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The amount of pathogenic bacteria in feces is an indicator characterizing the health status of animals. This is important to protect the environment and the health of farm workers. The aim of the present study is to determine the effect of probiotic Zoovit on the microbiome of the faecal mass of Holstein cattle. In the experiment, two groups were formed - experimental and control, 30 each. A probiotic was added to the combined feed of the experimental group, but not to the control group. The food ration with the participation of probiotic is placed twice a day. To determine the microbiome in fecal matter in cattle, PCR analyzes were used for the detection of Lactobacillus delbrueckii subsp. bulgaricus, a species-specific PCR assay for the presence of total DNA in L. delbrueckii and L. delbrueckii subsp. bulgaricus and methods for determination of sulfite-reducing colonies. No representatives of Staphulococccua aureus and Clostridium sp were isolated in the faecal mass of the animals from both groups. For Escherichia coli and Coliforms, a significant decrease in the faecal mass of cows fed with feed containing probiotics was found. The presence of L. delbrueckii subsp. bulgaricus in animals receiving a probiotic. Colonies with a colonial characteristic typical of L. delbrueckii subsp. bulgaricus were isolated in the animals receiving probiotic Zoovit and in the control group, no colonies typical of Lactobacillus bulgaricus were found. [ABSTRACT FROM AUTHOR]

The present study detected Coxiella burnetii (C. burnetii) infection in patients with endocarditis with a negative bacterial culture from January, 2022 to February, 2023. For this purpose, 312 patients with endocarditis who were diagnosed and operated were included in the study after obtaining the consent of each study subject and approval from the hospital ethics committee. Following surgery, the blood samples from the patients were cultured to identify bacteria using an automatic system. In 52 cases for which the result of this culture was negative, PCR was also performed using C. burnetii primer pairs. As shown by the results, from the 52 blood samples from patients with endocarditis who were found to be negative for bacteria, 13 samples were found to be positive for C. burnetii by PCR. Among these 13 patients, all patients with a fever lasting 14 days after surgery, 6 patients with emaciation and pneumonia, and 2 patients with multi-organ failure, who then succumbed. [ABSTRACT FROM AUTHOR]

The salinity environment is one of the biggest threats to Glycyrrhiza uralensis Fisch. (G. uralensis) growth, resulting from the oxidative stress caused by excess reactive oxygen species (ROS). Flavonoids are the main pharmacodynamic composition and help maintain ROS homeostasis and mitigate oxidative damage in G. uralensis in the salinity environment. To investigate whether endophytic Bacillus cereus G2 can improve the salt-tolerance of G. uralensis through controlling flavonoid biosynthesis, the transcriptomic and physiological analysis of G. uralensis treated by G2 in the saline environment was conducted, focused on flavonoid biosynthesis-related pathways. Results uncovered that salinity inhibited flavonoids synthesis by decreasing the activities of phenylalanine ammonialyase (PAL) and 4-coumarate-CoA ligase (4CL) (42% and 39%, respectively) due to down-regulated gene Glyur000910s00020578 at substrate level, and then decreasing the activities of chalcone isomerase (CHI) and chalcone synthase (CHS) activities (50% and 42%, respectively) due to down-regulated genes Glyur006062s00044203 and Glyur000051s00003431, further decreasing isoliquiritigenin content by 53%. However, salt stress increased liquiritin content by 43%, which might be a protective mechanism of salt-treated G. uralensis seedlings. Interestingly, G2 enhanced PAL activity by 27% whereas reduced trans-cinnamate 4-monooxygenase (C4H) activity by 43% which could inhibit lignin biosynthesis but promote flavonoid biosynthesis of salt-treated G. uralensis at the substrate level. G2 decreased shikimate O-hydroxycinnamoyltransferase (HCT) activity by 35%, increased CHS activity by 54% through up-regulating the gene Glyur000051s00003431 encoding CHS, and increased CHI activity by 72%, thereby decreasing lignin (34%) and liquiritin (24%) content, but increasing isoliquiritigenin content (35%), which could mitigate oxidative damage and changed salt-tolerance mechanism of G. uralensis.

Abstract Background After the commercialization of insect-resistant transgenic Bt cotton Bollgard I & II, India ranks first in the world in cotton production. Cotton insecticide consumption was drastically reduced as nearly 95% of the cotton area was replaced with Bollgard II. However, the benefits of transgenic cotton appear to have been diminished as the pink bollworm, Pectinophora gossypiella (Saunders) (Lepidoptera: Gelechiidae) developed field resistance to Cry1Ac expressing Bt cotton in India in 2008. In 2015, there was an unusual survival of P. gossypiella on Bollgard II cotton in some parts of Gujarat and Maharashtra, which is a cause for concern. Results In the present study, PCR analysis and gel visualization of BGII resistant field population of P. gossypiella showed that the presence of r1, r2, r3, r1s, r2s, r3s, and ss mutated cadherin alleles, which produced amplicon sizes of 750 bp, 700 bp, 730 bp, 12,700 bp, 750 bp, 980 bp, 600 bp and 1600 bp, respectively, when seven different types of cadherin allele(s) specific primers were used. The r1 mutant allele was detected in Amaravati, Wardha, Yavatmal, and Nagpur, but not in Akola and Buldhana, using primers r1disfor and Int-540. The r2 mutant allele was detected in Akola, Wardha, and Nagpur, but not in Buldhana, Amaravati, and Yavatmal, using primers r2disback and Cad2366. The r3 mutant allele was detected in all locations using primers r3disback and Cad3221. Amplicons of sizes 750 bp, 700 bp and 730 bp were obtained for r1, r2, and r3 mutant alleles, respectively. The sizes of the amplicons were 1270 bp for r1s, 750 bp for r2s, and 980 bp for r3s.The absence of bands for r1, r2, and r3 cadherin alleles in individuals indicated the possibility of having the ss allele, which was confirmed using primers Cad3324 and Int-651. The presence of ss mutant allele was observed in field-collected P. gossypiella populations from BG II cotton in various locations, with a 600 bp and 1600 bp size amplicons produced using the same primers. Conclusions PCR analysis revealed the presence of r1, r2, r3, r1s, r2s, r3s, and ss mutated cadherin alleles in field-collected populations of Vidarbha which provide evidence to field-evolved resistance of P. gossypiella to BGII cotton.

