Achromobacter xylosoxidans na fibrose cística: infecção crônica e disseminação clonal

A mortalidade na Fibrose Cística (FC) é decorrente de infecções pulmonares causadas comumente por: Pseudomonas aeruginosa, Staphylococcus aureus e espécies do Complexo Burkholderia cepacia (CBc). Mais recentemente, tem sido observada a emergência de BGN-NF raros, como Achromobacter xylosoxidans, porém, sua prevalência, potencial de transmissão e significado clínico são desconhecidos. O objetivo deste trabalho foi verificar a ocorrência de colonização crônica por A. xylosoxidans e avaliar a possibilidade de transmissão cruzada entre os pacientes acompanhados em dois centros de referência na cidade do Rio de Janeiro. Foram incluídos 39 pacientes com FC, com pelo menos uma cultura positiva para o gênero Achromobacter spp., em um total de 897 analisadas, do período de janeiro de 2003 a dezembro de 2008. A frequência de isolamento de Achromobacter spp. nas culturas analisadas foi de 14,5% (130 em 897 culturas). A maioria (n=122; 93,8%) foi identificada como A. xylosoxidans por testes fenotípicos e pelo sequenciamento do gene rrs que codifica o 16S rRNA. A análise do polimorfismo genético dos isolados de A. xylosoxidans pela técnica de PFGE, mostrou 22 grupos clonais. Destes, sete foram compartilhados entre pacientes distintos sugerindo transmissão cruzada. Apenas o clone G foi amplamente disseminado entre 56,4% dos pacientes estudados, sugerindo a possibilidade de um surto. Os 15 clones restantes constituíram-se em clones exclusivos por pacientes. Os cinco pacientes colonizados cronicamente por A. xylosoxidans mostraram a prevalência de clones únicos. Até o momento, este é o primeiro caso da ocorrência de surto por A. xylosoxidans em pacientes com Fibrose Cística. A. xylosoxidans é um microrganismo que vem se destacando em frequência e como um possível patógeno pulmonar nesses pacientes. Entretanto, até o momento os dados são insuficientes para avaliar a sua contribuição para a evolução da doença pulmonar. Estudos que busquem elucidar as características de A. xylosoxidans que o permitem colonizar persistentemente o pulmão dos pacientes com FC, bem como seu potencial de virulência, são necessários.

Studies on fish epidemics from Assam

Investigations conducted since July 1988 on ulcerative fish epidemics in Assam, India, indicated that mainly four species of fishes belonging to the genera Puntius, Channa, Macrognathus and Mystus were widely affected by the disease. Results indicated that outbreak of the disease may not be due to organic pollution of water or radioactive and heavy-metallic contamination. Bacterial culture revealed colonies of Escherichia coli and Pseudomonas aeruginosa in the surface muscular lesions and gill tissues while preliminary electron microscopic studies indicated the presence of viruses in the muscles and gills of diseased fishes.

Bacterial flora of marine fish doma (Sciaenoids) (Small spp.) from Bombay coast

Marine fish doma (Sciaenoids) (Small spp.) from Bombay coast was studied for total bacterial counts on the surface and gut. Large number of Micrococcus species (77.4%) was found whereas few species from Achromobacter, Bacillus, Bacterium, Flavobacter, Pseudomonas and Sarcina were noted.

Quantitative and qualitative studies on the bacterial flora of fresh sardines

Plate counts at R T and 8 C on the skin with muscle and the gut contents of absolutely fresh sardines (Sardinella longiceps) caught off Cochin showed a seasonal variation when sampling was done over a period of 12 months. The counts of the gut contents ran parallel with those of the skin with muscle, but at a higher level of magnitude. Qualitatively, the analysis of 360 strains of bacteria isolated from the skin with muscle and 100 strains from the guts during a year's study revealed a very high preponderance of Gram negative rods, mainly of Achromobacter, Vibrio, and Pseudomonas groups. The percentage of Gram positive organism was very low or nil at times in the ocean fresh sardines.

Investigation on bacterial flora of farmed freshwater prawn (Macrobrachium rosenbergii de Man) in Bangladesh

Total bacterial load in the haemolymph of freshwater prawn Macrobrachium rosenbergii varied from 6.2x10⁴ to 1.9x10⁷ CFU/ml whereas in the hepatopancreas, bacterial load varied from 1.9x10³ to 2.9x10⁵ CFU/g. The total bacterial load in the pond water varied from 2.6x10² to 4.1x10⁵ CFU/ml. The isolated bacterial genera in the haemolymph and the hepatopancreas of prawn were Streptococcus, Acinetobacter, Micrococcus, Aeromonas, Vibrio, Flavobacterium, Staphylococcus and Pseudomonas, whereas the detected bacterial genera in pond water were Micrococcus, Streptococcus, Vibrio, Flavobacterium, Staphylococcus, Pseudomonas and Aeromonas. Among the detected genera, Vibrio and Staphylococcus were found to be dominant genera in the haemolymph of the sampled prawn throughout the study period whereas Staphylococcus and Pseudomonas were dominant in pond water.

