Agricultural University Research Progress Proceedings of the workshop of Bangladesh

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Volume 28

Proceedings of the workshop of Bangladesh Agricultural University Research Progress

15 & 18 March 2018

Bangladesh Agricultural University Research System (BAURES)

Bangladesh Agricultural University, Mymensingh, Bangladesh

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Published by:

Professor Dr. M.A.M. Yahia Khandoker Director

BAU Research System (BAURES) Bangladesh Agricultural University Mymensingh-2202, Bangladesh

Correct citation:

BAU Res. Prog. No. 28, 2017 Bangladesh Agricultural University Mymensingh-2202, Bangladesh

Address of correspondence:

Director

BAU Research System (BAURES) Bangladesh Agricultural University Mymensingh-2202, Bangladesh

Cable:

Agrivarsity Mymensingh Bangladesh

Phone : (091) 67417

(091) 67401-7 Ext. 2036, 2037 Fax : 88-091-67417

Email : [email protected]

Printed by:

Mohona Printing & Publication Chhoto Bazar

Mymensingh

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EDITORIAL BOARD

Professor Dr. M.A.M. Yahia Khandoker Chief Editor Director, BAURES

Professor Dr. Md. Alamgir Hossain Executive Editor Associate Director, BAURES

Dr. Paresh Kumar Sarma Managing Editor

Senior Scientific Officer (SG), BAURES

Professor Dr. S.M. Lutful Kabir Member

Department of Microbiology & Hygiene

Professor Dr. Mohammad Jiaul Hoque Member Department of Agricultural Extension Education

Professor Dr. Md. Azharul Hoque Member

Department of Animal Breeding & Genetics

Professor Md. Moniruzzaman Member

Department of Agribusiness & Marketing

Professor Dr. Chayan Kumar Saha Member

Department of Farm Power & Machinery

Professor Dr. Zakir Hossain Member

Department of Fisheries Biology & Genetics

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FOREWORD

Bangladesh Agricultural University Research System (BAURES) is coordinating, organizing and monitoring all non-degree oriented research programmes undertaken by the teachers of this university. At present 339 projects are being carried out by the teachers and researchers in different disciplines of agricultural sciences. The annual review workshop is a regular process to present, discuss and evaluate the progress of researches carried out in the preceding year. I am very happy to know that the publication of the 28 volume of the Bangladesh Agricultural University Research Progress in the form of proceedings of the workshop held on 15 & 18 March 2018.

This volume contains 166 abstracts of on-going/completed research projects conducted in the fiscal year 2016-2017. I am confident that the research findings documented in this report would be much helpful to the researchers working in the field of agricultural sciences.

I am taking the opportunity to congratulate the Principal Investigators/Project Directors and all other investigators/researchers of different research projects for presenting their research findings to make the workshop a success. My heartfelt thanks are due to the former Director of BAURES, Editor and the Editorial Board for their sincere and painstaking efforts in publication of this volume.

Special appreciation and thanks are expressed to the national and international donor agencies for funding and implementing the research projects under BAURES.

Finally, the encouragement and supports extended by the honorable Vice-Chancellor of BAU are gratefully acknowledged.

Professor Dr. A.M.A. Yahia Khandoker Chief Editor

&

Director

Bangladesh Agricultural University Research System (BAURES) Bangladesh Agricultural University, Mymensingh, Bangladesh

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EDITORIAL NOTE

Bangladesh Agricultural University Research System (BAURES) is responsible for the overall administration, supervision and execution of the project based researches conducted by the teachers and researchers of different faculties of Bangladesh Agricultural University (BAU), Mymensingh. BAU is providing fund for the research projects. In addition, various research projects are received from different national and international donor agencies. The findings of these researches are presented in the annual workshop held every year and the progresses of the projects are evaluated as part of the monitoring and evaluation of BAURES. The present volume of the proceeding includes abstracts of 325 research projects presented in the workshop held on 15 & 18 March 2018, where 172 projects are funded by BAURES and the rest are funded by the national and international donor agencies.

The aim of publishing this proceeding is to provide information generated through these projects based researches by the teachers, researchers and extension workers engaged in agricultural development of Bangladesh. This volume of the proceeding is edited by an editorial board consisted of intellectual expert members from six faculties of BAU. We have tried our best for quality publication of the proceeding. However, suggestions are always highly appreciated regarding improvement and standard of the publication and our best effort will be continued for improving the standard of the proceedings.

I would like to extend my sincere thanks and appreciation to the contributors and to the learned members of the editorial board for their sincere cooperation and help in preparing, improving and proof reading of the manuscripts. I would also like to acknowledge and appreciate the help and efforts of Mr. Md. Abul Hashem, Assistant Director, Dr. Paresh Kumar Sarma, Senior Scientific Officer and Mr. Md. Nazrul Islam, Computer Operator, BAURES for their sincere cooperation in publishing this proceeding.

I wish you all the best.

