Year 2021
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Item MOLECULAR AND HISTOPATHOLOGICAL STUDIES OF WHITE SPOT SYNDROME VIRUS INFECTION IN TIGER SHRIMP (Penaeus monodon)(DEPARTMENT OF BIOTECHNOLOGY, SHER-E-BANGLA AGRICULTURAL UNIVERSITY, DHAKA-1207, 2021-12) HASAN, MD. JUWELSince the 1980s, shrimp farming has been rapidly expanded and has become the third largest export earnings source, contributing about 3.57% to national GDP of Bangladesh. Now this sector facing a serious problem with a disease named White Spot Disease (WSD) caused by dreadful White Spot Syndrome Virus (WSSV), which leads to 100% mortality in the shrimp farm within 3-5 days. This study showed PCR detection and prevalence rate of WSSV infection in Penaeus monodon and further confirmed by histopathological study to observe the pathological changes in the tissue. The prevalence of this virus has been studied in six selected farms located in the Bagerhat and Satkhira districts of Bangladesh. Both one-step and two-step PCR were used to detect using two pairs of primers, namely, 146F1/146R1 and 146F2/146R2, amplifying the 1447 bp and 941 bp fragments, respectively. Out of 36 randomly selected shrimp samples, 25 samples were WSSV-positive other 11 samples were negative by one-Step PCR. The negative eleven samples were further subjected to two- step PCR and resulted only 3 samples being negative. The prevalence rates of WSSV infection were 83.33% and 55.56% in Satkhira and Bagerhat district, respectively, confirmed by one-step PCR and the prevalence rate of WSSV infection was 83.33% and 100% in Satkhira and Bagerhat district, respectively, confirmed by two-step PCR. However, the overall prevalence rate increased from one-step to two-step PCR from 69.44% to 91.67%. Between two methods two-step PCR was more efficient than one-step PCR. The shrimp specimens showing clinical signs of WSSV were positive by both one-step and twostep PCR. Some of the apparently healthy samples were also found to be positive for WSSV by two-step PCR. Histopathological studies of hepatopancreas of two-step PCR confirmed WSSV-positive samples resulted in degenerated tubule lumen, degenerated and sloughing hepatopancreas with necrosis and infected muscle tissue resulting in fragmentation and separation of muscle fibers. Furthermore, it is suggested that more molecular studies about WSSV and disease management is required to increase the shrimp production in Bangladesh.Item EFFECTS OF DIETARY SUPPLEMENTATION OF Sargassum vulgare ON GROWTH AND HEMATOLOGICAL PARAMETERS OF ORNAMENTAL FISH KOI CARP (Cyprinus rubrofuscus)(DEPARTMENT OF BIOTECHNOLOGY, SHER-E-BANGLA AGRICULTURAL UNIVERSITY, DHAKA-1207, 2021-12) HOSSAN, MOHAMMAD IQBALA research was conducted to assess different formulated diets supplemented with different doses of seaweed (Sargassum vulgare) on ornamental fish Koi carp (Cyprinus rubrofuscus) in the aquaculture laboratory of Department of Aquaculture, Sher-e-Bangla Agricultural University to evaluate the growth and hematological parameters. Total four treatments were applied including Treatment- 1 (Control, 0% seaweed), Treatment-2 (5% seaweed), Treatment-3 (10% seaweed) and Treatment-4 (15% seaweed) each with three replicates. Four iso-proteinase feed was formulated where average percentage of protein, lipid, fiber and carbohydrate in formulated diet were 27.59%, 6.1%, 7.59 and 48.96%. The initial mean weight of fishes was 14.4±2.12, 14.5±0.75, 13.1±2.4 and 12.8±1.43 g in T-1, T2, T-3 and T4 respectively. The experiment was concluded in 100L glass aquarium with mechanized water filter system for each for 8 weeks and weekly sampling were done. Feed was applied twice a day. Temperature, pH, TDS, NH 3, NO 2 - ii and NO 3 - data were collected daily during the experimental period and there was no significant difference in water parameters among treatments. Mean weight (g) of fish, length (cm) and width (cm) data were collected weekly basis. The mean