Identification of DNA sequence responsible for 5-bromodeoxyuridine-induced gene amplification

Bromodeoxyuridine (BrdUrd) treatment of the prolactin nonproducing subclone of GH cells (rat pituitary tumor cells) induces amplification of a 20-kilobase DNA fragment including all of the prolactin gene coding sequences. This amplified DNA segment, which is flanked by two unamplified regions, thus designates a unit of BrdUrd-induced amplified sequence. Cloned DNA segments, 10.3 kilobases long, from the 5' end of the rat prolactin gene of BrdUrd-responsive and -nonresponsive cells, were ligated to the thymidine kinase gene of herpes simplex virus type 1 (HSV1TK), and the hybrid DNA was transferred to thymidine kinase-deficient mouse fibroblast cells by transfection. The HSV1TK gene and the rat prolactin gene were amplified together in drug-treated transfectants carrying the hybrid DNA HSV1TK gene and rat prolactin gene of BrdUrd-responsive GH cells. These results suggest that the 10.3-kilobase DNA segment at the 5' end of the rat prolactin gene of BrdUrd-responsive GH cells carries the information for drug-induced gene amplification (amplicon) and that another gene, such as the HSV1TK gene, is also amplified when the latter is placed adjacent to this segment.     

Exploitation of SSR,SRAP and CAPS-SNP markers for genetic diversity of Citrus germplasm collection

The present study was to assess informativeness and efficiency of three different molecular markers for genetic diversity among 24 Citrus and its relative species. Sixty one SSR, 33 SRAP and 24 CAPS-SNP markers were used to evaluate the level of polymorphism and discriminating capacity. A total of 596, 656 and 135 polymorphic amplicons were observed in SSR, SRAP and CAPS-SNP markers with average polymorphism information content (PIC) of 0.97, 0.98 and 0.89, respectively. High levels of polymorphism were recorded for SSR and SRAP compared with CAPS-SNP markers. The highest correlations (r = 0.930) were obtained between SSR and SRAP markers, whereas SSR and CAPS-SNP were poorly correlated (r = 0.833). Cluster analysis was performed to construct dendrograms using UPGMA. And the dendrogram from SSR data was most congruent with the general dendrogram. These findings provide basis for future efficient use of these molecular markers in the genetic analysis of Citrus and its relatives.     

Prevalent Serotypes of Pasteurella multocida Isolated from Different Animal and Avian Species in India

Identification and estimation of the prevalence of Pasteurella multocida organisms in different animal and avian species in India during November 2000 to July 2003 was carried out. Out of 418 samples collected from different outbreaks suspected to be caused by P. multocida, a total of 206 bacterial cultures were identified as P. multocida on the basis of cultural, morphological and biochemical characteristics. All the 206 cultures were isolated from different domestic animal species (cattle, buffalo, sheep, goat, pig and rabbit), avian species (chicken, duck, quail, turkey, goose) and wild animals such as leopard and deer. Serotyping of P. multocida cultures revealed the presence of various serotypes (A:1, A:3, A:1,3, A:4, B:2, D:1 and -:1) among the livestock population. P. multocida polymerase chain reaction (PCR) assay applied on different forms of bacterial cultures (bacterial culture lysate, direct bacterial colony and mixed bacterial culture lysate) yielded an amplified product of approximately 460 bp specific for P. multocida. The results of PCR assay correlated well with conventional methods of identification. The present investigation revealed the presence of varied serotypes among livestock and PCR assay was found to be useful for rapid, sensitive and specific diagnosis of pasteurellosis in animals and avian species.     

Comparative study of different surface decontaminants on chicken quality

(1) A comparative study on the effect of different surface decontaminants: hot water at 70 degrees C for one minute; 2% lactic acid for 30 s; 1200 p.p.m. acidified sodium chlorite (ASC) solution for 5 s and 50 p.p.m. chlorine solution for 5 min in the form of dips and sprays on the surface of dressed broilers for 0, 24 and 48 h of storage was conducted. (2) The variables studied were, total plate count (TPC), presumptive coliform count (PCC), pH and extract release volume (ERV). All treatments reduced TPC and PCC. (3) Lactic acid dip and hot water dip were the most effective for reducing TPC (1.36 and 1.28 log/cm2, respectively) with no significant difference between them. (4) ASC and hot water in dip could diminish PCC (1.37 and 1.34 log/cm2, respectively) and did not vary significantly. (5) No treatment affected muscle pH, water holding capacity (WHC), ERV, appearance, smell, tenderness and overall acceptability of treated broilers significantly. (6) Hot water treatment is the cheapest, most convenient and simplest decontamination technique for hygienic and wholesome poultry production.     

