Effects of probiotics and antibiotics on biofilms

Experiments were conducted to study the effects of probiotics and antibiotics on 3-months old biofilms formed on three different substrates, namely glass, granite and fiberglass reinforced plastic. The variations in heterotrophic bacterial populations associated with biofilms were monitored for a period of one month after one-time application of the probiotic Biogreen (Gee Key Marine Pvt. Ltd, Chennai) at 0.2 mg lˉ¹ and the antibiotic tetracycline at 1.0 mg lˉ¹. The variations in heterotrophic bacterial populations associated with biofilms were compared with that of control to analyse the effects of probiotics and antibiotics on biofilms. The observations showed that the biofilm formation and succession in aquatic environment is substrate dependent and that the resistance to antibiotics also depends on the substrate. The probiotics did not show significant effect on biofilms.

Studies on the chemical control of psychrophilic bacterial spoilage of fish. 2 - The effect of antibiotics on the growth of psycrophilic bacteria isolated from marine fish

Among the various antibiotics tried, tetracyclines particularly chlorotetracycline (CTC), chloramphenicol and chlorostrep were found to be fairly effective at 8 and 10 p.p.m. levels. The order of sensitivity to CTC among the six genera studied was found to be Achromobacterantibiotics than species belonging to other genera. Among the resistant organisms, Achromobacter superficialis was found to be most resistant and only CTC at 10 p.p.m. level could partially inhibit this culture. The useful combinations of antibiotics for commercial application are discussed.

Use of antibiotics in the preservation of prawn

Effect of incorporating chlorotetracycline (CTC) in ice up to 5 ppm level on the keeping quality of prawns has been studied. A shelf life extension by nearly six days is obtained for the CTC-iced sample over the control. The headless prawns absorbed greater amounts of CTC than whole prawns during storage in CTC-ice. Traces of the antibiotic are found in the muscle of the CTC-iced prawns even after cooking for one hour. The cause of destruction of CTC when used for prawn preservation is discussed.

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 Pseudomonas

The trouble with antibiotics and pesticides is...

The paper discusses the output of the meeting on the use of chemicals in aquaculture in Asia. The effects of chemical use on cultured stocks in the farm, the immediate environment through discharges and effluents, surrounding areas, farm staff, consumers and drug resistance organisms are also discussed. It also shows how an antibiotic-resistant microorganism develops as the result of indiscriminate use of antibiotics.

Live-box experiments with anchovetas, Cetengraulis mysticetus, in the Gulf of Panama

ENGLISH: Extensive live-box experiments were conducted during 1958, 1959, and 1960 to test the efficacy of various tags and marks for use with anchovetas, and to test the effects of the use of anesthetics and antibiotics with the tagging. A total of 12,767 fish was involved in 72 experiments during the three years. Daily records of the mortalities and shedding were kept. Slightly over 7 months was the longest period of time any of the experiments was maintained. SPANISH: Durante 1958, 1959 Y 1960 se hicieron extensos experimentos en viveros para probar la eficacia del uso de varias marcas en las anchovetas y para ensayar los efectos del empleo de anestésicos y antibióticos en el momento de la operación de marcación. En estos tres años se utilizó un total de 12,767 peces en 72 experimentos. Se hicieron anotaciones diarias de la mortalidad y del desprendimiento de las marcas. El período más largo de duración de un experimento sobrepasó ligeramente los 7 meses.

