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Ten Tips for a Career in Psychology
"Any psychologist is in a privileged position and this brings a duty to think and act at the highest ethical level."
Written by Liz Blagrove
1.Psychology as a science
Although seen by many as a social science, psychology involves many of the techniques and concepts associated with natural sciences.
2.Educational background
It is not necessary to study psychology previous to embarking on a psychology degree, rather it is useful to have an understanding of a range of subjects, particularly sciences and maths.
If you are considering a career in psychology, consider the many paths that such a career could take and don't limit yourself to the competitive field of clinical psychology alone.
4.Word experience and volunteering
Experience in psychology-related positions can not only give you a valuable edge in applying for jobs, but it can also help you to decide what type of role you would prefer to pursue.
Working in psychology often involves facing problems to which there is no simple solution - you will need the confidence to decide how best to respond in such situations.
The desire to know and learn more is key in the field of psychology.
Psychologists must be able to to persist in their work and find ways to move forward when faced with resistance, which can come from a variety of sources.
8.Big picture? Small picture?
It is important to constantly keep in mind the details of the specific case at hand while considering its relevance at a more general level.
9.Independence and collaboration
Psychologists need to be able to work alone and motivate themselves, but must equally be able to function well in a team.
10.Rewards and responsibilities
Being a psychologist is a hugely rewarding job, as well as a career that brings with it many responsibilities and a need for a very high level of integrity.
Liz Blagrove is a lecturer and researcher in the Department of Psychology at the University of Warwick. She came to psychology as a mature student, having previously trained as a lawyer. She was "bitten by the psychology bug" during her legal studies, becoming fascinated by the way human psychology impacted on the legal system and harboured a long-standing wish to explore this interest further. She returned to higher education as an undergraduate, working as a researcher before undertaking her PhD. Outreach forms a major part of her professional role; in other words, Liz has the enjoyable task of explaining to children and young people why psychology is such an amazing field!


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Today around 5 pm, Kaden started getting fussy. He's been doing a lot better in the evenings, but he was starting to get fussy this afternoon. Our friend Stacey is in from Oklahoma for the weekend, and we had been talking about going to the pool all day. I know how much Kaden enjoys the pool, so we decided to load up and walk over. Like always, it did the trick. He was a little fussy when we first got there, and took about a 10 min. power nap. After that, we hung out in the pool and he was perfectly content. I love my little water baby!!
Friday, August 5, 2011
Cool Dude!
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The Learners reports show tons of information when you export them, like Supervisor, Group, custom fields, etc. This makes it very useful for our clients who prefer to do things in Excel, or need to gather email addresses to contact people, or even just have an export for their own records.
The newer compliance reports have filters for these fields, but they don't show up when exported. In fact, the exports really only show what's on the screen, making them much less useful for people who do use Excel to do more advanced things, as well as for recordkeeping.
I'd love it if the export showed more info in the CSV than what you see on the screen, making them just as useful as the old reports when exported.


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Engagement of Act Apprentices under Apprentice Act 1961 for the year 2018-19 in North Central Railway Wagon Repair Workshop, Jhansi
|Post Name
|Act Apprentice
|No. of Vacancy
|Stipend will be paid as per instructions issued by Railway Board from time to time
|Educational Qualification
The candidate must have passed 10th class examination or its equivalent (under 10+2 system) with minimum 50% marks with ITI passed certificate in related trade from a recognized Industrial Training Institute affiliated to NCVT
|Age Limit
|15-24 yrs
Application Fee : Application fee of Rs 100/- may be deposited in the form of DD in favour of "Workshop Accounts Officer, Jhansi". DD should necessarily be attached with the application form. The SC, ST, Physically handicapped and Women candidates are relaxed from application fee.
How to Apply : The application and call letter must be filled by candidate in his/her own hand writing in the given format on A4 sheet by ballpoint pen and sent to the following address by ordinary post only:
Recruitment Section, personnel Barnch, Office of the Chief workshop Manager, Wagon Repair Workshop, North Central Railway, Jhansi U.P. 284003.
The last date for receiving of the complete application form in all respects in the above mentioned address is 31.12.2018 by 17:45 PM


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Welcome to the Waste page.
Remember the 5Rs of waste.
Refuse - to buy things you don't need.
Reduce - don't be tempted by BOGOF offers on things you will not use.
Reuse - before throwing things away, can it be reused? Have a look at our eco wall for some ideas.
Repair - learn how to repair broken items instead of throwing them away.
Recycle - our school has collections boxes for a variety of products including ink cartridges, pens and batteries.


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Thiruvananthapuram Zoo and Museum
The historical center building itself is an engineering quality. The Napier Museum, the Natural History Museum, the Reptile House and the Sree Chitra Art Gallery are all inside the Thiruvananthapuram Zoo complex in the midst of a well laid-out garden and stop in the downtown area. There is a lake and a vessel club mutually worked by the Department of Museums and Zoos and the District Tourism Promotion Council. The zoo offers an uncommon open door for the guests to encounter an unparalleled, pleasant and all encompassing sylvan scene with an extensive variety of creature accumulations. The zoo has 75 types of creatures from India as well as from abroad. It has a few types of creatures and winged creatures from Ethiopian and Australian zoological districts. The lion-followed macaque, Nilgiri langur, Nilgiri tahr, Manipur deer, Indian rhino, Asiatic lion and the regal Bengal tiger are conspicuous among the indigenous jeopardized fauna, while giraffe, hippo, zebra and Cape bison are visitors from the African district. Another period will be introduced with the consummation of the continuous modernisation work in the zoo which will give for exciting experiences the creatures out in their outside scenes. The city additionally has the Kerala Science and Technology Museum which incorporates the Priyadarsini Planetarium.


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It's been a busy and fruitful week with family visiting from Sydney and Oregon, USA, and a break from cooking for me. Come Monday, the new school term kicks off again and it's that time again for more soulful home-cooking and easy-to-prepare practical dishes for the family.
One of the easiest and best 'one pot dishes' we love making at home would be this Claypot Chicken Rice. As Adam Liaw wrote in his other scrumptiously appealing and simple-to-cook 'Asian After Work' (Simple Food For Everyday)—the only trouble with claypot rice is the usual struggle with perfecting that 'balance of liquid'. As you know, there is the liquid released by the ingredients versus the liquid required to cook the claypot rice. In his recipe, 'Cheat's Claypot', Liaw shares his solution by way of a 'cheat's recipe' to avoid the problem of the right amount of liquid to add. He does this by pre-cooking the white rice.
The recipe I'm sharing from my own experience is similar in technique from his, yet cooked from the rice cooker, one of the most indispensable cooking utensils in my kitchen which I use on an almost daily basis. Yes I love my rice and the last time I had gone on a 'no carbs' Dukan Diet I had severe withdrawal symptoms (meaning I nearly ate my own arm…) from this source of starch that I cannot live without, i.e. beyond a few days. If you love rice as much as me then there is no better investment that a rice cooker just to make your life that much easier and your time spent preparing food, more of a pleasure than a chore.
One of the reasons I use the rice cooker for my claypot rice, aside from its convenience, is that I for one, don't actually appreciate the crusty texture of the rice that some might love about claypot rice. I love the flavour yet not the hard crust of rice that forms and sticks around the rim and edges of the claypot. I pre-cook my rice as well as the chicken, mushrooms and Chinese sausage, (separately) then serve it in a claypot to retain its hot temperature and to allow the flavours to infuse before tucking in.
4 to 6 chicken thighs, (skins off, sliced)
5 large dried shiitake mushrooms (soaked in 1 cup warm water; reserve the liquid)
Chinese cabbage (wombok or xiao bai cai)
1 chinese sausage (lap cheong)
1 small red shallot, sliced
1 inch knob of ginger (grated)
For pre-cooked rice:
Chicken stock
2 C jasmine rice grains
1 TB peanut oil
1 TB dark caramel sauce
2 TB oyster sauce
1 tsp salt
Claypot sauce marinade for chicken:
1 inch knob of ginger (grated or pressed)
2 cloves garlic (finely diced or pressed)
1/2 TB chicken seasoning powder
1 tsp white pepper
2 tsp sesame oil
1 TB light soy sauce
1 TB oyster sauce
1 TB dark soy sauce
1 TB sugar
1 TB shaoxing wine
2 tsp cornflour
Directions for Claypot Chicken Rice:
Soak dried shiitake mushrooms in about 1-2 cups of warm water till they become soft. Squeeze the excess water but reserve aside for cooking the claypot rice. Slice up the mushrooms with stems removed.
Toss and mix the sliced up chicken thigh pieces in a large mixing bowl with the claypot sauce marinade, adding the shiitake at the last. Leave refrigerated overnight for the marinade to infuse.
Once ready to cook, slice up the Chinese sausage. Heat up a wok with 2 TB peanut oil. Add grated ginger and sliced small red shallots and sauté slightly till fragrant. Add the chicken pieces, mushrooms and Chinese sausage and stir-fry for a bit, adding the remaining marinade and a little of the reserved shiitake mushroom liquid to simmer the chicken thighs, sausage and mushrooms till chicken pieces are no longer red(but need not be fully cooked at this stage). Turn off the heat.
Cooking the rice
Wash and rinse 2 cups of white jasmine rice grains. Cook according to the instructions for your rice cooker using readymade chicken stock to the appropriate level for the amount of rice (in this case it wold be for 2 cups of rice) with 1 TB peanut oil. Add 1 TB of dark caramel sauce for colour, 2 TB oyster sauce, 1 tsp salt (if desired) to the rice. Add the vegetables of your choice.
Start the rice cooker. Once rice is done, toss in the cooked chicken pieces, mushrooms and Chinese sausage into the rice and stir-through. The texture should be just right and the rice should not be wet. Add any extra liquid sauce from the cooked ingredients.
To serve, garnish with coriander, spring onions and deep fried shallots. Best with some chilli sauce. My favourite is some funky sambal belachan!


