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Research Title Prevalence and antibiotic sensitivity of enterohaemorrhagic Escherichia coli O157: H7 in cattle on smallholdings in Dhaka, Bangladesh Submitted to Government of the People’s Republic of Bangladesh Ministry of Science and Technology Dhaka, Bangladesh 1

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Page 1: Prevalence and antibiotic sensitivity of enterohaemorrhagic

Research Title

Prevalence and antibiotic sensitivity of enterohaemorrhagic Escherichia coli O157: H7 in cattle on

smallholdings in Dhaka, Bangladesh

Submitted toGovernment of the People’s Republic of Bangladesh

Ministry of Science and Technology Dhaka, Bangladesh

Submitted byAsst. Prof. Dr. Md. Kamrul Hassan

Principal Investigator and Chairman, Department of Microbiology & Parasitology

Sher-e-Bangla Agricultural University, Dhaka

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ANNEXURE- A

MINISTRY OF SCIENCE AND TECHNOLOGYGovernment of the People’s Republic of BangladeshBangladesh Secretariat, Dhaka – 1000Tel: 880-2-7164594, Fax: 880-2-7169606 www.most.gov.bd

RESEARCH CONTRACT PROPOSALCapacity Utilization Programme under Special Allocation for Science and Technology

(Additional Annexure should be submitted wherever necessary)

PART – I: GENERAL INFORMATION

1. NAME AND ADDRESS OF THE CONTRACTING INSTITUTE:

Sher-e-Bangla Agricultural University, Dhaka-1207, BangladeshTel : + 88029180921 Fax: +88028155800 E-mail : [email protected]: +8801723710845

Money receipt attached (Give Tick or Cross):

2. DEPARTMENT WHERE RESEARCH IS TO BE PERFORMED:

Department of Microbiology & Parasitology, Sher-e-Bangla Agricultural University, Dhaka 1207, Bangladesh

3. TITLE OF THE PROPOSED PROJECT:

Prevalence and antibiotic sensitivity of enterohaemorrhagic Escherichia coli O157: H7

in cattle on smallholdings in Dhaka, Bangladesh.(A) Name of Coordinated Research Programme (if applicable):

(1) Name and Designation of the authority of the Organization/Institutes /University forwarding the research contract proposal:

(2) Professor Dr. Alok Kumar Paul, Director, SAURES, Sher-e-Bangla Agricultural University, Dhaka-1207, Bangladesh

(3) Area of Research: Agriculture/ Medical Science/ Environmental Science/ Biotechnology / Engineering & Applied Science/ ICT/Animal Science/ Aquaculture/ Marine Science/ Microbial and Industrial/ Basic Sciences/ Others (Specify): Animal Science and Veterinary Medicine (Biology)

4. DURATION (in year): 01 year

5. TOTAL COSTS (in Taka): 22, 52,000 (Twenty Two lac Fifty Two Thousand Tk only)

.

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PART – 11: INFORMATION ABOUT PROJECT PERSONNEL

1. PROJECT PERSONNEL

A. Principal Investigator

Name Date of Birth

Sex: M/F Position held (since)

Dr. Md. Kamrul Hassan 01/02/1983 M

Assistant Professor and Chairman, Department of Microbiology & Parasitology, Sher-e-Bangla Agricultural University (April, 2014)

Present Address Permanent Address e-mail Address MobileDepartment of Microbiology & Parasitology,Sher-e-Bangla Agricultural University

Vill: Dhamachama, Post:Nimgachi, Upazilla: Dhunat, Dist: Bogra

[email protected] 01723710845

Academic degrees held

Subject Name of Degree University Country Class

Year of Graduati

on

Veterinary Medicine

MS in Microbiology

Bangladesh Agricultural University

Bangladesh

Grade; A CGPA: 3.859

(out of 4)1stclass(77.18%)

2009

Veterinary Science DVM

Bangladesh Agricultural University

Bangladesh

Grade; A CGPA: 3.591

(out of 4)1stclass(71.82%)