weight gain was found 7.13±1.84, 8.06±1.58, 6.98±1.84 and 4.72±0.52 in T-1, T-2, T-3 and T-4 respectively. The highest weight gain was found in T-2 (5% seaweed) and the percentage of weight gain 55.86% found in the same treatment. There was no mortality found in T-2 and T-4. Survival rate were found 96.6% in Control. The lowest FCR 1.43±0.26 found in T-2 and SGR also found highest in the same treatment. Hematological parameter showed that blood hemoglobin and glucose level was higher in T-2 than others treatments which were 11.6±0.17gm/dl and 6.33±0.26 mmol/l. Serum protein was found 6gm/dl in T-2 and lowest value was found in control. Spleen Somatic Index (SSI) percentage of experimental fishes was also found higher in T-2 and lowest one was recorded in T-1. Refractive index (RI) of blood serum was found better in T-2 fishes compared to other treatments. It can be concluded that feed supplemented with 5% seaweed (T-2) diet showed better growth performance and hematological parameters compared to other treatments. Feed supplemented with brown seaweed Sargassum vulgare as a dietary ingredient can be used for better immunity and growth performance of ornamental fish.Item BIOINFORMATIC ANALYSIS OF MITOCHONDRIAL GENOME OF HUMPED FEATHERBACK CHITAL FISH, Chitala chitala (HAMILTON, 1822)(DEPARTMENT OF BIOTECHNOLOGY, SHER-E-BANGLA AGRICULTURAL UNIVERSITY, DHAKA-1207, 2021-12) IRTIZA, MD. HASINThe present study was about the bioinformatic analysis of mitochondrial genome of an endangered featherback fish, Chitala chitala. Although Chitala chitala plays significant role in inland fishery, but its’ evolution has received little attention. In the current study, the mitogenome of Chitala chitala was retrieved and investigated. Some of the bioinformatics tools were applied to measure important gene parameters including AT/GC-skewness, codon adaptation index (CAI), the effective number of codons (ENc) and GC percentages of each protein coding gene. The majority of AT-skew and GC-skew values of all protein coding genes of the species were negative, and the amplitude of the GC-skew was larger than the AT-skew. On the basis of comparative selective strength analysis with the PCGs of two related mitogenomes, I discovered that most of the PCGs retained the Ka/Ks ratios less than one implying that they evolved under strong negative or purifying selection. Moreover, I analyzed the codon frequency and relative synonymous codon usage (RSCU). Then, I identified a total of 22 RSCU values (>1) and revealed 6 codons as “over-presented” that implied for codon usage bias to engage in highly expressed genes for efficient protein synthesis via translational selection. The phylogenetic tree and time tree were constructed by Maximum Likelihood (ML) method, providing further supplement to the evolution of the fish. Regarding on the facts of the presence of codon usage biasness rolling in translational selection and the signs of purifying selection identified in PCGs indicate obvious conservation of this endangered fish species.Item ASSESSMENT OF FECUNDITY, HATCHING & SURVIVAL RATE OF NILE TILAPIA (Oreochromis niloticus) APPLYING FEED WITH GARLIC AND GINGER EXTRACT(DEPARTMENT OF BIOTECHNOLOGY, SHER-E-BANGLA AGRICULTURAL UNIVERSITY, DHAKA-1207, 2021-12) NAYEEM, DIDER HAIDERThe study was conducted at commercial hatchery at BRAC Tilapia Hatchery, Magura, Bangladesh and the analytical studies were conducted at Aquaculture Laboratory, Sher-e-Bangla Agricultural University to evaluate the effect of Garlic and Ginger extract in Nile tilapia diets on their reproductive performance (fecundity and hatching), survival rate on produced fry and hematological parameters of fry. A treatment was used as control without any extract. Nile tilapia (O. niloticus) with average weight of 267 g ± 3.16 were stocked for spawning into nine hapas at a density of 13 fish (10 females and 3 males) per 10 sq meter hapa, Three hapas with same diameters were