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|Roland Weber <ossf...@dubioso.net>
|Re: Jakarta [was: Effects on corporate backing withdrawals [was: Incubator Proposal: Pig]]
|Sun, 30 Sep 2007 16:17:09 GMT
Niclas Hedhman wrote: > I don't know what to suggest, but perhaps recruiting one or more veteran > ASFer, either just off the member's list or some experienced Incubator > mentor, feeling this being important could just join the PMC and at least > ensure process with 3 pairs of eye balls. Yeah, we'll try to get a veteran (though not a member) to help us out as the chair for the initial phase. (speaking for HttpComponents, not JMeter) If anyone here feels like keeping an eye on us too, you're most welcome. We know our way through the code and public processes up to PMC, but we currently don't have an ASF member on board who is familiar with what's going on beyond PMC. (speaking for both) thanks for taking the time, Roland --------------------------------------------------------------------- To unsubscribe, e-mail: email@example.com For additional commands, e-mail: firstname.lastname@example.com
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Will not print black. Started printing top of page only. Replaced ink cartridge. Self test diagnostic page printed all colors but gave 40% ink level despite brand new HP ink cartridge.
I am the person who asked the question. New information - the black printed perfectly using Quicken "print checks"


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What To Do When Your Dog Has An Accident In The House
Coming home after a long day away from your dog is one of the best parts of having a pet. You open the door and there's a happy, furry friend waiting there. You look forward getting home, greeting your best friend and kicking off your shoes. But kicking off your shoes and stepping in something wet...not so much.
Whether you're potty-training a brand new puppy or have an old dog that refuses to learn new tricks, your carpets are at risk. Check out this infographic and learn how to combat pet odors and accidents before they ruin your floors and your day. Or for more serious incidents call a Chem-Dry professional and schedule your pet urine removal service.
*Based on studies conducted by independent laboratories of Chem-Dry's HCE (Hot Carbonating Extraction), P.U.R.T. (Pet Urine Removal Treatment), Granite Countertop Renewal, and Tile, Stone & Grout cleaning processes. Allergens tested were dog and cat dander and dust mite allergen. Pet odor results based on testing with the most common odor sources found in dog and cat urine. Pet urine bacteria results based on Chem-Dry's HCE cleaning process and a sanitizer, combined with P.U.R.T. All bacteria results include use of sanitizer. Figures are an average across multiple tests.


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See, it started off pretty simple - I used to have a work nemesis. Now I don't, mostly because I don't get enough juice out of having a nemesis (I can see how it works for some people) to justify the energy expenditure in maintaining one. (But it sounds so cool! Narrowed eyes, hissing "my nemesis," when people ask who you're narrowing your eyes at...) Once I realized that the only reason she was my nemesis was because I was jealous, I couldn't do it anymore. There should be a better reason! (I was jealous because she was hired after me and got a permanent job right away, while I still labor in the On Call salt mines.) ANYWAY. I'm over all that now. It's not like we're best pals or anything, but she's pretty loopy and I enjoy talking to her.
Today's misunderstanding came about when she said that she was talking to a patron "about our age," which is hilarious because I've got to be at least 15 years older than she is. (10 at the very least.) I don't think it was some ham handed attempt to flatter me, because she's a very blurty thinks it/says it kind of person. (example, to patron: "why are you wearing those rubber gloves?" a question I'd never ask in 10000 years because the answer could be anything from "severe OCD, thanks for asking" or "I've been treating the chickens for mad chicken disease" or etc. etc. you get the picture.) ANYWAY. Maybe the real answer is that she's a lot older than she looks, like a vampire or a creature who sleeps for 100 years and then is awake for a year, or maybe she simply meant an age greater than 10 and less than 80, in which case we are totally the same age.
The point of the story (if it has one! I can't even tell. I've got to start these posts earlier in the day) is that I didn't correct her even though it would have been relatively easy to say "but I'm ages older than you are!" but then I think it once again becomes about me and my insecurities, so maybe it's just as well I said nothing on that particular subject.
In related news, I thought for a minute that I had a new work nemesis lined up, but it turns out that the person is fine apart from an appalling work ethic.


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A L2 reading lesson around the optimism of Spring, and Easter and pagan traditions, especially suitable for the start of the Summer term, as a springboard to setting intentions for the rest of the course. Suitable for ESOL or Functional Skills courses.
Editor's note
Fully mapped to the reformed FS content - and GCSE English. Also includes a vocabulary consolidation task for the next lesson.
An answer sheet will be available shortly - for resource contributors only.
Reading comprehension and writing tasks designed to prepare adults who are returning to education and taking Functional English at Level 1 or Level 2.
Level 1 reading comprehension is based on two texts about returning to education. This is followed by two writing tasks. Level 2 includes all the L1 texts and questions plus an additional lengthy text and questions that involve summarising, and comparing and contrasting two or more texts.
A series of question sheets for learners to count, compare, add and subtract numbers of different fruits.
Learners' reference a front sheet displaying a collection of pictures of fruits to answer a series of questions covering; counting and comparing numbers to 10, words describing quantity, addition and subtraction. Question texts are supported with Makaton symbols and with Numicon shapes to support pre-entry learners' understanding.
A role play to encourage learners to join their schools' PTA. A parent asks at school about joining the PTA. Later she goes to a meeting and offers to help on a face painting stall at the Christmas fete.
An ESOL lesson embedding numeracy, diversity and British values, centred on the Spring Equinox and three festivals which happen around / on this day - Purim, Holi and Shunbun No Hi. Learners collect unfamiliar words on a vocabulary sheet and ask and answer questions to elicit meaning / explanation. There is a powerpoint to introduce the topic, using acronyms eg 7DIAW - Seven Days In A Week to elicit key words, and lead to an explanation of what the Vernal Equinox is and when it is.
Topical set of discussion prompt for World Book Day 2019 - but could be used at any time.
One set of general prompts and a second slightly different set of prompts for learners who are also parents (ideal for family learning situations).
Includes a full list of the revised Functional English speaking, listening and communicating content (for E2, E3 and L1) which comes into effect September 2019.
Short reading comprehensions on the 2019 New Year, one at L1 and one at E3. Includes some embedded maths and some E&D (references to Brexit - sorry, folks!).
The focus is on identifying, extracting and understanding the main points and detail. However there are also questions on fact & opinion, purpose of text, specialist words, using a dictionary and using knowledge of punctuation. Fully mapped to the new revised Functional English descriptors. With answers.
Entry Level 1-2 Maths questions based on a shopping poster from the BBC 2018 Children in Need appeal but can be used at any time. Main focus is positional vocabulary, money and basic number (+, -. x, /) skills
Fully mapped to the new 2018 Functional Maths content which takes effect from September 2019 (note that times tables to 12 x 12 and basic division skills are now expected at E2). Image below shows , as an example, the detailed E2 mapping.
Two sets of Functional English resources based on various texts from the set of seven How We Remember texts (follow link right, under See also to download the texts).
L1 tasks - reading questions based on Texts B ( Poppies web site) and E (A Nation's Thank you - the People's Procession) and a related writing task.
L1-2 tasks - reading questions based on Texts E (as above) and G (Nurse Mary Borden - forgotten female poet of WWI) and a related writing task that focuses on presentation devices and persuasive techniques.
A set of Functional English tasks that underpin the revised Functional Skills English content, 2018 [1]. Resources will be released gradually (as they are written!) from Oct 21st until Nov 11th 2018. All tasks are based on one or more texts from a graduated set of seven, all focusing on the 2018 Armistice Centenary commemorations. Five sets of tasks, based on Texts A, B, C, D and E are now available.