This article presents the results of research on the identification of informative DNA marker systems providing reliable control of the presence of the resistance gene to fusarium wilt Foc1 in white cabbage breeding material. At the beginning of the work, 14 molecular markers taken from the VegMarks database and published sources were tested on isogenic white cabbage lines with contrasting resistance to fusarium wilt (resistant line DT-46 and susceptible line Kb1P). InDel marker M10 and SSR markers Frg13 and Ol10-D01 have been ascertained to show polymorphism between white cabbage forms with contrasting resistance to fusarium wilt. Also, PCR analysis of segregating F2 population plants of hybrid combination DT-46 × Kb1P using these markers and phytopathology testing have been conducted. As a result of statistical analysis of segregation, it has been found that only SSR marker Ol10-D01 is co-segregated with the trait of resistance to fusarium wilt as expected segregation of F2 plants by genotype 1 : 2 : 1 according to Mendel's law; the least recombination frequency between the resistance gene Foc1 and the marker (1.6%) has just been observed only by this locus and it is highly polymorphic (PIC = 0.51). [ABSTRACT FROM AUTHOR]

The research is aimed at studying economically valuable traits and genetic control of resistance to leaf-stem diseases, photoperiodic reaction and short stemming of soft spring wheat variety Semenovna, created by scientists of the Omsk Agrarian Scientific Center (Russia) and the North Kazakhstan Agricultural Experimental Station (Kazakhstan). Using methods of state variety testing, molecular genetics and in vitro methods, morphological features of a new variety, features of its development have been described, the yield level at different ecological test points for three years (2015-2017) has been analyzed. The studies conducted at two ecological points made it possible to select a medium-sized promising line of soft spring wheat (Lutescens 354/04-6), which was transferred to the State Registration Service of the Republic of Kazakhstan and after two years of testing was included into the State Register of the Republic of Kazakhstan under the name Semenovna (patent No. 1023). In terms of grain quality, it was at the level of valuable wheat, exceeded the standard in protein content by 1-2 % and raw gluten by 3-4 %. The new medium-ripe Semenovna variety combines increased yield (2.73-4.40 t/ha) with resistance to drought (resistance index Ir = 0.57), brown and stem rust (IU = 0.00-0.23). The genotype of the variety contains wheat-rye translocation 1RS.1BL (with a cluster of Lr26/Sr31/Pm8/Yr9 genes). The medium-stem variety carries the Rht8b allele in its genotype (174 bp) and is photosensitive to the length of the day (allele 414 bp). The parameters of ecological plasticity of the new variety are determined: linear regression coefficient (bi = 1.08), stability index (σd2 = 0.27).

Mozzarella di Bufala Campana (MdBC) cheese is a Protected Designation of Origin (PDO) product that is important for the economy and cultural heritage of the Campania region. Food fraud can undermine consumers' trust in this dairy product and harm the livelihood of local producers. The current methods for detecting adulteration in MdBC cheese due to the use of buffalo material from foreign countries could exhibit limitations associated with the required use of expensive equipment, time-consuming procedures, and specialized personnel. To address these limits here, we propose a rapid, reliable, and cost-effective genotyping method that can detect foreign buffalo milk in a counterpart from the PDO area and in MdBC cheese, ensuring the quality and authenticity of the latter dairy product. This method is based on dedicated allele-specific and single-tube heminested polymerase chain reaction procedures. By using allele-specific primers that are designed to detect the nucleotide g.472G>C mutation of the CSN1S1Bbt allele, we distinguished an amplicon of 330 bp in the amplification product of DNA when extracted from milk and cheese, which is specific to the material originating from foreign countries. By spiking foreign milk samples with known amounts of the counterpart from the PDO area, the sensitivity of this assay was determined to be 0.01% v/v foreign to PDO milk. Based on a rough estimate of its simplicity, reliability, and cost, this method could be a valuable tool for identifying adulterated buffalo PDO dairy products. [ABSTRACT FROM AUTHOR]