Incidence of ulcer disease in African catfish (Clarias gariepinus Burchell) and trial for its chemotherapy

Young Clarias gariepinus cultured in an artificial tank were severely affected by an ulcer type of disease where 77% fish died within 5 weeks. From the lesions and kidney of affected fish Aeromonas, Pseudomonas, Flavobacterium, Micrococcus and Staphylococcus were isolated where Aeromonas was observed as the dominant bacteria. Among them, an A. hydrophila isolate AGK 34 was detected as a pathogen by the experimental challenge test. In order to find out a suitable remedial measure of the disease, four different chemotherapeutants were applied to the affected fish in 6 different ways under laboratory condition. Affected fish were recovered from the disease in different treatments. But the best result was obtained by a successive bath in 1-2% NaCl and subsequent oral treatment with commercial oxytetracycline at a dose of 75 mg/kg body weight of fish.

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.

Incidence of epizootic ulcerative syndrome (EUS) in freshwater fishes in the endemic area of Punjab, Pakistan

Incidence of Epizootic Ulcerative Syndrome (EUS) has been recorded for the first time in freshwater fishes in the endemic area of Punjab, Pakistan. Survey of private fish farms, hatchery and natural water bodies was conducted in a radius of 14 Km from around river Ravi near Lahore (Punjab Province) Pakistan. Out Of 1628 fishes belonging to 18 genera, 517 fishes of 10 genera were found affected with EUS. The incidence of EUS in culturable fishes was higher in Cirrhina mrigala (15.4%) moderate in Catla cat/a (13.3%) and lower in Labeo rohita (5.0%). Exotic fish, Chinese carp Ctenoparyngodon idella and Hypophthalmicthys molitrix were not affected with EUS. In non-culturable fishes the incidence of EUS was highest in Channa punctatus (72.8%) moderate in by C. straitus (65.45%) and comparatively lower Puntius ticto (43.7%). A slow growing temperature sensitive Saprolegnia spp. was isolated from all of EUS infected fish species. Aeromonas spp. and Pseudomonas spp. were isolated from the diseased fishes. Ectoparasites viz. Lernaea, Argulus and Triclwdina spp. were also isolated from the skin and gills of infected fish species. The disease was more severe in water having low alkalinity (70 mg/1), hardness (75 mg/1) and low temperature of 10-12 °C.

Incidence of ulcer type of disease in wild fishes of Bangladesh

Disease occurred in wild fish species investigated in different water bodies like canals, ditches, beel, haor, flood plain in 17 districts of Dhaka division. Haemorrhagic lesions were observed on the body surface of affected fishes. Incidence of the disease in the investigated water bodies ranged from 0 to 100%. In total 19 fish species were found to be affected and prevalence of infection ranged from 0.0 to 100.0%. Channa punctatus and Puntius ticto were severely affected in all locations. Percentage of infection in these fishes ranged from 0.0 to 100.0. The highest infection was observed in Netrokona, Kishoreganj, and Mymensingh districts. Bacterial genera isolated from the lesions of these affected fishes were Aeromonas, Pseudomonas, Flavobacterium, Micrococcus, and Staphylococcus. Among these isolates Aeromonas was the dominant. Abundance of Aeromonas in the lesions among the investigated bacteria ranged from 75 to 90%. Five identified Aero monas lrydrophila were examined for their pathogenicity and were able to infect the experimental fish, silver barb (Puntius gonionotus). The pathogen Aeromonas hydrophila was thus considered to have an association with the outbreak of ulcer type of disease in the investigated fish species.

Effect of delayed icing on quality changes and shelf-life of some freshwater fish from Bangladesh

The quality and shelf-life of three freshwater fish species of Bangladesh, catla (Catla catla), magur (Clarias batrachus) and tilapia (Oreochromis niloticus) stored at room temperature and ice were evaluated. Live fishes were killed by cranial spiking and stored at room temperature (27-30 °C), ice stored immediately after death, 5 hr after death and 10 hr after death. The shelf-life and quality of the fishes were evaluated by organoleptic method, rigor-mortis studies and bacteriological assessment. Fishes kept at room temperature became organoleptically unacceptable within 16-20 hr duration. Ice stored fishes showed considerable differences in their shelf-life when icing was delayed for different duration. Shelf-life of catla ice stored immediately after death was 20 days but shelf-life reduced to 12 days when icing delayed for 10 hr after death. Similar trend was observed for two other fish species magur and tilapia. Rigor-index of the fishes stored under different conditions also considerably varied among the three fish species, particularly effect of delayed icing was very much evident. Bacteriological study showed patterns of Aerobic Plate Count (APC) at the end of shelflife study when fishes became organoleptically unacceptable were more or less similar for all the three fish species stored under different conditions. No definite pattern was observed in the generic distribution of bacteria in different fish species under different storage conditions. Micrococcus, Coryneforms, Pseudomonas and Achromobacter were the dominant groups of bacteria isolated from the fishes spoiled at room temperature and ice stored condition.

