Paleosol studies of burial mounds in the Ilovlya River valley (the Privolzhskaya Upland)

Paleosols buried under kurgans of the Bronze (end of the fourth and the third millennia BC), Early Iron (1st–3rd centuries AD), and Medieval (13th century AD) epochs have been studied on the Ilovlya River (a tributary of the Don River) terrace. The evolution of chestnut soils in the south of the Privolzhskaya Upland during the last 5000 years has been traced. It is shown that the mean weighted contents and distribution of soluble salts, gypsum, and carbonates in the soil profiles have been subjected to cyclic changes. The total microbial biomass and its trophic structure in the A1, B1, and B2k horizons of paleosols of different ages have been determined. A comparative analysis of the morphological, chemical, and microbiological data on the paleosols of different ages has been used to reconstruct the climatic dynamics for the last 50 centuries. The aridity of the climate in the studied region increased at the end of the third-the beginning of the second millennia BC and in the second and third centuries AD. The humidization of the climate took place in the 1st and in the 12th–13th centuries AD.     

Estimation of the density of the protocatechuate-degrading bacterial community in soil by real-time PCR

The β-ketoadipate pathway is the major route for degradation of aromatic compounds by various soil microorganisms. Protocatechuate 3,4-dioxygenase, a key enzyme of this pathway and which is encoded by pcaGH genes, catalyses the ring cleavage of protocatechuate. Microorganisms harbouring pcaGH genes are widely distributed in the environment but little is known about their relative abundance within the total microflora. Hence, this paper reports the development of a real-time PCR assay to quantify the bacterial pcaH sequence that encodes the β sub-unit of the protocatechuate 3,4-dioxygenase. This real-time PCR assay was linear over seven orders of magnitude with a calculated efficiency of 99% and sensitive up to 10² copies of the pcaH sequence per assay. Real-time PCR analyses performed on six soils with different physico-chemical properties, revealed pcaH densities ranging from 10³ to 10⁴ copies of pcaH  ng⁻¹ of soil DNA, which corresponded to approximately 0.2–10.9% of the total bacterial community. The sequencing of real-time PCR amplicons yielded 48 deduced amino acid sequences that exhibited 44–100% identity to known bacterial PcaH sequences, thereby confirming the accuracy of this real-time PCR assay.     

Silver nanoparticles preplant bulb soaking affects tulip production

Nanomaterials and particularly silver nanoparticles (AgNPs) have increased great interest in many fields of science. The element may affect plant growth and development but so far research studies have been scarce and their results disparate. Tulip is one of the most important ornamental plants and its production technology is constantly being improved. The aim of this study was to investigate the effects of AgNPs on growth and yield attributes of tulip cv. ‘Pink Impression’. Before planting, the tulip ‘Pink Impression’ bulbs were soaked in the following concentrations of AgNPs: 25, 50, 100 and 150?mg?L^-1. Control bulbs were soaked in deionized water. The plants were forced in a greenhouse for two growing seasons. The tulips treated with 100?mg?L^?1 AgNPs flowered earlier, had longer cut-flower stem, larger petals, greater stem diameter and cut-flower fresh weight. Moreover, at this concentration of AgNPs, they showed increased leaf greenness index (SPAD), stomatal conductance, root fresh weight, root length and produced daughter bulbs of the greatest weight. The study outcomes indicate that AgNPs may be used as plant growth promoters in horticulture.     

Reliminary survey of ticks (Acari: Ixodidae) on cattle in Central Equatoria State, Southern Sudan

In a preliminary survey conducted in 2005, the species composition and seasonality of ticks infesting cattle in Central Equatoria State, Southern Sudan was determined. Three locations were selected (Gumbo, Khor Rumla and Nyaing) and surveyed every 3 months. Two cattle herds in each of the three locations were visited four times during the study period. Total body collections of ticks were made from each of five cattle (Nilotic Zebu breed) kept in six different herds. Four tick genera and ten species were identified. The tick species identified were Amblyomma lepidum, Amblyomma variegatum, Boophilus annulatus, Boophilus decoloratus, Hyalomma marginatum rufipes, Hyalomma truncatum, Rhipicephalus appendiculatus, Rhipicephalus evertsi evertsi, Rhipicephalus praetextatus and Rhipicephalus sanguineus group. The highest number of ticks was collected in October during the rainy season. A finding of great significance was that R. appendiculatus, vector of East Coast fever, has now firmly established itself throughout the year with possible implications for cattle production in Central Equatoria State.     

