Abstract: Marine Protected Areas can benefit from nature based
tourism, monitoring environmental impacts and also become target
for human presence. From more than 3 million tourists visiting
Cozumel Island every year, an average of 2,8 million arrive by cruise
ship, and 41% are estimated to have motivation for water activities.
The destination is relying so much on the tourism activity, that scuba
diving and snorkeling in the National Park Reef of Cozumel sustain
the major economic activity. In order to achieve the sustainable
development indicator designed for regional environmental
development, the PNAC offers a training course to tourism providers
to access the protected area. This way, the update of the last 5 years
of such training is directed to diving staff, boat crew and
professionals, making them able to assist in managing the natural
resource. Moreover, the case study is an example to be used for
raising awareness among tourists visiting protected areas.
Abstract: The biosynthesis of nanoparticles by microorganisms,
on the contrary to chemical synthesis, is an environmentally-friendly
process which has low energy requirements. In this investigation, we
used the microorganism Geobacillus wiegelii, strain GWE1, an
aerobic thermophile belonging to genus Geobacillus, isolated from a
drying oven. This microorganism has the ability to reduce selenite
evidenced by the change of color from colorless to red in the culture.
Elemental analysis and composition of the particles were verified
using transmission electron microscopy and energy-dispersive X-ray
analysis. The nanoparticles have a defined spherical shape and a
selenium elemental state. Previous experiments showed that the
presence of the whole microorganism for the reduction of selenite
was not necessary. The results strongly suggested that an intracellular
NADPH/NADH-dependent reductase mediates selenium
nanoparticles synthesis under aerobic conditions. The enzyme was
purified and identified by mass spectroscopy MALDI-TOF TOF
technique. The enzyme is a 1-pyrroline-5-carboxylate dehydrogenase.
Histograms of nanoparticles sizes were obtained. Size distribution
ranged from 40-160 nm, where 70% of nanoparticles have less than
100 nm in size. Spectroscopic analysis showed that the nanoparticles
are composed of elemental selenium. To analyse the effect of pH in
size and morphology of nanoparticles, the synthesis of them was
carried out at different pHs (4.0, 5.0, 6.0, 7.0, 8.0). For
thermostability studies samples were incubated at different
temperatures (60, 80 and 100 ºC) for 1 h and 3 h. The size of all
nanoparticles was less than 100 nm at pH 4.0; over 50% of
nanoparticles have less than 100 nm at pH 5.0; at pH 6.0 and 8.0 over
90% of nanoparticles have less than 100 nm in size. At neutral pH
(7.0) nanoparticles reach a size around 120 nm and only 20% of them
were less than 100 nm. When looking at temperature effect,
nanoparticles did not show a significant difference in size when they
were incubated between 0 and 3 h at 60 ºC. Meanwhile at 80 °C the
nanoparticles suspension lost its homogeneity. A change in size was
observed from 0 h of incubation at 80ºC, observing a size range
between 40-160 nm, with 20% of them over 100 nm. Meanwhile
after 3 h of incubation at size range changed to 60-180 nm with 50%
of them over 100 nm. At 100 °C the nanoparticles aggregate forming
nanorod structures. In conclusion, these results indicate that is
possible to modulate size and shape of biologically synthesized
nanoparticles by modulating pH and temperature.
Abstract: Food contamination occurs during post process
handling. This leads to spoilage and growth of pathogenic
microorganisms in the food, thereby reducing its shelf life or
spreading of food borne diseases. Several methods are tried and one
of which is use of antimicrobial packaging. Here, papain, a protease
enzyme, is covalently immobilized with the help of glutarldehyde on
polyurethane and used as a food wrap to protect food from microbial
contamination. Covalent immobilization of papain was achieved at a
pH of 7.4; temperature of 4°C; glutaraldehyde concentration of 0.5%;
incubation time of 24h; and 50mg of papain. The formation of -C=Nobserved
in the Fourier transform infrared spectrum confirmed the
immobilization of the enzyme on the polymer. Immobilized enzyme
retained higher activity than the native free enzyme. The modified
polyurethane showed better reduction of Staphylococcus aureus
biofilm than bare polymer film (eight folds reduction in live colonies,
two times reduction in protein and 6 times reduction in
carbohydrates). The efficacy of this was studied by wrapping it over
S. aureus contaminated cottage cheese (paneer) and cheese and
stored at a temperature of 4°C for 7days. The modified film reduced
the bacterial contamination by eight folds when compared to the bare
film. FTIR also indicated reduction in lipids, sugars and proteins in
the biofilm.
