Sequence polymorphism in the ribosomal DNA internal transcribed spacers differs among Theileria species

The genomic region spanning the two ribosomal RNA internal transcribed spacers (ITS1 and ITS2) and the 5.8S rRNA gene was cloned and sequenced from sixteen Theileria isolates. Each Theileria species possessed ITS1 and ITS2 of unique size(s) and species specific nucleotide sequences. Varying degrees of ITS1 and ITS2 intra- and inter-species sequence polymorphism were found among ruminant Theileria species. The spacers were most polymorphic in the agent of tropical theileriosis, Theileria annulata, and were more conserved in two benign species, Theileria buffeli and Theileria sergenti Chitose. Phylogenetic analysis of the rDNA ITS1-5.8S rRNA gene-ITS2 region clearly separated each taxon, placing them in three clusters. One held T. annulata, Theileria parva, and Theileria mutans, with the latter two most closely related. The second held T. sergenti Ikeda, T. sergenti Chitose, and T. buffeli, with the latter two most closely related. The third cluster held the Theileria ovis isolates.     

Comparison and phylogenetic analysis of the heat shock protein 70 gene of Babesia parasites from dogs

The heat shock protein 70 (hsp70) genes of Babesia gibsoni, B. canis canis, B. canis vogeli, and B. canis rossi isolated from infected dogs were cloned by polymerase chain reaction (PCR) and sequenced. In the nucleotide sequence and the predicted amino acid sequence of the gene, the parasites were very similar to each other. The nucleotide sequences of the hsp70 gene had more variety than those of 18S nuclear subunit ribosomal DNA (18S rDNA). A phylogenetic analysis of these sequences and comparisons with sequences from other Babesia and Theileria species revealed that all canine babesial isolates analyzed in the present study were closely related to each other and formed one cluster. Additionally, a phylogenetic analysis of Babesia and Theileria species showed that these parasites could be divided into three groups: group A including canine babesial isolates, B. divergens, B. odocoilei, B. bovis, B. caballi, and B. ovis; group B including Theileria annulata, T. orientalis, and T. cervi; and group C including B. microti and B. rodhaini. These results suggested that a phylogenetic analysis of the hsp70 gene sequence might be helpful in classifying Babesia and Theileria species, and that canine babesial isolates might be closely related to each other, indicating their evolution from the same ancestry.     

Comparative efficacy of standard AGID and precipitinogen inhibition test with monoclonal antibodies based competitive ELISA for the serology of <Emphasis Type="Italic">Peste des Petits Ruminants</Emphasis> in sheep and goats

This project was conducted to investigate the comparative efficiency of competitive ELISA (cELISA), standard Agar Gel Immunodiffusion Test (AGID) and Precipitinogen Inhibition Test (PIT) for the diagnosis of Peste des Petits Ruminants (PPR) in Pakistan. To deal with this, serum samples from 198 sheep and 82 goats were collected from three different government livestock farms and all the samples were run simultaneously with the three serological tests. The samples found positive for PPR antibodies through cELISA, AGID and PIT were 96 (34.2%), 60 (21.4%) and 72 (25.7%), respectively. Kappa statistics were applied to evaluate the concordance between the laboratory-based test (cELISA) and field-based tests (AGID and PIT). Kappa statistics scores for cELISA versus AGID and PIT were 0.6343 (95% Confidence Interval CI 0.5231–0.7456) and 0.7134 (95% Confidence Interval CI 0.5987–0.8281), respectively, which indicate a “substantial” agreement between cELISA and AGID and “significant” agreement between cELISA and PIT. AGID and PIT revealed relative diagnostic sensitivities with cELISA of 59.3% and 69.7% and relative diagnostic specificities of 98.3% and 97.2%, respectively. The data suggested that for mass screening and control of PPR, these serological tests proved practical in the absence of cELISA since they have high relative diagnostic specificities and a satisfactory relative diagnostic sensitivities.     

