Journal of Coastal Life Medicine

596-600

Authors: Alagarsamy Sakthivel, Periyasamy Selvakumar, Ayyaru Gopalakrishnan

Objective: To recored the infection of Acanthocephalan parasite, (Echinorhynchus sp.) in adult yellowfin tuna (Thunnus albacares). This is the first report of acanthocephalan Echinirhynchus sp attached to yellowfin tuna and moreover prevalence, mean intensity and mean abundance was observed. Methods: Totally 1 001 yellowfin tuna fishes were collected throughout the year of 2012 from the coast of Nagapattinam (10 °45' 36.25" N and 79 °50' 59.54" E), Tamil Nadu, India. The collected fishes were washed with fresh water to remove debris and immediately stored in ice, and brought to the laboratory. Intestines were placed in normal saline (0.85% NaCl). The worms were fixed in alcohol/ formalin /(AFA) fixative and in 70% ethanol until processed for identification. Results: Totally 1 001 yellowfin tuna was examined out of 387 (52.67%) hosts were infected with acanthocephalan (Echinorhynchus sp.). The proboscis Echinorhynchus sp. was long, cylindrical with cylindrical with a uniform width measured (0.41 ±0.02) mm in length and (0.12 ±0.03) mm in width. The endo parasite prevalence, intensity and abundance was recorded as 52.67%, 12.46%, and 5.43% respectively. Conclusions: This current phenomena was carried out to report of acanthocephalan Echinorhynchus sp. attached the new host of yellowfin tuna from Nagapattinam coast. Moreover, parasitological investigations are required to determine endo parasites of host due to its importance and numerous occurring among most important marine food fishes in world wild.

606-608

Authors: Maurizio Manera, Carla Borreca

Objective: To characterise vertebral aspects of a lordotic chub (Squalius cephalus L.) using texture analysis by radiograph and to verify possible alterations in the vertebral microarchitecture of the lordotic tract. Methods: Four vertebrae belonging to the lordotic tract were analyzed for their texture features by means of a software package (MaZda vers. 4.6) and were compared, for the same features, to other four vertebrae cranial (anterior) to the lordotic vertebrae. Results: Two representative features were selected out of 259 extracted texture features by means of the convex hull selection method. In particular, (5, 5) sum of squares (according to cooccurrence matrix method) and 45 ° short run emphasis (according to run-length matrix method) were used to classify, without classification errors, two selected spinal tracts (the lordotic one and the immediately cranial one, both corresponding to the anal fin pterigiophores vertebral portion) according to the following methods: linear discriminant analysis and hierarchical clustering. Conclusions: This is the first known application of texture analysis in medical imaging applied to fish species and represents a start point for further studies in aquariology and in aquaculture, where spinal deformities occur relative frequently and are cause of economic losses.

614-616

Authors: Caterina Faggio, Maria Pagano, Marina Morabito, Simona Armeli Minicante, Francesca Arfuso, Giuseppa Genovese

Objective: To study the activity of polysaccharides extracted from Undaria pinnatifida (U. pinnatifida) in vitro on red blood cell of Equus caballus, and compare it with heparin. Methods: Algal extracts was tested at two different concentrations 10 mg/mL and 20 mg/mL. In all studies, using horse red blood cells, control experiments were carried out without extract. We evaluated the toxicity of algal extracts through trypan blue test and haemolysis test and anticoagulant action measured by activated partial thromboplastin time, prothrombin time and fibrinogen test. Results: The polysaccharide extract of U. pinnatifida appeared to have no cytotoxic effect on the horse red blood cells. The values of prothrombin time, activated partial thromboplastin time and fibrinogen were significantly changed in the presence of the extract. Conclusions: This study suggests a possible exploitation of U. pinnatifida, thriving in the lagoon of Venice, as a source of anticoagulant drug, with the aim of transforming waste into a valuable biomass.

625-631

Authors: Ana D ’ors, María Carmen Bartolomé, Sebastián S ánchez-Fortún

Objective: To study mortality of copepod Artemia franciscana against the occurrence of harmful marine algae and possible toxicological changes exhibited by binary and tertiary combinations of these harmful algae toxins. Methods: Tweenty four hours acute toxicity assays were performed with selected concentrations of Alexandrium minutum, Prorocentrum lima and Nitzschia N1c1 living cells. Additionally, the results were analyzed using the median-effect/combination index (CI)-isobologram equation to assess possible changes in the toxic effect induced by phytoplankton functional groups. Results: Biotoxin equivalent values obtained by immunodetection were (2.12依0.10), (8.60依1.30) and (4.32依1.67) pg/cell for saxitoxin, okadaic acid and domoic acid, respectively. The 24-h LC50 values estimated to saxitoxin and okadaic acid equivalents were 4.06 and 6.27 µg/L, significantly below the value obtained for Nitzschia N1c1, which was established at 467.33 µg/L. CI analysis applied on phytoplankton assemblages showed that both ternary mixture as the binary combinations exhibited antagonic action on toxic effects in Artemia nauplii, which were significantly lower than the toxic effect exhibited by each species studied. Conclusions: These results show that, although these harmful algae represent a serious risk to estuarine zooplankton community, the presence of phytoplankton functional groups within the same bloom can reduce the potential risk compared to the expected risk when each of the phytoplankton groups are evaluated individually.