used as replicates for each treatment. Hapas were fixed in an earthen pond (5 decimals). Feeding was done at a rate of 0.5% from their total biomass. Water quality parameters (temperature, pH, dissolved oxygen, ammonia) were measured weekly. Production of seed (eggs and fry) was compared with the tested diets. The highest mean of egg number (132.05 pcs) & mean spawn number (98.29 ± 1.11 pcs) per gram were obtained with the diets contained 0.02% Garlic extract (Treatment 2). Survival rate (84.24±0.05%) of fry was also found best with Treatment 2. Tilapia diets supplemented with Garlic extract had best responded on their reproductive performance (Hatching 80.63% ±1.05) and highest value FCR (1.43±0.19) & SGR (1.50 ± .19). The garlic extract treatment showed excellent performance for all the parameter under studied. Hence, the feeding with (0.02%) garlic extract can be used for large scale in tilapia breeding.Item DNA BARCODING OF SILURIFORM FISHES OF THE MEGHNA RIVER ESTRUARY(DEPARTMENT OF BIOTECHNOLOGY, SHER-E-BANGLA AGRICULTURAL UNIVERSITY, DHAKA-1207, 2021-12) MOLLIK, NIAZ MORSHEDThe Meghna river estuary is the largest estuarine ecosystem in Bangladesh and the fish assemblage in this estuary is highly diverse. Estuaries are also very important as breeding and nursery grounds for a wide variety of fishes. Fish samples were collected from the different zone of the Meghna estuary around the year July 2020 to May 2021. In the present study, a total of 9 species were identified from the 7 seven genera and two families (Bagridae and Schilbeidae) under the order Siluriformes based on DNA barcoding approach along with morphology. The species were M. bleekeri, M. gulio, M. vittatus, R. rita, S. aor from the Bagridae family and A. coila, C. garua, E. vacha, S. silondia from the Schilbeidae family. The barcode sequences obtained from the present study showed high identity (> 99%) with the reference sequences deposited in the GenBank database. The assessment of species identities with previously known sequences and closely related species in NCBI BLAST generated 99% of identities indicating the effectiveness of COI sequences to provide species-level resolution. Sequences of the present study were clustered in a single clade with the sequences of conspecific species from GenBank which validate the genetic identification of the studied species. Genetic distances among the sequences of the present study and its compared sequences from GenBank ranged from 0.00 (0.2%) to 0.003 (0.3%) which also validates the genetic identification of this species. Fish diversity in the Meghna has long been declining due to the number of anthropogenic and natural factors. This study provides information on the species diversity of Siluriformes that will help in the management and conservation of the fish fauna of the Meghna river system.Item IN VITRO REGENERTION OF TEA [Camellia sinensis (L.) O. Kuntze](DEPARTMENT OF BIOTECHNOLOGY, SHER-E-BANGLA AGRICULTURAL UNIVERSITY, DHAKA-1207, 2021-12) KOLY, MOKARAM HANIFAThe present research was carried out at Biotechnology Laboratory, Department of Biotechnology, Sher-e-Bangla Agricultural University, Dhaka-1207 from the period of January 2020 to September 2021 to evaluate the effect of different plant growth regulators on in vitro callus induction, shoot regeneration and root formation for Tea. The experiment was conducted at Completely Randomized Design (CRD) with 3 replications. Four levels of BA (1.00, 2.00, 3.00 and 4.00 mg/L) alone and in combination with four levels of NAA (1.00, 2.00, 3.00 and 4.00mg/L) were used for shoot induction. The treatment 2, 4-D was used for callus induction. Different concentration of IBA (300, 400 and 500 mg/L) was used with different dipping time for root induction of regenerated shoot in natural open condition. The highest percentage (86.51%) of callus induction was observed in combined treatment of 2.00 mg/L BA + 1.50 mg/L 2, 4- D with minimum (2.12) weeks. Highest percentage (84.39%) of shoot induction was recorded with 2.00 mg/L BA+ 2.00 mg/L NAA as well as in minimum (32.43) days. Maximum number of shoot (1.42, 2.46 and 4.49) and the highest length of shoot (1.41, 3.28 and 4.61) at 60, 90 and 120 DAI were recorded from the treatment 2.00 mg/L BA + 2.00 mg/L NAA. The highest percentage (75.32%) of root induction with minimum (6.02) weeks was recorded when regenerated shoots were dipped in 500 mg/L IBA solution for 30 minutes. The highest number of roots (4.20 and 7.80) and highest length of roots (3.31 and 5.80 cm) were recorded in 500 mg/L IBA concentration in 30 minutes dipping time. No root induction was recorded in controlled treatment. The protocol developed from the present study might be useful for large scale production of healthy, disease free planting material for Tea.Item MOLECULAR DIVERSITY ANALYSIS OF AMAN RICE GERMPLASM USING SSR MARKERS(DEPARTMENT OF BIOTECHNOLOGY, 2021) HASAN, MD. ROKIBULAn experiment was carried out at the laboratory of Genetic Resources and Seed Division (GRSD), Bangladesh Rice Research Institute (BRRI), Joydebpur, Gazipur, during the period of January 2020 to August 2020 for genetic diversity analysis within 18 Aman rice germplasm. Eleven rice genome specific SSR Primers were used for molecular diversity analysis. All of the eleven primers were capable to produce polymorphic bands. A maximum (6) number of bands was observed in SSR primers RM206. Thirty Eight alleles were detected with an average of 3.45 alleles. The Polymorphic Information Content (PIC) of SSR markers ranged from 0.10 (RM455) to 0.75 (RM206) with an average value of PIC = 0.42. Gene diversity ranges from 0.10 (RM455) to 0.78 (RM206) with a mean value of 0.45. The RM206 marker can be considered as the best marker among the studied markers for 18 Aman rice germplasm. Four primers showed high value of genetic diversity, five primers showed moderate diversity and two primers showed low diversity value on the basis of PIC value. Nei’s genetic distance (D) among 18 rice germplasm was computed from combined data for the 11 primers, ranged from 0.09 to 0.82 with an average of 0.46. Dendrogram based on Nei’s genetic distance using Unweighted Pair Group Method of Arithmetic Mean (UPGMA) indicated the segregation of 18 genotypes into five main clusters. This result could be further used for molecular diversity analysis and identify parent for the development of high yielding variety of rice.Item REGENERATION OF POTATO (SOLANUM TUBEROSUM L.) BY UTILIZING A NEW COMPOSITION OF STOCK SOLUTION-1 FOR PLANT TISSUE CULTURE MEDIUM(DEPARTMENT OF BIOTECHNOLOGY, 2021) ALI, MD. ERSHADAn experiment was conducted at North Bengal Agro Farms Ltd., Thakurgaon, Bangladesh, during the period of January to September 2020 for plantlet regeneration of potato using different new composition of stock solution-1. Three chemicals viz. chemi-i, chemi-ii and chemi-iii were used in stock solution-1 as an alternate of ammonium nitrate. Eight different treatments were designed with those chemicals including Murashige and Skoog (1962) culture media as a check treatment (T ). Two potato varieties viz. Diamant and Asterix were used for in vitro regeneration. The Diamant variety showed the best regeneration potentiality in the T 3 , which consists of the chemi-iii. The check treatment (T 3 ) showed 2 nd 7 highest performance for all the parameters under studied. The Asterix variety gave the best performance in the check treatment (T 3 ) and the 2 nd highest result was found in the T . Different abnormalities were noticed in respect of media and plantlet regeneration. Brown, deep brown, off white and contaminated colour was found in the treatments T 4 1 , T 2 , T 5 , T 6 and T . Plantlet regeneration from those treatments showed aberrancy and it has stunted growth, dwarf size, small leaf formation, black layer at the root tips and finally plantlet died within 4 th week of regeneration. The T 7 8 treatment showed yellow colour and the plantlet regenerated from this treatment were thin, tall with less number of roots. The plantlet regenerated from check treatment (T ) showed good growth and development throughout the experimental period. Finally, it can be concluded that, the chemi-iii, can be used as a component of stock solution-1 which will be a good alternate of ammonium nitrate.Item IN VITRO REGENERATION OF GLADIOLUS (Gladiolus sp.)