and algal (Chlorella vulgaris) food density on the population growth of Brachionus calyciflorus (Rotifera: Brachionidae)

The combined effects of two food levels (0.5 x 10(6) and 1.5 x 10(6) cells ml(-1) of Chlorella vulgaris) and five concentrations (0, 0.000625, 0.00125, 0.0025, 0.005 mg l(-1) of HgCl2) of mercury on the population growth of the rotifer Brachionus calyciflorus was evaluated. The growth experiments were conducted for 18 days at 23+/-1 degrees C under continuous fluorescent illumination. For each food level - heavy metal combination, we maintained 3 replicates. Our data showed that regardless of food level, increase in the heavy metal concentration in the medium resulted in decreased population growth of B. calyciflorus. At any given heavy metal concentration, B. calyciflorus grown under higher food levels had higher population abundance. The rate of population increase was significantly influenced by both the heavy metal concentration and the algal level. The highest population growth rate (0.435+/-0.003 per day) was observed in controls at 1.5 x 10(6) cells ml(-1). The results of this study were discussed in relation to the protective role of algal density against heavy metal toxicity.     

投饵养鱼对潘家口水库藻类生长影响的围隔试验研究

设置投饵、投饵单养花鲢（Aristichthisnobilis）、投饵单养鲤鱼（Cyprinuscarpio）、投饵混养花鲢和鲤鱼以及空白对照5个围隔及水库背景水体,研究投饵养鱼对潘家口水库浮游藻类生长和群落结构的影响。结果发现,试验期间水库水体和空白对照围隔中总藻平均密度分别为2.60×10^6L^-1和3.38×10^6L^-1,均以蓝藻和绿藻为主且基本保持稳定状态;投饵能有效促进藻类生长,只投饵围隔中总藻平均密度达到1.17×10^8L^-1,藻种组成向单一鱼腥藻（Anabaena）方向演替;养殖花鲢抑制藻的生长,参照只投饵围隔,抑制率接近80%,绿藻占据极大优势,其中栅藻（Scenedesmussp）、芒锥藻（Errerellabornhemiensisconr）和空星藻（Coelastrumsp）成为优势藻种;投饵养殖鲤鱼也能有效降低藻生物量,总藻平均密度为4.07×10^7L^-1,绿藻占总藻的比例由27%增加到95%,其中空星藻和小球藻（ChlorellavulgarisBeij）为优势藻种;投饵混养花鲢和鲤鱼围隔中总藻密度稍高于单养花鲢和鲤鱼围隔,为4.37×10^7L^-1,栅藻和鱼腥藻为优势藻种。     

Characterization of protease from Alcaligens faecalis and its antibacterial activity on fish pathogens

Alcaligens faecalis AU01 isolated from seafood industry effluent produced an alkaline protease. The optimum culture conditions for growth as well as enzyme production were 37 degrees C and pH 8. The partially purified protease had specific activity of 9.66 with 17.77% recovery with the molecular weight of 33 kDa and it was active between 30-70 degrees C and optimum being at 55 degrees C and pH 9. The enzyme retains more than 85% activity at 70 degrees C and 78% even at pH 10. The enzyme inhibited the growth of fish pathogens such as Flavobacterium sp., Pseudomonas fluorescens, Vibrio harveyi, Proteus sp. and Vibrio parahaemolyticus. From the present study it can be concluded that Alcaligens faecalis AU01 has the potential for aquaculture as probiotic agent and other several applications.     

Infection and immunization trials of Asian seabass (Lates calcarifer) against fish pathogen Vibrio anguillarum

Vibriosis is one of the most prevalent fish diseases caused by bacteria belonging to the genus Vibrio sp. Fish disease will be controlled by proper vaccination trials and maintenance of fish form. Pathogenicity for Asian seabass (Lates calcarifer) against V anguillarum results in necrosis and haemorrhagic areas near the base of fins, exopthalmia and ulcers on the skin surface. Around 50, 100, 200 microl of formalin killed bacterial cells were injected (Intraperitoneal) to three different size (5-10, 20-30, 35-50 g) of seabass fishes respectively and control sere as saline were maintained separately. The Relative Percentage Survival (RPS) for vaccinated fishes was 60, 75, and 62.5 respectively and the vaccinations for 20-30 g fishes stay alive. These results stated that the vaccination for fishes with 20-30 g size may fabricate good immune response.     

DLL-1菌对甲基对硫磷农药的降解作用及其降解机理

研究了假单胞菌 (DLL - 1)在水溶液介质中降解甲基对硫磷的性能与影响因素及其降解机理。结果表明 ,当菌体浓度为 10 5个·ml-1时 ,即发生快速的降解作用 ,3h时 ,降解率达 88.5 %。在pH为 5 .0、7.0和 9.0条件下 ,DLL - 1菌均产生对甲基对硫磷农药的高效降解作用。采用气相色谱 -质谱联用技术与离子色谱法 ,测定了DLL - 1菌 -甲基对硫磷作用过程产生的中间产物和最终产物 ,表明对硝基苯酚为主要中间产物 ,且DLL - 1菌能将其进一步降解为NO2 -和NO3 -。     

Experimental evidence of chemokinesis in newly hatched cod larvae (Gadus morhua L.)

The swimming behaviour of laboratory-reared newly hatched cod larvae (Gadus morhua L.) was observed in a control solution of artificial seawater and in seven solutions, each with a different concentration of arginine (10⁹ to 10^-3 M). The behaviour of 20 larvae was analysed in each of the eight solutions; the individual observation time was 1 min. Individual movements were recorded on video and analyzed using a computer-assisted program. The larvae swam in straight lines (a trajectory), rested, moved and started swimming again. For the parameters analyzed, i.e., number of movements, angle between successive trajectories and straightness index, there was no significant difference between the behaviour of the larvae in the different solutions. However, for the larvae in 10^-5, 10^-4 and 10^-3 M arginine solutions, the analyzed parameters, i.e., time active, frequency of trajectories (number of movements exceeding body length), distance swum min^-1, length of individual trajectories and trajectory velocity, were all significantly lower than for the larvae in the control solution of artificial seawater and for larvae in the solutions of 10^-9, 10^-8, 10^-7 and 10^-6 M arginine. The results show that the mean distance swum by cod larvae min^-1 was two to five times longer in artificial seawater without arginine and in the four lower concentrations of arginine than in the three higher concentrations. Scanning micrographs show that newly hatched (pre-feeding) cod larvae possess olfactory organs. It seems reasonalbe to assume that the observed changes in swimming behaviour are mediated by the olfactory sense and are important in the feeding strategy of cod larvae. We suggest that the observed behaviour increases the probability of the larvae localizing patches of prey organisms and remaining in the patch once they have found it. The results show that chemokinesis is a mechanism by which the spatial distribution of fish larvae will be correlated with their prey.