HACCP based management system for improving quality of freshwater giant prawn, Macrobrachium rosenbergii

The study was conducted on the present status of HACCP based quality management system of golda, Macrobrachium rosenbergii farms in Fulpur region of Mymensingh. Information was collected on general condition of farms, culture systems and post-harvest quality management. In almost all farms, there is no or inadequate infrastructure facilities such as, road access, electric supply, telecommunications, ice, feed storage facility, vehicle for golda transportation, washing and toilet facilities. The problems associated with sanitation and hygiene was: widespread use of cow dung, poultry manure and construction of open toilet within the vicinity of prawn culture pond. Different grades of commercially available and locally prepared feeds were used for golda culture in the pond. Golda post-larvae (PL) of 40-50 days old were stocked with carp species. The price of golda PL ranged from Tk. 1.00 to Tk. 1.25/piece. The pond size varied from 50 decimal (0.2 ha) to 2.5 acre (1.0 ha) with an average depth of 2-2.5 m. The culture period of golda varied from April-May to November-December and survival rate ranged between 75 and 80%. Production of golda varied from 250-500 kg/acre (625-1,250 kg/ha). Harvested golda were transported to city market within 4 h. Two size grading were generally followed during pricing, e.g. Tk. 500 to 550/kg for >100 g size and Tk. 300/kg for <100 g size. The cost-benefit ratio was found to remain around 1:1.25 depending on availability of PL. Water quality parameters such as, water temperature, pH, dissolved oxygen, total alkalinity and chlorophyll a in five golda farms in Fulpur region were monitored. Water temperature ranged from 29°C to 33°C, dissolved oxygen from 2.28 to 4.13 mg/l, pH between 6.65 and 7.94, alkalinity from 44 to 70 mg/l and chlorophyll a concentration from 61.88 to 102.34 µg/l in the five investigated ponds. The Aerobic Plate Count (APC) of the water sample was within the range of 2.0x10^6 - 2.96x10^7 CFU/ml and of soil samples within the range of 6.9x10^6 - 7.73x10^6 CFU/g. Streptococcus sp., Bacillus sp., Escherichia coli, Staphylococcus sp., Pseudomonas sp. and Salmonella sp. were isolated from pond water and sediment. Different feed samples used for golda was analyzed for proximate composition. Moisture content ranged around 14.14-21.22%, crude protein 20.55-44.1%, lipid 4.67-12.54% and ash 9.7-27.69%. The TVB-N values and peroxide values of feeds used as starter, grower and fish meal were found within the acceptable ranges and samples were free from pathogenic organisms. A training was organized for the golda farmers on HACCP, water quality and post-harvest quality management of prawn.

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.

Study on regional differences in soil-water characteristics and their relevance to aquaculture productivity and fish disease

A total of 45 ponds used for fish polyculture were investigated in three zones of Bangladesh to identify the differences among the zones in respect to aqua-ecology, culture practices, fish productivity and health management. Four hundred and fifty fish from three zones were clinically examined by naked eye and histopathology. Out of total number of fish examined, 45 fish from Dhaka zones were examined for parasites and bacteria in addition to histopathology. Faded and haemorrhagic gill, skin, fin, scale loss and lesions were observed during fish examination. Aeromonas spp. Pseudomonas spp. and Streptococcus spp. were isolated respectively from 56%, 46% and 39% affected fish. Among the five water quality parameters analyzed, the highest average hardness and alkalinity respectively were recorded in Rajshahi (156 ppm and 142 ppm) followed by Dhaka (146 ppm and 132 ppm) and Chittagong (81 ppm and 90 ppm). The highest average pH was recorded in Mymensingh (7.52) followed by Rajshahi (7.13) and Chittagong (7.05). Water holding capacity of soil in Rajshahi zone was poor compared to other zones and farmers were found to be reluctant to fish farming.

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 AchromobacterPseudomonasPseudomonas were generally more resistant to antibiotics 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.

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 PseudomonasPseudomonas and Micrococci cultures. The significance of these results in fish preservation by CTC is discussed.