8. The following limitations were encountered into the study.

This objective had been accomplished by the method of a focus group, some questions were formed to generally access the information about money management generally speaking population as well as the role it may play within their monetary situation. This is done via open ended questions to give the participant power to get feedback and discuss in form of complex textual descriptions to access exactly how people feel the offered research issue. Sample here also included students, unemployed people, part time workers, regular workers and self employed people who have different sex and age ranges.

Objective 7: Suggest a fresh Theory on Money Management in hard times.

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Thus in previously listed method the objectives were addressed and data is going to be gathered and analyzed as well as the last objective to suggest a fresh Theory on Money Management in hard times to emerged due to the success of this previous research objectives.

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3320 words (13 pages) Essay

25th May 2018 Biology Reference this

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Tetrodotoxin is alkaloid based aquatic toxins. These toxins are one of the more potent non-proteinaceous toxins along with the best-known marine natural toxins. Diodon hystrix (porcupine fish) were collected from Chennai costal region and dissected under sterile conditions to have: liver, skin, gonads, intestine, eyes and kidney. 20g of each and every organ had been macerated in 200ml of Methanol:Acetic Acid [99:1]. The filtrate is then condensed in Rota-Vaccum evaporator to obtain crude extract. The focus with this study is always to confirm the clear presence of TTX (Tetrodotoxin) in six different organs of Diodon hystrix. Analytical techniques used were GC-MS and UV spectroscopy. Also, genotoxicity of this crude extract were analysed making use of human leukocyte culture and SCE assay making use of onion root recommendations. The outcome suggest the clear presence of TTX in major skin, liver and intestine and that, the organ extract won’t have any genotoxic effect but is with the capacity of increasing the sibling chromatid trade.

Keywords: TTX, Diodon hystrix, genotoxicity, root tip assay.

Tetrodotoxin (TTX) is a very powerful alkaloid neurotoxin that is non-proteinacious in nature. TTX can withstand extremely warm and is water soluble but is afflicted with extreme pH conditions, i.e., above 8.5 and below 3.0 [1, 2, 3, 4, 5]. These properties ensure it is a dangerous toxin capable to connect most readily useful with its environment [1, 2, 5]. It really is found in both aquatic in addition to terrestrial organisms and studies have proven it is synthesized by symbiotic microorganisms, bacteria correctly, present in the gut, initially acquired through the foodstuff chain or located on the skin of this animals but its biosynthesis pathway is still unknown [ 1, 2, 5, 6, 7, 8]. TTX acts as an ion pore blocker, binding

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to site 1 sodium channel receptor of this axon membrane hence inhibiting the influx of sodium ions and so resulting in the blockage of action potentials [1, 2, 3, 4, 5, 6, 7, 8, 9]. TTX is ten thousand times poisonous than cyanide and another of the very most fatal poisons on Earth.scholarships for minorities majoring in biology smart 2019 no essay The LD50 is approximately 0.2μg when injected in mice [2, 5]. On the other hand, combined with life-threatening faculties, clinical trials and clinical tests have demonstrated that TTX has remarkable therapeutic properties as an analgesic in cancer treatment process [2].

Puffer fish from the order Tetraodontiformes, was in fact identified to trigger many mortalities as a result of food poisoning as being a results of TTX intoxication. In several countries such as for example Japan and China, puffer fish is deemed a food delicacy provided it really is served by a licensed and well experienced chef many situations of poisoning still prevail [1, 3, 4, 5, 6, 7, 8, 9, 10]. It is often reported that just a very low dose of TTX

in blood is adequate for an immediate affect the host [5]. Studies have concluded that the absolute most toxic organs of this puffer fish are the liver followed closely by the intestine after which skin and ovary. As well as that, TTX can be found in low concentration in other organs including the eyes and muscles [3, 5, 8, 10].

