Isolation and identification of bubaline herpesvirus 1 (BuHV-1) from latently infected water buffalo (Bubalus bubalis) from Iran

In order to isolate buffaloes herpesvirus 1 (BuHV-1) from latently infected water buffalo (Bubalus bubalis), 16 buffalo heifers were selected from a herd. At first, animals were bled and their sera were tested by virus neutralization (VN) test, using bovine herpesvirus 1 (BoHV-1). According to the results of VN test and dexamethasone injection (0.1 mg/kg BW) for 5 consecutive days, the examined buffaloes were divided into 4 groups. Vaginal and nasal swabs were daily collected from all buffaloes from day 0 to 10 days later. Based on the cytopathic effects in cell culture, a herpesvirus was isolated only from nasal swabs of three seropositive buffaloes which they had received dexamethasone. The nasal swabs of these three buffaloes were also positive in PCR, using primers specific for ruminant herpesviruses gD gene. The identity of the isolated viruses was determined according to partial amino acid sequences of gD, deduced from the nucleotide sequences of the PCR products. On the basis of sequence alignment, phylogenetic analysis, and genetic distances, the three buffalo virus isolates were more closely related to BuHV-1 and BoHV-5 than to BoHV-1.

     

Salinity effects on osmoregulation and gill morphology in juvenile Persian sturgeon (<Emphasis Type="Italic">Acipenser persicus</Emphasis>)

The effect of abrupt and 5-day gradual salinity transfers from freshwater (FW) to 11 ‰ Caspian Sea brackish water (BW) was investigated in juvenile Persian sturgeon Acipenser persicus with three different weight groups: 1–2 g (1.62 ± 0.27 g), 2–3 g (2.55 ± 0.41 g) and 3–5 g (4.28 ± 0.76 g). Mortality rates, blood osmotic pressure, gill morphology and branchial Na⁺, K⁺-ATPase (NKA) activity were measured 4 and 10 days after abrupt transfer and 9 and 15 days after the initial gradual transfer (i.e. 4 and 10 days after reaching Caspian Sea salinity). Fish under 3 g could not survive increased salinity, and the blood osmotic pressure of the remaining surviving fish increased and remained elevated. However, heavier fish were able to survive and successfully acclimate, even to rapid salinity change with osmotic pressure reduced to Caspian Sea osmolality levels. At the gill level, the developmental increase in chloride cell volume and a higher NKA content most probably allow juveniles weighing more than 2 g to sharply increase NKA activity if the fish are transferred to BW. The rapid chloride cell proliferation occurring with increased salinity should strengthen this acclimation response. Therefore, a drastic physiological change occurs when fish weigh more than 2 g that allows migration to higher salinities. The triggering signal on chloride cells must be further investigated in order to optimize this functional step.     

Allelic segregation of sex‐linked microsatellite markers in Nile tilapia (Oreochromis niloticus) and validation of inheritance in YY population

Stocking of all‐male fingerling produced by direct administration of male hormone 17‐α‐methyltestosterone is the most preferred method for present‐day aquaculture of the Nile tilapia Oreochromis niloticus. However, due to the growing concern of negative impact of steroid hormone in food fish, production of ‘genetically male’ tilapia, which depends on the concrete and thorough understanding of sex determination, has long been a scientific curiosity. The objective of the present study was to identify reliable sex‐linked markers and to evaluate the applicability of those markers in terms of monosex production approach. ‘XY’ neofemales were produced by using synthetic oestrogen and identified through selective breeding and progeny testing. Three females with progeny not deviating from 3:1 sex ratio (male:female) were designated as ‘XY’ neofemales and were used subsequently to produce putative YY progeny. Among the fifteen microsatellite markers tested, marker ARO172 was most informative in differentiating male and female genotypes. Twenty‐seven F₂ fish from three families were identified as putative YY males based on marker genotyping, and four of them were crossed to produce F₃ to validate marker association by progeny testing. The YY males produced 86%–100% male progeny indicating ARO172 a unique sex‐linked marker applicable in marker‐assisted selection.     