2007

Science HSC Rajshahi Board

Bangladesh 1st class(76.70%) 2001

Science SSC Rajshahi Board

Bangladesh 1st class(79.90%) 1999

Previous experience

1. Assistant Professor and Chairman; From April 10, 2014 to till to date. Department of Microbiology & ParasitologyFaculty of Animal Science & Veterinary Medicine Sher-e-Bangla Agricultural University Dhaka-1207

2. Lecturer ; From 10 April,2012 to 09 April ,2014Department of Microbiology & ParasitologyFaculty of Animal Science & Veterinary Medicine Sher-e-Bangla Agricultural University, Dhaka-1207

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3. Any special Institution attended and course taken: (a) Training on “Operation and Maintenance of Laboratory Equipment” organized by

“Modernization of Animal Husbandry Lab of SAU” sub-project under Higher Education Quality Enhancement Program (HEQEP) Project, Bangladesh.

(b) Course on “Teaching Methods and Techniques” conducted by the Outreach Program, Sher-e-Bangla Agricultural University, Dhaka-1207, Bangladesh.

(c) Course on “Basics of MS Office” organized by Graduate Training Institute(GTI), Bangladesh Agricultural University, Mymensingh.

(d) Six months professional internship program in different Govt. and non Govt. organization in Bangladesh.

Publications(Please provide a complete list of publications in refereed International, regional as well as national journals):

1. Md. Kamrul Hassan1*, Mahfuzul Islam1, Jayedul Hassan2, J. K. Ghose3, Md. Alimul Islam2 and Md. Shahidur Rahman Khan. 2014. Isolation and Identification of bacteria associated with skin lesions of cattle rared in sadar upazilla of mymensingh. International journal of Bio research: volume 1, issue 4, April, 2014:01-06/Hassan et al.2. Islam M., Rahman M.M., Khan M.S.R, Hossain M.M. and Hassan M.K. 2011. Culture sensitivity patterns of bacteria associated with respiratory illness in human. J. Sher-e-Bangla Agric. Univ., 5(2): 30-35, Bangladesh3. JK Ghose1,

2 MM Rahman2, MSR Khan3 and MK Hassan3.2011. Life Style of Secondary School Students in two Selected Schools of Mymensingh Sadar Upazila of Mymensingh District. Bangladesh J. Environ. Sci., 21: 79-82.

B. Associate Investigator:

Name:

Dr. Md. Abdul Masum

Date of Birth:

01 January, 1985

Sex:

M

Position held (since):

Assistant Professor Department of Anatomy, Histology & Physiology, Sher-e-Bangla Agricultural University, Dhaka-1207 (April 2014)

Present Address

Department of Anatomy, Histology & Physiology, Faculty of Animal Science & Veterinary Medicine Sher-e-Bangla Agricultural University Dhaka-1207BangladeshE-mail Address: [email protected]

Permanent Address

Vill: Natun huzrapurP.O: Chapai NawabganjP.S: Chapai NawabganjDistrict: Chapai Nawabganj

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Academic degrees:

Subject Name of Degree University Country Class Year of

Graduation

Veterinary Medicine

MS in Anatomy

Bangladesh Agricultural University

Bangladesh

First Class Letter

Grade; A CGPA: 4 (out of 4)

2011

Veterinary Science DVM

Rajshahi University Bangladesh

First Class Letter

Grade; A CGPA: 3.883

(out of 4)

2009

Science HSC Rajshahi Board Bangladesh

First DivisionObtained 70.9%

marks2002

Science SSC Rajshahi Board Bangladesh

First DivisionObtained 76.7%

marks2000

Previous Scientific experience:

1. Assistant Professor; From April 10, 2014 to till to date. Department of Anatomy, Histology & PhysiologyFaculty of Animal Science & Veterinary Medicine Sher-e-Bangla Agricultural University Dhaka-1207

2. Lecturer ; From 10 April,2012 to 09 April ,2014Department of Anatomy, Histology & PhysiologyFaculty of Animal Science & Veterinary Medicine Sher-e-Bangla Agricultural University Dhaka-1207

Publications:(Please provide a complete list of publications in refereed International, regional as well as national journals):

Articles in journals: Total: 10 (National 07 and International 03).

