Optimization of callus induction and regeneration system for Pakistani wheat cultivars Kohsar and Khyber-87

Wheat is a member of family Poaceae. It is the major staple food of Pakistan. The present study was done to improve the regeneration of two commercially grown wheat varieties Kohsar and Khyber-87.Mature embryos were used as explants. Five different concentrations of 2,4-D; 2, 2.5, 3, 3.5 and 4 mg/L were used for callus induction. For regeneration, initially different concentrations (0.1 to 0.2) of IAA(indole-3-acetic acid) and BAP (6-benzylaminopurine) were experimented. The best combination of these hormones that is, 0.1 mg/L IAA and 0.5 mg/L BAP were further subjected to experimentation along with different concentrations of kinetin; 0.1, 0.2, 0.3, 0.4, 0.5 and 1 mg/L. Maximum calli of Kohsar (83.3%) was obtained at 3 mg/L 2,4-D whereas for Khyber-87 maximum callus induction (71.70%) was obtained at 3.5 mg/l 2,4-D. The maximum regeneration of both Kohsar and Khyber-87 (80.5 and 62.2%, respectively) were obtained at the combinations of 0.1 mg/L IAA, 0.5 mg/L BAP and 0.5 mg/L kinetin

PREPARATION AND EVALUATION OF VITAMIN E ADJUVANTED OIL EMULSIFIED INFECTIOUS BRONCHITIS EXPERIMENTAL VACCINE

The present study was conducted to prepare oil emulsified (OE) infectious bronchitis (IB) experimental vaccines. The vaccines were prepared using the vaccinal strain H-120 Infectious Bonchitis virus (IBV). The virus was cultivated in 9-day old embryonated eggs via allantoic cavity route. Allantoic-amniotic fluid (AAF) was collected and inactivated with formalin @ 0.12%. Water in oil emulsion was prepared by adding one part of AAF to four parts of mineral oil containing water phase (Tween 80) and oil phase (Span 80) surfactants. Hydrophile lypohile balance (HLB) of the emulsion was maintained at 7.0. Two oil emulsified experimental vaccines were prepared. Vaccine-I was prepared without vitamin E and Vaccine-II with vitamin E (300 mg/ml). A total of 120 day-old broiler breeder chickens were divided into 4 groups, A, B, C, and D, each having 30 birds. At the age of 21 days, experimental Vaccine-I, experimental vaccine-II and commercial IB killed (H-120) vaccine were inoculated @ 0.5 ml in the birds of groups A, B and C, respectively. Group D was maintained as nonvaccinated control. Efficacy of the vaccines was evaluated on the basis of humoral immune response (haemagglutination inhibition antibody titres) against IB in the four groups. The seven weeks cumulative mean antibody titres (CMT) of each group were calculated. The highest CMT was observed in group B (130), followed by group C (69), group A (58) and group D (17). Statistical analysis showed that haemagglutination inhibition (HI) antibody titres in group B (vaccine- II) were significantly higher than those of groups A, B and C (P< 0.05)

The response of sugarcane (Saccharum officinarum L) genotypes to callus induction, regeneration and different concentrations of the selective agent (geneticin -418)

Two commercial cultivars (CPF-245 and CPF-237) and three advanced lines (CSSG-668, S-2003US633, S-2003US114) of sugarcane (Saccharum officinarium) grown in Punjab, Pakistan were evaluated for their potential to induce callus, embryogenic callus and regeneration. Cultivar CSSG-668 was found to be the best genotype yielding maximum embryogenic callus and regeneration whereas cultivar CPF- 245 exhibited lowest callus induction frequency. Five different concentrations (0, 20, 40, 60, and 80mg/L) of the selective agent (geneticin-418) were used to optimize selection conditions with nontransformed embryogenic calli. The geneticin concentration 60 mg/L was found to be the optimal dose to select the embryogenic calli of genotypes CSSG-668, CPF-245 and S-2003US63, while 35 mg/L geneticin was found to be the best concentration for S-2003US-114. Similarly, 60 mg/L geneticin was optimum dose to select regenerated plantlets of the cultivars CSSG-668 and CPF-245 while it was 40, 25 mg/L for the cultivars S-2003US-114 and S-2003US-633, respectively. It is concluded from the present study that geneticin concentration in the range of 25 to 60 mg/L can be effectively used for the selection of transformed embryogenic calli and regenerants of different sugarcane cultivars.Keywords: Callus induction, embryogenic callus, regeneration, Saccharum officinarum L., selection, geneticin