638-644

Authors: Siti Suria Daud, Muhammad Taher, Deny Susanti, Mohamed Zaffar Ali Mohamed Amiroudine, Tengku Muhamad Faris Syafiq Tengku Zakaria

Objective: To evaluate antioxidant and antimicrobial activities of stembark of Shorea kunstleri (S. kunstleri) together with analysis of phytochemical and total phenolic contents. Methods: Extraction was conducted with different solvent polarity of n-hexane, dichloromethane (DCM) and methanol by using Soxhlet extraction. Total phenolic content was determined using Folin-Ciocalteu method. Free radical scavenging activity and inhibition of lipid peroxidation were evaluated with DPPH radical scavenging and ferric thiocyanate assays, respectively. Antimicrobial activities were performed using disc diffusion method, minimum inhibition concentration (MIC), minimum bactericidal concentration (MBC), and minimum fungicidal concentration. Results: S. kunstleri stembark extracts revealed presence of steroids, terpenoids, saponins, flavonoids, and phenolic compounds. Methanol extract exhibited the highest total phenolic content and free radical scavenging activity resulting in phenolic content of (8.340依0.003) g GAE/100 g of extract and (95.90依1.07)% DPPH inhibition (IC 50 value of 18.6 µg/mL), respectively. Ferric thiocyanate assay of n-hexane, DCM, and methanol extracts indicated lipid peroxidation inhibitory activity of (74.20依0.35)%, (74.00依0.10)%, and (72.80依0.27)%, respectively. In antimicrobial and antifungal tests, methanol extract showed inhibition against Staphylococcus aureus (S. aureus), Candida albicans, and Candida tropicalis with inhibition zones of 10-12, 18-22, and 18-19 mm, respectively. The MIC test of methanol extract showed highest inhibition against Candida albicans and S. aureus (0.04 and 0.08 mg/mL, respectively) while DCM extract exhibited the highest activity towards Candida tropicalis (MIC value of 0.63 mg/mL). Taken together, MBC test of methanol extract strongly demonstrated bactericidal effect against S. aureus with MBC value of 0.08 mg/mL. Conclusions: The study demonstrated that stembark extracts of S. kunstleri possessed antioxidant and antimicrobial properties.

651-655

Authors: Ghalem Bachir Raho

Objective: To examine the in vitro antimicrobial activities of essential oil of the needles of Pinus halepensis (P. halepensis). Methods: The antibacterial activity of essential oil of the needles of P. halepensis was determined using the agar well diffusion technique and disc diffusion method against Staphylococcus aureus ATCC 25923, and Escherichia coli ATCC 25922. Results: The diameter of zones of inhibition exhibited by the essential oil was between 6 and 17 mm. The essential oils was compared favorably with gentamycin used as a standard control. The minimum inhibitory concentration determined by the agar well diffusion method was 0.52 mg/ mL for Staphylococcus aureus and 2.15 mg/mL for Escherichia coli. The minimum bactericidal concentration of the oils against the two microorganisms was 4.17 mg/mL. Conclusions: The results obtained from this study reveal that P. halepensis essential oils possess antibacterial activities and can be used as antimicrobial agents in the search for new drugs.

662-664

Authors: Shahrzad Azizi, Seyed Razi Ghalebi, Reza Kheirandish, Neda Ghasemi, Fateme Akrami Mohajeri

Aspergillosis is the most common mycotic infection in a wide variety of bird and causes significant economic losses. The present study described concurrent occurrence of aspergillosis and proventricular impaction in a 4-year-old male ostrich. The bird had respiratory problems, coughing and anorexia. Postmortem examination revealed numerous greenish-white caseous foci, 0.5 to 1 cm in diameter distributed on the surfaces of the air sacs and throughout the lungs. In histopathological study, multifocal areas of caseous necrosis that surrounded by inflammatory cells including heterophils, lymphocytes and macrophages were present. Long branching septated hyphae were visible in the necrotic areas with hematoxylin and eosin and Periodic acid-Schiff staining. Thrombi were present in the blood vessels. The proventriculus was full of gravel.

666-672

Authors: Aderson de Souza Alcântara

This paper aims to discuss the current status of conservation of freshwater turtles of the Amazon and the absence of the genus Podocnemis the Official List of Species of Brazilian Fauna Threatened with Extinction. Amazonian turtles are used as food by indigenous people and fisherman communities. However, fishing of adult females, uncontrolled egg collecting, habitat degradation and trafficking in wildlife have caused the decline of these populations. Nevertheless, Podocnemis expansa and Podocnemis unifilis were not included in the Brazil’s official list of animals threatened. Therefore, the turtles remain at great risk, due to the intense pressure that they are suffering. It is recommended that the criteria and the conservation status are reviewed including those animals in the category of vulnerable and to ensure a thorough review and modification in the current Brazilian law to be covered studies and management of turtles for subsistence, respecting and adding value to way of life of Amazonian peoples.