1. M. A. Masum, M. Z. I. Khan, N. H. Siddiqi and M. Nasrin. Frequency of immunoglobulin containing plasma cells in ileum represents gut-associated-lymphatic tissues in the BCRDV-vaccinated broiler. Bangl. J. Vet. Med. (2012). 10 (1&2): 15-20. Bangladesh.

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2. M. A. Masum, M. Z. I. Khan, M. U. Ahmed, M. N. H. Siddiqi and M. Nasrin. Histological study of the central or primary lymphoid tissues of broiler chickens. J. Expt. Biosci. 4(1); January 2013. Bangladesh.

3. M. A. Masum, M. Z. I. Khan and M. U. Ahmed. Histological study of the secondary lymphoid tissues of broiler chickens. Int. J. Sustain. Agril. Tech. 8(11): 01-04, November 2012. Bangladesh.

4. M. Nasrin1, M. Z. I. Khan1, 2, M. N. H. Siddiqi1, M. A. Masum1. Mobilization of immunoglobulin (Ig)- containing plasma cells in Harderian gland, cecal tonsil and trachea of broilers vaccinated with Newcastle Disease Vaccine. Tissue and Cell 45 (2013) 191– 197. International.

5. Mohd Zahirul Islam Khan, Md. Abdul Masum, Md. Zubayer Ibna Khan, Abdur Rahman Bin Aziz, Morsheda Nasrin, Mohd Nazmul Hasan Siddique & Mohd Mokhtar Bin Arshad. Histomorphology of the Lymphoid Tissues of Broiler Chickens in the Kelantan State of Malaysia. Sains Malaysiana. Accepted. International.

6. N. Sultana, M. Z. I. Khan, M.A. Wares and M. A. Masum. Histomorphological study of the major lymphoid tissues in indigenous ducklings of Bangladesh. Bangl. J. Vet. Med. (2011). 9(1): 53 – 58. Bangladesh.

7. M. Nasrin, M. N. H. Siddiqi, M. A. Masum and M. A. Wares. Gross and Histological study of digestive tract during postnatal growth and development of broiler. Journal of Bangladesh Agricultural University. Volume 1, Number 10. Bangladesh.

8. M. Nasrin, M. N. H. Siddiqi, M. Z. I. Khan, M. A. Masum and N. Sultana. Gross and Histological Studies of Harderian Gland and MALT (mucosa associated lymphoid tissue) of Broilers Vaccinated with BCRDV. International Journal of Bio Research, Volume 1, Issue 2, February 2011: 1-4. Bangladesh.

9. Nasrin Sultana, Md. Rafiqul Islam and Md. Abdul Masum. Study on the innervaton of brachial plexus of Black Bengal goat of Bangladesh. International Journal of Bio Research, Volume 1, Issue 1, March 2011: 1-3. Bangladesh.

10. Md. Abdul Masum, Mohammed Zahirul Islam Khan, Morsheda Nasrin, M. Nazmul Hasan Siddiqi, Mohammed Zubayer Ibna Khan and Md. Nabiul Islam. Detection of immunoglobulins containing plasma cells in the thymus, bursa of Fabricius and spleen of vaccinated broiler chickens with Newcastle disease virus vaccine. International Journal of Veterinary Science and Medicine 2014. Accepted 2 June 2014.

C. Other Staff [which includes Ph. D/M.Phil/ MS or equivalent degrees (Research) student in the relevant field] (Applicable for Universities/Institutes which award-academic degrees):

Not Applicable

PART – III: TECHNICAL INFORMATION

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1. SCIENTIFIC BACKGROUND OF THE PROJECT:

A. Significance of the proposed research:

Escherichia coli were first described in 1885 by Theodor Escherich (Escherich, 1988). Most

Escherichia coli are harmless commensals which are part of the natural gastrointestinal flora

in the lower intestine of warm-blooded animals. There are over 225 serotypes (unique strains)

of E. coli, the majority of which are not dangerous to human health (CIDRAP, 2006).

However, Verotoxin producing Escherichia coli serotype O157:H7 (VTEC) can cause severe

disease on humans than others. On the contrary, animal particularly cattle are their reservoirs

causing no disease there because it does not bind to the walls of their GI (gastrointestinal)

track. In people, E. coli O157:H7 binds to the cells that line our GI tract, Therefore,

Escherichia coli O157:H7 (EC O157) is an important cause of human diarrheal disease.