Background: After the commercialization of insect-resistant transgenic Bt cotton Bollgard I & II, India ranks first in the world in cotton production. Cotton insecticide consumption was drastically reduced as nearly 95% of the cotton area was replaced with Bollgard II. However, the benefits of transgenic cotton appear to have been diminished as the pink bollworm, Pectinophora gossypiella (Saunders) (Lepidoptera: Gelechiidae) developed field resistance to Cry1Ac expressing Bt cotton in India in 2008. In 2015, there was an unusual survival of P. gossypiella on Bollgard II cotton in some parts of Gujarat and Maharashtra, which is a cause for concern. Results: In the present study, PCR analysis and gel visualization of BGII resistant field population of P. gossypiella showed that the presence of r1, r2, r3, r1s, r2s, r3s, and ss mutated cadherin alleles, which produced amplicon sizes of 750 bp, 700 bp, 730 bp, 12,700 bp, 750 bp, 980 bp, 600 bp and 1600 bp, respectively, when seven different types of cadherin allele(s) specific primers were used. The r1 mutant allele was detected in Amaravati, Wardha, Yavatmal, and Nagpur, but not in Akola and Buldhana, using primers r1disfor and Int-540. The r2 mutant allele was detected in Akola, Wardha, and Nagpur, but not in Buldhana, Amaravati, and Yavatmal, using primers r2disback and Cad2366. The r3 mutant allele was detected in all locations using primers r3disback and Cad3221. Amplicons of sizes 750 bp, 700 bp and 730 bp were obtained for r1, r2, and r3 mutant alleles, respectively. The sizes of the amplicons were 1270 bp for r1s, 750 bp for r2s, and 980 bp for r3s.The absence of bands for r1, r2, and r3 cadherin alleles in individuals indicated the possibility of having the ss allele, which was confirmed using primers Cad3324 and Int-651. The presence of ss mutant allele was observed in field-collected P. gossypiella populations from BG II cotton in various locations, with a 600 bp and 1600 bp size amplicons produced using the same primers. Conclusions: PCR analysis revealed the presence of r1, r2, r3, r1s, r2s, r3s, and ss mutated cadherin alleles in field-collected populations of Vidarbha which provide evidence to field-evolved resistance of P. gossypiella to BGII cotton. [ABSTRACT FROM AUTHOR]

Over-the-counter (OTC) limited ingredient canine diets could be reliable alternatives to veterinary therapeutic formulations for the diagnosis and management of adverse food reaction (AFR). However, the possibility of undeclared ingredients jeopardizes the efficacious use of OTC options for medical purposes. The objective was to determine the presence of undeclared ingredients in OTC canine dry diets marketed as limited or single protein source diets. Twenty-one OTC adult canine diets marketed as limited or single protein source diets were purchased. Multiplex PCR was used to screen for DNA of 10 mammalian species with species-specific primers that anneal to regions of the mitochondrial cytochrome b gene. The presence of DNA from one or more species not declared on the label was identified in all 21 diets: cow (Bos taurus), pig (Sus scrofa), sheep (Ovis sp.), goat (Capra hircus) and bison (Bison bison). Twenty diets were positive for the declared protein source and one diet was negative for the declared species. Cat (Felis catus), dog (Canis sp.), horse (Equus sp.), mouse (Mus musculus) and rat (Rattus norvegicus) DNA was not identified in any samples. The presence of undeclared mammal species in OTC canine dry diets marketed as having limited or single protein source ingredients may complicate AFR diagnosis and treatment. However, PCR can detect a miniscule amount of DNA which might not be clinically significant, because the amount needed to elicit a response is unknown. Quantification of the contamination was not determined in this study, precluding discrimination of intentional adulteration from unavoidable cross-contamination.

• Our research investigated the genetic variations among CL, VL, and CanL strains in northern Morocco. • Molecular and genetic analyses uncovered a complex array of genetic variants in the L. infantum and L. tropica species. The number of these variants is 7 and 9 for L. tropica and L. infantum species, respectively. • The Discriminant Analysis of Principal Components (DAPC) revealed significant genetic divergences among L. infantum strains. This suggests the influence of species origin on their genetic makeup. This study investigated nine provinces in northern Morocco and collected 275 skin scraping, 22 bone marrow aspirates, and 89 fine needle aspirations from suspected cutaneous leishmaniasis (CL) and visceral leishmaniasis (VL) patients and potentially infected dogs. Molecular analysis using ITS1 RFLP PCR and RT-PCR revealed a higher prevalence of L. infantum (66.18 %; χ2 = 28.804; df = 1; P-value = 8.01e−08) than L. tropica in skin scraping, with L. infantum being the sole causative agent for both VL and canine leishmaniasis. L. infantum was predominantly found in most provinces, while L. tropica was relatively more dominant in Taza Province. Discriminant Analysis of Principal Components (DAPC) revealed distinct clustering between L. tropica and the other three species. However, no small subset of SNPs could clearly differentiate between Infantum_CL, Infantum_VL, and CanL, as they likely share a significant genetic background. The high rate of L. infantum could be attributed to the abundance of sand fly species transmitting VL. In Taza Province, Phlebotomus sergenti, responsible for anthroponotic CL, is the most abundant species. DNA sequencing demonstrated sequence heterogeneity in L. infantum (variants 1-9) and L. tropica (variants 1-7). Phylogenetic analysis showed a distinct separation between L. tropica and L. infantum strains, with an overlap among L. infantum strains isolated from cutaneous, visceral, and canine cases, and dogs serving as the central population for L. infantum. [ABSTRACT FROM AUTHOR]

