Some physiological and morphological responses of Pyrus boissieriana to flooding

European pear is a flooding-sensitive species, and for its cultivation in lowland areas, it is necessary to carry out the grafting of scions of commercial pear varieties into rootstocks belonging to flooding-tolerant wild pear species. Flooding tolerance of Pyrus boissieriana—a type of wild pear—was studied as a promissory rootstock for commercial pear. For this purpose, 3-month-old plants of P. boissieriana were subjected for 30 days to control (C), well-irrigated treatment, short-term (15 days) flooding plus 15 days recovery (F + R) and long-term (30 days) continuous flooding (F). Physiological performance, plant morphological changes and biomass accumulation were assessed. Results showed that, although stomatal conductance, transpiration and photosynthesis were progressively decreased by flooding, when flooding was short term (i.e., 2 weeks, F + R treatment) plants were able to adequately recover their physiological activity (50–74 % with respect to controls). In contrast, when plants continued to be flooded (F treatment), the physiological activity became null and the plants died quickly after the water subsided. Adventitious rooting was the most conspicuous registered morphological response to flooding, despite that flooded plants had shorter shoots and roots than control plants. Leaf and root biomass were 63 and 89 % higher under short-term flooding (F + R) than under continuous flooding (F), condition in which plants did not survive. In conclusion, P. boissieriana appears to be a promising species for its use as rootstock of commercial pear in lowland areas prone to flooding of up to 2 weeks. However, if the flooding period is extended, plants of this species are at risk of perishing.     

Effects of Short-Term Over-supplementation of Copper in Milk on Hematology,Serum Proteins,Weight Gain,and Health in Dairy Calves

Thirty-six calves were used in the present study. The animals were divided equally into three groups (control, test 1, and test 2). The three groups of calves were homogeneous for parity of dams, sex, and month of birth. From 14 days of age, in the test 1 group copper as copper sulfate (Merck Co, Germany) was added to each meal of milk at a rate of 10 mg/kg of milk for 14 days and in test 2 group copper as copper sulfate was added to each meal of milk at a rate of 20 mg/kg of milk for 14 days. Blood samples were taken by jugular venipuncture using disposable syringes at 14 (before Cu supplementation), 30, 60, and 80 days of age. Anticoagulated blood was used for CBC determination. Plane tubes were used for harvesting of serum and the amounts of total serum protein, albumin, iron, and copper were measured. Calves were weighted at birth and at the end of trial (day 80) and total gain and mean daily gain were calculated. Days of treatment for ill calves were also recorded during experiment. Group (treatment) had no significant effect on the amounts of measured parameters except MCH values (p < 0.05) which were significantly lower in test 1 group than other trial groups. Age (sampling time) had significant effects on the values of most measured parameters (p < 0.05) except WBC, lymphocyte, total protein, and fibrinogen. Significant interactions between sampling time and group were not seen for any of measured parameters. No significant differences were seen for total weight gain and mean daily gain between trial groups. Chi-square test revealed no significant difference for the days of treatment between trials groups.     

Scaling up: A guide to high‐throughput genomic approaches for biodiversity analysis

The purpose of this review is to present the most common and emerging DNA‐based methods used to generate data for biodiversity and biomonitoring studies. As environmental assessment and monitoring programmes may require biodiversity information at multiple levels, we pay particular attention to the DNA metabarcoding method and discuss a number of bioinformatic tools and considerations for producing DNA‐based indicators using operational taxonomic units (OTUs), taxa at a variety of ranks and community composition. By developing the capacity to harness the advantages provided by the newest technologies, investigators can “scale up” by increasing the number of samples and replicates processed, the frequency of sampling over time and space, and even the depth of sampling such as by sequencing more reads per sample or more markers per sample. The ability to scale up is made possible by the reduced hands‐on time and cost per sample provided by the newest kits, platforms and software tools. Results gleaned from broad‐scale monitoring will provide opportunities to address key scientific questions linked to biodiversity and its dynamics across time and space as well as being more relevant for policymakers, enabling science‐based decision‐making, and provide a greater socio‐economic impact. As genomic approaches are continually evolving, we provide this guide to methods used in biodiversity genomics.     