Homegarden agroforestry systems: an intermediary for biodiversity conservation in Bangladesh

Biodiversity conservation is one of the important ecosystem services that has been negatively impacted by anthropogenic activities. Natural forests (NF) harbor some of the highest species diversity around the world. However, deforestation and degradation have resulted in reduced forest land cover and loss of diversity. Homegarden agroforestry (AF) systems have been proven to be an intermediary for biodiversity conservation. In this study, we evaluate the effectiveness of home garden AF practices to conserve tree species diversity in Bangladesh and compare them with tree species diversity in NF. A total of nine locations were selected for this synthesis from published literature which comprised of five AF sites and four NFs. Shannon?CWeiner Diversity Index (H) was similar for home-garden AF (3.50) and NF (2.99), with no statistical difference between them. Based on non-metric multi-dimensional scaling (NMDS) ordination analysis, the AF and NF plots showed distinct separation. However, Bray?CCurtis dissimilarity index ranged from 0.95 to 0.70 indicating nearly no overlap in species composition to significant overlap between AF and NF. Based on our results, we conclude that AF can serve as an important ecological tool in conserving tree species diversity, particularly on landscapes where NF fragments represent only a small fraction of the total land area. Creating and maintaining AF habitats in such human dominated landscapes should be part of the biodiversity conservation strategy.     

Survivability and causes of loss of broody-hen chicks on smallholder households in Bangladesh

We determined the flock sizes and rates of loss caused by different factors in broody-hen chicks (BHC) up to 60 days of age on 600 randomly selected smallholdings in Bangladesh. The smallholders were beneficiaries of a village poultry production chain called ‘Smallholder Livestock Development Project-2’ (SLDP-2) which was undertaken with the financial assistance of the Danish International Development Agency (DANIDA). For estimating survival time of BHC, we observed chicks in 80 smallholdings. SLDP-2 aims at ameliorating poverty among women by poultry rearing at village level; in total, 104,000 key rearers, constituting 96% of all of the beneficiaries of the SLDP-2 area, were enrolled in 26 upazilas (a lower administrative unit of Bangladesh). A key rearer is a smallholder who rears at least five ‘Sonali’ (RIR × Fayoumi) and some indigenous (desi) chickens in a semi-scavenging system. Sonali chickens are supplied from the development project, and have higher egg production while the broodiness of the desi hens is exploited to get chicks hatched for future stocks; thus, the chicks hatched and reared to 60 days old at key rearers’ households are called BHC. In this study 32% of the smallholders had BHC each month. At the beginning of a month, the median number of chicks in a flock was 8, and the mean survival time was 50.5 days. Incidence rates of loss of BHC from disease, predation, selling and slaughtering were 0.102, 0.086, 0.009 and 0.002 per chick-month at risk, respectively. The major predators were crows, mongooses and eagles with incidence rates of loss being 0.018, 0.016 and 0.010 per chick-month at risk, respectively. Colibacillosis (both single and mixed infections) contributed to the death of 21% of dead BHC collected; Newcastle disease and salmonellosis contributed to the next highest (14 and 12%) proportional mortalities.     

Effects of illite supplementation on in vitro and in vivo rumen fermentation,microbial population and methane emission of Hanwoo steers fed high concentrate diets

This study was conducted to evaluate the effects of feeding supplemental illite to Hanwoo steers on methane (CH₄) emission and rumen fermentation parameters. An in vitro ruminal fermentation technique was conducted using a commercial concentrate as substrate and illite was added at different concentrations as treatments: 0%, 0.5%, 1.0%, and 2.0% illite. Total volatile fatty acids (VFA) were different (P < 0.05) at 24 h of incubation where the highest total VFA was observed at 1.0% of illite. Conversely, lowest CH₄ production (P < 0.01) was found at 1.0% of illite. In the in vivo experiment, two diets were provided, without illite and with addition of 1% illite. An automated head chamber (GreenFeed) system was used to measure enteric CH₄ production. Cattle received illite supplemented feed increased (P < 0.05) total VFA concentrations in the rumen compared with those fed control. Feeding illite numerically decreased CH₄ production (g/day) and yield (g/kg dry matter intake). Rumen microbial population analysis indicated that the population of total bacteria, protozoa and methanogens were lower (P < 0.05) for illite compared with the control. Accordingly, overall results suggested that feeding a diet supplemented with 1% illite can have positive effects on feed fermentation in the rumen and enteric CH₄ mitigation in beef cattle.