In vivo morphological and antigenic characteristics of Photobacterium damselae subsp. piscicida

The present study was conducted to examine the morphology and antigenicity of Photobacterium damselae subsp. piscicida by culturing the bacterium in vivo in the peritoneal cavity of sea bass (Dicentrarchus labrax) within dialysis bags with either a low molecular weight (LMW) cut-off of 25 kDa or a high molecular weight (HMW) cut-off of 300 kDa. Differences were observed in the growth rate between the bacteria cultured in vivo or in vitro. Bacteria cultured in vivo were smaller and produced a capsular layer, which was more prominent in bacteria cultured in the HMW bag. Antigenicity was examined by Western blot analysis using sera from sea bass injected with live Ph. d. subsp. piscicida. The sera recognised bands at 45 and 20 kDa in bacteria cultured in vivo in the LMW bag. Bacteria cultured in vivo in the HMW bag did not express the 45 kDa band when whole cell extracts were examined, although the antigen was present in their extracellular products. In addition, these bacteria had a band at 18 kDa rather than 20 kDa. Differences in glycoprotein were also evident between bacteria cultured in vitro and in vivo. Bacteria cultured in vitro in LMW and HMW bags displayed a single 26 kDa band. Bacteria cultured in the LMW bag in vivo displayed bands at 26 and 27 kDa, while bacteria cultured in vivo in the HMW bag possessed only the 27 kDa band. These bands may represent sialic acid. The significance of the changes observed in the bacterium's structure and antigenicity when cultured in vivo is discussed

Prevalence of Gastro-Intestinal Nematodes in Goats in Hyderabad and Adjoining Areas

The study was conducted to determine the prevalence of gastrointestinal nematodes of goats (n=1065) in and around Hyderabad using qualitative and quantitative coprological examinations. Results revealed that 43.10% (459) goats were infected with different species of nematodes including Haemonchus contortus (14.65%), Trichuris ovis (8.17%), Trichostrongylus axei (7.61%), Trichostrongylus colubriformis (6.76%), Oesphagostomum columbianum (5.35%), Ostertagia circumcincta (5.35%), Chabertia ovina (4.79%) and Strongyloides papillosus (4.51%). Infections with mixed species of nematodes were recorded in 6.54% (n=30/459; T. ovis + H. contortus), 5.23% (n=24/459; C. ovina + H. contortus), 5.88% (n=27/459; S. papillosus + C. ovina), and 12.42% (n=57/459; O. circumcincta + T. ovis) goats. Of the total infected (n=459); 51.4, 38.3 and 10.2% goats had light, moderate and heavy infections, respectively. The prevalence, nature and intensity of the helminthiasis in goats warrant an immediate attention to devise strategies for its control to reduce the production losses

EPIDEMIOLOGY OF OVINE GASTROINTESTINAL NEMATODES IN HYDERABAD DISTRICT, PAKISTAN

An epidemiological study on gastrointestinal nematodes of sheep was carried out in farms of small farmers in Hyderabad district from May 2004 to April 2005. Faecal egg counts, pasture larval counts and worm counts from permanent grazing animals were recorded for 12 months. H. contortus (24.6%) was found to be predominant of gastrointestinal nematode parasites, Trichostrongylus spp. (18.0%) was the next most prevalent species, others, including: O. circumcincta, S. papillosus, T. ovis, Oe. columbianum and Chabertia ovina were found in varying percentages. The highest faecal egg counts (FEC) were recorded in September, whereas the lower FEC were in February. Statistical analysis revealed that the FEC were significantly (P<0.01) affected by months (seasons). The peak of pasture infectivity was in August and declined to lower level in January. The mean worm burden counts were the highest in September and declined toward the minimum level in February in necropsized animals. The worm counts was influenced significantly (P<0.01) by FEC and pasture larval counts. The results of this study could be used to design a programme to minimize and control gastrointestinal nematode infections in sheep

Molecular Diversity and Biochemical Content in Two Invasive Alien Species: Looking for Chemical Similarities and Bioactivities