A Statistical Comparison between Less and Common Applied Models to Estimate Geographical Distribution of Endangered Species (<Emphasis Type="Italic">Felis margarita</Emphasis>) in Central Iran

Species distribution in space is important in habitat conservation and biodiversity protection, so gaining knowledge about species range would be worthwhile to rescue endangered species and plan conservation policy. This study evaluates and compares the performance of an array of Species Distribution Models (SDMs), namely RF, SVM, MaxEnt, GLMNET, and MARS, in predicting rare sand cat distribution across a large unprotected sand dune area in central Iran. Due to absence of reliable data and difficulties in recording the species itself, the SDMs were challenged by limited data including 55 absence (background) and 40 presence points as well as nine climatic and geological parameters that influence on species distribution, including humidity, maximum, minimum and mean temperature, precipitation, amount of sunshine, ground water level, aspect, and DEM. Moreover, each model was replicated 20 times and the statistics including TSS, AUC, COR and Deviance were computed. Then, based on computed statistics, the model performances were evaluated by TUKEY and ANOVA. Finally, ensemble map was obtained by weighted approach using AUC. The results of this study showed that complex machine learning methods, like SVM, RF, and MaxEnt are more outperformed to predict the distribution of rare species.     

A Recombinant Probiotic, <Emphasis Type="Italic">Lactobacillus casei,</Emphasis> Expressing the <Emphasis Type="Italic">Clostridium perfringens</Emphasis> α-toxoid,as an Orally Vaccine Candidate Against Gas Gangrene and Necrotic Enteritis

The alpha-toxin is one of the virulence factors of Clostridium perfringens for gas gangrene in humans and animals or necrotic enteritis in poultry. The C-terminal domain of this toxin ( cpa _247-370 ) was synthesized and cloned into pT1NX vector to construct the pT1NX-alpha plasmid. This surface-expressing plasmid was electroporated into Lactobacillus casei ATCC 393, generating the recombinant L. casei strain expressing alpha-toxoid (LC-α strain). Expression of this modified alpha-toxoid was confirmed by SDS-PAGE, immunoblotting, and direct immunofluorescence microscopy. BALB/c mice, immunized orally by the recombinant LC-α strain, elicited mucosal and significantly humoral immune responses (p < 0.05) and developed a protection against 900 MLD/mL of the standard alpha-toxin. This study showed that this recombinant LC-α strain could be a promising vaccine candidate against gas gangrene and necrotic enteritis.     

Reassessment of inclusion body-based production as a versatile opportunity for difficult-to-express recombinant proteins

The production of recombinant proteins in the microbial host Escherichia coli often results in the formation of cytoplasmic protein inclusion bodies (IBs). Proteins forming IBs are often branded as difficult-to-express, neglecting that IBs can be an opportunity for their production. IBs are resistant to proteolytic degradation and contain up to 90% pure recombinant protein, which does not interfere with the host metabolism. This is especially advantageous for host-toxic proteins like antimicrobial peptides (AMPs). IBs can be easily isolated by cell disruption followed by filtration and/or centrifugation, but conventional techniques for the recovery of soluble proteins from IBs are laborious. New approaches therefore simplify protein recovery by optimizing the production process conditions, and often include mild resolubilization methods that either increase the yield after refolding or avoid the necessity of refolding all together. For the AMP production, the IB-based approach is ideal, because these peptides often have simple structures and are easy to refold. The intentional IB production of almost every protein can be achieved by fusing recombinant proteins to pull-down tags. This review discusses the techniques available for IB-based protein production before considering technical approaches for the isolation of IBs from E. coli lysates followed by efficient protein resolubilization which ideally omits further refolding. The techniques are evaluated in terms of their suitability for the process-scale production and downstream processing of recombinant proteins and are discussed for AMP production as an example.