VE和L-肌肽对大菱鲆幼鱼生长、抗氧化、非特异性免疫及血清生化指标的影响

实验以大菱鲆（Scophthalmus maximus）幼鱼[（14.00±0.02）g]为研究对象，采用2×4双因素设计，设2个VE水平（0和75 mg/kg）和4个L-肌肽水平（0、50、100和200 mg/kg），研究VE和L-肌肽对其生长、抗氧化、非特异性免疫及血清生化指标的影响。实验共分8组，每组3个重复，每个重复46尾鱼，实验周期为8周。结果显示:（1）在饲料中添加75 mg/kg VE显著提高了大菱鲆幼鱼增重率（WGR）和特定生长率（SGR）（P < 0.05），L-肌肽添加量≤100 mg/kg对实验鱼生长性能无显著影响（P>0.05），添加量为200 mg/kg时鱼体WGR、SGR和蛋白质效率（PER）显著降低，饲料系数（FCR）显著升高（P < 0.05）；（2）VE和L-肌肽对血清谷胱甘肽过氧化物酶（GSH-PX）、过氧化氢酶（CAT）活性和丙二醛（MDA）含量及肝脏总抗氧化能力（T-AOC）、超氧化物歧化酶（SOD）活性和MDA含量均具有显著的交互作用（P < 0.05），在VE 75 mg/kg水平下，L-肌肽添加量为50和100 mg/kg时血清GSH-PX活性最高，L-肌肽添加量为100和200 mg/kg时血清CAT活性最高且与添加量为50 mg/kg差异不显著（P>0.05），添加100 mg/kg肝脏T-AOC和SOD活性达到最高且50 mg/kg组的SOD与100 mg/kg组差异不显著（P>0.05），主效应结果显示，VE显著提高了血清T-AOC、SOD及肝脏CAT活性（P < 0.05），L-肌肽显著提高了血清T-AOC（P < 0.05）；（3）VE和L-肌肽对血清补体C3和LZM活性交互作用显著，在75 mg/kg VE水平下，L-肌肽添加量为50 mg/kg时，补体C3水平最高（P < 0.05），主效应显示，VE和L-肌肽对血清总蛋白（TP）影响均不显著（P>0.05）；（4）添加VE显著降低了血清总胆固醇（TCHO）和甘油三酯（TG）含量（P < 0.05），添加L-肌肽显著降低了血清TG含量，且在L-肌肽50 mg/kg时达到最低。综合考虑大菱鲆幼鱼[（14.00-39.43）g]的生长性能、抗氧化性能、非特异性免疫及血清生化指标得出，在实验配方条件下（鱼油70 g/kg，大豆卵磷脂10 g/kg），添加VE 75 mg/kg时，L-肌肽的适宜添加量为50 mg/kg。     

Evolutionary relationships and sequence-structure determinants in human SARS coronavirus-2 spike proteins for host receptor recognition

Coronavirus disease 2019 (COVID-19) is a pandemic infectious disease caused by novel severe acute respiratory syndrome coronavirus-2 (SARS CoV-2). The SARS CoV-2 is transmitted more rapidly and readily than SARS CoV. Both, SARS CoV and SARS CoV-2 via their glycosylated spike proteins recognize the human angiotensin converting enzyme-2 (ACE-2) receptor. We generated multiple sequence alignments and phylogenetic trees for representative spike proteins of SARS CoV and SARS CoV-2 from various host sources in order to analyze the specificity in SARS CoV-2 spike proteins required for causing infection in humans. Our results show that among the genomes analyzed, two sequence regions in the N-terminal domain “MESEFR” and “SYLTPG” are specific to human SARS CoV-2. In the receptor-binding domain, two sequence regions “VGGNY“ and ”EIYQAGSTPCNGV” and a disulfide bridge connecting 480C and 488C in the extended loop are structural determinants for the recognition of human ACE-2 receptor. The complete genome analysis of representative SARS CoVs from bat, civet, human host sources, and human SARS CoV-2 identified the bat genome (GenBank code: MN996532.1) as closest to the recent novel human SARS CoV-2 genomes. The bat SARS CoV genomes (GenBank codes: MG772933 and MG772934) are evolutionary intermediates in the mutagenesis progression toward becoming human SARS CoV-2.     

