Expression of the herpes simplex virus type 2 glycoprotein D in baculovirus expression system and evaluation of its immunogenicity in guinea pigs

Background: Herpes simplex virus type 2 (HSV-2) is highly prevalent and major cause of genital herpes in humans. The life-long nature of infection and the increasing prevalence of genital herpes imply that vaccination is the best strategy for controlling the spread of infection and limiting HSV disease. HSV glycoprotein D (gD) is one of the most important viral immunogen which has an essential role in virus infectivity and induction of immune responses. Methods: HSV-2 DNA was extracted and used as template in polymerase chain reactions to amplify gD2 gene. The PCR product was confirmed by restriction enzyme analysis, cloned into a cloning vector and then sequenced. The Bac-to-Bac expression system was used to express HSV-2 gD in insect cells. The expressed protein was used as subunit vaccine to immunize guinea pigs after confirmation. Results: The expressed protein was confirmed with SDS-PAGE and Western-blot analysis. In Western-blot analysis, two major protein bands, with approximate molecular weights of 52-55 and 41-43 kDa corresponding to the glycosylated and non-glycosylated forms of gD2 protein, were observed, respectively. Immunization with the recombinant gD2 could elicit humoral responses in guinea pigs as measured by neutralization test and ELISA, and offered high protection against induced HSV-2 genital disease. Conclusion: The baculovirus expression of heterologous genes permits proper folding, post-translational modification and oligomerization in manners that are often identical to those that occur in mammalian cells. Expression of proteins under the control of the strong polyhedrin promoter, allowing high level protein production, can be used as subunit vaccine.     

Identification of wood‐decay fungi and assessment of damage in log depots of Western Black Sea Region (Turkey)

The aim of this study was to determine and quantify the wood‐decay fungi found on logs of forest tree species (beech, oak, hornbeam, Scots pine and fir) stored in log depots located in six different provinces in the Western Black Sea Region of Turkey. Additionally, it was aimed to determine the natural durability of some important wood species against the most commonly detected wood‐decay fungi. Eighteen families, 31 genera and 45 species belonging to the division Basidiomycota were detected; Antrodia crassa was identified for the first time in Turkey. The abundance of Panus neostrigosus, Polyporus meridionalis, Trametes hirsuta, T. versicolor and Stereum hirsutumincreased significantly with the holding time of the logs (r = 0.99, 0.87, 0.53, 0.57 and 0.78, respectively, p < 0.05). The majority of the fungal species were detected on logs stored in depots for 4–6 years (66%). The percentage of fungal species found on the logs with a holding time of three years or less was 29%, whereas the percentage for those detected on logs stored for seven or more years was 31%. Among the wood species, the greatest number of fungal species (29) and highest amount of fungi (2,539) occurred on beech wood. Natural durability tests showed that T. versicolor caused the greatest loss of wood mass, with an average of 23%. Field studies and natural durability tests performed in the laboratory showed that beech wood lost the most mass among the timber species studied.     

新疆准东矿区土壤与降尘重金属空间分布及关联性分析

土壤重金属与大气降尘重金属之间的关联性可以反映重金属污染的来源、土壤-大气系统中重金属的传输、迁移和扩散特征。为了研究矿区表层土壤的污染状况及与降尘重金属质量分数间的关联性,该研究以新疆维吾尔自治区准东矿区为研究靶区,利用2014年采集的51个表层土壤和大气降尘样品的室内实测重金属质量分数数据,并基于此分析了样品中6种重金属(As、Cu、Cr、Hg、Pb、Zn)的空间分布特征、地积累指数以及潜在生态风险;利用Pearson相关性分析矩阵和灰色关联法对表层土壤与大气降尘中重金属浓度的相关性和关联度进行探讨。结果表明:1)准东矿区表层土壤重金属的分布状况存在着明显的空间差异,其中Hg的污染程度最严重,处于强-极强度污染,其潜在生态危害指数达到了较高生态风险;Zn和Cu基本处于无污染状态,属于轻微生态风险;2)大气降尘重金属空间分布存在着明显差异,降尘中的Zn处于极强度污染,Hg处于无污染状态;3)Pearson相关分析与灰色关联分析表明,准东地区表层土壤与大气降尘中6种重金属的相关系数大小顺序与关联度排序结果一致,其中Hg和As元素具有较强的一致性,且具有相同的来源,说明大气降尘对表层土壤中重金属的质量分数有一定影响。但因不同重金属元素沉降特性不同,导致各元素之间的关联度有所差异。     

Clutch size decisions of <Emphasis Type="Italic">Cotesia glomerata</Emphasis>, a gregarious parasitoid of <Emphasis Type="Italic">Pieris brassicae</Emphasis>

This work investigated the ability of the gregarious larval endoparasitoid Cotesia glomerata L. (Hymenoptera: Braconidae) to adjust the progeny sex ratio and clutch size with repeated oviposition experience and the effect of a honey-based diet on the clutch size. In the field-collected clusters many clusters were female-biased but some clusters (3.8%) produced only male wasps, suggesting that there is a low percentage of unmated females in the field. Superparasitism was common in the field, and females were believed to increase progeny sex ratio when attacking previously parasitized hosts. In the laboratory, the number of eggs laid in a day tended to decrease with increasing female age. For females that were offered two hosts per day and for those offered three hosts per day, this value became nearly the same at 9 days after the start of oviposition. Old females which attacked many hosts tended to lay fewer eggs in a day than young ones. However, the degree of this tendency was not the same for all the parasitoid females of all three groups because sperm remained viable throughout a female’s lifetime. The amount of sperm used in a single oviposition bout seemed fixed and was not dependent on the number of eggs laid. Over the 2 days of the clutch size response experiment, the number of hosts a female attacked per day was not affected by the presence or absence of honey.     