Severe manifestations include hemorrhagic colitis (i.e. bloody diarrhea) and hemolytic

uremic syndrome (HUS) (Griffin, 1998). Humans can get infected by this organism through

environmental pollution with cattle feces. As it is one of the most important zoonotic

pathogens, during the past 20 years, E.coli O157: H7 has become a world - wide public

health concern. As most cattle in Bangladesh are reared on smallholdings and farmers have

little knowledge about zoonotic disease transmission and how to handle cattle feces for

proper safety. This organism can easily enter into food chain of human to cause serious

disease problem, especially in children.

EC O157 was identified in different areas of the world. The occurrence of E. coli O157:H7

varies in different places of the world. Previous study showed that the prevalence rates

ranged from 0.3 to 19.7% in feedlot cattle and from 0.7 to 27.3% in grazing cattle in The

USA (Hancock et al., 1994), 0.4% of beef carcasses at slaughter and 0.83% of faeces from

live cattle in England and Wales and 15.4% faeces specimens from cattle for slaughter of one

British abattoir are positive to E. coli O157: H7 (Amstrong, 1996). Prevalence rates of E. coli

O157:H7 ranged from 0.3 to 19.7% in feedlots and from 0.7 to 27.3% on pasture have been

reported (Hussein, 2006).

The prevalence of E. coli O157 was also measured in different climatic conditions. In

Scottish beef its prevalence was found to be greater (P < 0.05) during the cooler months (i.e.,

11.2%) than during the warmer months (i.e., 7.5%) (Ogden et al., 2004).

Hussein and Bollinger (2005) reviewed published reports in the past 3 decades and

summarized EC O157 prevalence in beef cattle feces. In general, prevalence rates of E. coli

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O157 ranged from 0.3 to 19.7% in feedlot cattle, from 0.7 to 27.3% in cattle on irrigated

pasture, and from 0.9 to 6.9% in cattle grazing on rangeland forages.

The prevalence of VTEC in cattle in Bangladesh has not been demonstrated yet. Because

most cattle in Bangladesh raised in smallholdings and because smallholders have little idea

on its consequences to public health information on VTEC. Prevalence in cattle on

smallholding is to know to mass scale preventive measures to protect human health from the

organism. Another problem in Bangladesh is that use antibiotics to treat animal diseases with

suboptimal doses. This practice might have impact in emerging antibiotic resistance. VTEC

strains to zoonotic consequences farther. This study is aimed to examine the prevalence of

VTEC in cattle on smallholdings in Bangladesh and their antimicrobial susceptibility pattern

to some commonly used antibiotic in both medical and veterinary fields in Bangladesh

B. Related work already performed or in progress at the contracting institution / organization

Still there is no work in contracting institute.

C. Related work already performed or in progress at other Institutes in the country (If known)

Very few information are available only for Prevalence and antibiotic sensitivity of

enterohaemorrhagic Escherichia coli O157: H7 in cattle on smallholdings in Dept. of

Microbiology, Chittagong Veterinary and Animal Sciences University, Chittagong.

D. References to important related literature relevant to the project (including own

publication):

1. Amstrong L.G., Hollingsworth J., Glenn Morris J. 1996. Emerging Foodborne Pathogens: Escherichia coli O157: H7 as a model of entry of a new pathogen into the food supply of the developed world- Epidemiol. Rev., vol. 18 (1), Pp. 29.

2. Escherich, T. 1988. The intestinal bacteria of the neonate and breast-fed infant. 1884. Rev Infect Dis 10(6), Pp. 1220-5.

3. Griffin, P. M. 1998. Epidemiology of shiga toxinproducing E. coli infections in humans in the United States. In Escherichia coli O157:H7 and other Shiga toxin-producing E. coli strains, A. D. O’Brien and J. B. Kaper (Eds.). American Society for Microbiology, Washington, D.C., Pp. 15–21.

4. Hancock, D. D., Besser, T. E., Kinsel, M. L. P. I. Tarr, D. H. Rice, and M. G. Paros. 1994. The prevalence of Escherichia coli O157:H7 in dairy and beef cattle in Washington State. Epidemiol. Infect. Vol. 113. Pp. 199-207.