Environmental impact assessment of agrochemicals for fish production - a review paper

The environmental impact of agro-chemicals for fish production was extensively reviewed. The positive contribution of agro- chemicals and the devastating effect on aquaculture was x-rayed to alert users to this obvious environmental problem. Lime and fertilizers are commonly used in fish farming to increase pH of pond soil and water and to increase alkalinity and hardness, reduce humic acid content and to initiate primary and secondary productivity. Devastating effect of lime on environment is likely to be minimal. In the case of fertilizers, over utilization of this agro-chemical could impair water quality as phytoplankton bloom become excessive which consequently raises BOD. The use of Therapeutants in aquaculture was discovered to be more popular in Europe and North America than in the tropics (Africa). Commonly used therapeutants include antibiotics and antimicrobials. For fish pathology chemicals like formalin, potassium permanganate, Dipterex and malachite green are widely in use. Effluent from farms where these chemicals are commonly in use can distort the aquatic ecosystem. The changes in water quality, aquatic community structure and productivity caused by intensive aquaculture are typical of the impacts of pollution from a wide variety of sources like sewage, agricultural run-off and effluent discharges from industry

The role of probiotics in aquaculture in Nigeria - a review

Aquaculture is beset by many problems especially diseases caused by bacteria as the major deteriorating factors. The use of vaccines and antimicrobial agents have been centered on disease control, but are associated with problems The development of antibiotic resistance among the microorganisms have become a global concern as a result of indiscriminate use of antibiotics. Several alternative suggestions for disease prevention have been on probiotics for its efficacy, low cost, less side effects and accessible to farmers. Probiotics is gaining a high priority in the developed countries with the aim of replacing conventional drugs. The principal bacterial groups tested as probiotic bacteria in culture of shrimps, crabs, oysters, fish and humans are Vibrio, Pseudomonas, Bacillus, Bifidobacteria and several Lactobacilli. Experiments have mainly been conducted with fish larvae, adult fish, crustaceans and animals where significant reduction in mortalities has been obtained. The purpose of this review is to create awareness of the role of probiotics in disease control in aquaculture as alternative to antibiotics.

The potential for crop rotation in controlling diseases in shrimp culture

The use of antibiotics and other chemicals in controlling shrimp pathogens become ineffective as the strains grow more resistant to these chemicals. Moreover, the bacterial pathogen (Vibrio harveyi) produced biofilm coating that protects it from dying and disinfection procedures that are followed during pond preparation. Biological control is being considered as an alternative means of preventing shrimp disease outbreak. The main principle behind biological control is to enhance the growth of beneficial microorganisms which serve as antagonists or target pathogens. The paper discusses shrimp and tilapia crop rotation as a form of effective biological control, a technique which is already being practiced in Indonesia and the Philippines.

Saline-saturated DMSO-EDTA as a storage medium for microbial DNA analysis from coral mucus swab samples

The mucus surface layer of corals plays a number of integral roles in their overall health and fitness. This mucopolysaccharide coating serves as vehicle to capture food, a protective barrier against physical invasions and trauma, and serves as a medium to host a community of microorganisms distinct from the surrounding seawater. In healthy corals the associated microbial communities are known to provide antibiotics that contribute to the coral’s innate immunity and function metabolic activities such as biogeochemical cycling. Culture-dependent (Ducklow and Mitchell, 1979; Ritchie, 2006) and culture-independent methods (Rohwer, et al., 2001; Rohwer et al., 2002; Sekar et al., 2006; Hansson et al., 2009; Kellogg et al., 2009) have shown that coral mucus-associated microbial communities can change with changes in the environment and health condition of the coral. These changes may suggest that changes in the microbial associates not only reflect health status but also may assist corals in acclimating to changing environmental conditions. With the increasing availability of molecular biology tools, culture-independent methods are being used more frequently for evaluating the health of the animal host. Although culture-independent methods are able to provide more in-depth insights into the constituents of the coral surface mucus layer’s microbial community, their reliability and reproducibility rely on the initial sample collection maintaining sample integrity. In general, a sample of mucus is collected from a coral colony, either by sterile syringe or swab method (Woodley, et al., 2008), and immediately placed in a cryovial. In the case of a syringe sample, the mucus is decanted into the cryovial and the sealed tube is immediately flash-frozen in a liquid nitrogen vapor shipper (a.k.a., dry shipper). Swabs with mucus are placed in a cryovial, and the end of the swab is broken off before sealing and placing the vial in the dry shipper. The samples are then sent to a laboratory for analysis. After the initial collection and preservation of the sample, the duration of the sample voyage to a recipient laboratory is often another critical part of the sampling process, as unanticipated delays may exceed the length of time a dry shipper can remain cold, or mishandling of the shipper can cause it to exhaust prematurely. In remote areas, service by international shipping companies may be non-existent, which requires the use of an alternative preservation medium. Other methods for preserving environmental samples for microbial DNA analysis include drying on various matrices (DNA cards, swabs), or placing samples in liquid preservatives (e.g., chloroform/phenol/isoamyl alcohol, TRIzol reagent, ethanol). These methodologies eliminate the need for cold storage, however, they add expense and permitting requirements for hazardous liquid components, and the retrieval of intact microbial DNA often can be inconsistent (Dawson, et al., 1998; Rissanen et al., 2010). A method to preserve coral mucus samples without cold storage or use of hazardous solvents, while maintaining microbial DNA integrity, would be an invaluable tool for coral biologists, especially those in remote areas. Saline-saturated dimethylsulfoxide-ethylenediaminetetraacetic acid (20% DMSO-0.25M EDTA, pH 8.0), or SSDE, is a solution that has been reported to be a means of storing tissue of marine invertebrates at ambient temperatures without significant loss of nucleic acid integrity (Dawson et al., 1998, Concepcion et al., 2007). While this methodology would be a facile and inexpensive way to transport coral tissue samples, it is unclear whether the coral microbiota DNA would be adversely affected by this storage medium either by degradation of the DNA, or a bias in the DNA recovered during the extraction process created by variations in extraction efficiencies among the various community members. Tests to determine the efficacy of SSDE as an ambient temperature storage medium for coral mucus samples are presented here.