(DEPARTMENT OF BIOTECHNOLOGY, 2021) BAIDYA, DRISTYThe present research was carried out in Biotechnology Laboratory of the Department of Biotechnology, Sher-e-Bangla Agricultural University, Sher-e-Bangla Nagar, Dhaka -1207 from the period of July 2020 to June 2021 for in vitro rapid regeneration of Gladiolus. The healthy, disease free cormel of 1.5 cm- 2.0 cm were used as explants, was sterilized by 0.1% Bavistin followed by 0.1% HgCl2 mixing with few drops Tween-20. The explant were inoculated in MS media supplemented with different combination of Benzyl Adenene (1.0, 2.0, 3.0, 4.0, 5.0 mg/L) and Naphthalene Acetic Acid (0.5, 1.0, 1.5, 2.0 mg/L). The minimum days (11.00) to shoot (73.80%) induction with the highest percent of shoots induction were achieved in 4.0 mg/L BA. The highest number of shoot (1.60, 2.20, 2.40) and maximum shoot length (0.74 cm, 1.18 cm and 1.32 cm) were also observed in 4.0 mg/L BA and at 21, 35 and 45 Days After Induction (DAI) respectively. The combine effect of auxins and cytokinins were also investigated where 4.0 mg/L BA + 0.5 mg/L NAA performed the best incase highest frequency of shoot induction (74.80%) with minimum days (10.60) and gave the maximum shoot number (1.80, 2.60, 2.80) and shoot length (1.40 cm, 2.32cm, 2.82 cm) at 21, 35 and 42 DAI respectively. Moreover, the treatment 4.0 mg/L BA + 2.0 mg/L NAA showed the highest frequency (79.80%) of root induction with minimum days (8.40) and gave the maximum number (4.20, 6.00, 7.20) of root with optimum root length (1.02 cm, 1.26 cm, 1.34 cm) at 21, 35 and 42 DAI. In shaded condition the survival rate of plantlet was 83.33% and in open atmospheric condition the survival rate was 76%. Overall, a reliable and feasible protocol had been developed for in vitro multiplication of Gladiolus which can be a useful tool for tissue culture of GladiolusItem IN VITRO REGENERATION OF ORCHID (Dendrobium sp.)(DEPARTMENT OF BIOTECHNOLOGY, 2021) SULTANA, RAZIAThe experiment was conducted at the Biotechnology Laboratory of Department of Biotechnology, Sher-e-Bangla Agricultural University, Dhaka from January 2021 to June 2021 to investigate the effect of different plant growth hormone for shoot proliferation and root regeneration of Dendrobium sp. where shoot tips were used as explant. The experimental data were taken on different parameters of shoot and root growth. Different doses of individual shooting hormone(BA), individual rooting hormone(NAA) and the combination of these two hormones were experimented here. The minimum days to shoot induction were recorded 15 days from treatment T 8 (2.0 mg/L BA + 1.50 mg/L NAA). Maximum percentage (73.4%) of shoot induction was noticed in treatment T 8 (2.0 mg/L BA + 1.50 mg/L NAA) .The maximum days to root induction was 44 days and it was recorded in control treatment .The minimum days to root induction was 17 and it was observed from treatment T 8 (2.0 mg/L BA + 1.50 mg/L NAA) days. In the following study significant variation was observed for the use of BA and NAA on the percent of shoot induction explants. Significant variation was observed for the use of BA and NAA on the percent of root induction of explants . The treatment of 2.0 mg/L BA response excellent in shoot induction and 1.0 mg/L NAA showed better performance in root development in Dendrobium sp. Combined effect of BA and NAA was very significant. Overall a reliable protocol was developed for shoot and root proliferation which can be used for further crop improvement program.