Development of a real-time PCR method for the differential detection and quantification of four solanaceae in GMO analysis: potato (Solanum tuberosum), tomato (Solanum lycopersicum), eggplant (Solanum melongena), and pepper (Capsicum annuum)   总被引：3，自引：0，他引：3  

Chaouachi M  El Malki R  Berard A  Romaniuk M  Laval V  Brunel D  Bertheau Y 《Journal of agricultural and food chemistry》2008,56(6):1818-1828

The labeling of products containing genetically modified organisms (GMO) is linked to their quantification since a threshold for the presence of fortuitous GMOs in food has been established. This threshold is calculated from a combination of two absolute quantification values: one for the specific GMO target and the second for an endogenous reference gene specific to the taxon. Thus, the development of reliable methods to quantify GMOs using endogenous reference genes in complex matrixes such as food and feed is needed. Plant identification can be difficult in the case of closely related taxa, which moreover are subject to introgression events. Based on the homology of beta-fructosidase sequences obtained from public databases, two couples of consensus primers were designed for the detection, quantification, and differentiation of four Solanaceae: potato (Solanum tuberosum), tomato (Solanum lycopersicum), pepper (Capsicum annuum), and eggplant (Solanum melongena). Sequence variability was studied first using lines and cultivars (intraspecies sequence variability), then using taxa involved in gene introgressions, and finally, using taxonomically close taxa (interspecies sequence variability). This study allowed us to design four highly specific TaqMan-MGB probes. A duplex real time PCR assay was developed for simultaneous quantification of tomato and potato. For eggplant and pepper, only simplex real time PCR tests were developed. The results demonstrated the high specificity and sensitivity of the assays. We therefore conclude that beta-fructosidase can be used as an endogenous reference gene for GMO analysis.  相似文献   

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

V. A. Demkin  N. N. Kashirskaya  T. S. Demkina  T. E. Khomutova  M. V. El’tsov 《Eurasian Soil Science》2008,41(2):115-127

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.  相似文献   

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

N. El Azhari  D. Bru    A. Sarr  & F. Martin-Laurent 《European Journal of Soil Science》2008,59(4):665-673

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.  相似文献   

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

Salih DA  Julla II  Hassan SM  El Hussein AM  Jongejan F 《The Onderstepoort journal of veterinary research》2008,75(1):47-53

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.  相似文献   

The effect of local anaesthesia on anaesthetic requirements for feline ovariectomy     

Zilberstein LF  Moens YP  Leterrier E 《Veterinary journal (London, England : 1997)》2008,178(2):214-218

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.  相似文献   

A comprehensive evaluation and first molecular report of Theileria ovis infection in small ruminants in Saudi Arabia     

Alanazi  Abdullah D.  Said  Ashraf E.  Ghoneim  Ahmed M.  Alyousif  Mohamed S.  Alanazi  Ibrahim O. 《Tropical animal health and production》2019,51(1):89-98

A total of 1000 clinically healthy small ruminants comprising 500 sheep and 500 goats from five districts within Riyadh Province in Saudi Arabia were investigated by routine Giemsa staining for hematozoan parasites. Out of these, 100 sheep and 95 goat samples were investigated by PCR using three pairs of hemoprotozoan-specific primers. Based on microscopic examination, 33.2% of sheep and 25.2% of goats were found infected with hemoprotozoan parasites, while PCR detected hematozoan infection in 46% of sheep and 33.7% of goats. Extensive molecular characterization of hematozoan infection using six pairs of species-specific primers revealed the dominance of Theileria ovis, rather than any other species, which is recorded for the first time in small ruminants in Saudi Arabia. Prevalence of T. ovis in sheep and goats was found to be the highest in Riyadh (32, 48%) followed by AL-Kharj (31, 35%), Ad-Dawadimi (31, 33%), AL-Majmaah (15, 27%), and Rumah (17, 23%). The highest parasite prevalence was recorded in the 3 years of age and >?4 years of age ruminants, while the lowest prevalence was recorded in <?1 year of age ruminants. No noticeable differences in parasite prevalence between male or female ruminants were recorded. Partial sequencing of 18S rRNA gene revealed the infection of the studied ruminants with four new isolates of T. ovis. Further characterization of the pathogenicity and the clinical effects of these T. ovis isolates in sheep and goats is highly needed. The current results can be helpful in protecting and improving livestock industry in the countries that depend on a high number of small ruminants.

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Evaluating the public health response to a mass bat exposure—Wyoming, 2017     

Andrea Cote  Ryan M. Wallace  David A. Jackson  Maria A. Said  Karl Musgrave  Cuc H. Tran  Clayton Van Houten  Alexia Harrist  Danielle Buttke  Ashley Busacker  Veronica Pickens  Sarah Anne J. Guagliardo 《Zoonoses and public health》2019,66(5):504-511

Mass bat exposures (MBEs) occur when multiple people are exposed to a bat or a bat colony, often over an extended period. In August 2017, a public health investigation was started in response to an MBE that occurred during May–August 2017 at a national park research station in Wyoming. We identified 176 people who had slept primarily in two lodges (Lodges A and B) at the research station, and successfully contacted 165 (93.8%) of these individuals. Risk assessments (RAs) were administered to all 165 individuals to determine degree and type of exposures to bats (e.g., biting or scratching). Exposure status for research station guests was classified as “non‐exposed,” “low risk” or “high risk,” and counselling was provided to guide post‐exposure prophylaxis (PEP) recommendations. Prior to public health notification and intervention, 19 persons made the decision to pursue PEP. The healthcare‐seeking behaviours of this group were taken to represent outcomes in the absence of public health intervention. (These persons received a RA, and their risk classification was retrospectively assigned.) Approximately 1–2 weeks after conducting the RAs, we conducted a follow‐up survey to determine whether recommendations regarding PEP were ultimately followed. The proportion of individuals that unnecessarily pursued PEP was higher among the 19 individuals that sought health care prior to receiving the RA (p < 0.00001). Among those receiving the RA first, all persons classified as high risk followed public health guidance to seek PEP treatment. Despite this, upon re‐interview, only 21/79 (26.6%) of guests could accurately recall their risk classification, with most people (55.7%) overestimating their risk. Study findings demonstrate that early public health interventions such as RAs can reduce unnecessary use of PEP and that messaging used during rabies counselling should be clear.  相似文献   

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

Youssef A. Attia  Fulvia Bovera  Abd El‐Hamid Abd Elhamid  Sameer A. Nagadi  Mohamed A. Mandour  Saber Sh. Hassan 《Journal of animal physiology and animal nutrition》2019,103(3):959-968

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.  相似文献   

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

Boujenane  Ismaïl  El Khattaby  Nadia  Laghouaouta  Houda  Badaoui  Bouabid  Piro  Mohammed 《Tropical animal health and production》2019,51(6):1367-1373

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...  相似文献   

Biodiversity in Mediterranean buffalo using two microsatellite multiplexes     

A.R. Elbeltagy  Salah Galal  A.Z. Abdelsalam  F.E. El Keraby  M. Blasi  Mahasen M. Mohamed 《Livestock Science》2008,114(2-3):341-346

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.  相似文献