Abstract: Immunomodulators are substances that alter immune
system via dynamic regulation of messenger molecules. It can be
divided into immunostimulant and immunosuppressant. It can help to
increase immunity of people with a low immune system, and also can
help to normalize an overactive immune system. Aim of this study is
to investigate the effects of in vitro exposure to low and high doses of
several immunomodulators which include caffeine, kaloba and
quercetin on antigen-stimulated whole blood culture cytokine
production. Whole blood samples were taken from 5 healthy males
(age: 32 ± 12 years; weight: 75.7 ± 6.1 kg; BMI: 24.3 ± 1.5 kg/m2)
following an overnight fast with no vigorous activity during the
preceding 24 h. The whole blood was then stimulated with 50 μl of
100 x diluted Pediacel vaccine and low or high dose of
immunomodulators in the culture plate. After 20 h incubation (5%
CO2, 37°C), it was analysed using the Evidence Investigator to
determine the production of cytokines including IL-2, IL-4, IL-10,
IFN-γ, and IL-1α. Caffeine and quercetin showed a tendency towards
decrease cytokine production as the doses were increased. On the
other hand, an upward trend was evident with kaloba, where a high
dose of kaloba seemed to increase the cytokine production. In
conclusion, we found that caffeine and quercetin have potential as
immunosuppressant and kaloba as immunostimulant.
Abstract: The objective of the study was to select the survival of
probiotic strains when exposed to acidic and bile salts condition. Four
probiotic strains Lactobacillus casei subsp. rhamnosus TISTR 047,
Lactobacillus casei TISTR 1500, Lactobacillus acidophilus TISTR
1338 and Lactobacillus plantarum TISTR 1465 were cultured in
MRS broth and incubated at 35ºC for 15 hours before being inoculated
into acidic condition 5 M HCl, pH 2 for 2 hours and bile salt 0.3%,
pH 5.8 for 8 hour. The survived probiotics were counted in MRS agar.
Among four stains, Lactobacillus casei subsp. rhamnosus TISTR 047
was the highest tolerance specie. Lactobacillus casei subsp.
rhamnosus TISTR 047 reduced 6.74±0.07 log CFU/ml after growing
in acid and 5.52±0.05 log CFU/ml after growing in bile salt. Then,
double emulsion of microorganisms was chosen to encapsulate before
spray drying. Spray drying was done with the inlet temperature 170ºC
and outlet temperature 80ºC. The results showed that the survival of
encapsulated Lactobacillus casei subsp. rhamnosus TISTR 047 after
spray drying decreased from 9.63 ± 0.32 to 8.31 ± 0.11 log CFU/ml
comparing with non-encapsulated, 9.63 ± 0.32 to 4.06 ± 0.08 log
CFU/ml. Therefore, Lactobacillus casei subsp. rhamnosus TISTR 047
would be able to survive in gastrointestinal and spray drying condition.
Abstract: Fibrin degradation is an important part in prevention
or treatment of intravascular thrombosis and cardiovascular diseases.
Plasmin like fibrinolytic enzymes has given new hope to patient with
cardiovascular diseases by treating fibrin aggregation related diseases
with traditional plasminogen activator which have many side effects.