Background. The knowledge of genetic control of vernalization response in the ultra-early accessions can facilitate bread wheat breeding for a high adaptive capacity. Materials and methods. The study involved the ultra-early lines Rico (k-65588) and Rimax (k-67257) as the earliest maturing lines in the VIR bread wheat collection, as well as 10 Rifor lines (k-67120, k-67121, k-67250-67256) with a high rate of development before heading. A late ripening accession ‘Forlani Roberto’ (k-42641) and ‘Leningradskaya 6’ variety (k-64900), regionally adapted to Northwestern Russia, were also studied. The alleles of the Vrn and Ppd genes were identified by the PCR analysis using the allele-specific primers published in literature sources. The response to vernalization (30 days at 3°C) and a short 12-hour day were determined using a methodology accepted at VIR. Results. The ultra-early lines respond to a short 12-hour day and 30-day vernalization very poorly. The genotype of ultra-early wheat lines is mainly represented by three genes, Vrn-A1, Vrn-B1a, and Vrn-D1, which ensure insensitivity to vernalization alongside with the expression of Ppd-D1a, which controls the response to photoperiod. The ultra-early lines Rifor 4 and Rifor 5 have a recessive allele vrn-A1a, like the original ‘Forlani Roberto’ accession. The lines Rifor 4 and Rifor 5 are vernalization-insensitive under the long day and have a very weak response under the short day (3.5±0.42 days and 4.0±0.61 days, respectively). However, ‘Forlani Roberto’ with the vrn-A1a gene responds to vernalization in the same way under any photoperiod (12.3±1.58 days and 12.2±0.74 days). Conclusion The ultra-early lines of bread wheat Rifor 4 and Rifor 5 with the vrn-A1a gene can have no response to vernalization or have a low level response. This effect can be a reason for the formation of a complex of modifier genes along with the dominant gene Vrn-D1, which forms during the hybridization of F7-8 Rico × Forlani Roberto. The ultra-early lines of bread wheat Rico, Rimax and Rifor (k-67120, k-67121, k-67250-67256) can serve as effective sources of genes for earliness in common wheat breeding.

The genetic transformation of Simarouba glauca will help researchers across the globe in developing high medicinal value and high biomass varieties in a short period. Although genetic transformation methods have been trailed by many research groups for several years, the variation in transformation efficiency and difficulties in handling tissue culture plantlets made it difficult tooptimize a reliable protocol. In this study, we established an alternative protocol of genetic transformation by introducing GUSPlus reporter gene into the genome of an S. glauca through the Agrobacterium-mediated gene transfer method which is the first report in India. Leaf explants co-cultivated with Agrobacterium LBA4404 strains harbouring pCAMBIA 1305.1 vector have shown the presence of GUS Plus and HptII gene by PCR analysis as well as GUS assay. The outcome of our research provides fundamental information on efficient gene transformation in S. glauca which will help to enhance future research on the genetic transformation of S. glauca. [ABSTRACT FROM AUTHOR]

The applicability of the telomerase reverse transcriptase (TERT) gene Rs2736100 polymorphism in cancer research has been well documented for various malignancies except for head and neck cancers, where data is sparse. This study aimed to analyze this polymorphism with the pathological characterizers of laryngeal squamous cell carcinoma (LSCC) patients. Genetic testing was performed using the Real-time PCR technique on 56 paired samples of biological material (blood and tissue). Data were analyzed using Epi info 7 software. The subjects were predominantly male (95% vs. 5%), with a median age of 62 years, and smokers (89%). The primary tumor origin site was the glottic region (34%), and the advanced clinical stages III-IV were more common (46% vs. 18%). Results show high frequencies for the mutated variants of Rs2736100 (CC 36%>AC 34%>AA 30%), while distribution according to tumor classification criteria leaned towards moderately differentiated carcinoma specimens in T3-T4 stages for the AC/CC variants (P-value without statistical significance) but positively favored the relationship between the AA variant and lack of lymph node metastasis (P=0.0106). The genotypes tend to associate themselves with a better histological presentation regarding the pattern of tumor invasion and, thus, better prognostic values for LSCC. Results suggest that the wild-type genotype of TERT Rs2736100 may be a protective factor for lymph node metastasis and histological pattern of tumor invasion in LSCC. Results regarding the synergistic relationship between cancer and smoking corroborate literature data for moderate to severe smokers regardless of the genetic variant. [ABSTRACT FROM AUTHOR]

Human papillomaviruses (HPVs) are extremely widespread throughout the world. There are more than 100 types of HPVs, of which at least 14 types represent high oncogenic risk viruses (World Health Organization, 2020). Numerous attempts were made to analyze various water sources in order to (i) reveal the presence of DNA of pathogenic human papillomaviruses in them and (ii) assess the potential risks of occurrence of epidemics caused by HPV. With time, the necessity to solve these important problems stimulated the formation of a new direction in the world medical and environmental investigations. This paper contains the investigation of the presence of DNA of highly dangerous types of human papillomaviruses (HPV6, HPV11, HPV16 and HPV18) in water bodies of the Baikal natural territory, in particular in the water reservoirs in and near the villages of Listvyanka, Bolshiye Koty, Kultuk and the cities of Baikalsk and Slyudyanka. In course of our work, the conditions good for the study of the biological material obtained from water samples by the PCR technique to reveal the presence of DNA of HPV6, HPV11, HPV16 and HPV18 papillomaviruses were chosen. PCR analysis was conducted with the aid of both the already well-known universal primers GP5 + /6 + and the primers developed by our team to be applied to the conservative domains of nucleotide sequences encoding the main capsid protein L1 of human papillomaviruses HPV6, HPV11 (these types of the virus contribute to the occurrence of anogenital condylomatosis and the development of respiratory papillomatosis) and HPV16, HPV16 (these types of virus contribute to the occurrence of cervical cancer). The analyzes conducted by our team have revealed the presence of DNA of the four types of HPVs (6, 11, 16 and 18) in the samples taken from various water sources of the Baikal natural territory. [ABSTRACT FROM AUTHOR]