PCR-based detection of Theileria ovis in Rhipicephalus bursa adult ticks

Tick-borne diseases in ruminants are common in tropical and subtropical regions and lead to meat and milk production losses. In this study, polymerase chain reaction (PCR) was used to assess the presence of Theileria ovis in Rhipicephalus bursa ticks. We have demonstrated that the PCR enabled detection of T. ovis in field isolates of R. bursa collected from naturally infested sheep and goats in eastern Turkey. The sampling was done in spring season (between May and June 2004). A total of 420 R. bursa were collected and randomly selected 192 number of them (97 female and 95 male) were dissected. Primers specific for 520 bp fragments small subunit ribosomal RNA (ssu rRNA) gene of T. ovis amplified products from 37 of the 192 (19.27%) samples. The parasite was detected in 17 (17.52%) female and in 20 (21.05%) male ticks. Two T. ovis amplicons from the tick samples were purified and sequenced. The resulting sequences were identical to the nucleotide sequence of the Turkish sheep strain of T. ovis. These results showed that R. bursa might play an important role in the field as a natural vector of T. ovis.     

Evaluation of sunflower (Helianthus annuus L.) single cross hybrids under heat stress condition

Sunflower is an important oilseed crop, which shows susceptibility to heat stress. In this study, 63 single cross hybrids were evaluated under heat stress condition for 2 years and compared with the two commercial hybrids. Genotype and genotype × environment (GGE) was used to differentiate single cross hybrids on the basis of multiple traits. GGE biplot showed that several single cross hybrids had higher seed yield potential than standard check. Moreover, seed yield per plant (SYP) was related to pollen viability percentage, showing that achene yield was the product of high gametophytic fertility under heat stress. Hybrids having high seed yield potential under heat stress had lower cell membrane injury. GGE biplot for SYP and its components showed that single cross hybrids were characterized into two major groups. Group I was further characterized into two sub group. Group Ia included hybrids with high 100-SW, while group Ib had the hybrids with high number of seeds per head and head diameter. Group II had the hybrids with high kernel weight and kernel to seed ratio. The hybrids could be recommended according to their potential utilization in the seed industry.     

Methylene blue increases the tolerance of tomato plants to abiotic stresses

Summary

Environmental stresses such as high temperature and high salinity are known to cause oxidative stress in plants by enhancing the generation of reactive oxygen species (ROS), which have a strong impact on plant development. Mitochondria constitute one of the major sources of ROS in roots. Recently, it has been shown in mammalian systems that methylene blue (MB) at very low concentrations can attenuate mitochondrial ageing by scavenging ROS. In the present study, we tested whether MB could be used to protect tomato (Lycopersicon esculentum L.) seedlings against the harmful effects of high salinity and high root temperature. Tomato seedlings were grown hydroponically and were subjected to two abiotic stresses: high salinity (150 mM NaCl) in the nutrient medium, or high root temperature (35ºC) for 14 d. These stress treatments were applied with or without the addition of 10^–8 M MB. The results showed that, under normal conditions, MB enhanced root and shoot growth, increased root respiration, decreased root H₂O₂ and malonyldialdehyde (MDA) concentrations, and enhanced root superoxide dismutase (SOD) activity. Both high salinity and high root temperature stress impaired root and shoot growth, and caused an enhancement of root oxidative stress. MB had a significant protective effect against both abiotic stresses and restored the levels of oxidative stress-related components (i.e., H₂O₂ and MDA) in root and leaf tissues almost to normal levels. It is hypothesised that MB could confer its protective effect not only through interactions with root mitochondria, but also via additional cellular targets in the root.     

