Structural and Functional Annotation of Hypothetical Protein of Fusobacterium nucleatum Strain MJR7757B: An in Silico Approach

Fusobacterium nucleatum is an anaerobic, commensal, gram-negative oral bacterium that is carcinogenic and causes a wide range of human diseases. The present study focused on the analysis of the hypothetical protein, HMPREF3221_01179, derived from F. nucleatum strain MJR7757B, employing various computational methods to anticipate both its structure and functional characteristics. NCBI conserved domain analysis, NCBI BLASTp and MEGA Phylogenetic tree study characterize the target protein as an outer membrane efflux protein (ToIC family) which facilitate the bacterial transmembrane transport. With a molecular weight of 52120.02 Da, an isoelectric point (pI) of 8.33, and an instability index of 29.47, the protein is anticipated to exhibit good solubility in the extracellular space and crucial stability for pharmaceutical applications. The protein’s structure meets quality standards during the construction and refinement of its 3D model. The efflux inhibitor Arginine beta-naphthylamide exhibits a significant binding affinity (-7.1 kcal/mol) to the binding site of the target protein. The in-silico analysis improves the understanding of the protein and facilitates future investigations into therapeutic medication.

Comparative study of haematological,serum electrolyte and nonelectrolyte parameters of male and female Persian sturgeon(Acipenser persicus)brood stocks

Sturgeons are the most important principal market species in the Caspian Sea. In the present study, measurement of blood parameters was carried out with the aim of evaluating the normal value of hematological and serum biochemical parameters of brood stocks Acipenser persicus during spawning season. Blood samples were collected from 36 brood stocks of A. persicus（18 males and 18 females） during the spawning season. Higher value of red blood cells, white blood cells, haemoglobin, haematocrit was observed in male breeders with significant differences between female breeders（p〉0.05）. Haemoglobin and haematocrit value in this study was within the range of 8.70 to 9.2 g/d L and 29.73% to 30.26%, respectively. The statistically significant differences between mean corpuscular volumes, mean corpuscular haemoglobin, lymphocyte and neutrophil percentages, aspartate aminotransferase, alkaline phosphatase and creatine kinase were observed in the male and female brood stocks.It is concluded from this study, some of the hematological and serum biochemical parameters of male and female A. persicus breeders were significantly varied from each other. In comparative investigation with earlier reports,the values of red blood cells, haemoglobin and haematocrit are highly varied due to age variations of fish.However, further studies are necessary to understand the impact of sexual, age, size, length, and season and habitat variation. Serum biochemical parameters can be used for confirming the maturity of A. persicus and monitoring any changes in the water quality parameters and soil.

The Effects of Letrozole in Transfer of Maternal Immunity against Lactococcosis to Eggs and Larvae in Rainbow Trout (<i>Oncorhynchus mykiss</i>, Walbaum)

This study evaluated transfer of maternal lysozyme and immunoglobulin (IgM) against Lactococcus garviea, the causative agent of lactococcosis to eggs and larvae in rainbow trout. Changes in circulating lysozyme and IgM during development of eggs and larvae were measured by a method based on the ability of lysozyme to lyse the bacterium Micrococcus lysodeikticus and enzyme-linked immunosorbent assay (ELISA) respectively. For doing this, twelve broodstocks were injected weekly with 2.5 mg&middotkg-1 letrozole (an endocrine disrupter component) two months before spawning season and vaccinated intraperitoneally (i.p) with a bacterin (inactivated L. garviae) one month before spawning. Twelve broodstocks for vaccination and twelve female rainbow trouts as control group were also immiunised (i.p) with the bacterin and injected (i.p) with PBS respectively. Results showed that at day 8 after hatching, lysozyme and IgM levels during pre-larval stages decreased gradually, as yolks were absorbed. Lysozyme and IgM levels were significantly higher in the letrozole injected immiunised parents 30 days after immunisation as well as their larvae compared to the control group (p L. garviae) (one month before spawning) in maternal transfer of lysozyme and IgM levels to eggs and larvae.

Immunobiochemical Characteristics of Purified Native Leptin Protein from Indian Major Carp, Rohu (<i>Labeo rohita</i>Ham.)

Information regarding molecular characteristics of leptin protein in different animal species in-cluding fish is scarce. With the aim of characterizing the native leptin protein of Indian major carprohu (Labeo rohita), at molecular level, the present study was designed to isolate rohu leptin from its hepatocytes (the prime source of leptin in fish) and immunobiochemical characterization of the same, subsequently. In the present study, chemical treatment and ultra-sonication technique was used for isolating leptin from rohu liver tissue. Purification of the protein was attempted using affinity column chromatography. The molecular, biophysical and serological characterization of rohu leptin was carried out by 2D-gel electrophoreis, SDS-PAGE, MALDI-TOF Mass spectroscopy and Western blot. The SDS-PAGE and 2D gel analysis revealed that rohu native leptin possesses molecular mass of 16 kDa. Western blot analysis showed that the fish hepatocytes possessed the sero-reactive leptin protein of 16 kDa. MALDI-TOF mass spectroscopy and peptide analysis showed the molecular mass of rohu leptin as 16283.38 Da. The serodiagnostic potential of native leptin of rohu was revealed for the first time while assessing its serological responses by ELISA using anti-leptin antibodies.