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Interested in writing for PAC (Power Affiliate Club) as a PAC Expert Author? Thar's great!
Because we believe it will elevate your visual presence as a blogger on the internet.
And we know you want to be more visible, have MORE engagement, and rank higher in the search engines, right?
Here's what being a PAC Expert Author offers:
- Authority in your area of interest or "niche"
- More visibility
- An added experience to your resume
- More exposure for your website
- Grow more confident and improve writing skills
- Part of a growing popular community
- Credibility and respect as a PAC Expert Author
- More contacts and instant friends
- Get paid for writing for PAC
- Be a mentor to others
- Represent PAC
We are a community one that likes to promote its members as much as possible. We know being online is competitive – meaning there are many bloggers who want to be noticed, attract followers, monetize their websites, and have a high ranking.
We also know that many bloggers like to be noticed and accepted as writers on other more notable websites like Huffington Post, for example,right? Well we're not that, but we ARE an opening, a starting point for bloggers to write for a community such as ours and build an online/offline reputation.
Here's some feedback from our writers:
Writing for PAC and getting PAID has been a huge advantage. First of all, I can put that money back into my blog and help with the upkeep expenses. Secondly, the reward of getting paid for something that I love is an incredible feeling. After blogging for free, for months, it can get quite discouraging. But knowing that writing here will help others through what I write and help me financially?!??, I call that a win:win!
PAC Expert Author, Johanna Galyen
"I became a PAC member sometimes in 2014. PAC grew into a family like, supportive community with a lot of opportunities for bloggers. It's a privilege to be an Expert Author for PAC. Not only that it helped me to position myself as an expert blogger and build my authority and credibility, but at the same time I get paid for doing so.
The top 7 reasons why I recommend PAC to any blogger are:
1. Grow your visitors and customers.
2. Get more comments and engagement.
3. Get higher ranking on Alexa.
4. Build your authority and credibility.
5. Connect with professional bloggers.
6. Get Paid.
7. Grow your blogging skills."
PAC Expert Author Emi Koul
"The icing on the cake is that you can get paid for being an "Expert Author" on PAC. You can actually tell people you are a paid freelance writer.
Any writer needs to write every day and for different reasons and topics. The more you write, the more your writing is seen by many others and there's no reason why your freelance writing career can include more and more blogs to write for.
There is also a bit of prestige when you're asked to write for another blog, aside from your own. "
PAC Expert Author, Monna Ellithorpe
It's a great opportunity! So. How does this work?
And how would you get paid.
First, it's important to understand and accept that PAC is an effort based community. Meaning, the more effort you put into the community, the more you get back – give and receive. You have to do this anyway having a blog online and if you participate in our community activities you'll experience the magic of this.
We require our PAC Expert Authors to contribute to the community blog by writing ONE post per month. Yes, you can write more if so inspired. All your posts are reviewed and only published by the blog editor. On acceptance as a PAC Expert Author, you'll be added to the Facebook PAC Expert Authors Group which has writing guidelines for the community blog.
As a PAC Expert Author you're accepted into the "Authors Pay Pool" where you're paid for your writing contributions. Posting for two consecutive months is required. If you skip a month
you don't get paid.
The current pay is $20.00 (us dollars). The more authors we have contributing the pay rate can increase.
Becoming a PAC Patron by donating $10.00/month is required to participate. It's members helping members through their writing contributions. And writers keep the community blog alive and well.
There are bloggers who pay $100.00 a month to guest blog! PAC recognizes the efforts of writers and gives back to them.
What to do now?
- Fill out the application below
- If your writing (content on your blog) is accepted … (we will review it and contact you)
- Become a PAC Patron
We appreciate your interest and look forward to you joining us.
Thank you
Lesly Federici
CEO/Power Affiliate Club


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Get a coveted Shirley Road address and build your dream home on this 3/4 acre lot. 132 x 325 x 110 x 234. A short walk to down town Birmingham and Linden Park. Surrounded by grand homes this spectacular lot offers a wonderful opportunity to renovate current home or build a new home.
Property Summary -- ASSRS REP OF CORYELL PARK:
Located in BIRMINGHAM in the 48009 zip code, this home was built in Unknown and is Unknown square feet
with Unknown bedrooms and Unknown bathrooms on
0.71 acres. It has been on the market for 500 days with an original
listing price of $1,285,000. 381 Shirley Rd. is part of the
Birmingham School District.
Property Location
IDX provided courtesy of Realcomp II Ltd., via Keller Williams Realty and MIREALSOURCE, Copyright 2019 Realcomp II Ltd. Shareholders. IDX information is provided exclusively for consumers' personal, noncommercial use and may not be used for any purpose other than to identify prospective properties consumers may be interested in purchasing. The accuracy of all information, regardless of source, is not guaranteed or warranted. All information should be independently verified.


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Happy Sunday! Just a quick card to show today. I still had two of the Blushing Bride butterflies cut from January and wanted to use up. Just matched up with Blackberry Bliss, Whisper White and a little Sahara Sand for a fast card.
Thanks for stopping by!
Stamp Sets: Butterfly Basics and Simply Wonderful (SAB)
Paper: Whisper White, Blackberry Bliss, Blushing Bride
Ink: Blackberry Bliss, Blushing Bride, Sahara Sand
Butterfly thinlits dies, White cotton twine from painted Bloom suite


