Molecular detection and characterization of ovine herpesvirus-2 using heminested PCR in Pakistan

BackgroundMalignant catarrhal fever (MCF) is a highly fatal lymphoproliferative disease of cattle, deer, bison, water buffalo, and pigs caused by the gamma-herpesviruses alcelaphine herpesvirus-1 (AlHV-1) and ovine herpesvirus-2 (OvHV-2).ObjectivesThis study aimed to determine the prevalence of OvHV-2 in sheep, goats, cattle, and buffalo in Rawalpindi and Islamabad, Pakistan, by applying molecular and phylogenetic methods.MethodsBlood samples were aspirated from sheep (n = 54), goat (n = 50), cattle (n = 46) and buffalo (n= 50) at a slaughterhouse and several farms. The samples were subjected to heminested polymerase chain reaction (PCR), followed by sequencing and phylogenetic analysis of the OvHV-2 POL gene and the OvHV-2 ORF75 tegument protein gene.ResultsThe highest percentage of MCF positive samples was in sheep (13%), whereas goat, cattle, and buffalo had lower positive percentages, 11%, 9%, and 6.5%, respectively. Four OvHV-2-positive PCR products obtained from sheep samples were sequenced. The sequences obtained were submitted to the NCBI GenBank database ({"type":"entrez-nucleotide","attrs":{"text":"MK852173","term_id":"1694854431","term_text":"MK852173"}}MK852173 for the POL gene; {"type":"entrez-nucleotide","attrs":{"text":"MK840962","term_id":"1686143390","term_text":"MK840962"}}MK840962, {"type":"entrez-nucleotide","attrs":{"text":"MK852171","term_id":"1694854427","term_text":"MK852171"}}MK852171, and {"type":"entrez-nucleotide","attrs":{"text":"MK852172","term_id":"1694854429","term_text":"MK852172"}}MK852172 for the ORF75 tegument protein gene). Phylogenetic analysis revealed a close similarity of study sequences with those of worldwide samples.ConclusionsThis study is the first cross-sectional study on the prevalence and molecular detection of OvHV-2 in apparently healthy cattle and buffalo that could be carrying OvHV-2 acquired from OvHV-2-positive sheep and goats. The results indicate that OvHV-2 is circulating in Pakistan. Further studies are needed to characterize OvHV-2 and elucidate further its prevalence.     

Shrub Encroachment Under the Trees Diversifies the Herb Layer in a Romanian Silvopastoral System

Rangelands with scattered trees are complex and dynamic systems with a long history in Europe. Generally referred to as “wood-pastures,” they are considered to have outstanding conservation value. Thorny shrubs are important for supporting the biodiversity of these wooded rangelands, as well as facilitating the regeneration of trees by acting as nurse species. We assess the direct effects of temporary shrub encroachment under the cover of mature sparse trees on overall plant and habitat diversity. We surveyed the herb layer of the main landscape features of a wood-pasture: open pasture, trees with a grass understory, trees with shrubs, and adjacent forest edges. The herb layer under trees with shrubs resembled that of forest edges more than open pastures and trees with grass. Trees with grass had a higher cover of ruderal species than trees with shrubs, while forest edges and open pastures had a lower cover of them. Forest species were absent from open pastures but were well represented in the other sites. The herb layer of trees with shrubs and forest edges had similar cover values, while trees with grass had a significantly lower cover of herbs than the other types. Trees with shrubs had higher species richness than any of the other three landscape features and had a much higher proportion of diagnostic species. We conclude that the scattered trees and shrubs of the studied silvopastoral system have additive facilitative effects on their understory, probably through modifying the microenvironment and grazing pressure, leading to the formation of temporary diversity hot spots with distinct vegetation. Thus maintaining a moderate level of shrub-encroachment under sparse trees is recommended for not only creating safe havens for tree recruitment but also increasing the overall species and habitat diversity of wood-pastures.     

Quantitative assessment of metals in local brands of tea in Pakistan

In present study, Mn, Fe, Zn, Cu, Co, Pb, Cr, Ni and Cd were analyzed by FAAS in green and black tea samples of locally available in the Pakistani market. Na and K were also determined by Flame Photometer. Tea leaves can be the source of mineral components and trace elements, as well as some undesirable substances due to exposure to the environment. Among the metals tested, K was the most abundant one followed by Na, Mn and Fe. Fortunately, toxic heavy metals, Pb and Cd, had the lowest contents in tea samples and also in tea aqueous extracts. Concentration of heavy metals in tea aqueous extract was markedly lower than their total contents except that of K. The solubility of studied metals in tea aqueous extract varied widely and ranged from 0.0-95%. The lowest ranges of solubility were listed for toxic heavy metals Pb and Cd. The possible uptake of metals by the human body from tea aqueous extract has also been determined. The amounts of metals that one may take up through consumption of tea aqueous extract were found to match the acceptable daily intake even taking into account exposure from air, food and drinking water.     