Methylation of KvDMR1 involved in regulating the imprinting of CDKN1C gene in cattle

The CDKN1C gene encodes a cyclin‐dependent kinase inhibitor and is one of the key genes involved in the development of Beckwith–Wiedemann syndrome and cancer. In this study, using a direct sequencing approach based on a single nucleotide polymorphism (SNP) at genomic DNA and cDNA levels, we show that CDKN1C exhibits monoallelic expression in all seven studied organs (heart, liver, spleen, lung, kidney, muscle and subcutaneous fat) in cattle. To investigate how methylation regulates imprinting of CDKN1C in cattle, allele‐specific methylation patterns in two putative differential methylation regions (DMRs), the CDKN1C DMR and KvDMR1, were analyzed in three tissues (liver, spleen and lung) using bisulfite sequencing PCR. Our results show that in the CDKN1C DMR both parental alleles were unmethylated in all three analyzed tissues. In contrast, KvDMR1 was differentially methylated between the two parental alleles in the same tissues. Statistical analysis showed that there is a significant difference in the methylation level between the two parental alleles (P < 0.01), confirming that this region is the DMR of KvDMR1 and that it may be correlated with CDKN1C imprinting.     

Diversity measures in macroinvertebrate and zooplankton communities related to the trophic status of subtropical reservoirs: Contradictory or complementary responses?

Macroinvertebrate communities have been widely used as a tool for assessing the environmental quality of freshwater ecosystems, whereas zooplankton communities have been to some extent neglected. However, the importance of using different indicators to achieve a more comprehensive framework of assessment regarding water quality has been recognized. This study compared estimates of species richness (number of species) and the Shannon–Wiener index for data on macroinvertebrate and zooplankton communities in tropical reservoirs and related them to their trophic state. The trop+hic classification was obtained by applying the Carlson index (1977) modified by Toledo et al. (1983), and the index of the Brazilian Society of the Environmental Technology Agency. The comparative response of the different indicators was analyzed using a series of bivariate correlations (Draftsman’s plot). The results illustrate that diversity measures, namely species richness, responded differently when related to the trophic classification of reservoirs, depending on the community considered. The species richness of zooplankton was positively related to hypereutrophic conditions, due to the higher number of rotifer species, including tolerant generalist species and at the same time, as a result of the exclusion of species from other groups, whereas for macroinvertebrates, species richness was negatively related to hypereutrophic conditions. Melanoides tuberculatus, which exhibits a high tolerance and competitive ability under such conditions, was the dominant species in macroinvertebrate communities, which excluded endemic species and reduced local richness and diversity. The same indicators applied to the zooplankton and macroinvertebrate communities might therefore provide contradictory responses regarding ecological quality assessment in tropical reservoirs, which suggest that zooplankton should be taken into account among the biological quality elements considered in the ecological quality assessment, management, and restoration of water bodies.     

Intra- and interbreed genetic variations of mitochondrial DNA major non-coding regions in Japanese native dog breeds [Canis familiaris)

Mitochondrial DNA (mtDNA) major non-coding regions were amplified from 73 dogs of eight Japanese native dog breeds and from 21 dogs of 16 non-Japanese dog breeds by the polymerase chain reaction and their DNA sequences were determined. A total of 51 nucleotide positions within the non-coding region (969–972 base pairs) showed nucleotide variations of which 48 were caused by transition. These nucleotide substitutions were abundant in the region proximate to tRNA^Pro. In addition to the nucleotide substitutions, the dog mtDNA D-loop sequences had a heteroplasmic repetitive sequence (TACACGTÀCG) involving size variation. The DNA sequences of the non-coding region were classified into four different groups by phylogenetic analysis and the deepest branchpoints of this dog phylogeny was calculated to about 100 000 years before the present. Phylogenetic analysis showed that Japanese native dog breeds could not be clearly delimited as distinct breeds. Many haplotypes found in members of some clustering groups were seen in each dog breed, and interbreed nucleotide differences between Japanese dog breeds were almost the same as the intrabreed nucleotide diversities.     

