Regeneration in sugarcane via somatic embryogenesis and genomic instability in regenerated plants

In the present study, embryogenic calli of sugarcane variety BL4 were induced using 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin in different concentrations and combinations. In contrast to earlier studies, embryogenic callus sectors were identified and isolated microscopically within 1–2 weeks. Subsequently, 51 media formulations were used for regeneration of proliferated embryogenic callus, using MS medium supplemented with three different cytokinins [kinetin, 6-Benzylamino purine (BAP), and thidiazuron (TDZ)] and auxins (IAA/NAA and IBA) in different combination and concentrations. After acclimatization, the genomic DNA of regenerated plants was studied to explore the insertion polymorphism of transposable elements in order to ascertain the variation among somaclones. Though low concentration of kinetin with 2,4-D was found supportive to embryogenic callus development, the highest number of regenerated plantlets was observed using BAP (1 μM), however the plantlets had very low fresh weight (2.2 g). Conversely, TDZ alone supported a significant increase in the number of plantlets regenerated (38–40) with higher fresh weight. The somaclones generated during this study showed considerable positional polymorphism of activator-like transposable elements possibly due to the stress associated with in vitro culture. This study provides a procedure to produce regenerated sugarcane plants from embryogenic callus in a relatively short time.     

The metabolism of DDT in vivo by the Japanese quail (Coturnix coturnix japonica)

Male and female Japanese quail (Coturnix coturnix japonica) were given intraperitoneal injections of [¹⁴C]DDT in ethanol at a rate of 13.4 mg/kg body wt. Fifty-six days later the tissues and droppings were analysed for total ¹⁴C and metabolites. The rate of loss of ¹⁴C in droppings was very similar in males and females. The maximal rate was reached on the third day, and 65–66% of the injected dose was voided by the fifty-sixth day. Ninety-three to ninety-four percent of the ¹⁴C in droppings and 83–90% of the ¹⁴C in tissues were extracted by solvents. Combined extracts from males and females were used for determination of DDT and its metabolites. Expressing all results as percentages of injected dose, the following were isolated from droppings: DDA (24%), DDT (3%), DDD (5.1%), DDE (11%), and uncharacterised polar metabolites (17%). Twenty-five percent of the dose was retained in the tissues and this was largely accounted for as DDT (10.4%) and DDE (10.5%). Of the total metabolites found 31% was DDE (almost equally divided between tissues and droppings) and 35% was DDA (almost entirely in droppings). Since DDD was not found in significant quantities in tissues, the substantial quantities in droppings were probably produced from DDT by the action of microorganisms.     

Assessing solid waste compost application as a practical approach for salt-affected soil reclamation

Abstract

A short-term pot experiment was made to evaluate the effectiveness of municipal solid waste compost amendment on salt-affected soil. Hordeum maritimum plants were cultivated in pots filled with a clay-loam soil containing 0 or 40 t ha^?1 of compost and irrigated with tap water at 0 or 4 g l^?1 NaCl. Soil properties and heavy metal (Zn^{2 +}, Pb^{2 +}, Cd^{2 +}) accumulation were investigated. Municipal solid waste compost application significantly increased the soil contents of carbon, nitrogen and potassium under both non-saline and saline conditions. Soil heavy metal concentrations increased substantially too, but the recorded values were below the toxicity limits. Interestingly, plants subjected to the salt–compost interaction were more vigorous, compared with those grown on non-amended soil. Altogether, our data indicate that short-term utilization of municipal solid waste compost at 40 t ha^?1 may be of potential interest in the perspective of the rehabilitation of salt-affected soils. Yet, it must be stressed that the present findings are preliminary and need to be further evaluated under field conditions before practical recommendations can be inferred.     

In vitro maturation and fertilization of prepubertal and pubertal black Bengal goat oocytes

Oocytes retrieval, in vitro maturation (IVM) and fertilization (IVF) efficiency are inevitable steps towards in vitro production of embryos. In the present study, these parameters were investigated in the ovaries of prepubertal (n = 31) and pubertal (n = 61) black Bengal goats obtained from a slaughterhouse. Nuclear maturation was evaluated upon aspiration and following IVM in TCM-199 (Earle''s salt with L-glutamine and sodium bicarbonate) for 27 h at 39℃ under 5% CO₂ in humidified air. The oocytes retrieval and efficiency (mean ± SD) per prepubertal and pubertal goats were 5.2 ± 0.6 and 6.8 ± 0.6, and 77.3 ± 0.1% and 80.5 ± 0.6%, respectively. Anaphase I - telophase I stages differed significantly (7.3 ± 0.8 vs. 2.6 ± 0.2, p < 0.05) between the two groups of goats. After IVM, the percentages of metaphase II were significantly higher (66.3 vs. 60.3, p < 0.05) in pubertal goats than in their prepubertal counterparts. The percentages of normal in vitro fertilization (IVF) in Fert-Tyrode''s albumin lactate pyruvate of pubertal goat oocytes did not differ between Percoll and swim-up sperm separation methods (36.7 ± 0.9% vs. 32.7 ± 1.3%, p > 0.05). Furthermore, sperm capacitation by heparin alone or in combination with ionomycin did not lead to a significant increase in the normal fertilization rate (34.8 ± 1.7 vs. 32.2 ± 1.5%, respectively) in the oocytes of pubertal goats. In conclusion, the ovaries of pubertal black Bengal goats obtained from the slaughterhouse could be used for in vitro embryo production. However, further optimization of the IVM and IVF techniques are necessary for satisfactory in vitro embryo production.     