Growth and yield of lowland rice as influenced by potassium application and cultivation method under alternate wetting and drying water regime

Besides being an essential macronutrient for plant growth, potassium (K) also acts as a stress-relieving agent against various biotic and abiotic stresses, especially water stress. An experiment consisting of three lowland Thai rice varieties (Pathumthani 1, RD57, RD41), two cultivation methods [dry direct seeding (DDS), transplanting (TP)] and four K doses (0, 80, 120,160?kg ha^?1) under alternate wetting and drying (AWD) water regime was conducted to investigate the impact of K and cultivation method on lowland rice varieties subjected to AWD. Pathumthani 1 had significantly higher grain yield at the 120?kg K ha^?1 compared with all other K doses. RD57 and RD41 had maximum grain yield at the 80 and 120?kg K ha^?1, respectively, which was significantly higher than the control; however, grain yield among different K doses was mostly similar for RD57 and RD41. K application at the rate of 120?kg ha^?1 for Pathumthani 1 and 80?kg ha^?1 for RD57 and RD41 could be a feasible option. The performance of DDS and TP was better at the 80 and 120?kg K ha^?1, respectively. Application of K at the optimum dose with proper selection of variety and cultivation method could help in sustainable rice production.     

Phytochemical investigation and in vitro antinociceptive activity of Clerodendrum indicum leaves

The crude ethanolic extracts of Clerodendrum indicum Linn. leaves were investigated for possible antinociceptive activity using acetic acid induced writhing model in mice. Phytochemical analysis was also carried out according to the standard procedures to identify the presence of different phytoconstituents in the ethanolic extract of the plant leaves. The study results showed 38.91 and 55.24% inhibition of writhings in the tested mice when ethanolic extract of Clerodendrum indicum Linn. leaves at doses of 250 and 500 mg kg(-1) body weight was given intraperitoneally, respectively. The study results were also compared with antinociceptive activity of the standard drug, Diclofenac sodium (68.37% inhibition) used at 25 mg kg(-1) body weight. At the above doses, the crude ethanolic extract of the plant showed significant antinociceptive activity in dose dependent fashion in acetic acid-induced writhing model in mice. The inhibition of writhings was calculated in respective to control group and it was found that p-values (<0.0001) obtained in all cases were extremely statistically significant. However, the phytochemical analysis showed the presence of alkaloid, steroid, saponin, tannin, reducing sugar and gum. The results suggest that crude ethanolic extracts of Clerodendrum indicum leaves possess significant antinociceptive properties justifying its folkloric use as analgesics and further research is necessary to isolate the principle phytochemical constituent(s) responsible for this activity.     

Alphaherpesvirus-vectored vaccines against animal diseases: Current progress

Recombinant virus-vectored vaccines are novel agents that can effectively activate specific and nonspecific immunity, are multivalent and multieffective, and have high safety ratings. Animal alphaherpesviruses have a large genome, contain multiple nonessential regions that do not affect viral replication and are capable of accepting the insertion of an exogenous gene and expressing the antigen protein. Furthermore, animal alphaherpesviruses have a wide host spectrum, can replicate in the host and continuously stimulate the animal to produce immunity to the corresponding pathogen, thus making them ideal carriers for recombinant virus-vectored vaccines. With the development of gene-editing technology, recombinant viruses capable of expressing foreign genes can be constructed by various methods. Currently, studies on recombinant virusvectored vaccines constructed based on animal alphaherpesviruses have involved poultry, pigs, cattle, sheep, and companion animals. Studies have shown that the construction of recombinant animal alphaherpesviruses enables the acquisition of immunity to multiple diseases. This article mainly summarizes the current progress on animal alphaherpesvirus-vectored vaccines, aiming to provide reference for the development of new animal alphaherpesvirus-vectored vaccines.     