511-514

Authors: Mohammad Forouhar Vajargah, Aliakbar Hedayati

Objective: To determine toxicity and safety margins of trichlorofon in different species as trichlorofon toxicity has a large variation in the susceptibility of different species. Methods: In this research, four viviparous aquarium fish were exposed to trichlorofon for 96 h. LC 50 of 24 h, 48 h, 72 h and 96 h were attained by probit analysis software SPSS Version 16. Fish samples were exposed to different concentrations of trichlorofon (5, 10, 20 and 40 mg/L) for 96 h and mortality were recorded, separately. Results: The 96 h LC 50 of Poecilia latipinna, Poecilia reticulata, Gambusia holbrooki and Xiphophorus helleri were 9.80, 9.80, 9.95 and 7.99 mg/L, respectively. Conclusions: According to the results of this research, LC50 values indicated Xiphophorus helleri was the most resistant and Gambusia holbrooki, with a few differences, was the most sensitive species compared with the fishes examined.

520-526

Authors: Zeliha Selamoglu Talas, Mehmet Fuat Gulhan

Objective: To show hemolymph apoptosis in imported shrimp Litopenaeus vannamei from Hawaii to Iran. Methods: One hundred and eighty shrimps [(7.98依0.54) g] which were collected from a research shrimp farm located in Heleh site in north of Bushehr Province were distributed equally to 6 glass aquariums (50 cm伊50 cm伊60 cm) as group A in triplicate (imported batch in 2011, without crossing with other generations) with well clean aerated sea water (100 L per aquarium), salinity of 40 ‰ and temperature of 29 °C. Shrimps of group B (produced by crossing the adults of imported batches in 2009 up to 2011) were distributed also among 6 aquariums with the same conditions. Both shrimp groups were injected with concentration of LD50=1伊105.4 white spot virus. Results: The results showed that in group A, the mortality began approximately 24 h after exposure and reached 100% after 36 h but no mortality was occurred up to 15 d in shrimps of group B. The slide evaluation of hemolymph of group B showed an increasing trend of apoptosis occurrence in all three types of hemolymph cells, hyalinocytes, semi-granulocytes and granulocytes from 24 h to 72 h in contrary to group A that not any apoptosis was observed during the course of the study (15 d). Conclusions: It is concluded that crossing among the specific pathogen free generations could induce the increasing immunity level through apoptosis to protect them against white spot disease.

535-542

Authors: Gihan Ahmed El Shoubaky, Essam Abd El Rahman Salem

Objective: To estimate the fatty acids content in the brown algae Padina pavonica (P. pavonica) and Hormophysa triquetra (H. triquetra) and evaluate their potential antimicrobial activity as bioactive compounds. Methods: The fatty acid compositions of the examined species were analyzed using gas chromatography-mass spectrometry. The antimicrobial activity of crude and fatty acids was assessed using the agar plug technique. Results: The fatty acids profile ranged from C8:0 to C20:4. Concentration of saturated fatty acids in P. pavonica was in the order palmitic>myristic>stearic whereas concentration of the unsaturated fatty acids was oleic acid>palmitoleic>9-cis-hexadecenoic>linoleic acid>α- linolenic>arachidonic> elaidic acid. H. triquetra contained high concentration of saturated fatty acids than those of P. pavonica which was in the order as follows: palmitic>margaric>myristic> nonadecyclic>stearic>caprylic>tridecylic>pentadecylic>lauric while the unsaturated fatty acids consisted of oleic>nonadecenoic>non adecadienoate>margaroleic. The crude and fatty acid extracts of H. triquetra and P. pavonica were biologically active on the tested pathogens. H. triquetra exhibited a larger inhibitory zone than P. pavonica. Conclusions: The brown algae P. pavonica and H. triquetra have high efficient amount of fatty acids and showed strong antibacterial activity, especially H. triquetra.

555-559

Authors: Abdulselam Ertaş, Mehmet Boğa, Yeter Yeşil

Objective: To evaluate the chemical compositions and biological activities of an endemic Ballota nigra L. subsp. anatolica P.H. Davis. Methods: Essential oil and fatty acid composition were determined by GC/MS analysis. ABTS cation radical decolourisation and cupric reducing antioxidant capacity assays were carried out to indicate the antioxidant activity. The anticholinesterase potential of the extracts were determined by Ellman method. Results: The major compounds in the fatty acid composition of the petroleum ether extract were identified as palmitic (36.0%) and linoleic acids (14.3%). The major components of essential oil were 1-hexacosanol (26.7%), germacrene-D (9.3%) and caryophyllene oxide (9.3%). The water extract indicated higher ABTS cation radical scavenging activity than α-tocopherol and BHT, at 100 µg/ mL. The acetone extract showed 71.58 and 44.71% inhibitory activity against butyrylcholinesterase and acetylcholinesterase enzyme at 200 µg/mL, respectively. Conclusions: The water and acetone extracts of Ballota nigra subsp. anatolica can be investigated in terms of both phytochemical and biological aspects to find natural active compounds.

566-570

Authors: Saravanakumar Kandasamy, Kathiresan Kandasamy

Objective: To test antioxidant property of the endophytic Trichoderma species isolated from the leaves of 12 mangroves of Andaman Nicobar Islands. Methods: Eight strains of Trichoderma species were found predominant and their crude extracts were assessed for antioxidant activity by using seven assays. Results: Total antioxidant activity varied with the strains and it was maximum in Trichoderma EMFCAS8 and other strains also showed considerable activity. Conclusions: This work concluded that mangroves are rich in endophytic Trichoderma species with potential for antioxidant activity.