The biochemical composition, molecular diversity, and two different bioactivities of Asparagopsis armata and Rugulopteryx okamurae (two alien species with different invasive patterns in the southern Iberian Peninsula) were analyzed through spectrophotometric methods and Fourier transform ion cyclotron mass spectroscopy (FT-ICR-MS). A total of 3042 molecular formulas were identified from the different extracts. The dH2O extracts were the most molecularly different. A. armata presented the highest content of nitrogenous compounds (proteins, CHON) and sulphur content, whereas R. okamurae was rich in carbonated compounds (total carbon, lipids, CHO, and CHOP). Antioxidant capacity and phenolic content were higher in R. okamurae than in A. armata. Antimicrobial activity was detected from both species. A. armata showed capacity to inhibit human and fish pathogens (e.g., Staphylococcus aureus or Vibrio anguillarum), whereas R. okamurae only showed inhibition against human bacteria (Staphylococcus aureus and Cutibacterium acnes). In R. okamurae, molecules with a great number of pharmaceutical activities (e.g., anti-inflammatory or antitumoral), antibacterial, biomaterial, and other utilities were found. The main molecules of A. armata had also pharmaceutical applications (e.g., antimalarian, antithrombotic, anti-inflammatory, or antiarthritis). The valorization of these species can help to counteract the environmental effects of the bioinvasions.This research was funded by the BLUEMARO Project from the Science and Innovation Ministry of the Spanish Government (grant number PID2020-116136RB-I00) and by the Start-up funding for junior research groups of the Carl von Ossietzky University Oldenburg, “pharmaDOM”. Partial funding for open access charge: Universidad de Málaga

Variation in the molecular weight of Photobacterium damselae subsp. piscicida antigens when cultured under different conditions in vitro

The antigenicity of Photobacterium damselae (Ph. d.) subsp. piscicida, cultured in four different growth media [tryptone soya broth (TSB), glucose-rich medium (GRM), iron-depleted TSB (TSB + IR-), and iron-depleted GRM (GRM + IR-)] was compared by enzyme-linked immunosorbent assay (ELISA) and Western blot analysis using sera obtained from sea bass (Dicentrarchus labrax) raised against live or heat-killed Ph. d. subsp. piscicida. The antigenic expression of Ph. d. subsp. piscicida was found to differ depending on the culture medium used. A significantly higher antibody response was obtained with iron-depleted bacteria by ELISA compared with non-iron depleted bacteria obtained from the sera of sea bass raised against live Ph. d. subsp. piscicida. The sera from sea bass raised against live bacteria showed a band at 22 kDa in bacteria cultured in TSB + IR- or GRM+ IR- when bacteria that had been freshly isolated from fish were used for the screening, while bands at 24 and 47 kDa were observed with bacteria cultured in TSB or GRM. When bacteria were passaged several times on tryptic soya agar prior to culturing in the four different media, only bands at 24 and 47 kDa were recognized, regardless of the medium used to culture the bacteria. It would appear that the molecular weight of Ph. d. subsp. piscicida antigens change in the presence of iron restriction, and sera from sea bass infected with live bacteria are able to detect epitopes on the antigens after this shift in molecular weight

Photodynamic Antimicrobial Chemotherapy in Aquaculture: Photoinactivation Studies of Vibrio fischeri

BACKGROUND: Photodynamic antimicrobial chemotherapy (PACT) combines light, a light-absorbing molecule that initiates a photochemical or photophysical reaction, and oxygen. The combined action of these three components originates reactive oxygen species that lead to microorganisms' destruction. The aim was to evaluate the efficiency of PACT on Vibrio fischeri: 1) with buffer solution, varying temperature, pH, salinity and oxygen concentration values; 2) with aquaculture water, to reproduce photoinactivation (PI) conditions in situ. METHODOLOGY/PRINCIPAL FINDINGS: To monitor the PI kinetics, the bioluminescence of V. fischeri was measured during the experiments. A tricationic meso-substituted porphyrin (Tri-Py(+)-Me-PF) was used as photosensitizer (5 µM in the studies with buffer solution and 10-50 µM in the studies with aquaculture water); artificial white light (4 mW cm(-2)) and solar irradiation (40 mW cm(-2)) were used as light sources; and the bacterial concentration used for all experiments was ≈10(7) CFU mL(-1) (corresponding to a bioluminescence level of 10(5) relative light units--RLU). The variations in pH (6.5-8.5), temperature (10-25°C), salinity (20-40 g L(-1)) and oxygen concentration did not significantly affect the PI of V. fischeri, once in all tested conditions the bioluminescent signal decreased to the detection limit of the method (≈7 log reduction). The assays using aquaculture water showed that the efficiency of the process is affected by the suspended matter. Total PI of V. fischeri in aquaculture water was achieved under solar light in the presence of 20 µM of Tri-Py(+)-Me-PF. CONCLUSIONS/SIGNIFICANCE: If PACT is to be used in environmental applications, the matrix containing target microbial communities should be previously characterized in order to establish an efficient protocol having into account the photosensitizer concentration, the light source and the total light dose delivered. The possibility of using solar light in PACT to treat aquaculture water makes this technology cost-effective and attractive