5. Hussein, H. S., and L. M. Bollinger. 2005a. Prevalence of Shiga toxin-producing Escherichia coli in beef cattle. J. Food Prot. Vol. 68. Pp. 2224-2241.

6. Hussein, H. S. 2006. Prevalence and pathogenicity of Shiga toxin-producing Escherichia coli in beef cattle and their products. J Anim Sci. Published online Oct 23.

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7. Ogden, I. D., M. MacRae, and N. J. C. Strachan. 2004. Is prevalence and shedding of E. coli O157 in beef cattle in Scotland seasonal? FEMS Microbiol. Lett. Vol. 233. Pp. 297-300.

8. Pablo, N., Stuart, W., Naylor, John, F., et al. 2008. Responses of Cattle to Gastrointestinal Colonization by Escherichia coli O157:H7. Infection and Immunity, Vol. 76. Pp. 5366–5372.

2. SCIENTIFIC SCOPE OF THE PROJECT

A. Research Objectives:

a) Broad objective: To detect the obscure Pathogenic E. coli O157: H7 from Small

holdings in Bangladesh.

b) Specific objective:

1. To screen the cattle on smallholding for the prevalence of E. coli O157: H7 in

Bangladesh.

2. To examine the sensitivity of E. coli O157:H7 against some commonly used

antibiotic in both medical and veterinary fields in Bangladesh.

3. Geographical distribution and Risk factor will be identified.

B. Relationship of these objectives to the present state of knowledge in the field

C. Research plan including proposed methods or techniques is going to be used

1) Plan of action:

The following plan will be executed to complete the project within one year Preparation of questionnaire Preparation of Sampling Sample Collection sheet Body score: Sample code: Experiment trial on the basis of research objectives Collection of fecal sample Purchase of research equipments Isolation and Identification E. coli O157:H7 Statistical analysis Record keeping Analysis of experiments

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Analysis of data Report writing

2) Methodology: Sampling: About 100 cattle smallholding will be randomly selected in an around the Dhaka

district. ≥ 1 cattle heads will be examined in each selected smallholdings. The GPS

coordinates of each selected smallholdings will be noted

Collection of fecal sample:

By swab rectal fecal sample will be collected from the animals on selected smallholding.

Microbiological on the collected samples will be conducted at the department of

Microbiology and Parasitology, Faculty of Animal Science and Veterinary Medicine, Sher-e-

Bangla Agricultural University, Dhaka-1207 (SAU).

Isolation and Identification E. coli O157:H7:

With swabs, fecal samples will be collected from the rectal mucosa of cattle (male and

female, various ages). Initial selection of E. coli isolates will be conducted by adding 1 g of

fresh feces from each animal to 9 ML of buffered peptone water (BPW) medium and

incubated at 37 C for 5 hours. Then the samples will be streaked on cefixime tellurite⁰

Sorbitol Mac Conkey (CT-SMAC) agar plates, aerobically incubated at 37 C for 18-24 h,⁰

and then evaluated for sorbitol fermentation. At the end of this incubation time, sorbitol-

fermenting (i.e., pink colonies) and non-sorbitol fermenting (i.e., white colonies) bacteria on

CT- SMAC plates will be transferred separately into 5-mL tubes containing 2 mL of tryptic

soy broth (TSB) with labeling and incubated at 37 C for 6 h with continuous shaking. At the⁰

end of the incubation time, the culture will be diluted with equal volume (i.e., 2 mL) of sterile

glycerol, mixed well, and stored at – 80 C. Positive isolates per rectal fecal sample will be⁰

tested for the presence of the rfbO157 gene by using polymerase chain reaction (PCR). All

the presumptive positive E. coli O157: H7 isolates will be tested for susceptibility to

antimicrobial agents.

Statistical analysis: All the collected data will be entered into a spreadsheet program

(Microsoft Excel 2007) and transferred to STATA 9.2 for Windows (Stata Corporation,

College Station, Texas, USA) to determine the prevalence and univariable analysis and

multivariable logistic regression to identify the risk factors associated with the presence of

the pathogen and its resistance to antimicrobials.