579

Authors: Nguyen Van Hoa, Viroj Wiwanitkit

No

426-430

Authors: Iruthayam Vijaya Kumar, Gurusamy Chelladurai, Thangapandi Veni, S. Syed Hussain Peeran, Jayaraj Mohanraj

Objective: To study the effect of three medicinal plants on growth, survival and immunoprotection in Mystus montanus fingerlings. Methods: The three experimental groups of fish were fed with a diet supplemented of Ocimum tenuiflorum, Zingiber officinale and Allium cepa (0.5 g each), and control group was fed without supplementation of herbal diet. After 45 d of feeding the experimental group fishes were injected with 0.5 mL of Aeromonas hydrophila and were supplemented with herbal diet. In the control group fishes were injected with 0.5 mL of physiological saline solution alone. Results: The blood samples were collected for haemotological analysis after 7 d of post infection. The results showed that the haematological parameters of haemoglobin, serum glutamicoxaloacetic transaminase, serum glutamic pyruvic transaminase, serum glucose, red blood cell and white blood cell were found to be more in challenged fish. The survival rate of pathogen challenged fish maintained on the herbal diet is better than those in the control group which is supplemented with non-herbal diet. Conclusions: These results indicated that herbal diet is useful as a growth promoter and for disease resistance of Mystus montanus against Aeromonas hydrophila infection.

438-441

Authors: Thangapalam Jawahar Abraham

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 104 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.

447-452

Authors: Youngdeuk Lee, Saranya Revathy K, Sukkyoung Lee, Ilson Whang, Chulhong Oh, Do-Hyung Kang, Gee-Wook Shin, Jehee Lee, Mahanama De Zoysa

Objective: To investigate molecular characterization and the immune responses of Manila clam hydramacin (Rp-hdmc). Methods: cDNA sequence of hydramacin was isolated from Manila clam transcriptome database. Molecular characterization of hydramacin cDNA was performed by BLAST and SWISS-MODEL bioinformatics programs. Tissue-specific expression and transcriptional regulation after Vibrio tapetis challenge was done by quantitative real time PCR. Results: Rp-hdmc has 291 bp open reading frame (ORF), encoding 97 amino acids with a mature hydramacin consisting of 77 amino acid residues. In un-challenged clam, Rp-hdmc was constitutively expressed in all tested tissues and the highest expression level was detected in gill. After pathogenic bacteria Vibrio tapetis challenge, Rp-hdmc mRNA was up-regulated in gill and hemocytes. Conclusions: We identified hydramacin cDNA (Rp-hdmc) from mollusk Manila clam that shows the characteristic features of hydramacin sequence. It has eight cysteine residues with four disulfide linkages, three helices and two β-strands in secondary structure. Expression results after V. tapetis challenges suggest that Rp-hdmc is involved in immune response against pathogenic bacteria.

458-464

Authors: Emiliano Felici, Cesar Almeida, Martin Fernández Baldo, Luis D. Martínez, Fanny Zirulnik, María R. Gomez

Objective: To analyze the effect of cadmium (Cd) on soybean seedlings growth and the relationship with the distribution and concentration of macro-microelements. Methods: The ions concentrations were determined by ICP-MS. The extraction efficiency and digestion time were optimized. Also, oxidative stress parameters were determined and related with metal content. Results: The accumulated amount of dry matter in roots and leaves was lower in the Cd-treated group. Regression analysis showed that the exposure to Cd affected the accumulated amount of dry matter as well as the content of mineral elements in the analysis samples. In Cd treated plants, electrical conductivity increased respect to the controls, indicating that ionic permeability became altered. A strong inhibition of the chlorophylls (chl) biosynthesis in the Cd-treated group was also demonstrated by a decrease of chla and chlb concentration. This result was related with the observed significant decrease in the Mg uptake at the roots and leaves level. Conclusions: The stress caused by Cd exposure, evidenced by significantly high hydrogen peroxide levels in roots and leaves after 24 h and the content of specific macro-microelements is a factor that affects the accumulation of dry matter, electrical conductivity and chlorophylls concentration.

471-477

Authors: Kidanemariam Girmay, Bemnet Fikre, Atsede Asmelash, Biskut Getachew, Emebet Tekle, Nagappan Raja

Objective: To evaluate larvicidal and pupicidal activities of aqueous and ethanol extract of different parts of Schinus molle against filarial vector, Culex quinquefasciatus (Cx. quinquefasciatus) in the laboratory. Methods: The mortality rate of third, fourth instar larvae and pupal stages were tested at 20, 40, 60, 80 and 100 mg/L of plant extract using WHO standard protocol with modifications. The mortality rate was recorded continuously for 24, 48 and 72 h post exposure period and percentage mortality was calculated. Results: Maximum percentage mortality of third instar was 83.3% in ethanol extract of mature fruit at 100 mg/L after 24 h exposure period. After 48 h exposure period, 93.3% percentage mortality was recorded in ethanol extract of immature fruit at 100 mg/L. After 72 h exposure period, 100% mortality was recorded in water extract of leaf at 100 mg/L. In fourth instar larvae, maximum percentage mortality of 63.3% was recorded in water extract of mature fruit and ethanol extract of immature and mature fruit at 100 mg/L after 24 h exposure period. After 48 h exposure period 86.6% mortality was recorded in ethanol extract of mature fruit at 100 mg/L. After 72 h exposure period, 93.3% mortality was recorded in ethanol extract of mature fruit at 100 mg/L. In general immature Cx. quinquefasciatus, percentage mortality was increased with increase in exposure time and concentration of the plant extracts tested. Conclusions: From this laboratory study, Schinus molle plant parts were proved to have larvicidal and pupicidal activity against immature Cx. quinquefasciatus.