Professor Dr. Md. Alamgir Hossain Executive Editor

&

Associate Director

Bangladesh Agricultural University Research System (BAURES) Bangladesh Agricultural University, Mymensingh, Bangladesh

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CHAPTER I

Faculty of Veterinary Science

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Investigation on Disease Resistance Capability of Newly Developed Chicken Strain ‘Shuvra’ in Climatic Change Prone Delta Region of Bangladesh

Sukumar Saha* and Md. Tanvir Rahman

Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh, *E-mail: [email protected]

Abstract

Bangladesh Livestock Research Institute (BLRI) claims that the newly developed chicken strain, “Shuvra” is capable to resist diseases and sustain at local environment. Considering this fact the project was designed to determine the disease resistance capability of newly developed chicken strain “Shuvra” against Newcastle disease (ND), Fowl cholera (FC), Colibacillosis, Salmonellosis and Infectious bursal disease with indigenous chickens in climate change prone area in Bangladesh. In addition it was also aim to determine the effect of climate change on health and production of newly developed chicken strain “Shuvra” in the climatic change and non non-climatic change prone area. Initially the study was conducted to investigate the disease resistance capability of Shuvra compared with Indigenous chickens.

For this day-old chicks of Shuvra and Indigenous chickens were collected from BLRI and local sources. Initially the chicks were divided into 4 groups. In the first group 10 Shuvra and 10 Indigenous chickens were vaccinated by BCRDV at day 7 (one week) through intra ocular route (i/o) and with RDV at day 35 (5 weeks) of age of birds through intramuscular (i/m) route.

In the second group 10 Shuvra and 10 Indigenous chickens were vaccinated twice intramuscularly with BAU Fowl cholera vaccine at day 56 (8 weeks) and 84 (12 weeks) following manufacturer instructions. In the third group 10 Shuvra and 10 Indigenous chickens were vaccinated with BAU Salmonella at day 56 (8 weeks) and 84 (12 weeks) following manufacturer instructions. 10 Shuvra and 10 Indigenous chickens were kept as unvaccinated control to each group. Sera were obtained from all vaccinated and control chickens before and after 28 days (4 weeks) of each vaccination. The immune responses of the vaccinated and control chicks were determined by HI antibody titers against NDV vaccines and by passive haemagglutination antibody titer (PHA) against Fowl cholera and Salmonella vaccines. The protective potential of the vaccinated and control chickens of different groups Shuvra and Indigenous chickens were done by challenge infection using virulent field isolates (NDV, Fowl cholera, Salmonella pullorum). Very comparable level of antibody titer and protection following challenge infection were observed both in Shuvra and Indigenous chickens against three common diseases of chickens. The research work to investigate the effect of climate change on health and production of newly developed chicken strain “Shuvra”

in the climatic change and non non-climatic change prone area is on progress.

Isolation, Identification and Characterization of Bacterial Pathogens in Bioslurry from Different Biogas Plants of Gazipur and Mymensingh Districts of Bangladesh

Sukumar Saha

Abstract

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farms and the bioslurry samples were collected from 16 different natural bioslurry pits and from experimental anaerobic digester after 60 days digestion at environmental temperature and 30 days digestion at different temperature (25⁰C, 27⁰C, 29⁰C and 45⁰C). The total viable count (TVC), E. coli, Salmonella spp. and Staphylococcus spp. in the samples were determined by spread plate technique method according to ISO-6579 (International Standards Organization, 2002). For isolation and identification, samples were inoculated into Nutrient agar and other selective media such as Eosin Methylene Blue (EMB) agar, Salmonella Shigella (SS) agar and Mannitol Salt (MS) agar and incubated at 37⁰C for overnight and CFU was calculated in per gram volume of bioslurry. Identification of E. coli, Staphylococcus spp. and Salmonella spp. was confirmed by colony characteristics, staining, biochemical tests and finally using PCR. The average TVC, E. coli, Salmonella spp. and Staphylococcus spp. were 9.77, 6.11, 6.23 and 7.68 log CFU/gm respectively in manure sample. In natural bio slurry pits total viable count of bacteria varied from 8.65 to 7.26 log, E coli, Salmonella spp. and Staphylococcus spp. were 5.203 to 3.82, 7.18 to 4.00 and 7.9 to 3.14 log CFU/gm respectively. In anaerobic digester after 30 days digestion, the presence of E. coli varied from 0 to 5.11 log CFU/gm, Salmonella spp. 0 to 4.84 log CFU/gm and Staphylococcus spp. 0 to 6.59 log CFU/gm at different temperature of digestion (25⁰C, 27⁰C, 29⁰C and 45⁰C). In bio slurry from anaerobic digester after 60 days digestion showed absence of common indicator bacteria. In both natural slurry pits and in experimental anaerobic digester ( 30 and 60 days digestion) the total number of viable bacteria, E. coli, Salmonella spp. and Staphylococcus spp. were significantly reduced (p<.05). Antibiogram profile of isolated bacteria from bio slurry samples were also tested with commonly used antibiotics in veterinary practice and our results revealed that multidrug resistant indicator bacteria were present in the bio slurry samples. Our findings conclude that the microbial load after treatment of animal manure via anaerobic digestion (Bio gas plant) was grossly reduced and the reduction of bacterial pathogen depends on the duration and temperature of digestion.