Survival of antibiotic resistant Pseudomonas strains in different types of water

A study was conducted to investigate the survival of five Pseudomonas strains resistant to antibiotics in different types of water. The selected Pseudomonas strains were designated as strain P1 (CT-29), strain P2 (CT-25), strain P3 (CT-36), strain P4 (CT-20) and strain P5 (CT-27) which were only recovered from farmed fishes. Six types of water viz., distilled water, saline water, tap water, deionized water, pond water and river water were used. Among these experimental waters, river water was found to be the most suitable for long-term survival of these strains. Deionized water did not support survival of all these Pseudomonas strains. Pond water, tap water and distilled water were moderately suitable for strain P1 and strain P4. Saline water was also found to be highly suitable for long-term survival in case of the strain P3 and moderately suitable for normal survival of strain P2 and strain P5.

Pathogenicity of Aeromonas sobria to Thai silver barb (Barbodes gonionotus Bleeker) and its sensitivity to some antibiotic agents

Five isolates of Aeromonas sobria, collected from the diseased fish were selected for detection the pathogenicity following water-born infection method on silver barbs (Barbodes gonionotus) at the selected exposure dose 2.5x10⁸ CFU/ml which was standardized by preliminary test. In the experimental condition lesion and mortality were found in fishes. Among the isolate, Ass17 Ass19, Ass31 and Ass36 were successfully infected 20-60% fishes. Another isolate Ass20 was found non-pathogenic. Drug sensitivity test was performed by six antibiotics viz. Oxytetracycline, Oxolinic acid, Chloramphenicol, Stilphamethozazole, Streptomycin, Erythromycin. All the isolates showed variable reaction patterns to antibiotics. Most of the isolates were found sensitive to Oxytetracycline (OT), Oxolinic acid (OA) and Chloramphenicol (C) but resistance to Erythromycin and Sulphamethoxazole (SXT). Isolate Ass31 found resistant to Oxolinic acid.