Food and feeding habits of Omobranchus sp. (Blenniidae: Omobranchini) larvae in the seagrass-mangrove ecosystem of Johor Strait, Malaysia

The stomach contents of Omobranchus sp. (family Blenniidae) larvae were investigated in a seagrass-mangrove based ecosystem in Johor Strait, Malaysia from October 2007 to September 2008. Specimens of larval fish were collected through subsurface towing of a Bongo net from five different stations. The stomach sacs of 267 Omobranchus sp. larvae were separated and observed, which comprised of 24 significant food stuffs belonging to 6 main groups viz. phytoplankton (62.45%), zooplankton (18.24%), algae (5.56%), plant-like particles (5.75%), debris (4.22%) and unidentified particles (2.03%). In situ water parameters were also measured throughout the sampling cruises. There was a strong and significant positive correlation between stomach phytoplankton and salinity (r = 0.658, p < 0.05).? Canonical correlation analysis indicated a weak relationship (29.8%) between stomach contents and physico-chemical parameters. Only salinity appeared to be the controlling factor for the stomach contents of Omobranchus sp. larvae in the investigated area. Based on the stomach content analysis, it could be concluded that Omobranchus sp. were mainly herbivorous during the larval stages

Impacts of gamete concentration, age and contact time on the fertilization success in the tropical species of white sea urchin, Salmaciella dussumieri

The globoid sea urchin (Salmaciella dussumieri) occurs abundantly in the Indo-West Pacific from the Island of the West Indian Ocean, East Africa, Magdagascar, Red Sea, South-East Arabia, Ceylon, Bay of Bengal, North Australia, Philippine, China and South Japan and Australia, and also has significant biological, ecological, aquaculture, conservational and medicinal importance. An experiment was carried out to assess the influence of sperm dilution, egg concentration, sperm-egg exposure time, and gamete aging on fertilization success of S. dussumieri in a captive laboratory condition. It has been found that dilution, age and contact time of sperm to egg were successively the most dominant factors effecting the success of fertilization, but concentration of eggs did not significantly so over the range investigated. The sperms retained their competency for more than two hours only in relatively dense sperm concentrations (> 10-4 dilution of `dry' sperm), although they exhibited lower capability and potency with increasing dilutions and age. In the trials of the egg-sperm exposure time, >80% eggs were fertilized within 10 sec of contact time at lower dilutions (10-3-10-2) of concentrated sperm, while some longer times were required to get the higher rates of fertilization. On the contrary, eggs remained in good quality for up to a period of 3 h and no abnormality or any adverse effects in fertilization were observed. The findings obtained from the present experiment reveal that limited longevity of diluted sperm have an important effect on fertilization rate of sea urchin eggs during natural spawning seasons in the field. Hence, the globoid sea urchin (S. dussumieri) is under extensive selective pressures to breed synchronously with the purpose of producing high sperm concentrations and higher sperm-egg interactions to maximize the success of fertilization in the water column

Sex ratio, spawning season and the size at maturity of Parapenaeopsis sculptilis (Heller 1862) in the coastal waters of Perak, Peninsular Malaysia

Parapenaeopsis sculptilis (Heller, 1862) locally referred to as "udang kulit keras" in Malaysia has profound biological, ecological, aquacultural and conservational significance. The reproductive biology of this important penaeid from the coastal waters of Perak, Peninsular Malaysia, was studied during the period between February 2012 to January 2013. Females outnumbered males with a sex ratio of M: F= 1:3 (P < 0.05). Four maturity stages of female gonads viz., immature, maturing, mature and spent stages were distinguished. The first stage of sexual maturity was attained at a length of 9.3 cm, and female P. sculptilis showed a peak gonadosomatic index during the month of April, August and October, indicating that P. sculptilis potentially breeds throughout the year. The findings of this study would greatly contribute towards the understanding of gonadal maturation, spawning season and breeding biology, which could be important for the effective population management of this prawn species

