On a method of determination of chlorophyll in cells of algae collected on a membrane filter. [Translation from: Trudy Instituta Biologii Vnutrennykh Vodnany 11(14), 198-202, 1966.]

The original method, proposed by Yentsch (1957), of determination of chlorophyll directly in the cells, attracts attention by its simplicity. In order to measure the content of chlorophyll by this method, a determined volume of suspension of algae is filtered through a membrane filter. The latter is dried a little, clarified by immersion oil, clamped between two glasses, and spectrophotometrized. Extinction is read off at , wavelengths equal to 670 millimicrons (around the maximum absorption of chlorophyll a in the cell) and 750 millimicrons (correction for non- specific absorption and dispersion of light by particles of the preparation). The method of Yentsch was employed by the authors for determination of chlorophyll-a in samples of phytoplankton. They conclude that in spite of the simplicity and convenience of determination the method must be applied sufficiently carefully. It is more suitable for analysis of cultures of algae, where, non-specific absorption of light is insignificant.

On the structure of the seminal receptacle in cyclopids (Copepoda, Cyclopoida). [Translation from: Informatsionnyi Byulleten Biologiya Vnutrennikh Vod (6) 26-31, 1970.]

The seminal bag, or seminal receptacle, forms a characteristic organ of cyclopids, serving for retention of the sperms discharged from the spermatophores. The structure of the seminal receptacle, more precisely its form, is fairly widely used in diagnosis and undoubtedly can be more widely applied in the systematics of the group. Within the limits of the family Cyclopidae it is possible to distinguish crustaceans with three basic types of seminal bag. The differences consist of the position which this organ occupies in the genital segment. of one species, we carried out a series of observations on its formation in ontogenesis and during the life of the adult stage. As material for observation the study used laboratory cultures of three species; Acanthocyclops americanus (Marsh) from the plankton of the Moscow River, Cyclops vicinus Uljan and Mesocyclops leuckarti Glaus from the plankton of the channel section of the upper part of the Gorkovsk reservoir. The author concluded that the irreversibility of the changes in the seminal receptacle presents the possibility of utilising this structure as one of the indicators of the growth of the individual.

Investigations on saprolegniosis. II. Study on the process of penetrations showing evidence of a proteolytic enzyme and histopathological aspects [Translation from: Riv.ital.Piscicolt.Ittiopatol. 11(4), 109-117, 1976]

The study of enzymatic activity is of great importance in the immunology of fungi. Indeed, knowledge of biological activity of antigenic structures is important for the elucidation of host-parasite relations as well as in the search for a taxonomic factor permitting differential diagnoses. The authors used Saprolegnia cultures to analyse soluble antigenic fractions arising from the mycelium of cultures of 4 species of Saprolegnia, which are found most frequently in the parasitic state on fish: S. parasitica, S. ferax, S. delica, S. diclina. The authors conclude that in the study of saprolegniasis, the enzymatic approach affords new elements for the examination of the etiology of fungi as well as an element of gravity concerning the biochemical modifications necessary to the change of saprophytism to parasitism.

Lethal Parasites in Oysters from Coastal Georgia with Discussion of Disease and Management Implications

Extensive mortalities of oysters, Crassostrea virginica, occurred from 1985 through 1987 in coastal waters of Georgia. Fluid thioglycolate cultures of oysters collected from 16 of 17 locations revealed infections by the apicomplexan parasite Perkinsus marinus. An ascetosporan parasite, Haplosporidium nelsoni, was also observed in histopathological examination of oysters from 4 of the locations. While the range of H. nelsoni currently is recognized as the east coast of the United States from Maine to Florida, this is the first report of the parasite in Georgia waters. This paper documents the occurrence of these two lethal parasites in oysters from coastal waters of Georgia, along with potential disease and management implications. Results of an earlier independent and previously unpublished survey are also discussed which document the presence of P. marinus in Georgia as early as 1966.

Increased toxicity of Karenia brevis during phosphate limited growth: ecological and evolutionary implications

Karenia brevis is the dominant toxic red tide algal species in the Gulf of Mexico. It produces potent neurotoxins (brevetoxins [PbTxs]), which negatively impact human and animal health, local economies, and ecosystem function. Field measurements have shown that cellular brevetoxin contents vary from 1–68 pg/cell but the source of this variability is uncertain. Increases in cellular toxicity caused by nutrient-limitation and inter-strain differences have been observed in many algal species. This study examined the effect of P-limitation of growth rate on cellular toxin concentrations in five Karenia brevis strains from different geographic locations. Phosphorous was selected because of evidence for regional P-limitation of algal growth in the Gulf of Mexico. Depending on the isolate, P-limited cells had 2.3- to 7.3-fold higher PbTx per cell than P-replete cells. The percent of cellular carbon associated with brevetoxins (%C-PbTx) was ~ 0.7 to 2.1% in P-replete cells, but increased to 1.6–5% under P-limitation. Because PbTxs are potent anti-grazing compounds, this increased investment in PbTxs should enhance cellular survival during periods of nutrient-limited growth. The %C-PbTx was inversely related to the specific growth rate in both the nutrient-replete and P-limited cultures of all strains. This inverse relationship is consistent with an evolutionary tradeoff between carbon investment in PbTxs and other grazing defenses, and C investment in growth and reproduction. In aquatic environments where nutrient supply and grazing pressure often vary on different temporal and spatial scales, this tradeoff would be selectively advantageous as it would result in increased net population growth rates. The variation in PbTx/cell values observed in this study can account for the range of values observed in the field, including the highest values, which are not observed under N-limitation. These results suggest P-limitation is an important factor regulating cellular toxicity and adverse impacts during at least some K. brevis blooms.