Probiotic supplementation influences the diversity of the intestinal microbiota during early stages of farmed Senegalese sole (Solea senegalensis, Kaup, 1858)

Ingestion of bacteria at early stages results in establishment of a primary intestinal microbiota which likely undergoes several stages along fish life. The role of this intestinal microbiota regulating body functions is crucial for larval development. Probiotics have been proved to modulate this microbiota and exert antagonistic effects against fish pathogens. In the present study, we aimed to determine bacterial diversity along different developmental stages of farmed Senegalese sole (Solea senegalensis) after feeding probiotic (Shewanella putrefaciens Pdp11) supplemented diet for a short period (10–30 days after hatching, DAH). Intestinal lumen contents of sole larvae fed control and probiotic diets were collected at 23, 56, 87, and 119 DAH and DNA was amplified using 16S rDNA bacterial domain-specific primers. Amplicons obtained were separated by denaturing gradient gel electrophoresis (DGGE), cloned, and resulting sequences compared to sequences in GenBank. Results suggest that Shewanella putrefaciens Pdp11 induces a modulation of the dominant bacterial taxa of the intestinal microbiota from 23 DAH. DGGE patterns of larvae fed the probiotic diet showed a core of bands related to Lactobacillus helveticus, Pseudomonas acephalitica, Vibrio parahaemolyticus,and Shewanella genus, together with increased Vibri o genus presence. In addition, decreased number of clones related to Photobacterium damselae subsp piscicida at 23 and 56 DAH was observed in probiotic-fed larvae. A band corresponding to Shewanella putrefaciens Pdp11 was sequenced as predominant from 23 to 119 DAH samples, confirming the colonization by the probiotics. Microbiota modulation obtained via probiotics addition emerges as an effective tool to improve Solea senegalensis larviculture.En prens

Bauran Kebijakan Fiskal-moneter dan Dampaknya Bagi Anggaran Pertahanan

Tulisan ini hendak mengulas fenomena yang sedang dihadapi pemerintah Indonesia, yakni adanya kesenjangan antara kebutuhan yakni permintaan sekaligus ekspektasi akan pembangunan kekuatan sektor pertahanan yang memerlukan peningkatan anggaran dengan kondisi kemampuan sumber daya/anggaran yang terbatas. Untuk dapat mengakomodasi kebutuhan tersebut maka pertumbuhan Produk Domestik Bruto (PDB) yang tinggi menjadi prasyarat utama dan pemerintah perlu merumuskan strategi kebijakan makro ekonomi yang tepat untuk menciptakan pertumbuhan PDB yang tinggi. Penelitian ini membahas kebijakan fiskal-moneter sebagai instrumen pendorong pertumbuhan ekonomi dan mengetahui dampak kebijakan tersebut terhadap anggaran sektor pertahanan. Adapun penelitian ini menggunakan pendekatan deskriptif kualitatif purposive sampling dengan metode Means-Ways-Ends dan kurva IS-LM sebagai alat analisis. Dari hasil penelitian, disimpulkan bahwa bauran kebijakan fiskal-moneter yang paling tepat adalah dengan mempertimbangkan situasi dan kondisi makro ekonomi yang sedang dihadapi sebagai dasar penentuan kombinasi kebijakan fiskal-moneter yang tepat guna mencapai pertumbuhan PDB yang tinggi. Bauran kebijakan tersebut harus disertai syarat adanya sinergi dan koordinasi antara kebijakan fiskal dan moneter

OVICIDAL AND LARVICIDAL PROPERTIES OF ADHATODA VASICA (L.) EXTRACTS AGAINST GASTROINTESTINAL NEMATODES OF SHEEP IN VITRO

The main objective of this study was to evaluate the anthelmintic activity of Adhatoda vasica (Acanthaceae) in vitro against the gastrointestinal nematodes of sheep. The aqueous and ethanolic extracts of Adhatoda vasica aerial parts were evaluated by egg hatching and larval development assays. The aqueous and ethanolic extracts at 25-50 mg/ml concentrations exhibited ovicidal and larvicidal (p<0.05) activity against gastrointestinal nematodes. The plant extracts showed dose-dependent inhibition (P<0.05). The ethanolic extract at the concentration of 50.0 mg/ml was more effective in inhibiting egg hatching and larval development of gastrointestinal nematodes. The effective dose (ED50) of aqueous and ethanolic extracts were determined graphically from linear regression equation with probit scale, y = 5. The results of this study suggested that Adhatoda vasica extracts may be useful in the control of gastrointestinal nematodes of sheep