Structured analysis of seepage losses in irrigation supply channels for cost-effective investments: case studies from the southern Murray-Darling Basin of Australia

Much of inland Australia has been in perpetual drought since 1997 except during 2010 when above average rainfall occurred. It has been the worst drought since 1788 when European settlement began. Water scarcity poses a serious threat to the sustainability of the irrigated agriculture in major irrigation systems across the Murray-Darling Basin (MDB). There is a need for water-saving measures and a structured approach to assess water loss in earthen supply channels. This paper presents such an approach to assess and reduce seepage losses for improving irrigation efficiencies. Main elements of this approach are the following: field measurements, hydrologic modelling, potential options for seepage reduction, economic analysis and financing water-saving investments. Using data from two irrigation systems in the southern MDB, a case is made for reducing seepage water losses in irrigation supply channels in a cost-effective manner using low-cost technologies. Increasing the level of security for investments in water-saving programs provides incentives to key stakeholders to achieve water-saving targets. Considering the value of water recovered from reducing seepage loss at irrigation system level, this study demonstrates how reducing just one component (seepage) from the total water losses in irrigation systems can help improve water supplies as well as the environmental flows. Potential options for financing infrastructure improvement for saving irrigation water are proposed and discussed.     

Comparison of sulfurous acid generator and alternate amendments to improve the quality of saline-sodic water for sustainable rice yields

The prolonged irrigation with marginal quality water can cause secondary salinization of soils, which necessitates for better understanding of water management alternatives. Relative performance of sulfuric acid and gypsum is still controversial to counter sodium hazards in soil/water system. As an alternative, sulfurous acid generators (SAG) are also being marketed. But up-till-now, there is not even a single field study published in scientific journals about their efficiency and economical viability for the treatment of saline-sodic water. Therefore, a field study was carried out to compare the effectiveness of SAG and alternate amendments applied on an equivalent basis to grow rice crop. SAG treatment of saline-sodic tube well water decreased only residual sodium carbonate (RSC) from 5.4 to 3.6 mmol_c l⁻¹, and had no beneficial effect on its sodium adsorption ratio (SAR) or electrical conductivity (EC). All the treatments kept soil EC and SAR around their respective threshold levels. For paddy yield, SAG, sulfuric acid, and gypsum treatments depicted nonsignificant differences. SAG and sulfuric acid treatments of water were about six times expensive than that of gypsum. It was concluded that soil-applied gypsum, to counter sodic hazards of irrigation water, is economical to sustain irrigated rice in dry regions.     

Microscopic, serologic and molecular surveys on Dirofilaria immitis in stray dogs, Turkey

The prevalence of Dirofilaria immitis infection was evaluated in stray dogs of Erzurum, Turkey. A total of 123 whole-blood and 93 sera samples were collected from stray dogs older than 6 months were lived in animal shelter. The PCR and direct microscopic examinations were used for the detection of microfilaria and indirect-ELISA was performed for the detection of anti-D. immitis antibodies. The prevalence of D. immitis in the canine population was 8.1% by PCR, 2.1% by ELISA. In addition, microfilaria burdens of Dirofilaria sp. was 4.8% by direct blood smear examination. There was a statistical difference (P=0.05) in the prevalence between males (10.5%) and females (2.3%) by direct blood smear examination. Similarly there was a statistical difference (P<0.05) in the prevalence between males (15.8%) and females (4.7%) by PCR. Dogs belonging to the 0.5-1 years old group showed the highest prevalence than 2-4 ages group with three tests. Among the 93 samples screened by the ELISA, two samples were positive for the D. immitis antibodies. Both positive dogs with ELISA were females.     

Preparation and antinutritional characteristics of dry bean (Phaseolus vulgaris L.) protein concentrates

Protein concentrates and starches were prepared by a wet extraction process from five dry bean (Phaseolus vulgaris L.) cultivars. The protein contents ranged from 69.7–76.4%. Concentrates prepared from dehulled beans under similar conditions had higher protein contents (80.6–87.9%). Each additional washing of the concentrates with distilled water increased their protein content. However, the protein recovery progressively decreased. The yield of starch ranged from 48.0–51.1% of the starting material. The solubility of bean proteins was minimal at pH 4.0, and under alkaline conditions, it was influenced by the tannin contents of the concentrates. Protein concentrates had lower trypsin, chymotrypsin, and amylase inhibitory activities as well as lower phytic acid and tannin contents compared to whole bean flours.