Microbial community analysis of Acropora palamata mucus swabs, water and sediment samples from Hawksnest Bay, St. John, U.S. Virgin Islands

Colonies of the scleractinian coral Acropora palmata, listed as threatened under the US Endangered Species Act in 2006, have been monitored in Hawksnest Bay, within Virgin Islands National Park, St. John, from 2004 through 2010 by scientists with the US Geological Survey, National Park Service, and the University of the Virgin Islands. The focus has been on documenting the prevalence of disease, including white band, white pox (also called patchy necrosis and white patches), and unidentified diseases (Rogers et al., 2008; Muller et al., 2008). In an effort to learn more about the pathologies that might be involved with the diseases that were observed, samples were collected from apparently healthy and diseased colonies in July 2009 for analysis. Two different microbial assays were performed on Epicentre Biotechnologies DNA swabs containing A. palmata coral mucus, and on water and sediment samples collected in Hawksnest Bay. Both assays are based on polymerase chain reaction (PCR) amplification of portions of the small rRNA gene (16S). The objectives were to determine 1) if known coral bacterial pathogens Serratia marcescens (Acroporid Serratiosis), Vibrio coralliilyticus (temperature-dependent bleaching, White Syndrome), Vibrio shiloi (bleaching, necrosis), and Aurantimonas coralicida (White Plague Type II) were present in any samples, and 2) if there were any differences in microbial community profiles of each healthy, unaffected or diseased coral mucus swab. In addition to coral mucus, water and sediment samples were included to show ambient microbial populations. In the first test, PCR was used to separately amplify the unique and diagnostic region of the 16S rRNA gene for each of the coral pathogens being screened. Each pathogen test was designed so that an amplified DNA fragment could be seen only if the specific pathogen was present in a sample. A positive result was indicated by bands of DNA of the appropriate size on an agarose gel, which separates DNA fragments based on the size of the molecule. DNA from pure cultures of each of the pathogens was used as a positive control for each assay.

The bacterial flora and their possible association with spoilage in a variety of fresh-water fish: Cyprinus carpio. var. communis

The bacteria from a variety of fresh-water fish, Cyprinus carpio. var. communis, showed the presence of micrococci, Gram positive and Gram negative rods. These have been characterized as far as was possible. Of thirty-eight strains of bacteria used, only six strains were considered as causing spoilage of fish flesh in experiments where flesh was incubated with individual cultures of the bacteria. These six strains had been found on the surface and/or intestine of the fish and support the suggestions that, after death, invasion of flesh by bacteria from the surface and intestine could be the cause of bacterial spoilage of fish.

Behaviour of marine microflora towards antibiotics 1. Sensitivity towards chlortetracycline (CTC)

The sensitivity of 61 cultures of bacteria isolated from fish towards chlortetracycline (CTC) at 5 ppm and 20 ppm levels has been determined on two solid media: sea water agar (SWA) and a distilled water based medium consisting of peptone, beef extract, glucose and NaCI (PBGA). The cultures employed consisted of (i) gram-negative rods of marine origin (Achromobacter, Pseudomonas, Vibrio and Flavobacterium) and (ii) gram positive organisms (Micrococci and Corynebacterium). Depending on the inhibition zone diameter, the order of CTC sensitivity was found to be Pseudomonasof 5 and 20 ppm. The SWA enhanced the development of resistance in majority of the cultures as compared to PBGA. PBG broth containing CTC at less than 10 ppm exerted a bacteriostatic effect only while that containing 10 ppm or more exerted a bactericidal effect on Achromobacter, Pseudomonas and Micrococci cultures. The significance of these results in fish preservation by CTC is discussed.