Prevalence of Ectoparasitic Infestation in Indian Major Carps During Winter at Different Blocks of South 24-Parganas District, West Bengal, India

The study was carried out to find the prevalence of ectoparasites in carp species specifically Indian Major Carps(IMC)during the post monsoon season(November’16 to March’17).Four groups of ectoparasites viz.myxozoan,ciliophoran,monogenean and crustacean were recorded from a total 500 number of carp species like,Labeo rohita,Catla catla,Cirrhinus mrigala and Labeo calbasu collected from different ponds of selected blocks of South 24-Parganas district of West Bengal.The highest prevalence(64.8%)of infestation had been recorded by Myxozoans and the lowest was by Monogeneans(4.8%).The highest and lowest ectoparasitic prevalence in carp was observed in L.rohita(32.9%)and C.catla(27.3%).Beside these,lower temperature(Average 19.3℃),low pH(Average 6.9)and marginal level of dissolved oxygen(Average 6.0ppm)were also created an unfavorable condition for parasitic infestation during this season.At the end of this experiment it was concluded that disease occurrences due to ectoparasites was high in winter with some key factors like temperature,pH and dissolved oxygen(DO).

Prevalence of Argulus sp. in Indian Major Carps from Bhangore Block of South 24 Parganas District, West Bengal, India

The study was carried out to find the prevalence and severity of Argulus sp.in Indian Major Carps(IMCs)collected from Bhangore block of South 24 Parganas district of West Bengal during November 2018 to August 2019.January(2019)recorded the highest parasitic prevalence(PFI,83.33%)and the month of August(2019)recorded the lowest parasitic prevalence(PFI,9%).The identities of selected parasites were further confirmed by molecular identification through 18S rDNA analysis.The study revealed that Argulus sp.infestations had great economic implications especially in the winter months and is one of the most prevalent problems in fresh water aquaculture systems.

Assessment of nitrogen and sulphur cycle bacteria and shrimp production in ponds treated with biological products

Objective:To study the influence of biological products on the levels of nitrogen and sulphur cycle bacteria in shrimp culture systems of West Bengal,India.Methods:The pond water and sediment samples were analyzed for physico-chemical parameters as per standard methods.The bacteria involved in ammonification,nitrification,denitrification,sulphate reduction and sulphur oxidation were enumerated by most probable number technique.Results:The semi-intensive and modified extensive shrimp farms used a variety of biological products during various stages of production.No biological products were used in traditional farms.The water and sediment samples of modified extensive system recorded significantly higher mean heterotrophic bacterial counts.The counts of ammonia,nitrite and sulphur oxidizers,and nitrate and sulphate reducers varied among the systems.The cycling of nitrogen and sulphur appeared to be affected with the intensification of culture practices.Conclusions:The application of biological products in certain systems helped to maintain the bacteria involved in nitrogen and sulphur cycles and safe levels of ammonia,nitrite and nitrate.An assessment of these metabolically active bacteria in shrimp culture ponds and the application of right kind microbial products would help ameliorate the organic pollution in shrimp aquaculture.

Efficacy of oxytetracycline and potentiated sulphonamide oral therapies against Aeromonas hydrophila infection in Nile tilapia Oreochromis niloticus

Objective: To assess the efficacy of feeds containing approved antibiotics, viz., oxytetracycline (OTC) or potentiated sulphonamide [(sulphamethoxazole-trimethoprim) SMZ-TMP] at 1 g, 2 g, 3 g and 4 g of respective antibiotics/kg feed at 2% body weight ration in preventing the Aeromonas hydrophila (A. hydrophila) infection in Oreochromis niloticus. Methods: Commercial pellet feed was top dressed with respective antibiotics using 5 mL vegetable oil as a binder. Fish were injected intramuscularly with A. hydrophila at ≈ 6.0 × 107–8.6 × 107 CFU/fish, and fed subsequently with OTC and SMZ-TMP feeds for 10 and 5 days, respectively. Fish mortalities were recorded during the pre-treatment, treatment and post-treatment periods. Results: Highest mortalities (7.42%–8.33%) were observed in challenged and untreated fish. The mortalities observed in fish fed with OTC or SMZ-TMP were 0%–6.66% with decreasing concentrations of antibiotics from 4 to 1 g/kg feed. Significant differences existed in the mortalities among fish fed with different doses of antibiotics (P < 0.05). The relative percent survival values were 20, 40, 40 and 60 for 1 g, 2 g, 3 g and 4 g OTC/kg feed groups, respectively;while in SMZ-TMP fed fish, the respective relative percent survival values were 10, 100, 100, and 100. Conclusions: The fish fed with feed containing 2 g antibiotic/kg at 2% body weight was the lowest concentration that recorded significantly lower mortality (P < 0.05), which could be the treatment of choice for the control of A. hydrophila in Oreochromis niloticus in tropical condition.