484-489

Authors: Marley García, Ramón Scull, Osmany Cuesta, Gaëlle Boulet, Louis Maes, Paul Cos, Lianet Monzote

Objective: To investigate the leishmanicidal effect of the Bixa orellana crude seed extract and its fractions against Leishmania amazonensis. Methods: Four main fractions (BO-A, BO-B, BO-C and BO-D) were obtained by exhaustion with solvent with increased polarity from the Bixa orellana crude seed extract and 28 sub-fractions. The antileishmanial activity was evaluated in intracellular amastigotes and the cytotoxicity was assessed in murine intraperitoneal macrophages. Results: The BO-A and BO-B fractions showed a good antileishmanial activity with IC50 values of (12.9±4.1) and (12.4±0.3) μg/mL, respectively. The sub-fractions BO-B1 (IC50=(11.8±3.8) μg/mL) and BO-B3 [IC50=(13.6±4.7) μg/mL] also proved to have a good leishmanicidal effect. In general, the sub-fractions showed a lower toxicity than the crude extract. A selectivity index of 9 indicated a moderate selectivity of the BO-A, BO-B and BO-C fractions and BO-B1 sub-fraction. Conclusions: Potential of this plant against cutaneous leishmaniasis should be further investigated.

496-499

Authors: Musa Abdel-Raouff, Mohamed Mobarak Elbasheir

Objective: To determine the sero-prevalence of Toxoplasma gondii (T. gondii) infection among pregnant Sudanese women. Methods: One hundred and sixty three pregnant women attending antenatal care in Omdurman Maternity Hospitals, Khartoum, Sudan during June to August in 201 3 were enrolled and screened for immunoglobulin G (IgG) and IgM antitoxoplasma antibodies using enzyme linked immunosorbent assay technique. Results: Among 163 pregnant women, 33 (20.2%) were positive for (IgG) antitoxoplasma antibodies, while 130 (79.8%) were seronegative. None of the examined women had IgM antitoxoplasma antibodies. The highest rate of infection (26.7 %) was detected among women aged 21-29 years. No statistically significant relation was observed between T. gondii sero-prevalence and the other variable of risk factors studied. Conclusions: Over 79% Sudanese women screened for antitoxoplasma IgG antibodies were seronegative and they were at risk of seroconversion during pregnancy. Moreover, the study showed that screening of T. gondii infections during antenatal care should be considered in Khartoum state as the main strategy to minimize congenital toxoplasmosis.

501-504

Authors: Angel Alberto Noda, Islay Rodríguez

Objective: To compare the value of leptospiral DNA extraction procedures from clinical samples for the early diagnosis of leptospirosis. Methods: Three DNA extraction procedures were applied for microbiological analysis, results of QIAmp DNA mini kit (QIAGEN, Germany), CLART HPV kit (GENOMICA, Spain) and Chelex-100 assay were compared concerning extraction efficiency, DNA purity and DNA suitability for amplification by specific polymerase chain reaction for pathogenic leptospires from blood, plasma and serum artificially infected. Results: The comparison of extraction methods highlighted the efficiency of Chelex-100 and QIAmp DNA mini kit. Chelex-100 achieved the isolation of the highest concentration of leptospiral DNA from the culture and the spiked samples, with acceptable purities and without inhibitors to PCR. Conclusions: Chelex-100 assay is a rapid and effective approach for DNA isolation in clinical samples having pathogenic leptospires and it could be useful in the early diagnosis of leptospirosis.

344-349

Authors: Hasna Kadri, Sondes Marouani, Mohamed Nejmeddine Bradai, Abderrahmen Bouaïn, Eric Morize

Objective: To investigate the length distribution, sex ratio, length-weight relationship, age and growth of Raja radula in the Gulf of Gabes. Methods: The age and growth characteristic of the rough skate (Raja radula) inhabiting in the Gulf of Gabes (Central Mediterranean Sea) was determined. A total of 1 250 specimens were sampled. According to the marginal increment band its growth was annual. Results: The growth parameters were derived by using the von-Bertalanffy method and were separately evaluated as follows: TL∞=97.2 cm, K= 0.15 y-1, to= 0.35, and W∞= 5.28 kg, and TL∞= 76.4 cm, K = 0.22 y-1, to= 0.16, and W∞= 3.77 kg, for females and males respetively. The maximum age was 12 years for females and 10 years for males. Conclusions: Results from this research will provide a starting point to develop a management plan for the rough skate in the Gulf of Gabes. Further investigations are necessary to quantify the impact of the existing regulations on the population dynamics and recruitment patterns of this species in the region.

357-361

Authors: Seyed Ali Johari, Mohammad Reza Kalbassi, Il Je Yu

Objective: To investigate the effects of powdered silver zeolite (SZ) on the in vitro growth of the fish pathogen Saprolegnia sp. Methods: The antifungal activity of SZ was evaluated by determining the minimum inhibitory concentrations using two-fold serial dilutions of powdered SZ in a glucose yeast extract agar at 22 °C. The growth of Saprolegnia sp. on the SZ agar treatments was compared to that on SZ-free agar controls. Results: The results showed that SZ had an inhibitory effect on the in vitro growth of the tested fungi. The minimum inhibitory concentration of SZ for Saprolegnia sp. was also calculated at 600 mg/L, which is equal to 0.06 percent. Conclusions: SZ is a potential good candidate to replace teratogenic and toxic agents, such as malachite green in aquaculture systems.