The analysis is targeted on Diodon hystrix which really is a form of puffer fish from the class Diodontidae and it’s also also referred to as Porcupinefish because of the sharp needle-like structures covering its system as being a defense system against predators. Presence of TTX was reported in Diodon hystrix worldwide [2, 4, 5] but studies on this animal from the sea of this eastern shore of India that is the Bay of Bengal is yet to be reported. The goal of this research is to spot TTX into the crude extract from Diodon hystrix obtained from Chennai Coastal line also to investigate the Genotoxicity of this crude extract from respective organs making use of human leukocyte culture and onion root tips.

The puffer fish had been collected from the coastal lines of marina beach, Chennai in early July 2014. The identification of this puffer fish was done by visual comparison having an online fish database –www.fishbase.org. The database parameters were set correctly to sample collection site as well as the possible species for sale in Bay-of-Bengal region aided by the matching morphology were only two forms of Diodon sp.. Out of which Diodon hystrix had the closest match, on the basis of the skin coloration pattern.

The collected puffer fish were dissected and visceral organs like liver, intestine, kidney, eye, and skin were removed and organs were weighed. The isolation for the tetrodotoxin[3] include from the samples 10 grams of organs were taken and Then suspended in 100ml of three level of 1% acetic acid in methanol without damaging the tissues then a whole materials were into the ice box all day and night at a sterile condition, as an incubation period next step the tissue were macerated in a mortar and pestle gently, if the tissues get dried up add required level of the chilled ethanol if needed. Then a slurry were filtered by making use of whatman no. 1 filter paper. Then a filtrate solutions were centrifuged at 12000 rpm for ten full minutes at 4 degree Celsius. Then a supernatant were separated not only that the samples were concentrated by making use of lyophilisation to have crude extracts for our intent behind study

To spot the clear presence of alkaloids [10] to 2mg of crude extracts 5ml of distilled water were added and then 2M hydrochloric acid had been added until an acid effect occurs. To the 1 ml of Dragendorff’s reagent was added. Formation of orange or orange red precipitate suggests the clear presence of alkaloids

Fuel chromatography (GC) and mass spectrometry (MS)[8][11][12]forms an effective combination for Chemical analysis. GC-MS analysis were an indirect solution to detect TTX in a crude extract,

that has been tough to purify in other higher level analysis practices. In this technique, we dissolved TTX as well as its derivatives in 2 ml of 3 M NaOH and heated in a boiling water bath for 30 min. After cooling to room temperature, the alkaline solution of decomposed compounds had been adjusted to pH 4.0 with 1N HCl as well as the resulting mixture had been chromatographed on a Sep- Pak C18 cartridge (Waters). After washing with H2O first after which 10% MeOH, 100% MeOH fraction were collected and evaporated to dryness in vacuo. To the resulting residue, an assortment of N, O-bis acetamide, trimethylchlorosilane and pyridine (2: 1: 1) had been included with generate trimethylsilyl (TMS) ‘‘C9-base’’ compounds. The derivatives were then put into A hewlett packard fuel chromatograph (HP-5890-II) built with a mass spectrometer (AutoSpec, Micromass Inc., UK). A column (φ 0.25 mm × 250 cm) of UB-5 had been used, as well as the column temperature is increased from 180 to 250°C at the rate of 5 or 8°C/min. The flow rate of inlet helium carrier fuel were maintained at 20 ml/min. The ionizing voltage is generally maintained at 70 eV aided by the ion source temperature at 200°C. Scanning was performed into the mass range of m/z 40–600 at 3s intervals. The total ion chromatogram (TIC) as well as the fragment ion chromatogram (FIC) were selectively checked.

In UV spectroscopy, TTX had been generally dependant on irradiating a crude toxin with UV light [11][12]. Handful of samples were dissolved in 2 ml of 2 M NaOH and heated in a boiling water bath for 45 min. After cooling to room temperature, samples were examined in UV spectrum and results were noticed in the product range 270nm to 280nm.