Planktonic indices in the evaluation of the ecological status and the trophic state of the longest lake in Poland

Due to the intensive mixing polymictic lakes should be homogenous. However, morphometric diversity and high water dynamics contribute to the differentiation of many parameters in various areas of the lakes. This study analyzes both phytoplankton and zooplankton to assess differences in water quality along the north–south axis of the longest lake in Poland. New phytoplankton indicators were applied for determining the lake's ecological status: the Q index based on functional groups and the PMPL (Phytoplankton Metric for Polish Lakes) index based on phytoplankton biomass. TSI_ROT index (Rotifer Trophic State Index), which comprises the percentage of species indicating a high trophic state in the indicatory group and the percentage of bacteriovorus in the Rotifera population, was used for zooplankton analysis.TP content was different at different sites – we observed its gradual increase from the south to the north. Spatial variation of phosphorus did not considerably affect plankton diversity. The phytoplankton was dominated by Oscillatoriales, typical of shallow, well-mixed eutrophic lakes. The ecological status of the lake based on the EQR (Ecological Quality Ratio) was poor or moderate. The zooplankton was dominated by rotifers (at almost all sites), which indicates a eutrophic state of the lake. The values of phytoplankton indices at the studied sites did not differ considerably; the differences resulted more from local conditions such as the contaminant inflow and the macrophyte development than water dynamics.We have demonstrated that in the lake dominated by filamentous Cyanobacteria the ecological status should be determined according to the PMPL index or other indices dependent on the dominant Cyanobacteria species. Since the Q index does not include the functional group S1, the results can lead to the false conclusion that water quality improves with an increased amount of phytoplankton. The high abundance of Cyanobacteria in the lake may have contributed to the poor growth of rotifers.     

Origin of the cultivated passion fruit Passiflora edulis f. flavicarpa and genomic relationships among species of the subgenera Decaloba and Passiflora

• Passiflora edulis f. flavicarpa is the most economically important species in the genus Passiflora. However, the origin of this yellow form of passion fruit remains unclear, being suggested as a hybrid (P. edulis f. edulis × P. ligularis) or wild mutant.
• Here, the origin and genomic relationships of P. edulis f. flavicarpa with some related species in the genus Passiflora (subgenera Decaloba and Passiflora) were investigated using genomic in situ hybridization (GISH). Genomic DNA of 18 species was used as probe, which was hybridized onto chromosomes of P. edulis f. flavicarpa.
• Of all genomic DNA probes tested, none allowed us to identify a specific chromosome set in P. edulis f. flavicarpa. Conversely, probes from the subgenus Passiflora, P. edulis f. edulis, P. alata, P. cincinnata, P. coccinea, P. nitida and P. vitifolia, produced intense and uniform hybridizations on all chromosomes of P. edulis f. flavicarpa. Moreover, probes from P. ligularis, P. foetida and P. sublanceolata produced more intense hybridizations in the terminal region of four chromosomes, corresponding to the DNAr 45S locus, and also dispersed, less intense, hybridization across all chromosomes. Probes from the subgenus Decaloba, P. biflora, P. capsularis, P. cervii, P. coriacea, P. micropetala, P. morifolia, P. rubra and P. suberosa, produced hybridizations restricted to the DNAr 45S sites.
• The hybrid origin of P. edulis f. flavicarpa could not be supported based on the GISH results, and it is suggested that this species is conspecific with P. edulis f. edulis, because the probe with DNA of this form hybridized strongly throughout the target genome. The other putative parent species, P. ligularis, showed only a distant relationship with the target genome. The results also suggest that species of the subgenus Passiflora share many repetitive sequences and that the relationship between subgenera Decaloba and Passiflora is very distant.