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Sample Collection sheet

Date: Sample code number:

GPS coordinates = X:

Y:

Name of the Place:

Name of the Thana:

Number of cattle in the smallholdings: calf: ………..; Adult: ………….

Clinical presence of any diseases on the day of sample collection:? Yes/No; if yes which disease suspected?

*Body score: +/++/+++/++++

Any antibiotic used in the past 15 days? Yes/no; if yes which antibiotic:

Duration of antibiotic used:

Is anybody in the family suffering from diarrhea? Yes/no, if yes, duration of illness.

Sample collected from diarrheic person? Yes/no

Sample code: Area/breed/age category/sex/sl.no.

Area = C or N or R; D= Dhaka Breed type = L or Cb; L= local, Cb= Crossbred Age category= C or A; C= Calf (≤ 1 year), A= Adult (≥ 1 year) Sex = M or F Sl. No. = 1, 2, 3, Example: C/L/A/F/15

*Body Score: (+) = Good body condition

(++) = Rough body condition and ribs are moderately visible

(+++) = cachectic, protruding rib with prominent pin bone, no diarrhea.

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(++++) = cachectic, protruding rib with prominent pin bone, presence of

diarrhea.

Result of the test: Positive / negative

3) Time schedule of activities with milestones

Sl. No.

Name of milestones Starting date

Completion date(Approx.)

1 Preparatory work and selection of study area.Literature review, acquiring equipments and reagents for methodology, preparing questioner and data record sheet, taking necessary hazard precaution and preparation for methodology.

November,2014

January,2015

2 Sample collection and laboratory investigation.Data and sample collection, Isolation and identification of organism and characterization of E. coli O157: H7 from positive isolates.

February,2015

June,2015

3 Cultural Sensitivity test and data accumulationCS test of positive sample, all the data will be accumulated for statistical analysis.

July,2015

August,2015

4 Draft report preparation Report writing

September,2015

October,

20155 Final Report Submission

Submission to the department and funding authority.

October,2015

November2015

Total duration 12 months

D. How is the project related to the stated objective of the Special Allocation for Science and Technology programme of the GOB?

It is well documented that cattle are the reservoir of E. coli O157: H7 which doesn’t harm in cattle but it is an important cause of human diarrheal disease. As it is one of the most important zoonotic pathogens, during the past 20 years, E.coli O157: H7 has become a world - wide public health concern. The prevalence of VTEC in cattle in Bangladesh has not been demonstrated yet. This study is very important to determine the prevalence in cattle on smallholding to know to mass scale preventive measures to protect human health from the organism and to conduct CS test to minimize the impact of emerging antibiotic resistance which is highly relevant in Bangladesh context. Moreover, the output of this research is directly related to the aim and objectives of DLS that is enhancing research and collaboration between private and public stakeholders. If a baseline data on prevalence of E. coli O157 in smallholdings are established, we can disseminate it to public health worker and farmers through DLS.

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E. How is the programme related to academic degree programme? (If applicable) (Only for universities/Institutes which awarded degree): Not applicable

F. What outputs from the project can be considered for the assessment of its success?

This research might reveal the existence of E. coli O157: H7 on cattle of small holdings in Bangladesh will be determined. These findings will be very important for further research regarding to disease control in Bangladesh as well as less biological hazard for the mankind.

G. How does the project contribute in the development of sustainable technology?

If we own the project it will enhance the lab basis education, specially it will be very helpful in molecular diagnosis, which are now the base of accurate disease diagnosis. Above all, both Ms and undergraduate student will avail the opportunity of using modern disease diagnostic equipments.

3. LIST OF FACILITIES AVAILABLE (Equipment and other facilities including laboratory space)

Laboratory with few equipments

PART – IV: BUDGET INFORMATION

1. BUDGET

A. Current year:

Item Cost (thousand Taka) Taka (Thousand)1. Minor equipment 4,00.002. Spare parts for major equipment 9,00.003. Consumables including chemicals, books, software etc. and sample

collection expenses3,00.00

4. Other essential expenses (maximum one fifth of the total budget) 652,00.00Total- Twenty Two lac Fifty Two Thousand Tk only

B. If the project is expected to last more than one year, please include budget estimates

for the total period: Not applicable

Item Cost (thousand Taka)1st year 2nd year 3rd year

1. Minor equipment

2. Spare for major equipment

3. Consumables

4. Other essential expenses

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Total

* Detailed list of items with approximate price must be included.