367-375

Authors: Rajasekar Thirunavukkarasu, Priyadharshini Pandiyan, Kumaran Subaramaniyan, Deivasigamani Balaraman, Sakthivel Manikkam, Balamurugan Sadaiyappan, George Edward Gnana Jothi

Objective: To isolate bioactive molecules from marine seaweeds and check the antimicrobial activity against the fish pathogenic bacteria. Methods: Fresh marine seaweeds Gracilaria edulis, Kappaphycus spicifera, Sargassum wightii (S. wightii) were collected. Each seaweed was extracted with different solvents. In the study, test pathogens were collected from microbial type culture collection. Antibacterial activity was carried out by using disc diffusion method and minimum inhibition concentration (MIC) was calculated. Best seaweed was analysed by fourier transform infrared spectroscopy. The cured extract was separated by thin layer chromatography (TLC). Fraction was collected from TLC to check the antimicrobial activity. Best fraction was analysed by gas chromatography mass spectrometer (GCMS). Results: Based on the disc diffusion method, S. wightii showed a better antimicrobial activity than other seaweed extracts. Based on the MIC, methanol extract of S. wightii showed lower MIC than other solvents. S. wightii were separated by TLC. In this TLC, plate showed a two fraction. These two fractions were separated in preparative TLC and checked for their antimicrobial activity. Fraction 2 showed best MIC value against the tested pathogen. Fraction 2 was analysed by GCMS. Based on the GCMS, fraction 2 contains n-hexadecanoic acid (59.44%). Conclusions: From this present study, it can be concluded that S. wightii was potential sources of bioactive compounds.

382-388

Authors: Samaresh Samanta, Tapas Kumar Das, Amalesh Choudhury, Susanta Kumar Chakraborty

Objective: To record the fractional components of lipid and polyunsaturated fatty acids of Lingula anatina (L. anatina), a Precambrian intertidal benthic brachiopod, giving emphasis on -ω series group especially eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) alongside assessing their biotransformation within the population and mangrove-estuarine associated community. Methods: Different biological samples after being collected from three contrasting study sites viz. SI, SII and SIII at Talsari (Longitude 87 °5′ E to 88 °5′ E and Latitude 20 °30′ N to 22 °2′ N) were stored at -20 °C until analyzed. Total lipids were extracted from each sample following Bligh and Dryer method. Identification and conformation of fatty acids were done by following Ackman method. Results: On analyzing different collected samples, muscles of L. anatina exhibited the highest amount of total lipids (2.95%) of which 54.03% belongs to phospholipid groups. Different body parts of studied species contained appreciable and greater amount of EPA and DHA than α-linolenic acid. Conclusions: Different collected samples exhibited variabilities in respect of total lipids and its fractional fatty acid components. The muscles of L. anatina showed maximum storage of lipids and fatty acids. Differential occurrences of EPA and DHA in different body parts of L. anatina are supposed to be due to the biotransformation process converting the α-linolenic acid from its primary food sources.

394-401

Authors: Md. Sarfaraj Hussain, Sheeba Fareed, Mohammad Ali, Md. Sarfaraz Alam, Md. Akhlakquer Rahman, A. K. Srivastava

Objective: To perform a simultaneous quantitative estimation of two biologically active triterpenoid compounds lupeol and a steroid compound, stigmasterol, in Abutilon indicum (A. indicum) using high-performance thin-layer chromatography (HPTLC). Methods: TLC aluminum plates precoated with silica-gel 60 F254 (20 cm×10 cm) were used with a mobile phase of toluene-methanol-formic acid (7.0:2.7:0.3, v/v/v) and densitometric determination of these compounds was carried out at 530 nm in reflectance/absorbance mode. Results: Compact bands for lupeol and stigmasterol were obtained at Rf 0.52±0.02 and 0.28±0.05. The limit of detection (45 and 18 ng/band), limit of quantification (135 and 54 ng/band), recovery (98.2%-99.7% and 97.2%-99.6%) and precision (≤ 2.18 and 1.91) were satisfactory for lupeol and stigmasterol respectively. Linearity range for lupeol and stigmasterol were 100-1000 (r2=0.999 4) and 50-500 ng/band (r2=0.994 1) and the contents were estimated as (0.59±0.10)% and (0.83依0.10)% w/ w respectively. The total phenolic, flavonoid, proanthocyanidin, alkaloidal and saponin contents of methanolic extract of A. indicum were also measured in this work. According to International Conference on Harmonization (ICH) guidelines, the method was validated for linearity, precision, accuracy, and recovery, limit of detection, limit of quantification, specificity, and robustness. Conclusions: The HPTLC method was found to be reproducible, accurate, and precise and could detect these two compounds at nanogram level from the A. indicum.