Various researches involving wide range of sources for production of
fibrinolytic proteases, from bacteria, fungi, insects and fermented
foods. But few have looked into endophytic fungi as a potential
source. Sixteen (16) endophytic fungi were isolated from Hibiscus sp.
leaves from six different locations in Shah Alam, Selangor. Only two
endophytic fungi, FH3 and S13 showed positive fibrinolytic protease
activities. FH3 produced 5.78cm and S13 produced 4.48cm on Skim
Milk Agar after 4 days of incubation at 27°C. Fibrinolytic activity
was observed; 3.87cm and 1.82cm diameter clear zone on fibrin plate
of FH3 and S13 respectively. 18srRNA was done for identification of
the isolated fungi with positive fibrinolytic protease. S13 had the
highest similarity (100%) to that of Penicillium citrinum strain TG2
and FH3 had the highest similarity (99%) to that of Fusarium sp.
FW2PhC1, Fusarium sp. 13002, Fusarium sp. 08006, Fusarium
equiseti strain Salicorn 8 and Fungal sp. FCASAn-2. Media
composition variation showed the effects of carbon nitrogen on
protein concentration, where the decrement of 50% of media
composition caused drastic decrease in protease of FH3 from 1.081 to
0.056 and also S13 from 2.946 to 0.198.
Abstract: The extracellular proteins secreted by bacteria may be increased in stressful surroundings, such as in the presence of antibiotics. It appears that many antibiotics, when used at low concentrations, have in common the ability to activate or repress gene transcription, which is distinct from their inhibitory effect. There have been comparatively few studies on the potential of antibiotics as a specific chemical signal that can trigger a variety of biological functions. Therefore, this study was carried out to determine the effect of Streptomycin Sulfate in regulating extracellular proteins secreted by Bacillus subtilis ATCC21332. Results of Microdilution assay showed that the Minimum Inhibition Concentration (MIC) of Streptomycin Sulfate on B. subtilis ATCC21332 was 2.5 mg/ml. The bacteria cells were then exposed to Streptomycin Sulfate at concentration of 0.01 MIC before being further incubated for 48h to 72 h. The extracellular proteins secreted were then isolated and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Proteins profile revealed that three additional bands with approximate sizes of 30 kDa, 22 kDa and 23 kDa were appeared for the treated bacteria with Streptomycin Sulfate. Thus, B. subtilis ATCC21332 in stressful condition with the presence of Streptomycin Sulfate at low concentration could induce the extracellular proteins secretion.
Abstract: The present study consisted of an applied test in meat
system to assess the effectiveness of three bio agents bacteriocinproducing
strains: Lm24: Lactobacillus sakei, Lm14and Lm25:
Pediococcus spp. Two tests were carried out: The ex situ test was
intended for three batches added with crude bacteriocin solutions at
12.48 AU/ml for Lm25 and 8.4 AU/ml for Lm14 and Lm24. However, the
in situ one consisted of four batches; three of them inoculated with
one bacteriocinogenic Lm25, Lm14, Lm24, respectively. The fourth one
was used in mixture: Lm14+m24 at approximately of 107 CFU/ml. The
two used tests were done in the presence of the pathogen
St. aureus ATCC 6538, as a test strain at 103 CFU/ml. Another batch
served as a positive or a negative control was used too. The
incubation was performed at 7°C. Total viable counts, staphylococci
and lactic acid bacteria, at the beginning and at selected times with
interval of three days were enumerated. Physico-chemical
determinations (except for in situ test): pH, dry mater, sugars, fat and
total protein, at the beginning and at end of the experiment, were
done, according to the international norms. Our results confirmed the
ex situ effectiveness. Furthermore, the batches affected negatively the
total microbial load over the incubation days, and showed a
significant regression in staphylococcal load at day seven, for Lm14,
Lm24, and Lm25 of 0.73, 2.11, and 2.4 log units. It should be noticed
that, at the last day of culture, staphylococcal load was nil for the
three batches. In the in situ test, the cultures displayed less inhibitory
attitude and recorded a decrease in staphylococcal load, for Lm14,
Lm24, Lm25, Lm14+m24 of 0.73, 0.20, 0.86, 0.032 log units. Therefore,
physicochemical analysis for Lm14, Lm24, Lm25, Lm14+m24 showed an
increase in pH from 5.50 to 5.77, 6.18, 5.96, 7.22, a decrease in dry
mater from 7.30% to 7.05%, 6.87%, 6.32%, 6.00%.This result
reflects the decrease in fat ranging from 1.53% to 1.49%, 1.07%,
0.99%, 0.87%; and total protein from 6.18% to 5.25%, 5.56%,
5.37%, 5.5%. This study suggests that the use of selected strains as
Lm25 could lead to the best results and would help in preserving and
extending the shelf life of lamb meat.