Prevalence and Molecular Detection of Quinolone Resistant E.

coli in Rectal Swab of Apparently Healthy Cattle in Bangladesh

Md. Tanvir Rahman

Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh, E.mail: [email protected]

Abstract

Emergence of antibiotic resistance is a serious health problem both in human and animal all over the world. In this study, we investigated the prevalence of quinolone-resistant E.

coli isolated from apparently healthy cattle in Mymensingh district, Bangladesh. A total of 137 rectal swabs was screened among which 95 was found positive for E. coli. Confirmation of isolation of E. coli was done by PCR targeting 16S rRNA gene of E. coli (prevalence 69.3%).

Resistance against quinolone is primarily due to activities of qnrS and qnrA gene products.

Among these E. coli quinolone-resistant gene qnrS was detected in 11 isolates. None of the isolates were found positive for qnr Agene. The overall prevalence of qnrS positive E.

coli was 8.0%. Many of these quinolone-resistant E. coli was multidrug-resistant. Nucleotide sequence analysis of qnrS gene showed homology with the qnrS gene detected in China, Nigeria, Taiwan, Russia, Turkey and USA. All isolates that were resistant to multiple quinolones were found highly sensitive to imipenem, ertapenem and meropenem. The results of this study indicated that apparently healthy cattle harbor quinolone-resistant E. coli which

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have both clinical and public health significance. If strict regulation on the use of quinolones in food animals is not maintained, these quinolone-resistant E. coli may be transmitted to humans and other animals and may cause serious health problems in future. To the best of our knowledge, this is the first report on the molecular detection of quinolone-resistant E.

coli in cattle in Bangladesh.

An Epidemiological Investigation of Campylobacter in Bangladesh Agricultural University Dairy Farm

S. M. Lutful Kabir

Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh, ^*E-mail: [email protected]

Abstract

This study was aimed for isolation, identification and characterization of Campylobacter species from Bangladesh Agricultural University (BAU) dairy farm during the period of January to May, 2016. A total of 80 samples (faecal samples of calves, heifers and cows;

milk samples of cows) were collected from Bangladesh Agricultural University dairy farm for isolation and identification of Campylobacter species by using cultural, biochemical and molecular methods. Moreover, the isolated Campylobacter species were subjected for antimicrobial susceptibility test. Campylobacter like organisms were presumptively identified in 20 samples. Isolates were biochemically positive to catalase and oxidase tests and in hippurate hydrolysis test some of the isolates (n=6) shown negative that indicated the isolates were C. coli and some of the test isolates (n=14) shown positive that indicated the isolates were C. jejuni. Campylobacter specific 16S rRNA genes were amplified from the isolates. C.

jejuni were resistant to amoxicillin, erythromycin, azithromycin and susceptible to gentamicin, ciprofloxacin, norfloxacin and streptomycin. Furthermore, C. coli were resistant to amoxicillin and erythromycin and susceptible to gentamycin, ciproflaxacin. Out of 20 Campylobacter isolates, 57.14% C. jejuni and 33.33% C. coli were identified as multidrug resistant. To the best of our knowledge, this study has brought the first report on the occurrence of Campylobacter species with their antibiogram profiles in any dairy farm of Bangladesh.

Keywords: Dairy farm, C. jejuni, C. coli, molecular identification, antibiogram profiles

Development of a Low Cost Probiotic Formulation for the Production of Organic Poultry in Bangladesh

S. M. Lutful Kabir

Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh, ^*E-mail: [email protected]

Abstract

The second year of this project was designed with a view to sample collection from different poultry farms of Bangladesh and from different environmental sources, isolation and identification of micoorganisms by conventional and molecular methods, molecular characterization of the isolated bacterial strains having probiotic potentiality. The above mentioned activities were performed successfully. In addition, a total of 750 samples were

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contents of chicken after culturing on MRS broth and agar. In caecal samples, Lactobacillus spp. was detected in 88 (29.33 %) out of 300 samples, Enterococcus spp. was 102 (34%) out of 300 samples, Streptococcus spp. was 49 (16.33 %) out of 300 samples, Bacillus spp. was 42 (14 %) out of 300 samples, Bifidobacterium spp. was 15 (5%) out of 300 samples. In cloacal swab samples, Lactobacillus spp. was detected in 82 (27.33%) out of 300 samples, Enterococcus spp. was 99 (33%) out of 300 samples, Streptococcus spp. was 48 (16%) out of 300 samples, Bacillus spp. was 46 (15.33%) out of 300 samples, Bifidobacterium spp. was 13 (4.33%) out of 300 samples. In litter sample (n=50), Lactobacillus spp. was detected in 09 (18%), Enterococcus spp. was 14 (28 %), Streptococcus spp. was 12 (24%), Bacillus spp.

was 10 (20%), Bifidobacterium spp. was 04(8%). In water sample (n=50), Lactobacillus spp.

was detected in 05 (10%), Enterococcus spp. was 05 (10 %), Streptococcus spp. was 02 (4%), Bacillus spp. was 02 (4%), Bifidobacterium spp. was 01(2%). In feed sample (n=50), Lactobacillus spp. was detected in 02 (4%), Enterococcus spp. was 00(00 %), Streptococcus spp. was 01 (2%), Bacillus spp. was 02 (4%), Bifidobacterium spp. was 00(00 %).