411-415

Authors: Suhaila Qari, Samia Moharram, Safaa Alowaidi

Objective: To determine the fatty acids profiles in female fish, Rhabdosargus sarba (R. sarba) from the Red Sea during the spawning season. Methods: Monthly individual R. sarba were obtained from Bangalah market in Jeddah, Red Sea and transported to the laboratory in ice aquarium. The total length, standard length and weight were measured, fishes were dissected. Ovaries were removed, weighed and 10 mL of concentrated hydrochloric acid were added to 10 g of the ovary in a conical flask and immersed in boiling water until the sample was dissolved and the fat was seen to collect on the surface. The conical was cooled and the fat was extracted by shaking with 30 mL of diethyl ether. The extract was bowled after allowing the layers to separate into a weighed flask. The extraction was repeated three times more and distilled off the solvent then the fat dried at 100 °C, cooled and weighed. Then 50 mg of lipid was put in a tube, 5 mL of methanolic sulphuric acid was added and 2 mL of benzene, the tube well closed and placed in water bath at 90 °C for an hour and a half. After cooling, 8 mL water and 5 mL petroleum were added and shaked strongly and the ethereal layer was separated in a dry tube, evaporated to dryness. The fatty acid methyl esters were analyzed by using a Hewlett Packard (HP 6890) chromatography, a split/splitless injector and flame ionization detector. Results: In female R. sarba, a total of 29 fatty acids were detected in ovaries throughout the spawning season. The main fatty acid group in total lipid was saturated fatty acid (SFA, 28.9%), followed by 23.5% of polyunsaturated fatty acids (PUFA) and 12.9% of monounsaturated fatty acids (MUFA). The dominant SFA were palmitic and stearic, the major MUFA were palmitoleic and oleic, and the major PUFA were C18:2 and C22:2. During spawning stages, there were no significant differences in total SFA, MUFA and PUFA. The highest value of SFA was in late spawning (36.78%). However, the highest value of MUFA and PUFA was in spawning (16.70% and 24.96% respectively). During spawning season there were significant differences in total SFA between March (late spawning stage) and December (nearly ripe stage), P<0.05. Conclusions: From the present study it has been shown that 29 fatty acids were detected in ovaries of R. sarba throughout the spawning season. The main fatty acid group in total lipid was SFA followed by PUFA and MUFA. The dominant SFA were palmitic and stearic, the major MUFA were palmitoleic and oleic, and the major PUFA were C18:2 and C22:2. The highest value of SFA was in late spawning. However, the highest value of MUFA and PUFA was in spawning.

259-263

Authors: Syed Ariful Haque, Md. Shaheed Reza, Md. Rajib Sharker, Md. Mokhlasur Rahman, Md. Ariful Islam

Objective: To observe the effectiveness of most widely used antibiotic, oxytetracycline (OTC) in reducing the bacterial load in rohu fish under artificial culture condition in the laboratory. Methods: The experiment was conducted in the Faculty Fisheries, Bangladesh Agricultural University, Mymensingh-2202. The fish were reared in 8 aquaria where fish in 5 aquaria were used for replication of the treatment (experimental group) and fish in remaining 3 aquaria were considered as a control (Control group). OTC was fed to the fish in the experimental aquarium at the rate of 2 g/kg through diet twice daily whereas fish reared under control condition was given feed without antibiotic for 20 d and bacterial content in the aquarium water, gills, skin and intestine of fish were estimated at every alternative day after onset of the experiment. Results: Rearing the fish with OTC treated feed resulted in gradual decrease of bacterial load in the aquarium water, gills, intestine and skin of the fish whereas the content remain unchanged or little increased in the control group. Water quality parameters such as dissolved oxygen, pH and total hardness were within the suitable range in the experimental aquarium but not in control aquaria throughout the experimental period. Conclusions: These results suggest that OTC could be a potential antibiotic to reduce the bacterial load in fish and can be used commercially for maintaining the fish health in aquarium conditions.

270-273

Authors: Milena Polifrone, Miquel Rosell-Fieschi

Objective: To monitor the distribution of Cymodocea nodosa (Ucria) Ascherson in Las Canteras Beach (Las Palmas de Gran Canaria, Spain), comparing the status in 2005 with the distribution observed in 1985 and 1995. Methods: Field observations by selfcontained underwater breathing apparatus diving records and cartographic report. Results: The seagrass meadow originally distributed between the beach and a bar of volcanic rocks, which sheltered it from the strong north-east winds and swells. Since 1985 a gradual reduction of its extent has been observed and more than 80% of the original canopy disappeared in twenty years time, mostly due to the anthropogenic impact and modification of the sedimentary dynamics of the beach. Conclusions: The degradation of this seagrass meadow determinates the loss of the only meadow in the north of the island of Gran Canaria and the absence of a management plan for its conservation.

281-286

Authors: Kumaran Subramanian, Deivasigamani Balaraman, Dhayanithi Nagarajan Balachandran3, Rajasekar Thirunavukarasu, Suresh Gopal, Pugazhvendan Sampath Renuka, Alagappan Kumarappan

Objective: To analyse experimental infection and immune system of shrimp (Penaeus monodon) against Vibrios furnissii (V. furnissii). Methods: Experimental animals were collected and acclimatized by maintaining specific temperature, pH and salinity to avoid mortality. Shrimps were experimentally infected with V. furnissii and their immune responses were monitored. After the infection all the shrimps were monitored for any symptoms, death rate in 0, 12, 24, 36, 48 h. Then haemolymph were collected and tetrahydrocannabinol, phenol oxidase, nitroblue tetrazolium and lysozyme were monitored in every 12 h at the interval of 48 h. Results: Shrimps infected by live V. furnissii had showed gradual increase in tetrahydrocannabinol, phenol oxidase activity, nitro-blue-tetrazolium and lysozyme activity comparing with the killed and control. Conclusions: The live V. furnissii had showed infection in the shrimp immune system. The live V. furnissii shows infection in experimental shrimps comparing with killed V. furnissii. So the V. furnissii in nature cause the infection in shrimp Penaeus monodon immune system. This report could be applied to control of the infection in shrimp hatchery.