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Original Research ARTICLE
Production of Cold-Active Lipase by Free and Immobilized Marine Bacillus cereus HSS: Application in Wastewater Treatment
- 1National Institute of Oceanography and Fisheries, Alexandria, Egypt
- 2City of Scientific Research and Technological Applications, Alexandria, Egypt
Lipases are enzymes that have the potential to hydrolyze triacylglycerol to free fatty acids and glycerol and have various applications. The aim of the present study was to isolate and screen marine bacteria for lipase production, optimize the production, and treat wastewater. A total of 20 marine bacterial isolates were obtained from the Mediterranean Sea and were screened for lipase production. All isolates were found to have lipolytic ability. The differences between the isolates were studied using RAPD-PCR. The most promising lipase producer (isolate 3) that exhibited the highest lipolytic hydrolysis (20 mm) was identified as Bacillus cereus HSS using 16S rDNA analysis and had the accession number MF581790. Optimization of lipase production was carried out using the Plackett–Burman experimental design with cotton seed oil as the inducer under shaking conditions at 10°C. The most significant factors that affected lipase production were FeSO4, KCl, and oil concentrations. By using the optimized culture conditions, the lipase activity increased by 1.8-fold compared with basal conditions. Immobilization by adsorption of cells on sponge and recycling raised lipase activity by 2.8-fold compared with free cells. The repeated reuse of the immobilized B. cereus HSS maintained reasonable lipase activity. A trial for the economic treatment of oily wastewater was carried out. Removal efficiencies of biological oxygen demand, total suspended solids, and oil and grease were 87.63, 90, and 94.7%, respectively, which is promising for future applications.
Lipases (triacylglycerol acylhydrolase, EC 126.96.36.199) catalyze the hydrolysis and synthesis of long-chain acylglycerols (Salihu and Alam, 2012; Lajis, 2018). They also catalyze the hydrolysis of triglycerides and reverse reactions (interesterification and esterification) (Saxena et al., 2003) and are involved in aminolysis, acidolysis, and alcoholysis. Lipases are produced by archaea, bacteria, fungi, animals, and plants (Cai-hong et al., 2008). Animal and plant lipases are less stable than those of microbial origin. The production of microbial lipases is more suitable, economic, and safer (Vakhlu and Kour, 2006; Treichel et al., 2010). Lipolytic enzymes secreted by Bacillus spp. are of considerable biotechnological importance. B. alcalophilus, B. licheniformis, B. pumilus, and B. subtilis are the common bacterial lipase producers (Lindsay et al., 2000; Awad et al., 2015).
The production of biodiesel and biopolymers using lipases has been successfully recognized as a novel biotechnological application; furthermore, they can be used in the assembly of new pharmaceuticals, agrochemicals, and flavor compounds in addition to their various industrial applications (Hasan et al., 2006; Chary and Devi, 2018; Sarmah et al., 2018).
Cold-adapted enzymes are considered as biocatalysts for industrial applications. They are used in the synthesis of fine chemicals, food additives, detergents, and in the bioremediation of contaminated water and soil (Margesin and Feller, 2010; Cavicchioli et al., 2011; Patel and Desai, 2018). The manufacture of lipases with cold-active properties is generally extracellular and is greatly affected by nutritional and physicochemical factors, including inducers, nitrogen source, carbon source, temperature, agitation, pH, inorganic sources, and dissolved oxygen (Joshi et al., 2006; Ephraim et al., 2014). It was stated that Bacillus spp. could easily use different nutrient sources. However, there were limited searches for the development of a suitable medium supplemented with natural nutrient sources such as vegetable oils for higher lipolytic capability (Ephraim et al., 2014). Optimization of lipase production was performed in many studies (Park et al., 2013; Anasontzis et al., 2014; Bharathi et al., 2018; Sahoo et al., 2018).
For long term enzyme activity, immobilization meets this requirement. Immobilization proposes several advantages such as recycle, ease in applications, stability, improved control of reactions, and simple removal of the immobilized object from the medium (Betigeri and Neau, 2002). Methods of immobilization can be divided into three categories: encapsulation, non-covalent attachment to a support, or covalent and carrier-free cross-linking (Gabrielczyk et al., 2018).
Presence of oily wastes in the aquatic environment prevents the diffusion of oxygen into water, which causes the death of many aquatic organisms. Lipases and microbial strains represent green alternatives for wastewater treatment and can have an effect on reducing the oil and fat content in the wastewater; they may assist in controlling severe pollution problems caused by wastewater. Many previous studies have reported the use of lipolytic enzymes for wastewater treatment (Sarac and Ugur, 2015; Silva-Bedoya et al., 2016; Hu et al., 2018).
Thus, the main goal of the present study is screening for lipase production by diverse bacteria isolated from marine environment and optimization of production setting by statistical means and immobilization techniques. Furthermore, an experiment for wastewater management using the most promising lipase producer will be investigated.
Materials and Methods
Isolation and Primary Screening for Lipase Production by Different Bacterial Isolates
Marine bacteria were isolated from different sites along the Mediterranean Sea, including Eastern Harbor, Al Shatby, and Abu-Qir, and were cultured on nutrient agar plates (pH 7) that were prepared with aged filtered seawater. Media used throughout this study were supplied from Oxoid LTD, England. All the chemicals used were of analytical reagent grade.
Lipolytic capability of the isolated bacteria was screened using modified Tween agar base plates that contained the following (gl−1): beef extract, 3; peptone, 5; (50% seawater and 50% distilled water). The medium was amended by 1% Tween 80 (v/v). The development of clearance zone (hydrolysis) was considered as a positive result. The size of the halo-forming zones was measured after 24 h of incubation.
Secondary Quantitative Screening for Lipase Production by the Different Isolates
Secondary screening was performed by the quantification of lipase activity (Kanwar et al., 2006). All isolates were inoculated in mineral based medium that was supplemented with 1% (v/v) cotton seed oil and were incubated at 10°C. Production of lipase was assayed for different time intervals (24, 48, and 72 h). Production medium contained the following (gl−1): yeast extract, 0.5; sucrose, 0.5; NaNO3, 0.03; K2HPO4, 0.01; MgSO4, 0.05; KCl, 0.05; and FeSO4.7H2O, 0.001, adjust pH to 7. Subsequently, centrifugation at 10000 ×g for 20 min at 4°C was carried out to collect the cell free supernatant for lipase assay (Jensen, 1983).
Lipase Assay
Estimation of lipase activity was resolute by the titrimetric technique using cotton seed oil as a substrate (Jensen, 1983). In brief, cotton seed oil was emulsified with Arabic gum in 0.05 mM Tris buffer (pH 7). A volume of 100 μl of the enzyme was added to the emulsion and incubated at 10°C for 30 min. The reaction was blocked by the addition of acetone: ethanol (1:1). The liberated fatty acids were predicted by titration with 0.05 M NaOH using phenolphthalein as an indicator. Lipase activity was articulated as the amount of enzyme essential to hydrolyze 1 mole of fatty acids from triglyceride per minute.
DNA Isolation and Random Amplification of Polymorphic DNA (RAPD-PCR)
Genomic DNA was extracted using the Thermo Fisher kit. Quantity and quality of DNA was determined by Nanodrop spectrophotometric method (Kirby, 1993). Quality of the DNA was checked by calculating the ratio between OD260 and OD280. RAPD-PCR was used to differentiate between the bacterial isolates to detect the dissimilarity between the isolates. Reaction mixture of RAPD-PCR contained the following: 50 ng genomic DNA, 1 μl of 1X buffer, 0.2 mM dNTPs, 1 μl of 10 picomole primer, 2 units of Taq DNA polemerase, and RNAse and DNAse free water. RAPD is a multiplex marker system that predictably uses single primer to amplify random DNA fragments. Two general oligonucleotide primers were used for the amplification: 16S F: AGAGTTTGATCMTGGCTCAG, 16S1100R: GGG TTG CGC TCG TTG. PCR consists of the following three steps: denaturation at 94°C for 1 min, annealing at 30°C for 1 min, and extension at 72°C for 1 min. After amplification by PCR, the products were checked in 2% agarose gel. Genetic miscellany was determined as the experimental number of differentiation.
Molecular Identification of the Most Potent Isolate (HSS)
DNA was isolated and purified using the standard procedure (Sambrook et al., 1989). The region of 16S rRNA was amplified using the universal primers (F: 5′AGAGTTTGATCMTGGCTCAG3′and R: 5′TACGGYACCTTGTTACGACTT3′). The reaction was performed using a template DNA. Sequences of the 16S rRNA genes were obtained from the NCBI database. Multiple alignments based on the most closely related sequences and similarity levels were carried out using the BLAST program1. A phylogenetic tree was reconstructed using the Bioedit software.
Effect of Temperature on Lipase Activity
The effect of temperature on lipase production was determined by incubating the culture flasks at different temperature (10, 25, and 30°C). Lipase assay was performed separately for each experiment.
Effect of Agitation and Different Inducers on Lipase Activity
Two sets of flasks were used to study the effect of aeration; one was incubated under static conditions and the other was kept under shaking condition at 120 rpm. For investigating the effect of different substrates on lipase production, 1% (v/v) of each sterilized oil, including coconut oil, olive oil, sesame oil, corn oil, and palm oil was added separately to the basal medium for 24 h at 10°C. Lipase assay was performed separately for each experiment.
Optimization of Culture Conditions for Lipase Production Using Plackett–Burman Experimental Design
Screening for the virtual consequence of different factors on lipase production was performed by applying the Plackett–Burman experimental design (Plackett and Burman, 1946; Yu et al., 2009). Low (−) and high (+) levels of each variable were tested as shown in Table 3. In this experiment, 11 independent variables were screened in 12 combinations that were organized according to the Plackett–Burman matrix (Table 3). Row no. 13 in Table 3 illustrates the control (basal). The following equation was used to estimate the main effect of each variable: Exi = (Mi + – Mi-)/N, where Exi is the variable main effect, Mi + and Mi- are the lipase activities (U/ml) in the trials where the independent variables were shown in low and high concentrations, respectively, and N is the number of trials divided by two. Microsoft Excel was applied to calculate the statistical t-values for equal unpaired samples and significance of each variable. Verification test was performed to confirm the validity of the optimized medium.
Effect of Immobilization on Lipase Production by B. cereus HSS
Immobilization by Entrapment in Calcium Alginate Beads
Entrapment was performed using 3% sodium alginate as previously described (Eikmeier and Rhem, 1987). Sodium alginate (0.75 g) was dissolved in 23 ml of distilled water, followed by autoclaving at 121°C for 10 min. A volume of 2 ml of B. cereus HSS suspension was added to the prepared sodium alginate solution. A volume of 10 ml of the bacterial-alginate combination was drawn by using a sterilized syringe and was introduced into sterile 2% CaCl2 solution, and the resultant calcium alginate beads entrapping the bacteria were left in CaCl2 solution for about 2 h to solidify. Later, the beads were washed several times with sterile distilled water and used as inoculum for 25 ml of the optimized medium.
Immobilization by Adsorption
Bacterial immobilization by adsorption was carried out on different solid supports (luffa pulp, clay, ceramic, sponge, and pumice). Bacterial suspensions (2 ml) were added to 25 ml sterile flasks containing the optimized culture medium and 5 g of each porous support. Both sponge and luffa pulp were cut to pieces of the size 0.5 cm and were washed with water before use. The flasks were incubated under slow shaking condition at 120 rpm for 24 h. Lipase activity was estimated each time and was compared with the free cells.
Recycling of Adsorbed B. cereus HSS
A total of 20 cubes of sponge were added to the optimized medium, inoculated with 2 ml of bacterial suspension, and incubated at 10°C for 24 h, followed by removal of the culture medium and addition of a new sterilized medium (25 ml); subsequently, a new cycle was run. This process was regenerated several times. At the end of each cycle, activity of lipase was determined.
Efficiency of B. cereus HSS in Removing Oil and Grease in Addition to Organic Load in Wastewater
Wastewater samples were collected from the drainage effluent of Misr Petroleum Company, Alexandria, Egypt. A volume of 250 ml of the wastewater sample was introduced into each conical flask, sterilized at 121°C for 20 min, and allowed to cool. The sample was inoculated with B. cereus HSS containing 106 CFU/ml and was incubated under shaking conditions at 150 rpm. Later, the samples were aseptically drawn every 24 h for 72 h and analyzed for oil and grease, total suspended solids (TSS), and biological oxygen demand (BOD) before and after bacterial inoculation. Samples without the bacterial inoculum were kept as control (Bala et al., 2014). The estimation of each parameter was done according to the method described by Clesceri et al. (1999).
The efficiency for organic load reduction was calculated as follows:
Efficiency % (BOD) removal = (Initial (BOD) of the raw sample -final (BOD) after treatment/ Initial (BOD) of the raw sample) ∗100
Efficiency % (TSS) removal = (Initial TSS) of the raw sample -final (TSS) after treatment/ Initial (TSS) of the raw sample) ∗100
Efficiency % (oil) removal = (Weight of crude oil before treatment (initial) – Weight of crude oil after treatment)/ Weight of crude oil initial)∗100
Analysis of Residual Oil by Gas Chromatography-Mass Spectrometry (GC-MS)
The residual oil was extracted by dichloromethane. The aqueous phase sample was discarded, and the crude oil containing the solvent was concentrated to approximately 0.1 ml using a rotary evaporator under reduced pressure. The residual oil was subjected to chemical analysis using gas chromatography-mass spectrometry (GC-MS; Agilent Technologies 7890A GC System with a flame ionization detector, a 5975C inert XL MSDTriple-Axis Mass Detector and Agilent 19,091S-433 Trace Analysis column). The GC experiments were conducted by injecting 1 μL of the sample, operating in the split less mode with an evaporation temperature of 250°C, pressure of 1.8 bar, and flow rate of 2.5 ml/min. Helium carrier gas temperature was maintained in gradients of 50°C/min, 40°C/min, 300°C/min, and 300°C/5 min. Compounds were characterized based on the similarities between their mass spectrum and those of the authentic data presented by Wiley Library.
Results and Discussion
Qualitative Screening for Extracellular Lipase Production by the Isolated Bacteria
The capability of all isolates to degrade Tween 80 was tested using Tween agar base medium. Lipase-producing potentiality was investigated based on the size of the clear zone around each colony (Ramle and Abdul Rahim, 2016). Results indicated that all isolates were positive producers of lipase with different efficiency as the lipolytic zone diameter range was 6–20 mm. The highest lipolytic activity was detected for isolate 3, which recorded 20 mm (Figure 1) followed by strain 10 (17 mm), whereas strains 15, 16, and 19 showed the lowest lipolytic activity (6 mm) as shown in Table 1. Tween has been used as a substrate in different studies and is recommended as it is more easy and convenient for the detection of lipase activity (Anbu et al., 2011). Lo Giudice et al. (2006) studied the effectiveness of different isolated bacteria for their lipolytic activity on Tween and tributyrin by hydrolytic plate assay, and they reported that among the strains tested for lipid hydrolysis, only 7 isolates were unable to utilize the media, while the remaining hydrolyzed at least one substrate, and the diameters of the halos were generally 9–29 mm using Tween. In a parallel study, Rasmey et al. (2017) reported that among the 56 tested isolates, only 24 isolates were recorded as lipase producers with lipolytic zone diameters of 18–32 mm.
Quantitative Screening of Lipase Production by the Isolated Bacteria
Secondary screening was carried out on the isolated bacteria to ensure the capability for lipase production in mineral based medium supplemented with 1% (v/v) cotton seed oil. Lipase production by the selected isolates was estimated at different time intervals (24, 48, and 72 h). As shown in Table 2, the highest production of lipase (225 U/ml) was recorded for strain 3 after 24 h; thus, it was selected for further study. In a parallel study by Rasmey et al. (2017), it was reported that the highest lipase activity (20.0 ± 0.29 U/mL) was recorded for Pseudomonas monteilii using olive oil as the substrate in the medium. Similarly, the highest lipase activity (2.33 ± 0.12 U) was indicated by Staphylococcus sp. in the presence of cotton seed oil as the carbon source (Patel et al., 2018).
Detection of Polymorphism Among Lipase Producers Using RAPD-PCR
The genetic analysis of the isolated lipase producers was carried out with the main goal of identifying duplications of the same isolates. RAPD-PCR is the most commonly used marker to detect polymorphism and variability (Shahid et al., 2012; Sobhakumari, 2012). It is easy, quick, and requires small quantity of template DNA. In the present study, isolates coded 1–20 were exposed to DNA extraction and RAPD-PCR with the aim to differentiate between the bacterial isolates. As shown in Figures 2A,B, there was clear polymorphism between the isolates, which was identified using different primers namely 16S1100R and 16S F. RAPD-PCR profiling dependent polymorphism was previously used to differentiate between lipase producing bacterial isolates (Tilwari et al., 2016).
Molecular Identification of the Most Promising Isolate
The comparison of 16S rRNA gene sequence of an isolate with the sequences of the different types of strains provides a powerful tool to classify and identify prokaryotes (Kim and Chun, 2014; Zuppa et al., 2014). In the current study, DNA of isolate 3 was extracted and 16S rRNA gene was amplified; the amplicons produced were analyzed using agarose gel electrophoresis. The sequence was deposited in GenBank and had the accession number MF581790; the isolate was identified as Bacillus cereus HSS with a similarity of 98%. Figure 3 represents the phylogenetic tree showing the incidental evolutionary relationships among different Bacillus species. Bacillus species were previously reported as lipase producers in various studies (Bharathi et al., 2018; Femi-Ola et al., 2018; Sahoo et al., 2018).
FIGURE 3. Phylogenetic tree showing the incidental evolutionary relationships between different biological Bacillus species. The dendrogram was generated using the MEGA 5 program.
Optimization of Fermentation Factors Affect Lipase Production
Effect of Temperature on Lipase Production
The culture media were incubated at different temperatures (10, 25, and 30°C) under static conditions for the optimum incubation time. Results indicated in Figure 4A showed that the most suitable temperature for the highest lipase activity (225 U/ml) was 10°C, whereas the lowest lipase activity (175 U/ml) was detected at 30°C. These findings were consistent with previous studies, where lower fermentation temperature had a positive effect on lipase production (Anasontzis et al., 2014) Among the tested range of temperature (15–35°C), the optimum temperature for cold-active lipase production by Pichia lynferdii NRRL Y-7723 was 20°C as previously reported (Park et al., 2013). The effect of temperature on enzyme activity is attributed to its effect on stability, rate of the reaction, substrate solubility as well as direct influence on esterification reaction (Musa et al., 2018).
FIGURE 4. Effect of temperature (A), agitation (B), and different inducer substrates (C) on lipase activity of B. cereus HSS.
Effect of Agitation on Lipase Production by B. cereus HSS
Lipase production was premeditated under the effect of agitation at 10°C for 24 h. Data presented in Figure 4B showed that lipase activity reached a value of 285 U/ml, which represented a 1.3-fold increase at shaking condition when compared with the (225 U/ml) static condition. Aeration has up-and-down effects on lipase production by different organisms (Gilbert et al., 1991). Enzyme activity had a significant elevation under agitation when compared with the stationary state. Enzyme production increased when the agitation rate increased up to 150 rpm, and, subsequently, a reduction in activity was reported (Sooch and Kauldhar, 2013). The decrease in enzyme activity at elevated agitation rates might be attributed to the depletion in the nutrient level and oxygen in the medium and the shared stress on bacterial cells in addition to enzyme structure (Calderbank and Moo-Young, 1959).
Effect of Different Inducers on Lipase Production by B. cereus HSS
Lipases are inducible enzymes. Glycerol, hydrolysable esters, fatty acids, bile salts, Tweens, and triacylglycerols take action as inducers for lipase production (Griebeler et al., 2011). In the present study, sesame oil, corn oil, olive oil, palm oil, coconut oil, and cotton seed oil were tested as inducers for lipase production by B. cereus HSS; the enzyme appeared most active toward cotton seed oil with an enzyme activity of 285 U/ml. In contrast, the lipase attacked olive oil, corn oil, palm oil, sesame oil, and coconut oil more slowly (Figure 4C). Cotton seed oil is a cheap, commercial, and available economic source for lipase production. Therefore, it was previously used as a carbon source for lipase production (Abd-Elnaby et al., 2015). The current study indicated that the use of different lipids caused variations in lipase activity. These results are in harmony with the findings of El-Sawah et al. (1995). It was reported in many published experimental data that natural oils stimulate lipase production (He and Tan, 2006; Kaushik et al., 2006). Similarly, elevated level of lipase production by Bacillus sp. was observed upon using olive oil as substrate in the custom medium (Eltaweel et al., 2005).
El-Sawah et al. (1995) tested the effect of different substrates that included olein, sunflower, coconut, olive oil, soybean, cotton seed, stearin, butter, castor, and tallow fat, which were appended to the basal medium at a concentration of 1%. It was shown that olive oil was the most favorable substrate for the increased production of lipase by Lactobacillus delbrueckii subsp. bulgaricus.
Divya et al. (2015) studied lipase production by cold-adapted Geotrichum sp. and Rhodotorula sp. with a wide range of commercial oils like sunflower oil, olive oil, coconut oil, vanaspati, and ghee at 30°C and reported that palm oil was the most preferable substrate for maximum lipase production.
Optimization of Culture Conditions Using Plackett–Burman Experimental Design
Plackett–Burman experimental design was used with 11 different fermentation conditions. The examined factors and their levels are presented in Table 3. The average results of duplicate experiments are given as the response (U/ml) (Table 3). The estimated main effect of each variable on lipase production and t-values were shown in Table 3 and Figure 5.
TABLE 3. Levels of examined factors and statistical analysis of the Plackett–Burman experimental design.
As shown (Figure 5), positive effects of sucrose, NaCl, MgSO4, KCL, FeSO4, and inoculum size on lipase production were observed, which means that high levels of these parameters will correspond to higher lipase activity, whereas yeast extract, K2HPO4, pH, culture volume, oil concentration had negative effects. Thus, the predicted optimum medium that resulted from this experiment included the following (g/l) sucrose, 0.8; yeast extract, 0.2; K2HPO4, 0.01; MgSO4, 0.08; and KCl, 0.08; pH, 6; culture volume, 25; oil concentration, 0.5%. Data analysis using t-test (Table 3) revealed that FeSO4, KCl, and oil concentrations were the most significant factors that affected lipase activity. Interactions among the three factors are illustrated (Figure 6). Many authors applied Plackett–Burman experimental design for the enhancement of enzyme production (Collaa et al., 2016; Kai and Peisheng, 2016; Vasiee et al., 2016; Ebrahimipour et al., 2017; Mazhar et al., 2017).
FIGURE 6. Response surface plots of the interaction between process variables in lipase production by B. cereus HSS.
A verification test was carried out to validate the results gained from the statistical analysis of Plackett–Burman design, using the predicted optimized media against the basal condition media. As shown in Figure 7, lipase activity reached a value of 500 U/ml with an 1.8-fold increase when grown in the optimized medium compared with the basal condition. In comparable studies conducted by Kai and Peisheng (2016) and Sharma et al. (2009), 1.85 and 1.6 fold increases, respectively, were achieved in lipase production upon the application of the Plackett–Burman experimental design for medium optimization.
FIGURE 7. Verification experiment: Lipase production by B. cereus HSS grown on basal versus optimized medium.
Another study by Maharana and Ray (2014) showed that the formulated medium predicted by the Plackett–Burman design enhanced lipase production by Pseudomonas sp. Akm-L5 at 15°C by 3.2-fold.
Effect of Immobilization on Lipase Production by B. cereus HSS
Immobilization of microbial cells could eliminate the time consuming, tedious, and costly steps involved in the isolation and purification of intracellular enzymes. It also intends to improve the stability of the enzyme (Chandorkar et al., 2014; Lee et al., 2017). To study the effect of entrapment on lipase activity, entrapment of B. cereus HSS cells was carried out in 3% sodium alginate. Results (Figure 8A) showed that entrapped cells exhibited lower lipase activity when compared with free cells. Adsorption of cells was carried out using luffa pulp, pumice, clay, ceramic, and sponge as support materials. The adsorbed cells were tested for their potentiality in the production of lipase. As shown in Figure 8B, lipase activity was decreased by 1.5, 3.3, 11 and 3.7 fold upon using ceramic, pumice, clay, and luffa, respectively. The productivity obtained by the immobilized cells on sponge was considerably higher than the free cells. A comparative study indicated that the dramatic variation and the efficiency of an immobilization process rely on the support used (Musa et al., 2018). Thus, cells adsorbed on sponge (Figure 8C) were chosen to complete the study, when these immobilized cells were reused for five successive cycles. As shown in Figure 8D, lipase activity increased in the first cycle and was found to be nearly constant from the second to the fourth cycles, followed by an increase in the fifth cycle to reach a value of 800 U/ml. Accordingly, recycling of B. cereus HSS cells on sponge revealed increases of 1.5-fold and 2.8-fold when compared with free cells grown in the optimized and basal conditions, respectively.
FIGURE 8. Production of lipase by entrapped cells (A) and adsorbed cells on different support materials (B), electron micrograph of sponge (C1) and cells adsorbed on sponge (C2), and recycling of adsorbed B. cereus HSS on sponge (D).
It was reported that in comparison with free suspended cells, immobilized cells exhibit tolerance to toxic substrates, enhance fermentation productivity, can adapt to a wide range of pH environments and high process temperature, and are reusable (Jaiswal and Joseph, 2011).
Efficiency of B. cereus HSS in Wastewater Treatment
The present experiment was a trial for the reduction of contaminants in wastewater by the marine lipase producer B. cereus HSS. To achieve this goal, BOD, TSS, and oil and grease were estimated before and after the treatment of wastewater with B. cereus HSS. Results mentioned in Table 4 showed that the highest reduction efficiencies of BOD, TSS, and oil and grease were 87.63, 90, and 94.7%, respectively, after 72 h. These results confirmed the reduction efficiency of the tested parameters at low temperature, which is considered to be feasible and economic, representing a low energy biological method compared with the traditional methods that require high temperature and energy levels. In a similar study, El Bestawy et al. (2005) confirmed the efficiency of biological treatment of wastewater using the lipase producer Pseudomonas spp. individually or in combination. The same finding was also reported in another study (Bala et al., 2014).
TABLE 4. Removal efficiency percentages of BOD, TSS, and oil and grease after treatment by the lipase producer marine B. cereus HSS.
GC-MS Analysis of the Residual Oil
The degradation of oily wastes by marine B. cereus HSS after 72 h of incubation was confirmed using GC-MS analysis and was compared with the control (untreated waste). Chromatographic profile of the extracted oil (Figure 9) showed a remarkable decrease in the concentration of fatty acids (n-C12-n-C23). On the other hand, complete removal of other compounds (n-C25-n-C32) was detected after 72 h. Shon et al. (2002) confirmed the removal efficiency of fat, oil, and grease by Pseudomonas sp. strain D2D3. Another study found that the biological addition of microorganisms to wastes showed efficiency in removing fat, oil, and grease from wastewater (Kang, 1988).
FIGURE 9. GC-MS chromatogram of the oily wastewater (A) control (untreated) and (B) treated with lipase producer B. cereus HSS after 72 h.
The current research explored our local environments, searching for marine bacterial isolates that produce lipase. It suggested that B. cereus HSS isolated from the local marine habitat had the potentiality for lipase production. Optimization of the fermentation conditions and the medium components was carried out using one factor at a time experiment, Plackett–Burman experimental design and immobilization, indicating a 2.8-fold increase in lipase activity. The marine isolate B. cereus HSS would be potentially useful for the removal of grease traps to treat oily wastewaters, which would represent an economic alternative for wastewater treatment. Large scale applications in waste treatment need to be conducted in the future work.
Author Contributions
SH conceived, designed, and coordinated the study. SH, HAEL, and SA carried out the experiments. All authors read and approved the final manuscript.
Conflict of Interest Statement
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
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Kaushik, R., Saran, S., Isar, J., and Saxena, R. K. (2006). Statistical optimization of medium components and growth conditions by response surface methodology to enhance lipase production by Aspergillus carneus. J. Mol. Catal. B Enzym. 40, 121–126. doi: 10.1016/j.molcatb.2006.02.019
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Mazhar, H., Abbas, N., Ali, S., Sohail, A., Hussain, Z., and Ali, S. S. (2017). Optimized production of lipase from Bacillus subtilis PCSIRNL-39 A. Afr. J. Biotechnol. 16, 1106–1115. doi: 10.5897/AJB2017.15924
Musa, N., Latip, W., Abd Rahman, R. N., Salleh, A., and Ali, M. S. (2018). Immobilization of an Antarctic Pseudomonas AMS8 lipase for low temperature ethyl hexanoate synthesis. Catalysts 8:234. doi: 10.3390/catal8060234
Park, S., Kim, J., Bae, J., Hou, C., and Kim, H. (2013). Optimization of culture conditions for production of a novel cold-active lipase from Pichia lynferdii NRRL Y-7723. J. Agric. Food Chem. 61, 882–886. doi: 10.1021/jf304919y
Patel, P., and Desai, P. (2018). Isolation, identification and production of lipase producing bacteria from oil contaminated soil. BMR Microbiol. 4, 1–7.
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Vakhlu, J., and Kour, A. (2006). Yeast lipases: enzyme purification, biochemical properties and gene cloning. Electron. J. Biotechnol. 9, 69–85. doi: 10.2225/vol9-issue1-fulltext-9
Vasiee, A., Behbahani, B. A., Yazdi, F. T., and Moradi, S. (2016). Optimization of the production conditions of the lipase produced by Bacillus cereus from rice flour through Plackett-Burman Design (PBD) and response surface methodology (RSM). Microb. Pathog. 101, 36–43. doi: 10.1016/j.micpath.2016.10.020
Yu, Y., Li, H., Zeng, Y., and Chen, B. (2009). Extracellular enzymes of cold-adapted bacteria from Arctic sea ice. Canada basin. Polar Biol. 32, 1539–1547. doi: 10.1007/s00300-009-0654-x
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Keywords: lipase, Plackett–Burman, immobilization, recycling, wastewater treatment
Citation: Hassan SWM, Abd El Latif HH and Ali SM (2018) Production of Cold-Active Lipase by Free and Immobilized Marine Bacillus cereus HSS: Application in Wastewater Treatment. Front. Microbiol. 9:2377. doi: 10.3389/fmicb.2018.02377
Received: 05 July 2018; Accepted: 18 September 2018;
Published: 23 October 2018.
Edited by:Pankaj Kumar Arora, Babasaheb Bhimrao Ambedkar University, India
Reviewed by:M. Oves, King Abdulaziz University, Saudi Arabia
Kumari Kavita, National Institutes of Health (NIH), United States
Copyright © 2018 Hassan, Abd El Latif and Ali. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
*Correspondence: Sahar W. M. Hassan, email@example.com