Since COVID-19, mpox was the first emerging pathogen to have spread globally in 2022. Wastewater-based surveillance (WBS) has proven to be an efficient early warning system for detecting potential resurgences. This report aims to provide insight into the development and implementation of WBS of mpox in Slovenia and to incorporate the surveillance results into the development of public health interventions. WBS of mpox was conducted during the period from 1 June 2023 to 30 September 2023 at the wastewater treatment plant (WWTP) Ljubljana and WWTP Koper. The selected detection method of the monkeypox virus (MPXV) in the wastewater sample was based on PCR analysis. The implemented laboratory method showed that the sample preparation and concentration method enables a stable procedure for MPXV detection in wastewater samples. The laboratory analysis of wastewater samples from the selected WWTPs did not detect the MPXV during the monitoring period. In the event of MPXV detection in a wastewater sample, targeted public health interventions would be implemented, focusing on increasing awareness among the groups of men who have sex with other men and searching for positive mpox cases. We recommend that the developed system be retained in the case of an emergency epidemiological situation.

Aflatoxin B1 is a carcinogenic and mutagenic mycotoxin mainly produced by Aspergillus flavus and A. parasiticus, and prevalent in food and feed. Microbial degradation is a promising strategy which can be performed in mild and environmental friendly condition. This work is a step towards identifying the enzyme responsible for biodegradation of AFB1 by P. putida. Experiments were performed with P. putida lysate and compared with commercial lipase to see the degradation efficiency and the temperature stability. The cell free lysate of P. putida efficiently degraded AFB1 in a range of temperature from 20 to 90 °C. The lysate is thermostable and could retain its activity on pre-incubation up to 90 °C. Highest rate of degradation was observed at 70 °C. These observations show that the P. putida lysate is not only stable at higher temperatures but its enzymatic activity increases after incubation. Similarly, the commercial lipase degraded AFB1 efficiently. However, both, the P. putida lysate and lipase ceased degradation in presence of a lipase inhibitor, HgCl2. The Hill function accurately predicted enzyme activity at various times and temperatures. Like lipase, the lysate also hydrolyses the p-nitrophenyl palmitate to p-nitrophenol. Kinetic parameters such as Vmax, Km and n values are good measures to characterize the lysate response with respect to changing paranitro phenyl palmitate levels. The substrate specificity test of lipase showed linear correlation between the absorbance at 410 nm vs amount of product paranitro phenol. The value of Km, Vmax and n are 0.62 mM, 355.7 μmol min−1 and 1.29, respectively. The lipase gene presence in P. putida was confirmed using PCR technique. These observations indicate that the main enzyme responsible for AFB1 degradation by P. putida is lipase. Thus, lipase as a multifunctional biocatalyst provides a promising future for a variety of industries and may also help to ensure the food safety by degrading the mycotoxins.

Mozzarella di Bufala Campana (MdBC) cheese is a Protected Designation of Origin (PDO) product that is important for the economy and cultural heritage of the Campania region. Food fraud can undermine consumers’ trust in this dairy product and harm the livelihood of local producers. The current methods for detecting adulteration in MdBC cheese due to the use of buffalo material from foreign countries could exhibit limitations associated with the required use of expensive equipment, time-consuming procedures, and specialized personnel. To address these limits here, we propose a rapid, reliable, and cost-effective genotyping method that can detect foreign buffalo milk in a counterpart from the PDO area and in MdBC cheese, ensuring the quality and authenticity of the latter dairy product. This method is based on dedicated allele-specific and single-tube heminested polymerase chain reaction procedures. By using allele-specific primers that are designed to detect the nucleotide g.472G>C mutation of the CSN1S1Bbt allele, we distinguished an amplicon of 330 bp in the amplification product of DNA when extracted from milk and cheese, which is specific to the material originating from foreign countries. By spiking foreign milk samples with known amounts of the counterpart from the PDO area, the sensitivity of this assay was determined to be 0.01% v/v foreign to PDO milk. Based on a rough estimate of its simplicity, reliability, and cost, this method could be a valuable tool for identifying adulterated buffalo PDO dairy products.

The identification of effective specialized DNA markers providing the clear control of target locus inheritance by the trait of submergence tolerance has been conducted. Among the studied set of microsatellite markers, two the most informative SSR-markers - RM 7481, PrC3 showed high efficiency in detecting intraspecific polymorphism of rice varieties and lines used in the work. With the use of these markers the clear genotype marking the obtained hybrid rice plants by this trait has been conducted and it is has been verified by phenotype evaluation as a result of laboratory trials. The plant samples carrying the target gene in heterozygous and homozygous state has been selected. About 400 backcrossed self-pollinated rice lines with introgressed and pyramided resistance genes Pi-1, Pi-2, Pi-33, Pi-ta, Pi-b to Pyricularia oryzae Cav. were obtained within the frameworks of program to develop genetic rice sources resistant to blast. The conducted testing for resistance to blast and the assessment by economically valuable traits have allowed to select the prospective rice samples. The plant samples of F2 and BC1F1 generations with combination of resistance to blast genes (Pi) and submergence tolerance gene (Sub1A) in homozygous and heterozygous state that is confirmed be the results of analysis of their DNA have been obtained. The obtained hybrid plants are being tested in breeding nurseries for a complex of economically valuable traits. The best plants will be selected and send to State Variety Testing system. Their involving in rice industry will reduce the use of plant protection chemicals against diseases and weeds, thereby increasing the ecology status of the rice industry. [ABSTRACT FROM AUTHOR]