Abstract: Muscid flies are known to be vectors of disease agents and species that annoy humans and domesticated animals. An example of these flies is Musca domestica (house fly) whose adult and immature stages occur in a variety of filthy organic substances including household garbage and animal manures. They contribute to microbial contamination of foods. It is therefore imperative to control these flies as a result of their role in Public health. The second and third instars of Musca domestica (Linn) were infected with varying cell loads of Bacillus subtilis in vitro for a period of 48 hours to evaluate its larvicidal activities. Mortality of the larvae increased with incubation period after treatment with the varying cell loads. Investigation revealed that the second instars larvae were more susceptible to treatment than the third instars treatments. Values obtained from the third instar group were significantly different (P
Abstract: The volatile organic compounds - BTEX (Benzene, Toluene, Ethylbenzene, and Xylene) petroleum derivatives, have high rates of toxicity, which may carry consequences for human health, biota and environment. In this directon, this paper proposes a method of treatment of these compounds by using corona discharge plasma technology. The efficiency of the method was tested by analyzing samples of BTEX after going through a plasma reactor by gas chromatography method. The results show that the optimal residence time of the sample in the reactor was 8 minutes.
Abstract: The goal of presented work is the development phytoremediation method targeted to cleaning environment polluted with organochlorine pesticides, based on joint application of plants and microorganisms. For this aim the selection of plants and microorganisms with corresponding capabilities towards three organochlorine pesticides (Lindane, DDT and PCP) has been carried out.
The tolerance of plants to tested pesticides and induction degree of plant detoxification enzymes by these compounds have been used as main criteria for estimating the applicability of plants in proposed technology. Obtained results show that alfalfa, maize and soybean among tested six plant species have highest tolerance to pesticides.
As a result of screening, more than 30 strains from genera Pseudomonas have been selected. As a result of GC analysis of incubation area, 11 active cultures for investigated pesticides are carefully chosen.
Abstract: IFRN – Mossoró is a Brazilian technical education institute that develops several activities to encourage entrepreneurship, such as a curricular discipline about enterprise management and the existence of a business incubator. Despite efforts, the business incubator does not produce the expected effects. Therefore, what predisposes students to start their own business? If literature review explores determinant factors like the family and personal characteristics, it can be sustained that entrepreneurship skills can be taught since primary level, until university level. This paper presents the results of research project “Empreende IFRN” to understand the entrepreneurial predisposition and intention of the students from technical level courses. Data from 365 students from technical level courses reveal an increased entrepreneurial intention of students during time (from a 2 years period to someday in the future). The entrepreneurial behavior of parents affects students’ perception about starting their own business. Students also present a cautions behavior, preferring bank deposit and investment fund instead starting a business.
Abstract: Seventy-nine accessions, including two local wild species (Chenopodium album and C. murale) and several cultivated quinoa lines developed through recurrent selection in Morocco were screened for their resistance against Peronospora farinose, the causal agent of downy mildew disease. The method of artificial inoculation on detached healthy leaves taken from the middle stage of the plant was used. Screened accessions showed different levels of quantitative resistance to downy mildew as they were scored through the calculation of their area under disease progress curve and their two resistance components, the incubation period and the latent period. Significant differences were found between accessions regarding the three criteria (Incubation Period, Latent Period and Area Under Diseases Progress Curve). Accessions M2a and S938/1 were ranked resistant as they showed the longest Incubation Period (7 days) and Latent Period (12 days) and the lowest area under diseases progress curve (4). Therefore, M24 is the most susceptible accession as it has presented the highest area under diseases progress curve (34.5) and the shortest Incubation Period (1 day) and Latent Period (3 days). In parallel to this evaluation approach, the accession resistance was confirmed under the field conditions through natural infection by using the tree-leaf method. The high correlation found between detached leaf inoculation method and field screening under natural infection allows us to use this laboratory technique with sureness in further selection works.