Reproductive performance of Holstein dairy cows in Iran

The objectives of the present study were to describe the current reproductive parameters of Holstein dairy cows in Iran and to determine factors which have a significant effect on reproductive performance with emphasis on high-producing dairy cows. Five large Holstein dairy herds from the major milk production areas of Iran were selected for this study. Reproduction data including parity of dam, calving date, days to first service, days open, number of services, dry period, and occurrence of diseases for cows that calved from 2004 to 2007 were collected from the herds' databases. To determine the effects of year and season of calving, disease status, level of milk production, and parity of dam on reproductive parameters, the proc-mixed procedure in SAS software was used. Overall, data about 8,204 lactation were collected for the study. Results showed that mean (±SD) days open, calving interval, and days to first service for study herds were 134 (±89), 403 (±86), and 67 (±38) days, respectively. Conception rates at the first service and the overall service-conception rate were 41.6% and 41%, respectively. The level of milk production and diseases had significant negative effects on days open and service per conception (P < 0.05); statistical analysis showed that for every 100-kg increase in milk yield, days open will increase by about 0.3 days. However, no significant effect of level of productivity was observed on days to first service. The results of this study indicate that high milk production is a risk factor for decreasing fertility in Iran, like many other parts of the world.     

Subclinical mastitis causes alterations in nitric oxide, total oxidant and antioxidant capacity in cow milk

The aim of this study was to investigate total antioxidant (TAC), and oxidant capacity (TOC) and nitric oxide (NO) levels in milk of cows with subclinical mastitis. Brown Swiss and Holstein breed cows were screened with California Mastitis Test (CMT) to determine mammary glands with subclinical mastitis. Moreover, somatic cell counts (SCC) were determined electronically in all milk samples. Mammary quarters were classified as healthy (n = 25) or subclinical mastitis (n = 35) based on CMT scores and somatic cell count (SCC: ?200,000/ml or >200,000/ml) in milk. Nitric oxide, TOC and SCC levels were significantly higher (p < 0.001, p < 0.005 and p < 0.001, respectively) in milk from mammary quarters with subclinical mastitis compared to those from healthy mammary quarters. In conclusion, subclinical mastitis results in higher NO concentrations, TOC and SCC, and NO and TOC were positively correlated with SCC. Moreover, alterations in NO levels and TOC in milk could be used as an alternative diagnostic tool to screen for subclinical mastitis.     

Structural characterization of carangid fish myoglobins

The primary structures of myoglobin (Mb) from the following five carangid species were determined: yellowtail Seriola quinqueradiata, greater amberjack Seriola dumerili, yellowtail kingfish Seriola lalandi, Japanese horse mackerel Trachurus japonicus, and silver trevally Pseudocaranx dentex. The sequences were of varying composition both in the coding and in the noncoding regions, but all contained the open reading frame of 444 nucleotides encoding 147 amino acids. Amino acid sequence identities of carangid Mbs were in the range of 81-99%. The similarity of the heme pocket and associated heme-binding residues of carangid Mbs were evidence of the conservative nature of Mbs. Similar to the other teleost Mbs, carangid Mbs did not contain a D helix and had mostly conserved A and E helices as well as E-F and G-H inter-helical segments. Hydropathy profiles of carangid Mbs showed species-specific variations where silver trevally Mb exhibited generally higher hydrophobicity. Phylogenetic analysis based on the primary structures was in agreement with conventional morphological taxonomy, establishing close proximity of carangid Mbs with those of cichlid and scombroid, the other members of the Perciformes order.     

Improving the Quality of Silver Carp Fish Fillets by Gamma Irradiation and Coatings Containing Rosemary Oil

The objective of this work was to study the combined effect of gamma irradiation and coatings containing 0.5% rosemary (Rosmarinus officinalis) essential oil (RO) on the chemical, microbiological, and sensorial qualities of silver carp (Hypophthalmichthys molitrix) fish fillets (SFF) during cold storage (4°C). SFF were divided into three groups: uncoated (control), coated with edible coating (without additives), and γ-irradiated (0, 1, 3, and 5 kGy) coated with coating containing 0.5% rosemary. Gamma irradiation at 1, 3, and 5 kGy with coating reduced the initial total bacterial count, psychrophilic bacteria, and lactic acid bacteria and prolonged shelf life of the samples. Coated samples irradiated at 1 kGy reduced the counts of Enterobacteriaceae, Staphylococcus aureus, and Bacillus cereus, as well as eliminating Vibrio spp. and Salmonella spp., while coated samples irradiated at 3 and 5 kGy completely eliminated these bacteria. Combined treatment showed a slight increase in thiobarbituric acid-reactive substances postirradiation during cold storage but had no effect on the total volatile basic nitrogen and trimethylamine contents, while a gradual increase in these chemical quality indices was observed during cold storage. Combined treatment had no adverse effects on the sensory properties of SFF. The increase in the bacterial inhibitory effect is caused by both rosemary oil and irradiation.