Chromosome preparations were obtained from PHA-stimulated peripheral blood lymphocytes[14][15]. Towards the fresh tubes 5ml of Hikaryo XL RPMI ready-mix media and 0.5ml of heparinized Blood (50drops) were added as well as the contents were mixed gently by shaking. Then Incubated for 72 hours in standing position in a incubator. By the end of 48th hour of incubation, the culture had been treated with TTX (0.5ug/ml) (10ul/ 5ml of culture) and once more kept it in incubator for another a day. By the end of 24th hour incubation, the culture had been thoroughly washed by centrifuging the information at 1500rpm for five full minutes, discard the supernatant and add 5ml of RPMI 1640 medium. To the content 60 microliter of colchicine had been added and tubes were kept for 20 mins incubation in incubator at 37oC while the content had been centrifuged at 1500 rpm for ten full minutes after incubation. The supernatant had been removed and 6ml of pre-warmed 0.075M hypotonic solution had been added. The content was mixed employing a Pasteur pipette and incubated at 37 oC in incubator for 6 mins. After incubation the content tube had been centrifuged at 2000 rpm for five full minutes. The supernatant had been discarded and 6ml of Carnoy’s fixative had been added and mixed vigorously. After fixation the information had been kept in room temperature for 1-2 hours. The information had been again centrifuged at 1500 rpm and supernatant had been removed and this step had been continued until pellet becomes white. For the preparation of slides the newest slides were first refrigerated and then cell button mix was dropped within the slides and dried straight away on a hot plate, after which had been kept in a incubator for proper drying. The slides were then put into a coplin jar containing Giemsa staining for 4 mins and destained in a coplin jar containing distilled water for 1 minute. The slides were dried and then viewed under microscope for stained chromosome. . The slides were then viewed under 100X power under oil immersion objective of this microscope to evaluate the chromosome aberrations.

The onion root tips[1], 2-3 cm long, were soaked in 100 µM 5-bromodeoxy uridine (BrdUrd) for pretty much 20 h followed closely by an hour treatment aided by the crude extract after having a brief wash, the roots were allowed to grow for another round in growing media. The treatments were terminated by washing the roots with distilled water after which 0.05% Colchicine was added then incubated for 2.5 h. Roots were washed, excised and fixed in Carnoy’s fixative, for 1-3hrs and preserved at 4°C. The roots were processed making use of cytology practices for SCE analysis.. The roots were then hydrolysed in 5 N HCI at 25°C for 92 min and stained with haematoxylin for at least 2hrs. The stained root[16] were washed in distilled water, squashed in a drop of 45% acetic acid and tapped for metaphase chromosome separation under coverslips. Regular water controls were within the assay. The slides were observed at 100X magnification in oil immersion making use of light microscope

Fig 1: Showing results of sample after Dragendorff’s test

The alkaloids present in the puffer fish was precipitated as a complex formation by dragendorff’s reagent. Dragendorff’s test results showed extremely high precipitation in skin and intestine, high precipitation in liver and extremely low precipitation or almost no precipitation had been noticed in kidney, gonads and eye.

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Characteristic peak was observed at retention time 8.33 and 8.66 in liver, intestine and skin after performing alkaline treatment and there was clearly no characteristic peak observed in kidney, eyes and gonads. After boiling of samples that incorporate TTX in alkaline solution (NaOH) the compound TTX present gets paid down to C9 base TMS (trimethysilyl). It really is noteworthy that all peak of selected ion monitored at m/z = 376, 392 and 407 appears at the same retention time into the Selected ion-monitored mass chromatogram of this TMS derivatives of alkali-hydrolyzed. From types of liver, kidney and intestine, mass fragments of ion peaks had been observed at ion M/z 376, 392 and 407, which are characteristic of this quinazoline skeleton (C9 base), that has been very nearly similar as those from the TMS-C9 Base derived authentic TTX

Fig 2: Showing GC-MS spectrum of the TMS derivatives of alkali-hydrolysed toxin from Diodon hystrix

In UV analysis method characteristic peaks were noticed in all samples. Shoulder peak had been noticed in liver, intestine and skin, Declining and Inclining Peaks were observed in kidney, eyes and gonads. The UV spectrum is analyzed for the characteristic of absorptions, connected with C9-base .The shoulder peaks were observed at 276 nm suggests the synthesis of C-9 base which were specific to TTX or related substances.