Investigation of the efficiency of direct and indirect regeneration in evening primrose (Oenothera biennis)

As a medicinal plant, the importance of evening primrose (Oenothera biennis L.) is due to its unsaturated fatty acids in the seeds and roots, and also oenotherine and comfarol in the leaves. Low germination and difficulties in seed production are the main problems encountered with growing this plant in the field. As an alternative approach, an in vitro experiment was set up for the evaluation of evening primrose production via direct and indirect regeneration of the cultivars NC-1 and VNK. For callogenesis and direct regeneration, the explants from the apical bud and petiole were cultured on MS medium supplemented with 0.25, 0.75, and 1.25 mg L^?1 of both BAP and Kinetin (KIN). Indirect regeneration was performed by placing apical buds, petioles, and leaf explants on MS medium supplemented with 0.5 and 1 mg L^?1 2,4-D and 0.5, 1, and 1.25 mg L^?1 of both BAP and KIN. The highest shoot induction from direct regeneration was obtained with apical bud explants of VNK treated with 0.75 mg L^?1 BAP. The highest callus weight (3.17 g) obtained from indirect regeneration was with petiole explants treated with 1 mg L^?1 2, 4-D and 1 mg L^?1 BAP in VNK cultivars. The highest number of torpedo embryogenic clusters (23.8) was obtained from the VNK petiole explants treated with 0.5 mg L^?1 2, 4-D and 1.25 mg L^?1 BAP. BAP had higher positive effects on in vitro production of evening primrose than KIN in both direct and indirect regeneration. In general, results indicated that VNK was more potent for regeneration than NC-1 and concentrations of 0.75 mg L^?1 BAP for direct and 0.5 mg L^?1 2, 4-D and 1.25 mg L^?1of BAP for indirect regeneration had a higher efficiency for increasing in vitro production of evening primrose.     

Molecular characterisation of <Emphasis Type="Italic">Turnip mosaic virus</Emphasis> isolates from Brassicaceae weeds

Eight provinces of Iran were surveyed during 2003–2008 to find Brassicaceae reservoir weed hosts of Turnip mosaic virus (TuMV). A total of 532 weed samples were collected from plants with virus-like symptoms. The samples were tested for the presence of TuMV by enzyme-linked immunosorbent assay using specific antibodies. Among those tested, 340 samples (64%) were found to be infected with TuMV. Rapistrum rugosum, Sisymberium loeselii, S. irio and Hirschfeldia incana were identified as the Brassicaceae weed hosts of TuMV, and the former two plant species were found to be the most important weed hosts for the virus in Iran. The full-length sequences of the genomic RNAs of IRN TRa6 and IRN SS5 isolates from R. rugosum and S. loeselii were determined. No evidence of recombination was found in both isolates using different recombination-detecting programmes. Phylogenetic analyses of the weed isolates with representative isolates from the world showed that the IRN TRa6 and IRN SS5 isolates fell into an ancestral basal-Brassica group. This study shows for the first time the wide distribution and phylogenetic relationships of TuMV from weeds in the mid-Eurasia of Iran.     

Evaluation of <Emphasis Type="Italic">Aloe vera</Emphasis> Gel as an Alternative Edible Coating for Peach Fruits During Cold Storage Period

Application of bio-based edible coatings, which are in line with sustainable agriculture practices, has been a hot topic in recent years. In order to evaluate the effect of Aloe vera L. gel coating, on the store ability of peach fruits, a 30-day postharvest experiment with 10-day sampling intervals was performed. A. vera gel coating had significant positive effects on the weight loss, color change, and sensory evaluation. The amount of weight loss, color change, Total Soluble Solids (TSS) and Titratable Acidity (TA) in coating was lower than control. Furthermore, our results showed that A. vera gel coating can enhance visual properties, and could also lead in more favourable taste and texture. Regarding the obtained results, it can be suggested that A. vera gel coating can enhance the qualitative and quantitative post-harvest traits of fruits, hence, it could be a suitable alternative for chemicals preservative in commercial storage of peach fruits.     

Potential of <Emphasis Type="Italic">Trichoderma</Emphasis> spp. and <Emphasis Type="Italic">Talaromyces flavus</Emphasis> for biological control of potato stem rot caused by <Emphasis Type="Italic">Sclerotinia sclerotiorum</Emphasis>

Sixteen isolates belonging to 11 species of Trichoderma (T. asperellum, T. ceramicum, T. andinensis, T. orientalis, T. atroviride, T. viridescens, T. brevicompactum, T. harzianum, T. virens, T. koningii and T. koningiopsis) were evaluated for biological control of potato (Solanum tuberosum) stem rot caused by Sclerotinia sclerotiorum. In dual culture tests, all antagonists significantly reduced sclerotia formation, and were able to inhibit radial growth of the pathogen. Growth inhibition by production of volatile and non-volatile inhibitors was also measured in in vitro tests. In screening the most efficient species of Trichoderma, establishment of mycelium on sclerotia and sclerotia lysis were also considered as important biocontrol qualities. Excluding T. asperellum, T. brevicompactum, T. andinensis and T. harzianum, all tested Trichoderma species were able to lyse sclerotia. The sclerotia-destroying species of Trichoderma and one isolate of Talaromyces flavus were tested in greenhouse tests and during 2 years of field experimentation during the 2007 and 2008 cropping seasons. After one aerial application of spore suspension in greenhouse trials, T. koningii, T. virens, T. ceramicum and T. viridescens were the most effective bio-agents and reduced significantly disease severity, and the least biocontrol efficacy was observed in T. flavus. Under field conditions and after five soil and foliar applications of spore suspension, all tested antagonists reduced significantly disease incidence. T. viridescens followed by T. ceramicum showed the best results. T. flavus and T. orientalis were less effective than other tested antagonists in both field trials.     