Salt tolerance of bacteria isolated from tropical marine fish and prawn

Salt tolerance of selected cultures of Pseudomonas, Moraxella, Vibrio, Micrococcus, Acinetobacter and Flavobacteria/ Cytophaga was determined. More than 80% of the cultures belonging to each of the above genera, were capable of growth in presence of 1.5 to 3.5% salt (NaCl) and at least 25 to 30% of the cultures in each group required 1.5 to 3.5% salt for growth. 40% each of Pseudomonas and Vibrio strains and 30% each of Moraxella, Micrococcus and Flavobacteria/Cytophaga strains tolerated 10% salt. Majority of the cultures belonging to the genera Pseudomonas, Vibrio, Moraxella, Micrococcus, Acinetobacter and Flavobacteria/Cytophaga were slightly halophilic (2 to 5% salt tolerant), about 25% especially of Micrococcus spp. moderately halophilic (5 to 20% salt tolerant) and none from Pseudomonas, Vibrio, Moraxella, Acinetobacter and Flavobacteria/Cytophaga spp. extremely halophilic (20 to 32% salt tolerant).

Effects of furanace on Brachionus

Tiger prawn P.monodon) larvae utilize Brachionus a rotifer, as food in the Zoea 3 and mysis stages when they change from an herbivorous to an omnivorous diet. The present work aims to show the effects of furanace on the population growth of Brachionus. Cultures of Brachionus were obtained and fed with Chlorella at a density of 1-2x10 SUP-6 cells/ml. Five liters of the culture water were placed in each of 4 white, circular, 152x304 mm plastic basins. The mean initial densities of the rotifer ranged from 26 . 5 to 38 . 5 individuals/ml. The concentrations of furanace were 0, 1, 2 and 3 mg /l. The cultures were vigorously aerated. Population growth was observed after 3, 6, and 9 hours of exposure. The cultures were thoroughly mixed before samples were taken to ensure an almost equal distribution of the rotifers in the water. To facilitate the counting of the rotifer, one drop of Lugol's solution was added to each sample. This immobilizes the rotifer as well as stops further reproduction. Individuals with only the lorica left or with badly deformed lorica were considered dead. Population counts were done using a Sedgwick-Rafter counting chamber. Among the different durations of exposure, the percentage survival of the populations in the furanace baths were highest after 3 hr. There were slight increases in the control and 2 mg/l and slight decreases in 1 and 3 mg/l. The differences in the mean densities are statistically insignificant at . 01 significance level. After a 6-hr exposure, the control population reached its peak density with a survival of 89%. Populations in furanace baths decreased to 88 . 5% in both 2 and 3 mg /l followed closely by 87% in 1 mg/l. Again, no statistical differences exist among all the levels. The mean percentage survival in 1 and 2 mg/l increased (89% and 91%, respectively) after a 9-hr expsoure, while those in the control and 3 mg/l decreased to 86 . 5% and 88 . 25%, respectively. There were no marked differences in appearance noted among the individuals in furanace baths and those in the control.

Effect of different light intensities on the growth of the diatom Chaetoceros calcitrans

Batch cultures of C. calcitrans were maintained indoors at a temperature range of 21 to 25 C and continuously illuminated by 40-watt daylight fluorescent lights. Cultures were exposed to 5 different intensities from 200 to 25,000 lux.Population counts show that light intensity affects growth and reproduction of the algae cultivated. A comparison of population peak growths showed cultures illuminated by 12,000 lux to have higher cell counts than those exposed to higher or lower light intensities.

Viability of frozen algae used as food for larval penaeids

Freezing with added chemicals as flocculants and protectants was assessed as a means of preserving stock cultures of 4 algal species used for larval penaeid food Chaetoceros calcitrans, Skeletonema costatum, Tetraselmis chuii and Isochrysis galbana . The maximum storage effectability of the preservation techniques for each species was also determined.

In vitro effects of fungicides on the fungus Haliphthoros philippinensis

Pure cultures of Haliphthoros philippinensis, isolated from infected Penaeus monodon larvae, were exposed for 24 hours to varying concentrations of antifungal agents. The efficiency of each agent to inhibit sporulation and mycelial growth was measured. Effects on P. monodon eggs and larvae were also investigated. It is concluded that preliminary bioassay of larval tolerance to the suggested effective doses should always be made prior to prophylaxix or therapeutic applications.

Viability of frozen algae used as food for larval penaeids

Freezing with added chemicals as flocculants and protectants was assessed as a means of preserving stock cultures of 4 algal species used for larval penaeid food Chaetoceros calcitrans, Skeletonema costatum, Tetraselmis chuii and Isochrysis galbana. The maximum storage effectability of the preservation techniques for each species was also determined.

The changes in various iron compounds in culture media for algae. [Translation from: Archiv fur Hydrobiologie Supplemente Band 38(1/2) 151-169, 1970.]

The purpose of this work is a contribution to the quantitative record of the use of iron by planktonic algae. Preliminary experiments with Chlorella to determine the rate of iron intake in the presence of inorganic sources of iron did not produce the desired result. The crucial point of this work is the investigation of the influence of various external factors on the stability of FeEDTA (FeEDTA = Ferric(III)-compound of ethylene-diamine tetra-acetic acid), since this compound appears to be particularly well-suited as a source of iron for planktonic algae (e.g. TAMIYA et al. 1953). Cultures of Chlorella fusca in a light thermostat were used in experimental research. Methods and results are discussed.