Furthermore, molecular characterization of the isolated bacteria is going on.

Prevalence, Isolation and Detection of Virulent Gene in Escherichia coli from Duck

Marzia Rahman

Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh, E-mail: [email protected]

Abstract

This study was conducted to determine virulent genes in Escherichia coli prevalent in duck population by multiplex Polymerase Chain Reaction. A total of 60 cloacal swab samples were collected from two duck farms of Bangladesh Agricultural University and Shamvuganj. Initially the samples were screened for the detection of E. coli on the basis of cultural, staining and biochemical properties, followed by molecular detection of E. coli using genus specific primers to amplify 16s RNA. According to the results, out of 60 samples, 26 (43.33%) were confirmed to be E. coli positive. Among the E. coli positive samples, 12 (46.15%) samples were found positive for Stx-1 and 11 for Stx-2. Among 26, 11 (42.31%) samples possess both Stx-1 and Stx-2 genes, whereas only one isolate had Stx-1 gene. The prevalence of both Stx- 1 and Stx-2 in Bangladesh Agricultural University Poultry Farm was 41.66%, and the prevalence of Stx-1 and Stx-2 in Shamvuganj was 50% and 42.86%, respectively. This is the first report on the detection of virulence genes in E. coli of duck origin in the context of Bangladesh. This study indicates that duck may play role for the transmission of Shiga-like toxin producing E. coli (STEC) to human or its environment through fecal contamination or eggs or meat.

Molecular Detection of STEC and Antibiotic Resistant Genes in E coli from Layer Birds

Marzia Rahman

Abstract

The present study was carried out to detect the shiga toxin producing and antibiotic resistant genes in Escherichia coli isolated from layer birds of three different farms located at Katiadi,

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Kishorganj district. The investigated genes includes 16SrRNA, Stx-1, Stx-2, tet(A)and bla_CMY.

According to cultural and biochemical and PCR results, 50% (25/5) isolates from layer birds were recognized as E. coli. The distribution of virulent gene Stx-1, 8.0% among positive isolates and all isolates were found negative for Stx-2 by PCR. The distribution of antibiotic resistant genes tet(A) and bla_CMY.were44% and 36% respectively. All isolates were found resistant to a t least two microbial agents. About 80% isolates were found resistant to three or more agents termed as multidrug resistant. The antibiotic sensitivity test revealed that 96%

isolates showed resistance to amoxicillin followed by ampicillin (92%), tetracycline (76%), erythromycin (56%) and gentamycin (36%). The highest sensitivity was found to streptomycin (98%). The results indicate that the presence of multidrug resistant E. coli in layer birds are alarming for human being and indicate increased risk of infection via meat and egg.

Epidemic Survey on Respiratory Bacterial Infection of Buffalo Reared in Some Selected Areas if Bangladesh for Their Effective Control Measures

Md. Shahidur Rahman Khan* and Md. Shafiqul Islam

Abstract

The study was performed with a view to proper control of respiratory bacterial infection in Buffalo. For the epidemiological study at first data were collected through a structured questionnaire at field level. On the basis of history and clinical signs and as per the information provided by the farmers the prevalence rate was recorded as 38.78% and the P value was calculated as 0.004 (p<0.01) which was noted as highly significant. The prevalence rate was 13.33% Pasteurella multocida, 8% was E. coli and 8.22% was Staphylococcus aureus on the basis of location. The organisms were characterized using cultural, biochemical and molecular techniques. The isolated Pasteurella multocida produced small, round, opaque colonies on blood agar. Staphylococcus aureus produced golden yellow colony in mannitol salt agar. E. coli produced black color colonies with metallic sheen on EMB agar. Pasteurella multocida showed Gram negative, bipolar rods. Staphylococcus aureus showed Gram positive, cocci shaped and E. coli showed Gram negative, small rod shaped. On the basis of their cultural and biochemical characteristics it was found that among 40 nasal samples 5 was found positive for Pasteurella multocida, 3 was E. coli & 4 was Staphylococcus aureus. The antibiogram study concluded that amoxicillin, gentamycin, &

ciprofloxacin should be first choice of treatment of respiratory bacterial infection caused by isolated 3 bacteria. Pasteurella multocida was further characterized by PCR where 5 isolates showed positive band at 620 bp. Based on the present findings it may be concluded that epidemiological parameters such as age, sex, season and location have great impact on respiratory bacterial infection of buffalo in some selected areas of Bangladesh. The present research work covering epidemiological study is a first and preliminary report in the context of Bangladesh.