297-301

Authors: Shirin Forouzan, Amir Rahimirad, Roya Seyedkhoei, Jafar Asadzadeh, Mahmoud Bahmani

Objective: To monitor antibiotic residues in pasteurized milk in West Azerbaijan province, North West of Iran. Methods: In this study, 848 pasteurized milk samples were collected from factories and tested for the presence of antibiotic residues using the Copan test kit based on the manufacturer’s instructions. Results: Results indicated that 30.14% of samples were contaminated with a variety of antibiotics based on the detection of associated residues and 3.19% of these samples were suspected. Given the current rise of antimicrobial resistance among microbial pathogens, these findings amplify the need to ensure continuous monitoring of pasteurized milk that intended for human consumption. Conclusions: Continuous monitoring of pasteurized milk may improve human health but also limit the development and transmission of antibiotic resistant strains in the environment.

308-312

Authors: Govindarajan Marimuthu, Rajeswary Mohan

Objective: To assess the larvicidal and ovicidal potential of the crude hexane, benzene, chloroform, ethyl acetate and methanol solvent extracts from the medicinal plant, Pithecellobium dulce (P. dulce) against filariasis vector mosquito, Culex quinquefasciatus (Cx. quinquefasciatus). Methods: Twenty five early third instar larvae of Cx. quinquefasciatus were exposed to various concentrations and were assayed in the laboratory by using the protocol of WHO (2005). The larval mortality was observed after 24 h of treatment. The ovicidal activity was determined against Cx. quinquefasciatus mosquito eggs to various concentrations ranging from 100-750 mg/L under the laboratory conditions. Results: The methanol extract of the leaves and seed of P. dulce was the most effective against the larvae with LC 50 and LC90 values 164.12 mg/L, 214.29 mg/L, 289.34 mg/L and 410.18 mg/L being observed after 24 h of exposure. The efficacy of methanol was followed by that of the ethyl acetate, chloroform, benzene and hexane extracts. The mean percent hatchability of the egg rafts were observed after 48 h of treatment. About 100% mortality was observed at 500 mg/L for leaf and 750 mg/L for seed methanol extracts of P. dulce. Conclusions: From the results, it can be concluded that the larvicidal and ovicidal effect of P. dulce against Cx. quinquefasciatus make this plant product promising as an alternative to synthetic insecticide in mosquito control programs.

319-323

Authors: Rohollah Dehghani, Hamid kassiri, Narges Mehrzad, Nilofar Ghasemi

Objective: To investigate the status of cutaneous leishmaniasis (CL) during 2008 through 2012 in Badrood City. Methods: This is a descriptive-analytical study that was conducted during 2008-2012. The statistical population consisted of all patients, who were under treatment and follow up with the diagnosis of CL through laboratory confirmation (amastigotes view in the lesion smear) and clinical symptoms. The impression smears were prepared from the sides of ulcers of all patients by scraping; then, they were fixed in methanol, stained by standard Geimsa method, and observed exactly under the microscope. For this purpose, a researcher-made questionnaire was completed; after extracting the information, the results were analyzed using the SPSS version 17. Results: Among the number of 499 patients studied, 297 cases (59.5%) were male and the 202 cases (40.4%) were female. Slides prepared from the lesions of these patients had a Leishmania parasite with large vacuole (the features of Leishmania major). The highest cases of disease were related to the year 2012 with 190 patients, and later 2010, 2009, 2008, and 2011 respectively had the highest cases. In total, 26 patients (5.2%) have had the disease history. The highest incidence of the disease was respectively in the months of September (143 cases), August (112 cases) and October (106 cases). The most ulcers (38.08%) were observed on the hand; then the foot and face had the maximum number of the ulcer. Also, in most patients (53.9%) over an ulcer were observed. Conclusions: The results of this study showed an increase in the prevalence of CL during the years 2008 to 2012. It can be said that the main reason for this increase is people's lack of awareness of the disease transmission method. The increased disease process can be controlled through education and appropriate prevention methods.

332-336

Authors: Satinder Kakar, Ramandeep Singh, Alok Semwal

Area of drug delivery is vast, and various advances have been made in the medical field. Besides the versatility in the dosage forms, various orders for the drug release are known, which includes zero order, first order, Higuchi model, Hixon Crowell model and Korsmeyer Peppas model. In vitro dissolution is recognized as an important element in the development of drug. The nature of the drug such as its shape, crystallinity, particle size and solubility reflects the kinetics of the drug. Various models are used to study the dissolution profiles of the new drug substances. Qualitative and quantitative changes in the drug alters the drug release and performance that is action of drug in the body, which is in vivo performance. Various model dependent methods and model independent methods have been taken into consideration for studying the drug release kinetics.