<Emphasis Type="Italic">Eimeria bovis</Emphasis> meront I-carrying host cells express parasite-specific antigens on their surface membrane

Host immune responses conducted against antigens of Eimeria bovis are key factors for the development of protective immunity against this protozoan disease. In this study we investigated the expression of E. bovis-derived antigens on the host cell surface membrane during E. bovis first merogony in vitro. Host cells carrying E. bovis-meront I stages expressed E. bovis host cell surface antigens (EbHCSAg) on their surface membrane which were recognised by hyperimmune sera of calves and by sera from rats immunized with E. bovis merozoites I, when tested by indirect immune fluorescent antibody test (IIFAT), laser scanning confocal microscopy (LSCM) and immune electron microscopy. Expression of EbHCSAg on permissive host cells was earliest detected 7 days p. i., thus coinciding with the onset of the parasite replication. Membrane-associated EbHCSAg were removed from infected host cells by proteinase K, partially by Triton X-100, Triton X-114 and Triton X-405, but not by 1 M NaCl, CHAPS or phospholipase C treatment. Antibodies, affinity-purified on paraformaldehyde/glutardialdehyde (PAGA)-fixed E. bovis meront I-infected bovine host cells bound to the surface meront I-carrying cells and to merozoites I (IIFAT, LSCM) but, in contrast to untreated sera, not to sporozoites. When tested on methanol-fixed merozoites I and sporozoites by IIFAT, affinity-purified antibodies bound to structures in the apical complex area of merozoites I, but not to sporozoites, whilst untreated sera caused diffuse labelling of internal structures of both parasite stages. Immune electron microscopy demonstrated binding of affinity-purified antibodies to micronemes and dense granules of merozoites I. Although the function of EbHCSAg is still unknown, results of this study might suggest an involvement in the development of protective immunity against E. bovis infections.     

Influence of Organic Manures on Soil Characteristics and Yield of Jerusalem Artichoke

ABSTRACT

The objective of this study was to study the influence of three organic manures, farm yard manure (FYM), poultry manure (PLM), and pigeon manure (PGM), on soil physical and chemical properties on tuber yield of Jerusalem artichoke in a newly reclaimed saline calcareous soil. A field experiment was conducted applying the three manures, alone and/or in different combinations. Soils were investigated at surface (0–30 cm) and subsurface (30–60 cm) layers before and after planting, and analyzed for physical and chemical properties. The results indicated that the application of 31.5 kg ha^?1 of PLM+10.5 kg of PGM T7 recorded highest available nitrogen, zinc, copper, and moisture content at the surface layer. The same results were obtained for iron and manganese at both layers. While, applying 21.0 kg ha^?1 FYM+21.0 kg ha^?1 PLM T10 recorded the best treatment for pH, phosphorus, zinc, copper, moisture content, and saturation percentage at the subsurface layer. Applying 21.0 kg ha^?1 PGM + 10.5 kg ha^?1 FYM+10.5 kg ha^?1 PLM T15 recorded the best treatment for organic matter content and bulk density at surface layer and reduced the electrical conductivity and inulin tuber content at both layers. On the other hand, calcium carbonate and sodium adsorption ratio were reduced in both layers by applying 21.0 kg ha^?1 PLM+10.5 kg ha^?1 FYM+10.5 kg ha^?1 PGM T14. The best treatment for tuber nitrogen content and total yield was obtained with applying 42.0 kg ha^?1 PLM T2 only and 31.5 kg ha^?1 FYM+10.5 kg ha^?1 PLM T4, respectively.     

Unusual manifestation of a concurrent demodectic and sarcoptic mange in a Zebu-Friesian cross-bred heifer

We describe a rare case of a concurrent demodectic and sarcoptic mange in a 2-year-old heifer in Khartoum, Sudan. The lesions were massive lumps of granulomatous tumour-like dermatitis with thick, nodular folds mainly covering the head, neck and shoulders. Histopathological examination of the lesions revealed the presence of both Demodex bovis and Sarcoptes scabiei. The animal died regardless of the anti-parasitic treatment it received.     

The Cytological Status of Plants Regenerated from Shoot-Meristem Culture of Pisum sativum L

The cytological status of plantlets regenerated from shoot apical meristems of Pisum sativum was investigated. Chromosome counts in root apices of in vitro regenerated plants showed a preponderance of diploid cells. Moreover, the karyotypes of root-tips from plants derived from culture and from normal plants were basically the same. Topics such as the treatment of chromosomal armlength data, simple statistical comparison of samples derived from normal and regenerated plants are discussed.