 

Fig 3: Showing chart of UV-spectroscopy of this crude extract from various organs of Diodon hystrix, peak at 276nm indicating the clear presence of TTX.

Metaphase plates were obtained while observing under 100X magnification in oil immersion making use of light microscope. It is often observed in most of the samples that there were no chromosomal aberration that is structural or numerical chromosomal modification are not observed. Using this result, it could be reported that the crude extract from Diodon hystrix does not have any clastogenic (breakage of chromosome) or aneugenic ( change in chromosomal number) results.

Fig4(left): Showing metaphase plate from control leukocytes. Fig5(right): Showing metaphase plate from crude extract leukocytes.

The Sister Chromatid Exchange (SCE) assay was reported to be one of the more sensitive and painful short-term genotoxicity assays due to its capacity to determine genotoxins at really low doses (Tucker et al.1993). It is often observed that the crude extract from Skin and intestine enhanced SCE notably over the control even though the Liver, Eye, Gonads and Kidney have very low results. In order that it is put forth that the crude extract from skin and intestine interfere to a good deal aided by the SCE and further studies have to be performed.

Fig6(left) : Showing results of SCE in control onion root tip. Fig7(right): Showing results of SCE in crude extract root tip.

From the study, it could be reported that Diodon hystrix from the eastern coastal region of India, observed to own accumulated TTX in its organs. Hence it could be toxic when ingested and also life-threatening towards the predators. Nonetheless further studies should be performed on this fish to ensure the clear presence of a homologue of TTX and acquire a purified sample of this TTX.

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5246 words (21 pages) Essay

1st Jan 1970 Economics Reference this

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Big Onion crop had been introduced to Sri Lanka by the British in 1855 and commercial cultivation had been introduced by the Department of Agriculture during the 1950’s and within the last years, the crop performance had been assessed in several places also it had been observed that big onions is grown economically during every Maha season in nearly all places.

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2. Nonetheless, at present the cultivation of big onion is confined simply to Matale, Anuradhapura, Puthalama, Pollonnaruwa, Mahawelli and Jaffna Districts. Significantly more than 50% of this total onion production in Sri Lanka is cultivated from the Matale District. [1] 

3. The Government strives to achieve a self enough stage in the production of big onions since Sri Lanka spends a substantial amount of money outflow every year regarding the importation of this big onions. Meanwhile, into the immediate past it was pointed out that the big onion production was affected in Sri Lanka and so customers may also be spending an increased price for the big onions. In particular the big onion production in Dambulla area was declining within the last few several years.

4. The Dambulla area plays a essential role in the big onion cultivation in Sri Lanka. The us Government was spending less attention and support on advertising the big onion production in Dambulla.

Therefore, it offers so happened that the onion production in Dambulla has declined into the immediate past as a consequence of the government’s less support because of this sector. Therefore, the key intent behind this study is always to promote the big onion cultivation into the Dambulla area.

5. This research is performed aided by the following certain and general objectives.

a. The key general objective with this study is always to determine the key dilemmas encountered into the onion cultivation of this Dambulla area.

6. The precise objective with this study is always to supply the recommendation to boost the Big onion cultivation into the Dambulla area and certain objectives are as follows.

a. To examine the present history of Big onion cultivation in Dambulla area also to compare the present situation associated with Big Onion cultivation.

b. To spot the main dilemmas encountered in big onion cultivation in Dambulla.

c. To spot the critical contributing factors.

d. To help make suggestions on the basis of the findings.

2. The Matale District plays a essential role in the big onion cultivation in Sri Lanka in particular Dambulla provides big onions for the Sri Lankans’ consumption. In the recent past as a result of lack of support from the government sector the big onion cultivation was declining.