<Emphasis Type="Italic">Fusarium ershadii</Emphasis> sp. nov., a Pathogen on <Emphasis Type="Italic">Asparagus officinalis</Emphasis> and <Emphasis Type="Italic">Musa acuminata</Emphasis>

Two Fusarium strains, isolated from Asparagus in Italy and Musa in Vietnam respectively, proved to be members of an undescribed clade within the Fusarium solani species complex based on phylogenetic species recognition on ITS, partial RPB2 and EF-1α gene fragments. Macro- and micro-morphological investigations followed with physiological studies done on this new species: Fusarium ershadii sp. nov can be distinguished by its conidial morphology. Both isolates of Fusarium ershadii were shown to be pathogenic to the monocot Asparagus officinalis when inoculated on roots and induced hollow root symptoms within two weeks in Asparagus officinalis seedlings. In comparison mild disease symptoms were observed by the same strains on Musa acuminata seedlings.     

Influence of zinc sulfate and municipal solid waste compost on chemical forms of zinc in calcareous soils

Zinc (Zn) is an essential micronutrient for crop growth. This metal can be found in chemical forms or fractions in the soil. The objective of this study was to investigate the distribution of Zn in special chemical forms using the sequential extraction method after treating eight calcareous soils by zinc sulfate and municipal solid waste (MSW) compost. Zn was separated in seven defined forms as exchangeable (Ex), carbonate (Car), organic (Om), manganese oxide bound (MnOX), amorphous iron oxide bound (FeAOX), crystalline iron oxide bound (FeCOX), and residual (Res). According to the results, the mean concentrations of Zn in chemical forms in untreated soils from higher to lower were 31.84, 8.13, 2.64, 2.57, 0.45, 0.39, and 0.16?mg?kg^?1 for the Res, FeCOX, Car, FeAOX, Om, MnOX, and Ex forms, respectively. The total applied Zn to the studied soils from both the sources of zinc sulfate and MSW compost after incubation for 30 days was converted to chemical forms in the following order: Car?>?Res?>?FeCOX?>?Om?>?FeAOX?>?MnOX?>?Ex. On average, 30.3% of total Zn from zinc sulfate and 28.8% of total Zn in the MSW compost were converted into the Car form. The high content of calcium carbonate (CaCO₃) in the studied soils is an important factor affecting the conversion of Zn forms, mostly in the Car form through the application of both the inorganic and the organic sources of Zn.     

Methoxyfenozide and pyriproxifen alter the cellular immune reactions of Eurygaster integriceps Puton (Hemiptera: Scutelleridae) against Beauveria bassiana

Effects of the two insect growth regulators (IGRs) methoxyfenozide, 20-hydroxyecdysone (20E) agonist, and pyriproxifen, Juvenile hormone (JH) agonist, were examined on the cellular immune responses of the Sunn pest, Eurygaster integriceps versus the entomopathogenic fungus Beauveria bassiana. The simultaneous treatment with the IGRs and the fungal spores altered haemocyte count (total and differentiate), nodulation response and phenoloxidase (PO) activity in a dose- and time-dependent manner. It was observed that different concentrations of methoxyfenozide increased total and differentiate haemocyte numbers as well as B. bassiana-induced nodulation response. In contrast with the JH agonist, pyriproxifen significantly decreased total and differentiate haemocyte numbers and inhibited nodule formation in E. integriceps adults. The 20E agonist displayed major effects when injected at the doses 2.79 and 5.59 μg/mg adult. In contrast, injecting adults by pyriproxifen significantly impaired their ability to raise an efficacious response against the fungal spores. The ability of the two IGR analogues to interfere with activity of the PO system in haemolymph of E. integriceps adults was also investigated 6 h after injection by fungal spores. Methoxyfenozide had an excitatory effect on PO activity when the 5.59 μg/mg concentration was used against adults. Conversely, pyriproxifen had an inhibitory effect on PO activity when used at 1.49 μg/mg adult concentration. These findings demonstrate that pyriproxifen may interfere with cell-mediated immunity of E. integriceps. So, pyriproxifen could be a good candidate for the integrated control of the Sunn pest.