This paper aimed to evaluate 50 wheat samples collected from different western Romanian locations based on microbiological, molecular, and toxicogenic assays to determine their correlation when species of the genus Fusarium were analyzed. The presence of toxins determined by biochemical ELISA (Enzyme-Linked Immunosorbent Assay), the DNA analysis based on PCR (Polymerase Chain Reaction), and even accurate species identification using specific gene sequencing were used to evaluate the fungal early infection. Considering that in Romania the prevalence of Fusarium graminearum, and Fusarium proliferatum infections is the most important, it can be stated that the screening with primers specific to fungal species ensures a preliminary test for fungal infection identification before performing the test for mycotoxins.

This article describes the occurrence, antibiograms, and detection of antibiotic resistance genes of Enterobacterales isolated from fresh vegetables commercialized in Riobamba, Ecuador. Escherichia coli isolates were screened to detect diarrheagenic pathotypes via PCR. Agar diffusion assay was performed to determine the phenotypic antibiotic resistance of the Enterobacterales strains. The presence of antibiotic resistance genes conferring resistance against beta-lactams, mobile colistin resistance, carbapenems, quinolones, tetracyclines, and sulphonamides was detected via PCR amplification.

• A case of Whipple's Disease presenting with enlarged intra-abdominal lymph nodes. • Whipple's disease masked as lymphoma. • Histopathologists can be the first to encounter signs suspicious of Whipple's disease. • Diagnosing Whipple's disease requires a high index of suspicion. [ABSTRACT FROM AUTHOR]

Context: Dental tissue remains are the toughest, and chemically, the most stable tissue in the body. Its high resilience in the events of fire and bacterial decomposition makes them vital for DNA analysis by PCR method. Aims: Determination of sex of children through molecular analysis of pulp tissue of exfoliated deciduous teeth stored in different media and analyzed after a different time period. Settings and Design: Sixty samples of deciduous teeth were divided into three groups. Group IA and Group IIA were stored in natural soil and wet clay for 1 month, respectively. Group IB and Group IIB were stored in natural soil and wet clay for 6 months, respectively. Group III was analyzed immediately after extraction. Methods and Material: Sex determination was carried out in five steps: Pulp tissue removal, DNA isolation, DNA quantification, PCR amplification, Sex determination. X and Y specific chromosomes from each sample were amplified and compared. Statistical Analysis Used: Kruskal-Wallis test, Dunn's test, and Wilcoxon signed rank test. Results: Group III revealed the highest amount of DNA quantified. Amount of DNA quantified after 6 months of storage in natural soil and wet clay decreased in both the groups with the samples stored in wet clay showing a maximum decrease. Results of the PCR analysis also showed 100% accuracy rate in the samples of Group III. Conclusions: Sex determination from pulp tissue depends a lot on the quality and quantity of DNA extracted. Sex could be effectively determined among the samples evaluated immediately after extraction. This ability decreases as the storage condition changes and the time period increases. Samples stored in wet clay were found to show the least sex identification ability than dry soil.

Background: Testing for SARS-CoV-2 is essential to provide early COVID-19 treatment for people at high risk of severe illness and to limit the spread of infection in society. Proper upper respiratory specimen collection is the most critical step in the diagnosis of the SARS-CoV-2 virus in public settings, and throat swabs were the preferred specimens used for mass testing in many countries during the COVID-19 pandemic. However, there is still a discussion about whether throat swabs have a high enough sensitivity for SARS-CoV-2 diagnostic testing, as previous studies have reported a large variability in the sensitivity from 52% to 100%. Many previous studies exploring the diagnostic accuracy of throat swabs lack a detailed description of the sampling technique, which makes it difficult to compare the different diagnostic accuracy results. Some studies perform a throat swab by only collecting specimens from the posterior oropharyngeal wall, while others also include a swab of the palatine tonsils for SARS-CoV-2 testing. However, studies suggest that the palatine tonsils could have a tissue tropism for SARS-CoV-2 that may improve the SARS-CoV-2 detection during sampling. This may explain the variation of sensitivity reported, but no clinical studies have yet explored the differences in sensitivity and patient discomfort whether the palatine tonsils are included during the throat swab or not., Objective: The objective of this study is to examine the sensitivity and patient discomfort of a throat swab including the palatine tonsils compared to only swabbing the posterior oropharyngeal wall in molecular testing for SARS-CoV-2., Methods: We will conduct a randomized controlled study to compare the molecular detection rate of SARS-CoV-2 by a throat swab performed from the posterior oropharyngeal wall and the palatine tonsils (intervention group) or the posterior oropharyngeal wall only (control group). Participants will be randomized in a 1:1 ratio. All participants fill out a baseline questionnaire upon enrollment in the trial, examining their reason for being tested, symptoms, and previous tonsillectomy. A follow-up questionnaire will be sent to participants to explore the development of symptoms after testing., Results: A total of 2315 participants were enrolled in this study between November 10, 2022, and December 22, 2022. The results from the follow-up questionnaire are expected to be completed at the beginning of 2024., Conclusions: This randomized clinical trial will provide us with information about whether throat swabs including specimens from the palatine tonsils will improve the diagnostic sensitivity for SARS-CoV-2 molecular detection. These results can, therefore, be used to improve future testing recommendations and provide additional information about tissue tropism for SARS-CoV-2., Trial Registration: ClinicalTrials.gov NCT05611203; https://clinicaltrials.gov/study/NCT05611203., International Registered Report Identifier (irrid): DERR1-10.2196/47446., (©Benedikte Hartvigsen, Kathrine Kronberg Jakobsen, Thomas Benfield, Niels Tobias Gredal, Annette Kjær Ersbøll, Mathias Waldemar Grønlund, Henning Bundgaard, Mikkel Porsborg Andersen, Nina Steenhard, Christian von Buchwald, Tobias Todsen. Originally published in JMIR Research Protocols (https://www.researchprotocols.org), 12.06.2024.)