3. As a result the big onion cultivation in Dambulla is going to be non existence in the extremely near future. Also, many farmers be determined by the big onion cultivation as their livelihood in Dambulla. Ergo, if the big onion cultivation in Dambulla is affected many families will totally lose their income and it surely will influence the survival of several families. Hence having less support from the government as well as the consequent less onion cultivation are believed while the research problem because of this study.

4. This clinical tests the declining stage of this onion cultivation in Dambulla. The scope covers only the Dambulla part of big onion farmers. Therefore, this research was limited by the onion farmers of this Dambulla area.

7. This clinical tests the factors influencing the decline of this big onion cultivation in Dambulla. Therefore, the responses were collected from the regional onion farmers from the Dambulla area. Hence, 100 big onion farmers were thought to be a sample because of this study since all farmers could not be accessible inside the limited time because of this study. These farmers were selected in a random basis. Therefore, the straightforward random sampling method had been sent applications for picking a the sample.

8. The following limitations were encountered into the study.

a. Time is limited, to ensure that within the limited time the research has to be finished this is why in-depth analysis can not be applied.

b. The researcher encountered limitation of resources.

c. The sample had been limited only to 100 farmers.

9. The big onion is an essential minor crop consumed by many Sri Lankans and possesses been believed that 34,000 metric a great deal of onion is imported annually and Sri Lanka spends around 300 million rupees on onion importation (Gunawardena, 2009). Also, it is often also believed that 45,000 labour units are used into the onion cultivation and production annually by Sri Lankans and so, it raises income and employment generation for several Sri Lankans. [2] 

10. Many countries worldwide are getting mixed up in big onion production. In particular they have been; Belarus, Russia, Lithuania, Poland, Ukraine, India, Pakistan etc (Research Institute for Vegetable crops, 2006).

11. In accordance with Shanmugasundaram (2001) you can find kinds of onion also it mainly includes the sweet, red, white, yellow, brown and green etc.

Source – Shanmugasundaram (2008)

12. Also, it is often identified that the big onion production brings several comparative benefits in comparison to with other crops (Autko & Moisevich, 2006). A number of the benefits are given just below.

a. Output can be had in a little while of time.

b. Initial costs such as for example; seeds costs, fertilizer costs are comparatively less.

c. It generally does not demand a set expense.

d. Less technology the machines are enough.

e. High employability of manual labourers.

f. No problem finding markets.

g. Less storage period.

13. The onion essentially was divided in to red onions and onions that are big each variety requires different eco-agricultural conditions, labour, fertilizer, weather and climatic conditions, temperature, etc.

14. The literature implies different demands for smooth growing of this big onion production. A number of the conditions suggested by Autko and Moisevich (2006) are given below.

a. Increase of fertile soil layers into the zone of plant root by 4-6 cm

b. Increase of aeration and warming of soil, excluding over wetting into the period of heavy precipitation

c. Decrease of fertilizer rate application by 30%

d. Decrease of seed sowing rates

e. Ensuring of looser soil state through the whole period of vegetation

f. Chance for soil surface copying by working organs of machines, during inter-row treatment, decreasing of plant protective zone 3-5 cm, mechanical weed destruction by 70-75% and band application of pesticides that ensures the decrease of their rates by 2-3 times

g. Increase of irrigation efficiency

h. Diminution of nitrate content into the production

j. Decrease of energy expense during harvesting by 20-40%.

15. Therefore, the aforementioned conditions can be viewed while the basic demands for the growth and survival of this big onion production.

16. The onion essentially was divided in to red onions and large onions and each variety requires different eco-agricultural conditions, labour, fertilizer, weather and climatic conditions, temperature, etc.

17. Shanmugasundaram, (2001) has identified the following diseases that affect the onion cultivation. He’s got divided these deceases into two.

a. Field diseases

b. Storage diseases

18. The field diseases comprises of Stemphylium blight , Purple blotch, Anthracnose, Botrytis leaf blight, Downy mildew, Pink root, Smudge, Smut and several Basal rots (Shanmugasundaram, 2001).