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Test post on my new-old Blogger site-all comments very welcome.Bit on the temperature challenged side at the Castle this morn, the kitchen is once again filling up with broken computers, but the first daffodil has opened in the garden.
Piss Poor Policies Dave C's hopes of following Tone into a Middle Eastern country have been dashed it seems.
PPP Dave was forced into an embarrassing retreat over military action against Libya yesterday as the Coalition announced the biggest cull of the Armed Forces in a generation.
Within 24 hours of announcing that British warplanes might enforce a 'no-fly zone' above the crisis-torn state, the Ministry of Defence revealed plans to axe two RAF Tornado squadrons and fire 170 trainee fighter pilots.
Defence Secretary Liam Fox's decision to axe 11,000 military personnel in four years cast doubt on Britain's ability to carry out such ambitious missions.
Just to make Dave's day, the Prime Minister was forced to downplay his assertion that he was prepared to arm rebels seeking to oust Colonel Gaddafi.
Even Libya's opposition told Mr Cameron they don't want his help, unveiling a banner in the rebel stronghold of Benghazi which read: 'No Intervention.'
Take the hint Dave-we don't want you either.
The Castle cupboards are getting empty, the reason being that the price of food seems to be linked to the price of oil.
Supermarket giants seem to be boosting their profit margins and blame rising inflation, a leading City bank warned today.
Analysts at UBS said of the seven leading world economies, prices in Britain were rising at the fastest rate - and by far more than is justified.
The sector could now come under intense political pressure or face a formal investigation, they suggested.
'UK food prices are rising more rapidly than most other developed economies' food prices, and have significantly outstripped food retailers' cost inflation,' they said.
'This could allow UK politicians to suggest that food price inflation is 'unfair' or 'excessive'.'
Analysts at the Swiss bank found prices in Britain were rising at an annual rate of 4.9 per cent - against 3.6 per cent in Germany, a euro zone average of 1.8 per cent and a U.S. increase of 1.5 per cent.
Won't be long before I can't even afford stale bread and gruel.
A London council has confiscated supplies of breast milk ice cream from a specialist parlour which launched the new flavour in the British capital last week on concerns the frozen treat may spread viruses.
Westminster Council said it had visited the Icecreamartists restaurant in London's Covent Garden and removed all ice cream containing breast milk for testing after being contacted by members of the public and the Food Standards Agency.
"Selling foodstuffs made from another person's bodily fluids can lead to viruses being passed on and in this case, potentially hepatitis," Brian Connell, Westminster Council's cabinet member for business, said on Tuesday.
He added that the owner had agreed to cease making and serving the ice cream while it was being tested.
Sod it! And I have already bought my train ticket.
A record 241 nominations were submitted for the 2011 Nobel Peace Prize and the Norwegian jury has now begun the secretive process to select a winner, the panel's spokesman said Tuesday.
Russian human rights activist Svetlana Gannushkina, secret-spilling organization WikiLeaks and Cuban dissidents are among the candidates who have been publicly announced by those who nominated them.
The Norwegian Nobel Committee does not reveal the names of nominees and doesn't discuss any candidates until the winner is announced in October.
Geir Lundestad, the permanent secretary of the committee, told The Associated Press that 188 individuals and 53 organizations have been nominated for the prestigious award.
Other nominees are-Tone Bliar, Gadaffi, Mugabi and Ahmadinejad-apparently.
"Och me dinners bad" has just seen his 1977 Peugeot 504 sold for $2.5 million.
The "company who purchased the car for 25 billion rials wanted to remain anonymous", it said after the auction in the south-western city of Abadan.
"My client bought the car because of a personal interest in president Ahmadinejad and his spirit of anti-arrogance and anti-Zionism," said lawyer Mahmoud Esari, who acted for the firm that bought the car.
SWO chairman Ahmad Esfandiari said more than 500,000 people registered on the bidding website, with most saying they were doing so because Mr Ahmadinejad leads a simple lifestyle.
He said there were bids from Europe, Asia and the United States.
On January 1, Mr Ahmadinejad launched a website to invite international bids for the car, with the proceeds funding a project to build 60,000 homes for disabled and needy women who are providing for their families.
Bit annoyed about that, according to "we buy any car" the Rover is only worth fifty quid; maybe I should change my name.
A Glendale man allegedly drove in reverse for more than three miles down North Glendale Avenue on Saturday, without crashing or hurting anyone.
An off-duty police officer saw Gurgen Gukasyan, 49, driving at about 30 miles per hour with his right arm on the passenger seat and his head turned, according to the Glendale News Press.
Gukasyan was arrested on a misdemeanor charge of reckless driving, according to the Los Angeles Sheriff's Department inmate report.
Gukasyan was not under the influence of drugs or alcohol and no injuries were reported, Glendale police Sgt. Tom Lorenz told the Glendale News Press.
He reportedly told police he was driving to a friend's home.
"This is just sheer disregard for public safety," Lorenz told the newspaper. "It appears that he did want to take that vehicle and have some fun with it."
But this isn't the first time Gukasyan has allegedly made the road his playground.
He was also arrested in December after he allegedly drove his car onto Verdugo Park, causing thousands of dollars of damage before stopping his car and blasting music.
Maybe he should have a word with PPP Dave C, he can reverse as much as he likes and get away with it.
And finally:
Ladies sauntering.
That's it: I'm orf to cheat the law of light
And today's thought: Some days you're a pigeon . . . and some days you're the statue.