Abstract: The aim of this study was to determine the activity of superoxide dismutase (SOD), glutathione peroxidase (GPx), total antioxidant status (TAS) and accumulation of Hsp70 in porcine ovarian granulosa cells after deoxynivalenol (DON) and zearalenone (ZEA) exposure in vitro. Porcine ovarian granulosa cells were incubated with DON/ZEA administrations as follows: group A (10/10 ng/mL), group B (100/100 ng/mL), group C (1000/1000 ng/mL), and the control group without any additions for 24h. In this study mycotoxins developed stress reaction of porcine ovarian granulosa cells and increased accumulation of Hsp70 what resulted in increasing activities of SOD and GPx in groups with lower doses of mycotoxins. High dose of DON and ZEA had opposite effect on GPx activity than the lower doses. Slight increase in TAS of porcine granulosa cells was observed after mycotoxins exposure. These results contribute towards the understanding of cellular stress and its response.
Abstract: Aldehyde oxidase is molybdo-flavoenzyme involved in the oxidation of hundreds of endogenous and exogenous and N-heterocyclic compounds and environmental pollutants. Uncharged N-heterocyclic aromatic compounds such phenanthridine are commonly distributed pollutants in soil, air, sediments, surface water and groundwater, and in animal and plant tissues. Phenanthridine as uncharged N-heterocyclic aromatic compound was incubated with partially purified aldehyde oxidase from rainbow trout fish liver. Reversed-phase HLPC method was used to separate the oxidation products from phenanthridine and the metabolite was identified. The 6(5H)-phenanthridinone was identified the major metabolite by partially purified aldehyde oxidase from fish liver. Kinetic constant for the oxidation reactions were determined spectrophotometrically and showed that this substrate has a good affinity (Km = 78 ± 7.6µM) for hepatic aldehyde oxidase, will be a significant pathway. This study confirms that partially purified aldehyde oxidase from fish liver is indeed the enzyme responsible for the in vitro production 6(5H)-phenanthridinone metabolite as it is a major metabolite by mammalian aldehyde oxidase, coupled with a relatively high oxidation rate (0.77± 0.03 nmol/min/mg protein). In addition, the kinetic parameters of hepatic fish aldehyde oxidase towards the phenanthridine substrate indicate that in vitro biotransformation by hepatic fish aldehyde oxidase will be a significant pathway. This study confirms that partially purified aldehyde oxidase from fish liver is indeed the enzyme responsible for the in vitro production 6(5H)-phenanthridinone metabolite as it is a major metabolite by mammalian aldehyde oxidase.
Abstract: Environmental pollution is a global problem and best possible solution is identifying and utilizing native microorganisms. One possible application of microbial product -biosurfactant is in bioremediation of hydrocarbon contaminated sites. We have screened forty two different petroleum contaminated sites from Oman, for biosurfactant producing spore-forming bacterial isolates. Initial screening showed that out of 42 soil samples, three showed reduction in surface tension (ST) and interfacial tension (IFT) within 24h of incubation at 40°C. Out of those 3 soil samples, one was further selected for isolation of bacteria and 14 different bacteria were isolated in pure form. Of those 14 spore-forming, rod shaped bacteria, two showed highest reduction in ST and IFT in the range of 70mN/m to
Abstract: Study production of tempeh inoculums powder by freeze-drying comparison with dry at 50°C and the sun bask for developing efficient tempeh inoculums for tempeh producing. Rhizopus oligosporus in PDA slant cultures was incubated at 30oC for 3-5 days until spores and mycelium. Preparation spores suspension with sterilized water and then count the number of started spores. Fill spores suspension in Rice flour and soy flour, mixed with water (in the ratio 10: 7), which is steamed and sterilized at 121°C 15min. Incubated at room temperature for 4 days, count number of spores. Then take the progressive infection and full spore dough to dry at 50°C, sun bask, and lyophilize. Grind