PART – IV: BUDGET INFORMATION

1. BUDGET

A. Current year:Item Cost (in taka)

1. Minor equipment 4,00,000.002. Spare parts for major equipment 9,00,000.00

3. Consumables including chemicals, books, software etc. and sample collection expenses

3,00,000.00

4. Other essential expenses 6,52,000.00

Total 22,52,000.00

Total- Twenty Two lac Fifty Two Thousand Tk only

B. If the project is expected to last more than one year, please include budget estimates for the total period: Not applicable

* Detailed list of items with approximate price included.

Detailed Proposed budget (in Taka)Sl Items Cost ('Tk)1    

Minor Equipment

Computer, printer, scanner, UPS, Volt Stabilizer, Computer table

Glass wares and equipmentsConical flaskMeasuring cylinderPostmortem boxSlide & slide boxInoculating loopMorter & pastleCollection vialPetridishesBunsen burnerSyringe & needleTest tube

 

200000.00

200000.00

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BeakerMortar and pastle

  Sub Total 4,00,000.002 Spare parts for major equipment    Parts for PCR machine and Gel documentation systems, Incubator,

Centrifuge Machine( High speed), Freeze, Dry air Oven, Hot water bath,

900000.00

  Sub Total 9,00,000.003 Consumables Expenses

Chemicals(10%formalin.,70% ethanol, absolute alcohol.5%propanol, nutrient agar media , selenite broth, EMB agar media, Blood agar media, Macconkey agar media)Reagents (crystalviolet stain, eosin stain, nigrosin stain, giemsa stain, malachite green stain, phosphate buffer solution), primer for PCR, antibiotic discs

300000.00 

Sub Total 3,00,000.00 4 Other Essential Expenses   

Traveling (Data collection and farm visit)Questionnaire preparation and printingPaper PurchasePhotocopySignboard, visualization and documentation BindingReport writingStationaryOther UtilitiesPublication and printingHonourium for PI (1; @ Taka 22000.00/ year) one month basicHonourium for Co PI (1; @ Taka 22000.00/ year) one month basicResearch Fellow (2; @ taka 10000/month)Guard (1; @ 5000.00 TK / month)Labors (1 person 180.00 TK/day, 300 days)

100000.0050000.00

50000.00

22000.00

22000.00240000.0060000.0054000.00

  Sub Total 6,52,000.00Grand Total 22,52,000.00

2. STATUS OF LEGAL PERSONALITY AND ACCOUNTING SYSTEM

As per GOB rule

PART – V: PREVIOUS FUNDING INFORMATION UNDER SPECIAL ALLOCATION FROM M/O SCIENCE AND TECHNOLOGY (MOST)

1. Did you get any funding under special allocation from MOST since 1997 – 98?

Yes No

(If your answer is no please escape the following sections)

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2. Funding year:

3. Amount of fund (in Taka):

4. What was the title of the project?

5. Project is completed or not?

6. If not what is the expected date of completion?

7. Already submitted working report or scientific report or not?

8. Expected date of submission of Scientific Report?

9. Any paper published in any International, regional / local journal from this research?

10. Quote the name of the journal, date of publication and title of the paper.

PART – VI: DECLARATION/CERTIFICATION

It is certified that –

(a) The same project has not been submitted to any other agency / agencies for financial support.

(b) The research work proposed in this project is not a duplicate work already done or being done in the field (i.e. area of research)

(c) We agree to accept the terms and conditions developed for the Special Allocation for Science and Technology as mentioned in the Guidelines.

(d) Associate Investigator assures the responsibility of the Project in case the Principal Investigator leaves the institution/Organization.

(e) Project will be provided with access to all available facilities in the Organization.

Signature and Name of thePrincipal Investigator

(With seal, Telephone number & Mobile number)

Signature and Name of the Head of the Organization / Institutes /University

(With seal, Telephone number & Mobile number)

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Signature and Name of the Associate Investigator:(With seal, Telephone number & Mobile number)

17