Aim. To investigate polymorphism of Gli-B1 locus in modern Ukrainian bread wheat cultivars, to analyze the distribution of alleles and to compare the received data with the "core-collection of wheat cultivars" presented by Dr. E. Metakovsky. Methods. Eighty one bread wheat cultivars and lines from different plant breeding institutions and stations of Ukraine were tested using allele-specific primers to Gli-B1 locus developed by Zhang et al. [2003]. PCR products were fractionated in polyacrylamide gel (PAG) and then were stained by silver nitrate. Allelic variants of gliadins were analyzed by electrophoresis in acid polyacrylamide gel (APAGE). Results. Nine allelic variants of gliadins were revealed by APAGE and six alleles of Gli-B1 locus were detected by PCR-analysis. In 52 % of modern Ukrainian bread wheat cultivars we revealed Gli-B1b allelic variant, according to PCR - Gli-B1.1 allele with a 369 bp amplification fragment. In the genotypes of Ukrainian wheat cultivars, the 1RS.1BL translocation, which carries resistance genes, is frequent, as was detected by the absence of any amplification fragments with Gli-B1 primers. The correspondence between allelic variants of gliadins and alleles of Gli-B1 locus is discussed. Conclusions. DNA polymorphism of Gli-B1 locus examined in our research coincides with the diversity of allelic variants of gliadins, which were detected by APAGE method for Ukrainian bread wheat cultivar. However, PCR-analysis with applied primers carried out in this study does not distinguish the alleles that correspond to the Gli-B1c, Gli-B1g and Gli-B1e allelic variants of gliadins. The most common allele (52 %) for the investigated Ukrainian wheat varieties is Gli-B1.1 allele, which was characterized by the amplification fragment of 369 bp, and the presence of 1RS.1BL translocation, which corresponds to Gli-B1b and Gli-B1l allelic variants of gliadins obtained by APAGE, respectively. [ABSTRACT FROM AUTHOR]

Currently, efforts in sugar beet selection are aimed mainly at the creation of highly productive hybrid combinations which could be repeatedly derived from source line material. Naturally, the important step in this process is the development of constant homozygous source lines with high combining ability. Traditional approach in development of uniform sugar beet lines is based on repeated selection of selfpollinated lines. However, biennial lifecycle, cross-incompatibility and inbreeding depression all make breeding and maintaining the genetic homogeneity of sugar beet a challenging endeavor. To increase efficiency of development and preservation of genetic homogeneity of sugar beet lines used in hybrid production at the LLP "KazSRIA&PG", our biotechnology lab started working on introduction of DNA markers into the selection process. Twenty sugar beet lines of various origins and levels of ploidy from LLP "KazSRIA&PG" collection have been studied, to assess their genetic homogeneity, using three SSR markers: Bvv21, Bvv53 and Bvv 155. The produced data showed that the majority of lines can be characterized as having average level of uniformity. The highest homogeneity among individual plants belonging to one line, based on data from all three markers, was estimated for CMS line MS 1631 (Ukraine) and by data from a set of two markers for CMS lines MS-7 and MS-1949 (Russia). [ABSTRACT FROM AUTHOR]

The results of the population genetic and taxonomic analysis of the critical species of Orchids, viz. Anacamptis morio s.l. in the flora of Ukraine are presented. The genetic analysis was carried out using PCR analysis (RAPD-, ISSR amplification) and DNA sequencing (matK, atpF-atpH, rbcL) and genetic differentiation of populations of these plants from Transcarpathian Ukraine (Zakarpattia), Ukrainian Polissia, and Crimea. A significant level of genetic difference between the Crimean populations and populations from Zakarpattia and Ukrainian Polissia, as well as a high level of genetic similarity between all of the studied populations of A. morio subsp. caucasica from Crimea, was established. The highest level of genetic polymorphism of the studied populations was detected by the RAPD-markers OPA-11, OPP-10, OPA-18, and UBC-297 as well as the ISSR-marker (AG)8TC. The main indices of genetic polymorphism of the five A. morio populations were as follows: the proportion of polymorphic loci (P) was 40.0–74.8%, the effective number of alleles (ne) was 1.276–1.479, the genetic diversity (h) was 0.156–0.276, the Shannon information index (I) was 0.229–0.408, and Nei's genetic distances between populations (DN) was 0.047–0.325. For a more complete genetic characterization of populations, the following characteristics were also determined: expected total genetic diversity (Ht), intrapopulation genetic diversity (HS), population differentiation coefficient (Gst), and the gene flow value (Nm). Based on the results of molecular genetic studies and the analysis of morphological and biological data in the flora of Ukraine, it is proposed to delimitate the taxa as follows: A. morio subsp. morio (L.) R.M. Bateman, Pridgeon et M.W. Chase, which grows in the Ukrainian Carpathians and the Polissia as well as A. morio subsp. caucasica (K. Koch) H. Kretzschmar, Eccarius et H. Dietr. distributed in the Crimea and the Northern Black Sea region on the territory of Ukraine. [ABSTRACT FROM AUTHOR]