The effect of local anaesthesia on anaesthetic requirements for feline ovariectomy

A dose of supplementary ketamine was used to evaluate the anaesthetic sparing effect of adding local anaesthesia to general anaesthesia in cats undergoing ovariectomy. Fifty-six healthy cats were randomly assigned to receive lidocaine 2% (group L) as skin infiltration (1 mg kg(-1)), topical application (splash block) on both the ovaries (2 mg kg(-1), each) and on abdominal muscular layers (1 mg kg(-1)), or an equal volume of NaCl 0.9% at the same sites (group S). Anaesthesia was induced with a mixture of 20 microg kg(-1) medetomidine and 5 mg kg(-1) ketamine administered intramuscularly. Rectal temperature, ECG, heart rate and respiratory rate were measured continuously. Ketamine supplemental boli (1 mg kg(-1), intravenously) were administered in response to movements during surgery. Local lidocaine significantly reduced the need for supplementary ketamine. All animals were returned to their owners without complications. With this protocol, local anaesthetics reduced the need for injectable anaesthetic during feline ovariectomy.     

Bee pollen and propolis as dietary supplements for rabbit: Effect on reproductive performance of does and on immunological response of does and their offspring

To evaluate the effect of bee pollen (BP) and/or propolis (Pro) supplementation on rabbit does, 64 nulliparous NZW rabbits does were distributed among eight groups (eight animals/group). One unsupplemented group was the control; the other seven groups were supplemented, respectively, with zinc bacitracin (ZnB) at 100 mg, BP at 150 and 300 mg, Pro at 150 and 300 mg, BP+Pro at 150 and 300 mg of each three times/week, day after day continuously along eight parities. The BP300, Pro300 and BP+Pro150 groups had higher body weight of litter at birth and number of kids born alive. The BP supplementation at 150 mg increased plasma total protein and albumin than the control group. The BP or Pro at 150 mg decreased plasma T₃ than the other groups except for BP+Pro150. The ZnB group had significantly greater T₃/T₄ ratio compared to BP, Pro and BP+Pro at 150 mg. The BP+Pro150 group had less ALT than the control; BP300 and Pro 300 mg resulted in lower plasma AST than the groups Pro150 with or without BP and the control group. The plasma alkaline phosphatase of BP at 150 or 300 mg and BP+Pro150 was significantly greater than that of the Pro150 group. The BP+Pro300 group had higher WBCs than the other groups. In contrast, the lymphocytes were greater in the Pro and BP+Pro300 groups than in BP, Pro and BP+Pro at 150 mg. The groups supplemented with BP and BP+Pro at 150 and 300 mg had significantly greater SRBCs of doe rabbits and their offspring compared to the control and the ZnB group. The BP at 300 mg increased the serum albumin and α₁‐globulin than the control group. The Pro300 group had greater serum α₂‐globulin and β‐globulin than the control group. The total globulin was significantly greater for the 300 mg propolis‐supplemented groups than the control.     

Morphological diversity of female camel (Camelus dromedarius) populations in Morocco

Tropical Animal Health and Production - Fourteen body measurements of 132 adult female camels belonging to three populations Guerzni (60), Khouari (28), and Marmouri (44) reared in 38 herds of 8...     

Biodiversity in Mediterranean buffalo using two microsatellite multiplexes

The present study was carried out to investigate genetic diversity in Nile-Delta and Southern-Egypt buffalo populations in comparison with the Italian buffalo utilizing two microsatellite multiplexes. A total of 104 animals classified into three groups were used, 28, 38 and 38 representing the Nile-Delta, Southern-Egypt and Italian buffalo, respectively. The 15 studied microsatellites were CSSM38, CSSM70, CYP21, CSSM42, CSSM60, MAF65, BM0922, CSSM19, INRA006, ETH02, BM1706, BMC1013, CSSM47, INRA026 and CA004. All studied microsatellites showed allelic polymorphism. Number of polymorphic alleles ranged between 4 alleles (CSSM38, CSSM70 and CYP21) and 11 alleles (CA004). Pairwise Chi-square test for Nile Delta and Southern Egypt showed significant differences in allelic distribution at five loci, CSSM70, CSSM38, BM0922, ETH02 and BM1706. Italian buffalo showed the lowest percentage of observed heterozygotes (65%), while the Southern Egypt showed the highest (71%). Both the Italian and the Delta populations deviated significantly (P < 0.05) from HW equilibrium. Italian buffalo is relatively the most inbred population while the Southern-Egypt buffalo is the only outbred population. High level of genetic differentiation (F_ST estimates) between the Italian group and each of the Delta and Southern-Egypt group (0.083 and 0.076, respectively) was observed while Southern-Egypt group showed a lower level of genetic differentiation with the Delta group (0.014). Italian buffalo had the greatest genetic distance values with the two Egyptian groups (0.25 and 0.23) while much lower values between the Southern-Egypt and the Delta groups (0.06) was observed. Genetic variation between the Italian buffalo and the Egyptian buffalo was detected in 14 (out of 15) microsatellite loci. While a lower level (than that between Egyptian and Italian) of genetic variation between Southern-Egypt and Delta buffalo populations was expressed by 5 loci. It was concluded that the Southern-Egypt buffalo could be considered as distinct population from the Delta buffalo. In addition, Southern-Egypt group, being the only group with non-significant global deviation from HW equilibrium, the most heterozygous, and the only outbred population as well, is expected to respond more favorably to selection.     