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Isolation and Molecular Detection of Fish Viruses Responsible for Cat Fish (Heteropneustes fossilis) Mortality in Cultured Fish Farm of Trishal and Bhaluka of Mymensingh Md. Alimuyl Islam

Abstract

Stinging catfish, Shing (Heteropneustes fossilis) is been cultured in Bangladesh which is one of the most popular and highly valuable fish species. A total of 150 diseased H. fossilis were collected from twenty fish farms of Trishal and Bhaluka Upazila of Mymensingh district, Bangladesh between June 2016 to February 2017. Affected fishes revealed variable clinical symptoms like loss of equilibrium and, erosions at the bases of fins and tail, hemorrhages and skin lesions on body surface, profuse mucous secretion and congestion along with enlargement of most of the internal organs. Some of them showed only white sported lesion on the body surface and enlargement of liver and spleen only. Three different kinds of selective media were used for the isolation of total population of bacteria from the affected fishes. A number of biochemical tests were performed for the identification of the isolated bacteria. Commercially available panel of antibiotics were used for the determination of sensitivity of the isolated bacteria. A total of 85 bacterial strains were isolated and the isolated were Aeromonas hydrophila, A. salmonicida, A. sobria, Pseudomonas angulliseptica, P.

fluorescens and Vibrio anguillarum. Aeromonas spp. appeared to be the main pathogen in the diseased fishes. Among the isolated spp of bacteria distribution of Aeromonas species was as follows: A. hydrophila 34 (40.00%), A. salmonicida 14 (16.47%), A. sobria 8 (9.41%) as well as 5 (5.88%) unidentified Aeromons strains. The other isolated of Pseudomonas anguliseptica, P. fluorescens, Vibrio anguillarum and Vibrio spp were distributed as 12 (14.11%), 4 (4.70%), 3 (3.52%) and 5 (5.88%), respectively in infected H. fossilis. However, results of antibiotic sensitivity test revealed that all the bacterial isolates of H. fossilis were found more or less resistant to all most all the antibiotics with the exception of ciprofloxacin and levofloxacinn which were found highly sensitivity against all the strains of Aeromonass pp. and other species of bacteria. Fish viruses of the family Nodaviridae, Herpesviridae and Orthomyxoviridae were failed to isolate from the fishes showed ulceration and erosions on their body surface. None of the above mentioned strains of bacteria were able to isolates from the fishes showed white sported symptoms. These fishes were preserved at -86⁰C for the detection and isolation of fish viruses.

Prevalence of Human Pathogenic Vibrio Species (Vibrio cholerae, V. parahaemolyticus and V. vulnificus) in the Coastal Regions of Bangladesh with Their Specific Detection by Multiplex PCR

Muhammad Tofazzal Hossain* and Md. Tanvir Rahman

Abstract

Among more than 70 identified Vibrio species, Vibrio cholerae, V. parahaemolyticus and V.

vulnificus are of major concerns as they are pathogenic to animals, including humans. All

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three species are commonly associated with seawater, sediment, shellfish, and the intestinal contents of fish. Multiplex polymerase chain reaction (PCR) has been proven to provide rapid and highly sensitive methods for the specific detection of microorganisms and can be easily performed in diagnostic laboratories. The objective of this study was to study the prevalence of three important Vibrio species, Vibrio cholerae, V. parahaemolyticus and V. vulnificus in the coastal regions of Bangladesh by multiplex PCR using the groEL gene, a potential phylogenetic marker. A total of 50 different samples (fish, shrimp, seawater, stool of diarrhoeic patient) were collected from the coastal regions, especially from Chittagong and Cox’s bazar. The samples were collected and transported maintaining cold temperature.

Each sample was processed and inoculated in TCBS agar media. Positive colonies were purified by subculture and chromosomal DNA from each isolate was extracted by simple boiling method. Multiplex PCR was performed using primer sets designed from GroEL gene for the specific detection of Vibrio cholerae, V. parahaemolyticus and V. vulnificus from the samples. All the collected samples were found negative for the target three species except four for Vibrio cholerae. Only 3 human samples and 1 seawater sample collected from beach were found positive for Vibrio cholerae. This indicates that beach might be a source of contamination.

Isolation and Identification of Pathogenic Bacteria and Virus Causing Duck Diseases from Recent Outbreak Areas of Mymensingh Division

Muhammad Tofazzal Hossain* and Md. Tanvir Rahman

Abstract

Duck farming is getting popular day by day among the small landless people of Bangladesh.

Ducks are considered as resistant birds compare to chicken in relation to disease because the morbidity and mortality rate in ducks due to disease is very limited. But sometimes, some bacterial and viral diseases may cause havoc to the duck population. Among the diseases, duck cholera, duck septicemia, colibacillosis, duck viral hepatitis and duck plague are mainly associated with high morbidity and mortality in ducks worldwide including Bangladesh. The present study was targeted to isolate and accurately identify different causal agents (bacteria and virus) of duck diseases prevailing in Mymensingh division. Samples were collected from 15 dead ducklings and 35 dead ducks of different regions of Mymensingh, Netrokona and Kishoregonj districts. Liver and intestine were aseptically collected for the isolation of causal agents. Samples were processed and inoculated in bacteriological media for bacteria isolation and duck embryo for virus isolation. Among the collected samples, 50 were found positive for E. coli, 2 for Pasteurella multocida, 4 for Staphylococcus spp., 4 for Salmonella spp. and 8 were found positive for duck plague virus. All the pure isolates were preserved;

chromosomal DNA was extracted and stored at -20°C for further study.