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(Adds details, share movement)
Oct 4 (Reuters) - Pfizer Inc , which is seeking a wider use of its multi-billion dollar vaccine Prevnar, said a late-stage study of the Prevenar 13 vaccine met the main trial goal and the data will help the company expand a European regulatory application for the vaccine.
Prevnar 13 - known as Prevenar 13 in Europe - fights pneumonia, meningitis and other diseases caused by the pneumococcus bacteria, and was approved in the United States in December 2011 for use in people aged 50 years and over.
It protects against 13 strains of the streptococcus pneumoniae, while Prevnar - one of Pfizer's top-selling products - protects against seven strains.
A competing vaccine from GlaxoSmithKline , Synflorix, protects against 10 strains of the bacteria.
Pneumonia, caused by the pneumococcal organism, is one of the biggest causes of death in older people and its incidence begins to increase after age 50.
Pfizer said Prevenar 13 met the main goal of a late-stage study by showing that the immune response to the vaccine in the 18 to 49 year old age group was not inferior when compared with the response in the 60 to 64 year group.
Pfizer's shares were up marginally at $25.40 on the New York Stock Exchange on Thursday.
(Reporting by Balaji Sridharan in Bangalore; Editing by Roshni Menon)
((email@example.com; within U.S. +1 646223 8780, outside U.S. +91 80 4135 5800; Reuters Messaging: firstname.lastname@example.com))
Keywords: PFIZER STUDY/