Heterogeneous Relational Databases for a Grid-enabled Analysis Environment

Grid based systems require a database access mechanism that can provide seamless homogeneous access to the requested data through a virtual data access system, i.e. a system which can take care of tracking the data that is stored in geographically distributed heterogeneous databases. This system should provide an integrated view of the data that is stored in the different repositories by using a virtual data access mechanism, i.e. a mechanism which can hide the heterogeneity of the backend databases from the client applications. This paper focuses on accessing data stored in disparate relational databases through a web service interface, and exploits the features of a Data Warehouse and Data Marts. We present a middleware that enables applications to access data stored in geographically distributed relational databases without being aware of their physical locations and underlying schema. A web service interface is provided to enable applications to access this middleware in a language and platform independent way. A prototype implementation was created based on Clarens [4], Unity [7] and POOL [8]. This ability to access the data stored in the distributed relational databases transparently is likely to be a very powerful one for Grid users, especially the scientific community wishing to collate and analyze data distributed over the Grid

Length-weight relationships and relative condition factor of Parapenaeopsis sculptilis (Heller, 1862) from the coastal waters of Perak, Peninsular Malaysia

Length-weight relationship (LWR) parameters and relative condition factor (K n) of marine shrimp, Parapenaeopsis sculptilis (Heller, 1862) were estimated using length-weight data collected between February 2012 and January 2013 from the coastal waters of Terong, Perak, Peninsular Malaysia. The estimated length-weight relationship of P. sculptilis for both sexes was W = 0.00027TL2.80. Meanwhile, the estimated relative growth coefficient (b) was 2.80 for both sexes, indicating a negative allometric growth pattern of P. sculptilis in the investigated area. Relative condition factor (Kn) values ranged from 0.99 to 1.064 (1.013±0.005, mean ±SD). Kn value changes in various months: the highest peak was in March-April, indicating the spawning period and the trough and small peaks indicating the cycle gonadal development

STUDIES ON SOME BIOCHEMICAL CONSTITUENTS OF OVARIAN FOLLICULAR FLUID AND PERIPHERAL BLOOD IN BUFFALOES

This project was designed to study some biochemical constituents of ovarian follicular fluid (FF) and peripheral blood in buffaloes. For this purpose, ovaries were collected from adult buffaloes immediately after slaughter, FF was aspirated and stored at -4°C. Blood samples were also collected from these buffaloes before slaughter, plasma was separated and stored for further analysis. Samples were classified into two groups according to the follicle diameter i.e. small (3-10 mm) and large (11-20 mm). The FF and plasma samples were analyzed for various biochemical constituents, including glucose, cholesterol, total proteins, albumin, globulin, sodium, potassium, calcium, magnesium, zinc, triiodothyronine and thyroxine contents, using commercial kits.The results showed that small follicles had significantly lower (P<0.05) glucose contents than large follicles, while blood plasma had significantly higher (P<0.05) glucose contents than fluid from both classes of follicles. The differences in concentrations of cholesterol, total proteins, albumin and globulin between small and large follicles were non-significant. The concentrations of these compounds were higher in the blood than in FF, except albumin, which was higher in FF than in the blood. Contents of electrolytes and trace elements did not vary between the two follicle classes. However, the plasma levels of these electrolytes and trace elements, except potassium and zinc, were significantly higher (p<0.05) than their levels in FF. The level of potassium was significantly higher (P<0.05) in FF than in the plasma, while serum zinc level did not differ from FF. The differences in concentrations of triiodothyronine and thyroxine in fluid from small and large sized follicles were non-significant. Likewise, the levels of these hormones were non-significantly higher in blood plasma than in FF. It was concluded that FF levels of glucose differed between small and large follicle groups, while blood levels of most of constituents were higher than their levels in FF

Population dynamics of sergestid shrimps Acetes japonicus in the estuary of Tanjung Dawai, Kedah, Malaysia

Population parameters of male and female A. japonicus were studied using the monthly length frequency data to evaluate the mortality rates and its exploitation level. The sex ratio (male: Female) was found at 1: 0.94. Asymptotic length (L∞) was 25.20 mm and 28.88 mm for male and female, respectively. Growth co-efficient (K) for males and females was estimated at 1.80 and 1.30 year-1, respectively. Total mortality (Z) was calculated at 5.98 and 4.44 year-1 for male and female of A. japonicus respectively. Natural mortality (M) was 2.82 and 2.19 year-1 for the male and female shrimps. The fishing mortality (F) was 3.16 year-1 for male and 2.25 year-1 for female. Exploitation level (E) for male and female of A. japonicus was calculated at 0.53 and 0.51. The exploitation level was slightly over (E>0.50) the optimum level of exploitation (p = 0.50). The stock of A. japonicus was found to be slightly over exploited in Tanjung Dawai estuarine waters