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Physio-Pathological Role of Curcumin on High Fat Induced Hyperlipidemia in Mice

Siddhartha Sarker, Md. Iqramul Haque and Mohammad Alam Miah*

Department of Physiology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

*Email: [email protected]

Abstract

Curcumin is an extract of turmeric plants. It has been shown to possess potent antioxidant and hypolipidemic properties. The experiment was designed to observe the effects of curcumin on butter fat induced hyperlipidemia in mice. A total of 40 Swiss Albino mice (Musmusculus), age of 28-30 days with an average body weight of 27g were randomly divided into 2 groups. Non-treated control group A(n=8)and butter group B (n=32), fed on 10% butter. After 5 weeks, butter fed mice were subdivided into 4 equal groups (n=8): Group B, C, D andE fed on varying concentration of curcumin: 1%, 2% and 3% extracts in drinking water respectively along with butter. The experiment was carried out for period 10 weeks.

Maximum weight gain was recorded in butter fed mice. Supplementation of curcumin to butter fed mice prevented additional weight gain. Butter fed mice had significantly (p<0.01) higher hemoglobin (Hb) concentration, total erythrocyte count (TEC) and packed cell volume (PCV).

Addition of different concentration curcumin on butter fed mice didn’t enhance the hematological values. In case of lipid profile, increased total cholesterol, triglycerides and LDL cholesterol values were observed in butter fed mice. However, addition of curcumin extracts dramatically reduced or prevented the lipid profile and more prominent result was found in 2% curcumin group. Histo-pathological studies revealed that butter altered the normal structures of liver and aorta. There were few fatty changes found along with pyknoticnuclei in the hepatocytes and thickening of the wall of aorta with disorganized lining cells of aorta. No detectable changes were found in the liver and aortic tissues of curcumin treated mice. It could be predicted that the butter fats have harmful effects on weight gain, on lipid profile and on organ structures without bad effect on hematology. These harmful effects could be prevented by addition of curcumin extracts.

Molecular Characterization of Bovine Rotavirus Causing Severe Diarrhea in Calves in Selective Areas of Bangladesh M.M. Alam

^1*

, N.U. Ahmad

¹

, and A. Khair

¹

1Department of Medicine, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh, *E-mail: [email protected]

Abstract

Rotavirus is distributed worldwide as an enteric pathogen in man, animal and poultry. Bovine rotavirus (BRV) has been detected in both dairy and beef cattle herds worldwide. A total of 200 faecal samples were collected from diarrheic calves of the dairy region of Barisal, Madaripur and Sirajgonj districts during the period from July 2015 to June 2017. These 200 faecal samples were consisted of 68 from Barisal, 92 from Madaripur and 40 from Sirajgonj.

The collected stool samples were screened using rapid test- strips BioK 152 to determine the presence of BRV. Group A BRV was detected in 22.50% of faecal samples (45/200). The highest rate of BRV infection was detected in the Barisal (26.47%) and Madaripur (26.08%)

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while the lowest detection rate was in Sirajgonj (7.5%). The detected positive rotavirus samples by rapid test- strips BioK 152 were also examined by Polyacrylamide Gel Electrophoresis and Silver Staining (PAGE-SS) technique for the detection of presence of rotavirus dsRNA. About 70% of positive rotavirus samples by rapid test- strips BioK 152 was found positive in PAGE. All the electropherotypes were long and single pattern. RT-PCR and sequencing was done for 30 positive BRV samples to determine the G and P serotypes of bovine rotaviruses are circulating in Bangladesh. Genotyping analysis of rotavirus positive samples indicated that G6P[11] was the most prevalent genotype (56.67%) followed by G10P[11] (3.33%). Seven samples (23.33%) could not be G and/or P typed. P[11] genotype was predominantly detected as 76.67%. This study demonstrates that BRV is a contributing pathogen to diarrhoeal disease in calves of different three district of Bangladesh. In the present study, it was not investigated that bacteria, parasites or any other pathogens responsible for causing diarrhea or not.