A simple and efficient CRISPR/Cas9 platform for induction of single and multiple,heritable mutations in barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L.)

Background

Genome editing of monocot plants can be accomplished by using the components of the CRISPR/Cas9 (clustered regularly interspaced short palindromic repeat/CRISPR associated Cas9) technology specifically optimized for these types of plants. Here, we present the development of RNA-guided Cas9 system for simplex and multiplex genome editing in barley.

Results

We developed a set of customizable RNA-guided Cas9 binary vectors and sgRNA modules for simplex and multiplex editing in barley. To facilitate the design of RNA-guided Cas9 constructs, the pBract derived binary vectors were adapted to Gateway cloning and only one restriction enzyme was required for construction of the sgRNA. We designed a synthetic, codon optimized Cas9 gene containing the N terminal SV40 nuclear localization signal and the UBQ10 Arabidopsis 1st intron. Two different sgRNAs were constructed for simplex editing and one polycistronic tRNA-gRNA construct (PTG) for multiplex editing using an endogenous tRNA processing system. The RNA-guided Cas9 constructs were validated in transgenic barley plants produced by Agrobacterium-mediated transformation. The highest mutation rate was observed in simplex editing of the cytokinin oxidase/dehydrogenase HvCKX1 gene, where mutations at the hvckx1 locus were detected in 88% of the screened T₀ plants. We also proved the efficacy of the PTG construct in the multiplex editing of two CKX genes by obtaining 9 plants (21% of all edited plants) with mutations induced in both HvCKX1 and HvCKX3. Analysis of the T₁ lines revealed that mutations in the HvCKX1 gene were transmitted to the next generation of plants. Among 220 screened T₁ plants we identified 85 heterozygous and 28 homozygous mutants, most of them bearing frameshift mutations in the HvCKX1 gene. We also observed independent segregation of mutations and the Cas9-sgRNA T-DNA insert in several T₁ plants. Moreover, the knockout mutations of the Nud gene generated phenotype mutants with naked grains, and the phenotypic changes were identifiable in T₀ plants.

Conclusions

We demonstrated the effectiveness of an optimized RNA-guided Cas9 system that can be used for generating homozygous knockout mutants in the progeny of transgenic barely plants. This is also the first report of successful multiplex editing in barley using a tRNA processing system.

     

Associations of Leptin and Pituitary‐Specific Transcription Factor Genes’ Polymorphisms with Reproduction and Production Traits in Dairy Buffalo

This study aimed to detect the genetic variability in Leptin and Pit‐1 genes using polymerase chain reaction–restriction fragment length polymorphism and DNA sequencing also to explore their possible associations with reproductive and productive traits of Egyptian buffaloes. Regarding Leptin gene, three genotypes (AA, AG and GG) were identified with frequency of 0.54, 0.40 and 0.06, respectively, and the genotypes were distributed according to the Hardy–Weinberg equilibrium. Allele A was comparatively higher than G with frequency of 0.74 and 0.26, respectively. For Pit‐1 gene, the association could not be performed due to the monomorphism (BB). The results showed that AA genotypes were found to be superior in most of production and reproduction traits. AA genotypes yielded more milk (2332.34 kg, p = 0.04) with higher fat% (6.10, p = 0.004) and fat yield (155.75 kg, p = 0.06), reach peak milk production at 42.19 days and required 2.19 services for conception in comparison with GG genotypes. Birthweight of animals with AA genotype was lesser than with GG genotype (39.35 and 43.67 kg, p = 0.02, respectively). The days open is numerically better in AA genotype animals (99.35 days), but the difference between the three genotypes was non‐significant. The distinct significant associations reported in this study suggested that Leptin is reputable candidate genetic marker, which might be used to enhance animals’ genetic potential for milk production in conjunction with reproduction.