to powder. Then pack in plastic bags, stored at 5°C. To investigate quality of inoculums which use different methods, tempeh was fermented every 4 weeks for 24 weeks of the experiment. The result found that rice flour is not suitable to use as raw material in the production of powdered spores. Fungi can growth rarely. Less number of spores and requires more time than soy flour. For drying method, lyophilization is the least possible time. Samples from this method are very hard and very dark and harder to grind than other methods. Drying at 50°C takes longer time than lyophilization but can also set time use for drying. Character of the dry samples is hard solid and brown color, but can be grinded easier. The sun drying takes the longest time, can’t determine the exact time. When the spore powder was used to fermented tempeh immediately, product has similar characters as which use spores that was fresh prepared. The tempeh has normal quality. When spore powder stored at low temperature, tempeh from storage spore in weeks 4, 8 and 12 is still normal. Time spending in production was close to the production of fresh spores. After storage spores for 16 and 20 weeks, tempeh is still normal but growth and sporulation were take longer time than usual (about 6 hours). At 24 week storage, fungal growth is not good, made tempeh looks inferior to normal color, also smell and texture.
Abstract: Water pollution is a major concern for the pulp and paper industry due to the large quantities of effluents generated. Biodegradation of industrial Lignin and AOX by a fungal isolate identified as Aspergillus flavus, white rot fungi which was isolated from Pulp and Paper effluent was studied in batch flask system with industrial effluent and synthetic solution. The flasks were operated at temperature 32°C at 200rpm for eight days in continuous mode. The average overall pH, Temperature, DO, C.O.D, T.D.S, T.S.S, Lignin, AOX were up to 4.56, 32oC, 4.2mg/l, 104mg/l, 6000 mg/l, 4000mg/l, 575.5mg/l, 2195 mg/l respectively after treatment. The Aspergillus flavus sp was the most effective in the biodegradation of Lignin of pulp industry for 94% at 480nm, AOX for 62% at 510nm and of chemical oxygen demand levels for 45% after 8 days of incubation. The optimal conditions found were 4 pH and 32oC temperature for lignin and AOX degradation.
Abstract: A total of 6 isolates of Bacillus subtilis were isolated from oil mill waste collected in Namakkal district, Tamilnadu, India. The isolated bacteria were screened using lipase screening medium containing Tween 80. BS-3 isolate exhibited a greater clear zone than the others, indicating higher lipase activity. Therefore, this isolate was selected for media optimization studies. Ten process variables were screened using Plackett–Burman design and were further optimized by central composite design of response surface methodology for lipase production in submerged fermentation. Maximum lipase production of 16.627 U/min/ml were predicted in medium containing yeast extract (9.3636g), CaCl2 (0.8986g) and incubation periods (1.813 days). A mean value of 16.98 ± 0.2286 U/min/ml of lipase was acquired from real experiments.
Abstract: L-asparaginase was extracted from pathogenic
Escherichia coli which was isolated from urinary tract infection
patients. L-asparaginase was purified 96-fold by ultrafiltration, ion
exchange and gel filtration giving 39.19% yield with final specific
activity of 178.57 IU/mg. L-asparaginase showed 138,356±1,000
Dalton molecular weight with 31024±100 Dalton molecular mass.
Kinetic properties of enzyme resulting 1.25×10-5 mM Km and
2.5×10-3 M/min Vmax. L-asparaginase showed a maximum activity
at pH 7.5 when incubated at 37 ºC for 30 min and illustrated its full
activity (100%) after 15 min incubation at 20-37 ºC, while 70% of its
activity was lost when incubated at 60 ºC. L-asparaginase showed
cytotoxicity to U937 cell line with IC50 0.5±0.19 IU/ml, and
selectivity index (SI=7.6) about 8 time higher selectivity over the
lymphocyte cells. Therefore, the local pathogenic E. coli strains may
be used as a source of high yield of L-asparaginase to produce anti
cancer agent with high selectivity.