Keywords: Bovine Rotavirus, G and P typing, RT-PCR & Sequencing, Severe Diarrhea &

Calves

Indiscriminate Use of Antimicrobial Drugs in Poultry Farms and Analysis of Antibiotic Residues in Poultry Products of Bangladesh

M.M. Alam

¹

*, A.K.M. Anisur Rahman

¹

and A.K.M. Humayun Kabir

¹

1Department of Medicine, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh, *E-mail: [email protected]

Abstract

Indiscriminate use of antimicrobial drugs in poultry causes the drug residue in edible poultry tissues which have an adverse effect on public health as consumers. There is paucity of information about antimicrobial drugs usage by the meat producing (broiler, cock, shonali, grameen types) farmers and qualitative detection of antibiotic residue. Hence a cross sectional study was undertaken through interviewing the farm owners or caretakers and by physical investigating of the farms including other secondary documents to measure the frequency of the antimicrobial drugs using in poultry farms of different upazila in six districts of Bangladesh and qualitative detection of drug residue in broiler sample from December 2015 to December 2017. The current study on 416 investigated poultry farms revealed that majority farmers reared broiler (91.1%) and the half (50.0%) of them were more than 1000 to 2000 nos. of birds. A total of 28 different kinds of antimicrobials were identified those used by the farmers and of them at least 16 antimicrobials used to treat the human illness. The mostly used antimicrobials were Oxytetrtacycline (72.4%), Amoxicillin (72.4%), Ciprofloxacin (70.4%), Enrofloxacin (67.1%), Sulfadrugs (61.1%), Anticoccidial (61.1%), Doxycycline (58.4%), Trimethoprim (55.8%), Colistin Sulfate (56.0%), Metronidazole (55.5%), Tylosin Phosphate (37.7%), and Erythromycin (36.1%). It was also revealed that more than 61.0%

farmers were used antimicrobials at least 04 to 06 times in each batch. For treatment, around 95% farm owners depend on unskilled and unauthorized personnel like company personnel, dealer and quack. On laboratory investigation of 2000 sample (Breast meat, thigh meat, Liver, kidney) from 500 live meat producing birds through Thin Layer Chromatography (TLC),it was found that about 100.00% samples indicated the presence of antibiotics. Based

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Identification and Characterization of Potential Food-borne Pathogens from Milk, Meat and Eggs in Relation to Food Safety in Bangladesh

M.M. Alam

^1*,

A.K.M.A. Rahman

¹

and M.A. Rahman

¹

1Department of Medicine, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh, *E-mail: [email protected]

Abstract

Food (milk, meat and egg) is never being free of risk because even sterile food can be recontaminated. Some toxins released from bacterial pathogen survive cooking, and some individuals may be sensitive to certain foods (e.g., allergy). Food-borne diseases causing infections and intoxications can affect everyone. The study was conducted to investigate the presence and characterization of food borne pathogens in milk, meat and eggs sold to consumer at a retail market or farm level of Mymensingh district. A total of 169 samples including milk (n=108), chicken meat (n=51) and beef (n=10) were collected from BAU dairy farm and retail market of municipal area during two year period from July 2015 to June 2017.

In this study the food-borne pathogens were isolated and identified by colony characteristics on selective agar media, gram staining, catalase test, coagulase test and biochemical test and PCR finally. It was found that out of 108 milk samples a total of 25 (23.1%) samples were contaminated with Staphylococcus spp. including S. aureus & S. epidermidis, 32 samples (29.63%) were E. coli and 02 samples were Salmonella spp. Out of 51 chicken meat samples 39 samples (76.47%) were contaminated with Staphylococcus spp., 25 samples (49.02%) were E. coli and 29 samples (56.86%) were Salmonella spp. including S. pullorum. Out of 10 beef samples only 06 samples were contaminated with Salmonella spp., and 07samples were contaminated with E. coli and Staphylococcus sp.. Among 64 samples of E.coli identified by conventional method only 24 samples (37.5%) were confirmed and out of 71 Staphylococcus sp., positive sample through conventional method only 39 samples were confirmed by PCR. Multi-drug resistant (MDR) E. coli were detected by disc diffusion test using 10 commonly used antibiotics. Overall 50% of E. coli isolates of food were found MDR.

About 28.13%, 57.14% and 76% of the E. coli isolates originated from milk, beef and chicken meat respectively were MDR. Out of 37 about 31 (83.78%) samples of Salmonella sp., were MDR. Out of 71 samples of Staphylococcus sp. about 23 (32.39%) samples were multidrug resistant, no MDR were in beef sample. Antibacterial resistant gene mecA gene (Methicillin–

Resistant Staphylococcus aureus, MRSA) were detected in 10 (25.64%) Staphylococcus sp.

out of 39 positive samples, strA gene in 13 (54.17%) samples and tet A gene in 9 samples were detected in 24 (37.5%) E. coli positive sample. Antibiogram study showed that E. coli isolated from chicken meat were resistant to oxytetracycline (92%), sulphonamide- trimethoprim (84%), amoxycillin (76%) and erythromycin (60%). E. coli isolated from beef sample were resistant to erythromycin (85.71%) and oxytetracycline (71.43%) and sensitive to ciprofloxacin (100%), gentamicin (100%) and neomycin (100%). However, all isolates of E.

coli were found sensitive to amikacin (100%). E. coli isolated from milk sample were 100%

sensitive to gentamicin followed by neomycin, ciprofloxacin, azithromycin, oxytetracycline and erythromycin. In case of S. aureus originated from chicken meat were highly resistant to oxytetracycline (71.79%) and highly sensitive to neomycin, amikacin and colistin sulphate; in milk sample S. aureus highly resistant to colistin sulphate (92%), amikacin (56%), erythromycin (44%) and sensitive to enrofloxacin(100%), neomycin(100%), ciprofloxacin (92%), gentamicin (92%); in beef sample S. aureus were resistant to enrofloxacin and sensitive to ciprofloxacin, gentamicin, erythromycin, doxycycline, azithromycin. In case of Salmonella sp. originated from chicken meat highly resistant to erythromycin(100%), colistin