Water status and drought stress in experimental gaps in managed and semi-natural silver fir--beech forests

The effect of gap characteristics, including gap age, vegetation cover, and stage of regeneration, on water balance and drought stress was studied over a four-year period in natural and experimental gaps of various sizes in Dinaric silver fir--beech forest in Slovenia. A medium gap, two experimental clear-cut gaps, and a closed stand were selected in a traditional irregular shelterwood managed forest, and an irregular-shaped natural gap and part of the stand were selected in a semi-natural forest. Soil moisture, throughfall, and stemflow were measured in the gaps and stands, and incident precipitation was monitored in the open area. Evapotranspiration and drainage water fluxes were estimated using the water balance model BROOK90. To estimate drought stress, the ratio between simulated actual and potential evapotranspiration was used. Precipitation in the 2001 and 2003 growing seasons was considerably less than that in 2002 and 2004. The highest drainage fluxes were in the newly created clear-cut gaps with sparse ground vegetation cover, followed by the natural gap and medium gap, which had older and denser natural tree regeneration. On average, the clear-cut gap drainage fluxes were 18% (164?mm) higher than those in the forest stands. Evapotranspiration was lower in the natural gap and higher in the managed gap compared with the experimental clear-cut gaps. Water supply stress was greater for the forest stands than for the vegetation in the gaps. Drought stress was indicated to be lowest in the natural gap, which had patches of varying development phases and the highest water storage capacity of the soil and vegetation.     

Influence of crop residue management, cropping system and N fertilizer on soil N and C dynamics and sustainable wheat (Triticum aestivum L.) production

Management of N is the key for sustainable and profitable wheat production in a low N soil. We report results of irrigated crop rotation experiment, conducted in the North West Frontier Province (NWFP), Pakistan, during 1999–2002 to evaluate effects of residue retention, fertilizer N application and mung bean (Vigna radiata) on crop and N yields of wheat and soil organic fertility in a mung bean–wheat sequence. Treatments were (a) crop residue retained (+residue) or (b) removed (−residue), (c) 120 kg N ha⁻¹ applied to wheat, (d) 160 kg N ha⁻¹ to maize or (e) no nitrogen applied. The cropping system was rotation of wheat with maize or wheat with mung bean. The experiment was laid out in a spit plot design. Postharvest incorporation of crop residues significantly (p < 0.05) increased the grain and straw yields of wheat during both years. On average, crop residues incorporation increased the wheat grain yield by 1.31 times and straw yield by 1.39 times. The wheat crop also responded strongly to the previous legume (mung bean) in terms of enhanced grain yield by 2.09 times and straw yield by 2.16 times over the previous cereal (maize) treatment. Application of fertilizer N to previous maize exerted strong carry over effect on grain (1.32 times) and straw yield (1.38 times) of the following wheat. Application of N fertilizer to current wheat produced on average 1.59 times more grain and 1.77 times more straw yield over the 0 N kg ha⁻¹ treatment. The N uptake in wheat grain and straw was increased 1.31 and 1.64 times by residues treatment, 2.08 and 2.49 times by mung bean and 1.71 and 1.86 times by fertilizer N applied to wheat, respectively. The soil mineral N was increased 1.23 times by residues, 1.34 times by mung bean and 2.49 times by the application of fertilizer N to wheat. Similarly, the soil organic C was increased 1.04-fold by residues, 1.08 times by mung bean and 1.00 times by the application of fertilizer N. We concluded that retention of residues, application of fertilizer N and involvement of legumes in crop rotation greatly improves the N economy of the cropping system and enhances crop productivity in low N soils.     

Biovolatilization of Arsenic by Different Fungal Strains

The quantification of arsenic biovolatilization by microscopic filamentous fungi Aspergillus clavatus, A. niger, Trichoderma viride and Penicillium glabrum under laboratory conditions is discussed in this article. The fungi were cultivated on a liquid medium enriched with inorganic arsenic in pentavalent form (H₃AsO₄). Filamentous fungi volatilized 0.010 mg to 0.067 mg and 0.093 mg to 0.262 mg of arsenic from cultivation systems enriched with 0.25 mg (5 mg.l^?1 of arsenic in culture media) and 1.00 mg of arsenic (20 mg.l^?1 of arsenic in culture media), respectively. These results represent the loss of arsenic after a 30-day cultivation from cultivation systems. The production of volatile arsenic derivatives by the A. niger and A. clavatus strains was also determined by hourly sorption using the sorbent Anasorb (CSC) on the 29th day of cultivation.