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The fall 2016 transportation workshop focused on the city of Charlotte's Blue Line Extension light rail and evaluated the successes and shortcomings of transit-oriented development (TOD) around the stations. The workshop was facilitated by Carolina Planning PhD Candidate Bill Bishop and supported by City of Charlotte Planning and regional transit staff.
The City of Charlotte planning staff and city council desired vibrant, mixed-use development along the Blue Line corridor, which to the dismay of both city officials and residents wasn't consistently happening. In some areas, development occurred with significant intensity, surpassing expectations. However, in most areas, new development was slow or even nonexistent. There were also concerns voiced about the "monoculture" nature of the projects along the corridor: development was almost exclusively four to five story apartments with 80-120 units.
The goal of the workshop was to establish a framework to guide the City of Charlotte's efforts to maximize value along the Blue Line corridor. This required reconsidering what TOD means for Charlotte and the Blue Line – a complex and nuanced task. The final report recommends the following four strategies to aid certain types of development surrounding the stations as well as tools to help the City of Charlotte implement the new strategies in areas not receiving private sector investment.
- Promote Residential Diversification: provide a more diverse array of residential product types that will appeal to families, aging households, and lower-income households through increased density, lax parking requirements, communal amenities, and affordability.
- Improve Multimodal Connectivity and Accessibility: incentivize multimodal access to destinations and take advantage of opportunities to benefit from pedestrian and bicycle amenities for transportation and recreation.
- Pursue Explicit Placemaking Strategies: promote placemaking to foster community ownership and create desirable destinations by putting function before form and proactively create places that are collaborative and context-specific.
- Engage Community in Station Area Planning: engage neighborhood organizations in identifying project opportunities, reducing risk, and supporting value creation as successful TOD requires understanding and support from the community
To learn more about Charlotte's light rail, visit http://www.lynxcharlotte.com/
Or to learn more about the Fall 2016 Workshops, visit https://carolinaangles.com/2016/12/02/fall-2016-workshop-review/.