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sulphate(80%), doxycycline (79.31%) and sensitive to gentamicin (86.21%), ciprofloxacin (83.33%), neomycin (82.76%); in milk sample Salmonella sp. were equally resistant to erythromycin, amoxycilin, doxycycline, colistin sulphate (100%) and equally sensitive to ciprofloxacin, gentamicin, neomycin (100%); in beef sample Salmonella sp. were resistant to erythromycin (83%), azithromycin (83%) and sensitive to gentamicin (100%),, neomycin (100%), ciprofloxacin (83%) and amikacin (83%). Higher prevalence of E. coli, S. aureus and Salmonella spp. in chicken meat, beef and milk indicates unhygienic production and processing of these foods. Presence of multi-drug resistant E. coli, Salmonella spp., and Staphylococcus spp. in these foods might pose serious public health threats. The antibiogram profile of the isolates will help therapeutic decision making in the treatment of food borne diseases in cattle and poultry in Bangladesh. Further study is needed to identify more antibacterial resistant gene and their genomic diversification in these organisms. All samples were examined for detection of Campylobacter spp., but not a single of isolate was detected from those samples. The obtained result showed that the presence of Staphylococcus aureus, E. coli ,Salmonella spp. and in retail chicken meat, beef and milk may represent a threat to public health.

Real Time PCR for Identification of Brucella Species in Cattle and Buffaloes

M. S. Rahman*, M. A. S. Sarker, Sayra Tasnin Sharmy, F. Melzer

¹

and H. Neubauer

¹

Department of Medicine, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh 2202, BANGLADESH; ¹ OIE Reference Laboratory for Brucellosis, Institute of Bacterial Infections and Zoonoses, Federal Research Institute for Animal Health, Friedrich-Loeffler-Institut, Naumburger Str. 96a, 07743 Jena, Germany, *E-mail: [email protected]

Abstract

According to the Food and Agriculture Organization (FAO), the World Health Organization (WHO) and the World Organization of Animal Health (OIE), brucellosis is considered to be the most widespread zoonosis throughout the world. The objectives of the present study were to identify the common species of Brucella in cattle and buffaloes in Bangladesh and determination of the characteristics and diversity of Brucella genotypes circulating in Bangladesh and to develop recommendations for agricultural and public health polices to control and prevent brucellosis and to improve the livelihoods of farmers, abattoir workers and consumers in Banglade

Agricultural University Research Progress Proceedings of the workshop of Bangladesh

Volume 28

Correct citation:

EDITORIAL BOARD

FOREWORD

EDITORIAL NOTE

Contents

Chapter II : Faculty of Agriculture

Chapter V : Agricultural Engineering and Technology

Chapter VI : Fisheries

CHAPTER I

Abstract

Isolation, Identification and Characterization of Bacterial Pathogens in Bioslurry from Different Biogas Plants of Gazipur and Mymensingh Districts of Bangladesh

Prevalence and Molecular Detection of Quinolone Resistant E.

An Epidemiological Investigation of Campylobacter in Bangladesh Agricultural University Dairy Farm

Development of a Low Cost Probiotic Formulation for the Production of Organic Poultry in Bangladesh

Prevalence, Isolation and Detection of Virulent Gene in Escherichia coli from Duck

Molecular Detection of STEC and Antibiotic Resistant Genes in E coli from Layer Birds

Epidemic Survey on Respiratory Bacterial Infection of Buffalo Reared in Some Selected Areas if Bangladesh for Their Effective Control Measures

Isolation and Molecular Detection of Fish Viruses Responsible for Cat Fish (Heteropneustes fossilis) Mortality in Cultured Fish Farm of Trishal and Bhaluka of Mymensingh Md. Alimuyl Islam

Prevalence of Human Pathogenic Vibrio Species (Vibrio cholerae, V. parahaemolyticus and V. vulnificus) in the Coastal Regions of Bangladesh with Their Specific Detection by Multiplex PCR

Isolation and Identification of Pathogenic Bacteria and Virus Causing Duck Diseases from Recent Outbreak Areas of Mymensingh Division

Physio-Pathological Role of Curcumin on High Fat Induced Hyperlipidemia in Mice

Molecular Characterization of Bovine Rotavirus Causing Severe Diarrhea in Calves in Selective Areas of Bangladesh M.M. Alam

Indiscriminate Use of Antimicrobial Drugs in Poultry Farms and Analysis of Antibiotic Residues in Poultry Products of Bangladesh

Identification and Characterization of Potential Food-borne Pathogens from Milk, Meat and Eggs in Relation to Food Safety in Bangladesh

Real Time PCR for Identification of Brucella Species in Cattle and Buffaloes