Bacterial flora associated with diseased fish and their antibiogram

Bacterial flora associated with tail rot/fin rot of Carassius auratus, Xiphophorus helleri and hemorrhagic ulcers of Clarias spp were studied. Sensitivity pattern of 33 isolates comprising Aeromonas spp, Pseudomonas spp and Gram-positive rods from diseased C. auratus, X. helleri and Clarias spp were screened against six broad-spectrum antibiotics viz. ciprofloxacin, chloramphenicol, co-trimoxazole, gentamycin, nitro-furantoin and oxytetracycline. Ciprofloxacin was the most effective in inhibiting bacteria at 0.05-0.10 µg/ml level. About 44% of Pseudomonas spp. was resistant to nitrofurantoin. Resistance to oxytetracycline was seen in 27% of Aeromonas spp Gram-positive rods were comparatively more resistant to antibiotics. The multiple antibiotic resistances were seen in 21% of the bacterial isolates of diseased fish.

Improved handling and preservation of freshwater giant prawn, Macrobrachium rosenbergii for producing safe and wholesome products

Studies were undertaken to evaluate the quality changes in freshwater giant prawn, Macrobrachium rosenbergii during various storage conditions of handling and preservation and producing safe and quality products. The samples kept in ice immediately after catch with head-on and head-less condition were found to be acceptable for 6 days and 7 days, respectively. Delaying of icing considerably shortened the shelf-life. The pH value increased from 6.36 to 8.0 after 10 days in ice. The initial average TVB-N value of sample increased from below 10 mg/100 g to 25 mg/100 g with the lapse of storage period. The Ca++ ATPase activity in presence of 0.1M KCl slightly decreased at the end of 10 days of ice storage. Immediately after harvest, initial aerobic plate count (APC) was 2.88x10^6 CFU/g which gradually increased to 1.12x10^8 CFU/g after 6 days in ice storage and showed early signs of spoilage. Initial bacterial genera in the prawn iced at 0 hours were comprised of Coryneform (22.21 %), Bacillus (7.40%), Micrococcus (11.11 %), Achromobacter (48.14%), Flavobacterium/Cytophaga (7.40%), Pseudomonas (3.70%) and Aeromonas (3.70%). During ice storage Coryneforms and Bacillus were always dominating along with less prominent ones - Micrococcus, Achromobacter and Flavobacterium. Studies were conducted on the stability of myofibrillar protein of M. rosenbergii under different storage and pH conditions. The influence of a wide range of pH on the remaining Ca++ ATPase activity of M. rosenbergii muscle myofibrils after storage at -20°C for 2 days, at 0°C for 2 days and at 35°C for 30 minutes demonstrated that ATPase activities were lower in acidic and alkaline pH regions and the activity remained relatively high. Mg++ ATPase activities both in presence and absence of Ca++ remained high at neutral pH compared to those of acidic and alkaline region. The solubility of myofibrillar protein decreased gradually both in acidic and alkaline pH regions. The study also examined the bacteriological quality of freshly harvested M. rosenbergii, pond sediment and pond water from four commercial freshwater prawn farms at Fulpur and Tarakanda upazilas in the district of Mymensingh. The study included aerobic plate count (APC), total coliform count, detection, isolation and identification of suspected public health hazard bacteria and their seasonal variation, salt tolerance test, antibiotic sensitivity test of the isolates and washing effect of chlorinated water on the bacterial load in the prawn samples. APC in sediment soil and water of the farm and gill and hepatopancreas of freshly harvested prawns varied considerably among the farms and between summer and winter season. The range of coliform count in water, gill and hepatopancreas ranged between 6 - 2.8x10^2 CFU/ml, 1.2x10^2 - 3.32x10^2 CFU/g and 1.43x10^2 - 3.89 x10^3 CFU/g, respectively. No coliform was detected in pond sediment sample. Suspected health hazard bacteria isolated and identified from pond sediment, water, gill and hepatopancreas included Streptococcus, Bacillus, Escherichia coli, Klebsialla, Salmonella, Staphylococcus, Pseudomonas and Aeromonas. Bacillus, Salmonella and Staphyloccus [sic], and were found to be highly salt tolerant and capable of growing at 10% NaCl. The antibiotic discs with different concentration of antibiotics were used for the sensitivity test. The organisms were found to be most sensitive against Tetracyclin and Gentamycin.