Ecology of antibiotic resistant vibrios in traditional shrimp farming system(bhery)of West Bengal,India

Objective:To study the ecology of antibiotic resistant bacteria with emphasis on sucrose negative vibrios in water and sediments samples of traditional shrimp farming system(bhery)in West Bengal,India.Methods:The vibrios were isolated from traditional shrimp farm samples on thiosulphate citrate bile salt sucrose agar and sucrose negative bacterial strains were used as biomarkers to assess the frequency of antibiotic resistance.Results:The incoming water brought presumptive vibrios ranging from 5.50×10^(1)to 1.00×10^(3)mL in to the bhery,and there appeared to build up vibrios in the culture system with days of culture,as there was about 9 fold increase in vibrios.The levels of vibrios were observed to be moderately higher in outlet water and ranged between 4.15×10^(2)and 4.15x10^(3)mL.The counts of vibrios in pond sediment was found to be 1.00x10^(2)-4.90×10^(3)g;while in inlet(2.00×10^(2)-4.20×10^(4)g)and outlet(3.00×10^(2)-6.85×10^(3)g)their levels were observed to be higher than the pond sediment.Thirteen different Vibrio species were encountered in traditional shrimp culture system and all vibrios were sensitive to chloramphenicol,followed by ciprofloxacin and gatifloxacin(98.24%),gentamicin(95.61%)and other antibiotics.The multiple antibiotic resistance(MAR),i.e.,resistance to at least two antibiotics,was noticed among 43.85%of the sucrose negative vibrios and 41.86%of the sucrose negative non--vibrios.All vibrios harveyi strains exhibited MAR.Although no antibiotic was used in the bhery,the prevalence of MAR in 44%of the sucrose negative vibrios and nonvibrios is a cause of concern.The MAR index was higher in inlet water and sediment samples.The MAR observed in biomarker strains of pond water and sediment(40%)was comparable to those of inlet samples,thus confirming the fact that incoming water was the major source of antibiotic resistant bacteria.Conclusions:It seems that the shrimp culture in bhery does not favour the proliferation and spread of antibiotic resistant bacteria.

Effect of polyculture of shrimp with fish on luminous bacterial growth in grow-out pond water and sediment

Objective:To study the distribution of marine luminous bacteria in shrimp culture systems of West Bengal and the effect of polyculture of shrimp with fish to reduce luminous bacteria.Methods:Luminous bacterial counts were enumerated by spread plating on seawater complex agar from shrimp grow-out pond water and pond sediment samples of West Bengal,India.Results:About 31.16%and 51.44%of pond sediment and pond water samples respectively had detectable levels of luminous bacteria.It was noticed that in normal ponds a shift happened in bacterial profile of water from the day of flooding up to 60 d,with the dominance of luminous bacteria among vibrios,reaching counts 10^(4) cells/mL or more.While in diseased ponds,luminous bacterial abundance within the ponds was noticed in the first 6 weeks of culture.Marked reduction in luminous bacterial counts of water and sediment was observed through out the culture period in polyculture ponds compared to monoculture ponds.There was no incidence of white spot syndrome viral disease and luminous vibriosis in both controlled and experimental ponds.Conclusions:The results suggest vigilant monitoring of ponds for luminous bacteria abundance and polyculture of shrimp with fish in ecofriendly sustainable aquaculture can reduce the impact of shrimp disease outbreak.

Prevalence of Edwardsiella tarda in commercially important finfish and shellfish of Bihar and West Bengal,India

Objective:To study the prevalence of Edwardsiella tarda(E.tarda)in finfish and shellfish of West Bengal and Bihar,India and their antibiogram.Methods:Fish samples were enriched overnight in Edwardsiella ictaluri broth and plated onto Edwardsiella ictaluri agar.Typical colonies were identified conventionally and by VITEK 2 compact system.Antibiogram was done by agar disc diffusion assay.Results:Of the 118 fish samples screened,only 14.41%had E.tarda.The incidence was marginally high in wild fish(15.30%)than in cultured fish(13.26%).Maximum incidence was in intestine(12.98%)followed by gills(6.60%)and skin(2.38%).All E.tarda strains were sensitive to ciprofloxacin and exhibited varying degrees of resistance to other antibiotics.Multiple antibiotic resistance was seen in 84.00%–87.50%of the E.tarda strains.Majority of them had high minimal inhibitory concentration values(200μg/mL)for oxytetracycline and gentamycin.Conclusions:The results suggested that considerable proportions of commercially important finfish are carriers of multiple antibiotic resistance E.tarda.This calls for proper sanitary measures to eliminate this pathogen in fish and fishery products.