19. The storage diseases covers common field rots, botrytis neck rot, black mold and bacterial soft rot (Shanmugasundaram, 2001).

20. Meanwhile it is often learned that into the immediate past the onion cultivation was reducing due to many factors. Some factors identified by Kulatunga (2006) are presented below.

a. Lack of quality seeds

b. Lack of advice offered for application of seeds

c. Insufficient loan facilities offered to purchase quality seeds

d. Long durations taken for harvesting from seeds

e. Lack of government support in providing fertilizer facilities towards the onion production

f. Lack of quality fertilizers designed for the onion producers

g. Lack of option of fertilizer at outside and private outlets

h. Absence of counselling and advice offered on the best way to apply the fertilizers for the new variety

j. Lack of storage facilities to store the onion production.

21. Though these problems are encountered into the onion production it could be divided in to two major categories. These are given just below.

a. Lack of government support in offering seeds towards the onion cultivators.

b. Lack of government support to deliver fertilizer to onion cultivation.

22. It is often observed that big onion cultivation was affected to greater level by having less government motivation in finding necessary seeds. Hence; lack of quality seeds, lack of counselling and advise on applying seeds, lack of new selection of seeds, insufficient government monetary support to get seeds, absence of assurance on harvesting timeframe etc are encountered under seeds (Kulatunga, 2006).

23. Kulatunga (2006) in addition has identified there is no enough fertilizer support to encourage the big onion production. In Sri Lanka it is often learned that the onion farmers lack government money and subsidies to get fertilizers. Also, fertilizer is sold at a fairly high price in the surface outlets. In addition the efficient and harvest stimulating fertilizers are not designed for the onion farmers. Also the quality and different selection of fertilizers may also be not available to boost the big onion cultivation into the Dambulla area.

24. It is essential that the onion production is increased so that you can protect the big onion industry also to ensure the livelihood of several Sri Lankans. Ergo the literature implies that the following measures can raise the onion production.

a. Involving in research and development activities so that you can raise the onion production.

b. Government providing support to find quality seeds.

c. Government has to offer seeds of this new varieties.

d. Government has to deliver seeds at subsidized rates.

e. Government has to deliver constant counselling and advice on maneuvering seeds.

f. Government has to give the fertilizer subsidy.

g. Providing high quality fertilizer.

h. Monitoring fertilizer distribution.

j. Counselling on handling diseases.

(Source – Formed because of this Research Study)

26. The aforementioned figure depicts two sets of factors that determine the decline in the onion cultivation; having less seed accessibility as well as the lack of fertilizer accessibility. This is produced from Kulatunga (2006). Each pair of the major factors have sub factors. Therefore, those two are believed while the independent variables. The decreasing onion cultivation is defined as the dependent variable. Ergo, this figure establishes links involving the factors as well as the decreasing onion cultivation. Through this research study one need to know which factor(s) cause for the decreasing onion cultivation, on the list of farmers into the Dambulla area.

Factors determining the onion cultivation

Lack of seeds accessibility

Receiving quality seeds

Likert

Q1

Distribution of seeds by the us government

Likert

Q2

Provision of subsidy by the us government to buy seeds regularly

Likert

Q3

Seeds offering the expected harvest

Likert

Q4

Purchase seeds from the Government Agricultural Department

Likert

Q5

Provision of training and counselling concerning the new seeds by the us government

Likert

Q6

I could get new kinds of seeds

Likert

Q7

I could get regular counselling and advice of this diseases regarding the seeds

Likert

Q8

Lack of fertilizers accessibility

Fertilizer subsidy from the government

Likert

Q9

Purchase of fertilizer from the Government Agricultural Department

Likert

Q10

Purchase of fertilizer from the private outlets at a less price

Likert

Q11

Getting quality fertilizer

Likert

Q12

Getting advice and counselling for the effective use of fertilizers

Likert

Q13

Getting different selection of fertilizers

Likert

Q14

Getting fertilizer that will maximize the harvest