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Veronica is based in Melbourne, Australia. She has recently completed a light art based PhD at Monash University, having previously completed a MVA at the VCA, Melbourne University. She has exhibited extensively with 12 solo and more than 30 group exhibitions in Melbourne and Perth. In 2015 she won the Contemporary Sculptors Association Annual Award and was finalist in the Mary & Lou Senini Award, McClelland Sculpture Gallery, Langwarrin. She attended an Associate Artist Residency, ACME Studios, London based around the Hayward Gallery, Light Show, supported by the Post Graduate Research Travel Grant, Monash University and the Australian Postgraduate Award.
"During my PhD project I became interested in ideas about passive solar houses inspired by Vastu Indian Architecture as a way of connecting with nature and natural light. I used the placement of my works to compare how disconnected our buildings are from nature and natural light. I aligned my works with true cardinal points of north, south, east and west with an aim to subvert the studio or gallery spaces. I began to use my own height as a unit of measurement to be compared with the architecture around me and natural light fields in extended space. I continue to be interested in connection between my practice, Colour Field Painting and Post-minimalism."
Above left: Light in plinth 4, 2014, Aluminium frame, LEDs, show saver, painted wooden plinth (72 × 72 × 72 cm)
Above right: Light in plinth 5 (just taller than me) 2015, LEDs, Perpsex and painted wood (160 × 66 × 66 cm).


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Saturday, November 24, 2012
According to the three peaks crowd, target and timing have hit for the great plunge to 1100 over the next six weeks. It have been extremely reliable for the past 18 months, and we must give it it's due.
The cycle guys see right translated moves and now a press to make new highs over the the next 20 trading days.
I'll follow my signals, but my gut says we overshoot the three peaks scenario, and when it is clear no deal before year end on taxes, a mad rush for the door, to 125 area. This will take down the miners I follow, as well as energy. Gold and silver will largely ignore this move in my opinion, and a the very least be neutral, just like the last sell off.
Now some charts
2 comments:
After you take your positions, defensively, do you place hard stops, and stick with those, hell or high water?ReplyDelete
As if it matters, I agree with you and the cycles boys who expect a quick run up and then a tank in gold and silver. I can't see anything myself in the S&P, so that money can't belong to my wards. They're long within 3 days of the lowest low in gold, because they're them and all I can do is say, "here". Here was 1685-90 in gold, 100 points off the high.
They're not hedged, though, as I can't get them to place defensive stops.
They'll be out shortly for a quick skim of the market, if lucky, equivalent to 75 - 100 points in the gold market, and wherever the S&P panic entry gets us.
The problem I see is a gap opening to the upside on Monday which spells as an average just a typical 2-5 trading day rise before all those chasing learn the sting of the retracement lash.
That retracement is so obvious, it's embarrassing to see it resolve that way, but that's I guess how the major players can play the psychology of the weak handed and emotional gamblers.
And after the retracement, a second clock cleaning with a month, the pattern's not clear... my sense is there's work to do to get the recent longs who just wouldn't release on this last drop "out" of the market, and that takes time, time and more time.
So, I'm up for guessing only up through this fast retracement back to the opening gap (and unfilled), the day over day gap, on Monday.
I'm pretty sure there was a gap in this area in the recent past, also, and to me that means that gap's gonna get marched all over.
Thx for the succinct welcoming words. ;-)Replies
Based on market movement today, and in the night session, gold will spring upwards with a mad tail of wannabee's chasing it for the next 2 days, chase and consolidate, or edge up and chase up, and then it will retrace downward quickly. Exit on the close of the chase day, or for the more aggressive, short the end of the chase day... whenever it's obvious that the short squeeze has is in motion. Exit 1800-1780 range, for a re-entry 50 points below the high, or back to 17485-0, where we are today. This is a gap fulfillment from a prior week, about 3-4 weeks ago, day over day in the ny pit.Delete


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Liberia mystery as 'banknotes worth $60m vanish
The Liberian Government says it is investigating what has happened to two huge consignments of money that were printed abroad and brought into the country between November 2017 and August this year.
The money, said to be local banknotes worth about $60m (£45.6m), has allegedly disappeared.
One media report says containers packed with the money left the port in the capital, Monrovia, under security escort in March and were thought to be headed for the Central Bank, but they vanished.
The consignment that came through the international airport cannot be accounted for either.
The Justice Ministry is asking the public to remain calm while a panel of security officers investigates the matter.
Minister of Information Eugene Nagbe has told state radio that President George Weah, who took office in January, is "unhappy" that he was not informed about the situation until now.
"This is alarming," Mr. Nagbe said.
No arrests have been made so far, but the Information Minister says he is certain they will get to the bottom of the suspected scandal.
But in line with the recent discovery, the Liberian government has banned 15 people, including the son of former President Ellen Johnson Sirleaf, from leaving the country while it investigates the alleged disappearance of tens of millions of dollars intended for the Central Bank, the
Ministry of Information has said in a statement.
Border posts have been informed that Charles Sirleaf, the deputy governor of the Central Bank, and Milton Weeks, the former governor of the bank, are among those barred from travelling abroad, the statement added.
Mr. Sirleaf and Mr. Weeks have not yet commented.
However, Mrs. Sirleaf was quoted by the Front Page Africa news site as saying that the bank had given "full evidence and clarification" to refute allegations that the money had disappeared.
The government has ordered an investigation into the alleged disappearance of the newly printed bank notes that had reportedly come from abroad between November last year and August this year.
"The government... takes the ongoing investigation seriously because it has national security implications," the statement added.
The statement did not say how much had allegedly gone missing. Some reports put the number at around $60m (£45.6m) and others at around $100m.
The Central Bank had flatly denied that containers packed with the money had disappeared from the port in the capital, Monrovia, Liberia's Daily Observer news site reported.
Mrs. Sirleaf, a former Nobel Peace Prize winner, stepped down as president in January after her two terms ended.
She was succeeded by former football star George Weah, following his stunning victory in elections a month earlier.


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Trail Conditions
Snowmobile Trail Reports
January 31st, 2019
1-31-19 The conditions are EXCELLENT! Get out and ride. Today we were out and opened the Sugarloaf Loop and the Robert's Mountain Loop. Tonight we're out grooming Shin Pond south to Whetstone.
January 30, 2019
Tonight I'm out flattening ITS 114 and 85 North and South out of Shin Pond so it's flat before tonight/tomorrow's expected snow, which is just starting to fall. We'll be back out there beginning Wednesday night and Thursday packing in the new snowfall. The snowpack is...
January 25, 2019
1-25-19 Today Johnny groomed 114/85 South and 71D from Shin Pond up onto Cunningham lookout and 85 North to the junction of 81&85 just North if Shin Pond. I also got work that Patten came in 81 and went up 81/85 to connect with Oxbow. I'm just headed out to...
January 21st, 2019
1-21-19 Tonight we're grooming ITS114/85 South towards Whetstone Bridge and ITS114/85 North to the junction of 81&85 and 85 South towards Matagamon to the Junction of 71D all shown highlighted in yellow on the map below. Also take note that the Huber Road has been...
January 12, 2019
Today Les groomed Shin Pond Sugarloaf loop and Club Trail 64 to ITS81 and Katahdin Views off 64. Rick groomed 114/85N to junction of 81 and 85 and 85South towards Matagamon to 71D junction. I'm grooming 114/85South to Whetstone Bridge (PT13). With the packing done...
Ski Trail Reports
January 3rd, 2019
We have not yet heard of any updates from the groomers at the National Monument. We will be posting updates as soon as grooming begins!


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Product Description
Dog Desktop Perpetual Calendar Block , a lovely decorative calendar block perfect for teaching kids to understand numbers , days and months of the year.
Perpetual Calendars can be used year after year and are an easy way to keep track of the date. Part of our Doggy Collection for young Children by award winning designer Gisela Graham.
This lovely doggy calendar block displays the day of the week, month and date of the month!
Consists of 2 number blocks and 3 month blocks, all inside a cute wooden holder. It is charming and will look gorgeous in the home. Clear and easy to see, yet compact to fit easily on any desk, shelf or windowsill. Simple and easy to change. Perfect for a child's Bedroom, Playroom or in the Kitchen. Simply change wooden blocks to the desired date.
Perfect for home or for a nursery or playgroup! Lovely Birthday or Christmas Gift or use as a prize!
Dimensions: 18 x 10.5 x 5.5 cm
Designed in London for Gisela Graham
Gisela Graham has built her reputation on becoming one of Europe's leading giftware and home ware design companies, she has become particularly well known for her wonderfully ornate seasonal item.


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