The salinity environment is one of the biggest threats to Glycyrrhiza uralensis Fisch. (G. uralensis) growth, resulting from the oxidative stress caused by excess reactive oxygen species (ROS). Flavonoids are the main pharmacodynamic composition and help maintain ROS homeostasis and mitigate oxidative damage in G. uralensis in the salinity environment. To investigate whether endophytic Bacillus cereus G2 can improve the salt-tolerance of G. uralensis through controlling flavonoid biosynthesis, the transcriptomic and physiological analysis of G. uralensis treated by G2 in the saline environment was conducted, focused on flavonoid biosynthesis-related pathways. Results uncovered that salinity inhibited flavonoids synthesis by decreasing the activities of phenylalanine ammonialyase (PAL) and 4-coumarate-CoA ligase (4CL) (42% and 39%, respectively) due to down-regulated gene Glyur000910s00020578 at substrate level, and then decreasing the activities of chalcone isomerase (CHI) and chalcone synthase (CHS) activities (50% and 42%, respectively) due to down-regulated genes Glyur006062s00044203 and Glyur000051s00003431 , further decreasing isoliquiritigenin content by 53%. However, salt stress increased liquiritin content by 43%, which might be a protective mechanism of salt-treated G. uralensis seedlings. Interestingly, G2 enhanced PAL activity by 27% whereas reduced trans-cinnamate 4-monooxygenase (C4H) activity by 43% which could inhibit lignin biosynthesis but promote flavonoid biosynthesis of salt-treated G. uralensis at the substrate level. G2 decreased shikimate O-hydroxycinnamoyltransferase (HCT) activity by 35%, increased CHS activity by 54% through up-regulating the gene Glyur000051s00003431 encoding CHS, and increased CHI activity by 72%, thereby decreasing lignin (34%) and liquiritin (24%) content, but increasing isoliquiritigenin content (35%), which could mitigate oxidative damage and changed salt-tolerance mechanism of G. uralensis. [Display omitted] • G2 regulated the relative flow of phenylpropanoids between lignin and flavonoid biosynthesis. • G2 enhanced plant flavonoid content at substrate level in G. uralensis. • G2 inhibited the downstream branches of lignin biosynthesis, thus indirectly promoted flavonoid biosynthesis. • G2 increased CHS and CHI activities, thus directly promoted flavonoid biosynthesis. [ABSTRACT FROM AUTHOR]

The Coronavirus disease 2019 (COVID-19) pandemic demonstrated the threat of airborne pathogenic respiratory viruses such as the airborne Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). The ability to detect circulating viruses in a workplace or dormitory setting allows an early warning system that can alert occupants to implement precautions (e.g. masking) and/or trigger individual testing to allow isolation and quarantine measures to halt contagion. This work extends and validates the first successful detection of SARS-CoV-2 virus in dormitory Heating, Ventilation, and Air Conditioning (HVAC) systems and compares different air sampling methods and media types combined with optimized quantitative Reverse-Transcription PCR (qRT-PCR) analysis. The study was performed in two environments; large dormitories of students who underwent periodic testing for COVID-19 (unknown environment) and the HVAC air from a suite with a student who had tested positive for COVID-19 (known dorm). The air sampling methods were performed using Filter Cassettes, BioSampler, AerosolSense Sampler and Button Sampler (with four media types with different pore sizes of 5 μm, 3 μm, 3 μm (gelatin), and 1.2 μm). The SARS-CoV-2 positive air samples were compared with the positive samples collected by individual student campus track tracing methods using PCR testing on saliva and nasopharyngeal samples. The results show a detection rate of 73% in the unknown environment and a 78% detection rate in the known dorm. Our data show that the virus was detectable with all the sampling methods we employed. However, the AerosolSense sampler and BioSampler performed the best at 63% and 61% detection rates, compared to 25% for the Filter Cassettes and 23% for the Button Sampler. Despite the success rate, it is not possible to definitively conclude which method is most sensitive due to the limited number of samples. These results show that with careful sampling and optimized PCR methods, pathogenic respiratory viruses can be detected in large buildings using HVAC return air. [Display omitted] • SARS-CoV-2 detection was possible by sampling air from the HVAC system inside. Dorms. • Four different air sampling methods were successful in SARS-CoV-2 detection. • Frequent air sampling led to a high detection rate of infected COVID-19 students. • The SARS-CoV-2 detection rate was 22–63% based on the air sampling method. [ABSTRACT FROM AUTHOR]

Abstract Over‐the‐counter (OTC) limited ingredient canine diets could be reliable alternatives to veterinary therapeutic formulations for the diagnosis and management of adverse food reaction (AFR). However, the possibility of undeclared ingredients jeopardizes the efficacious use of OTC options for medical purposes. The objective was to determine the presence of undeclared ingredients in OTC canine dry diets marketed as limited or single protein source diets. Twenty‐one OTC adult canine diets marketed as limited or single protein source diets were purchased. Multiplex PCR was used to screen for DNA of 10 mammalian species with species‐specific primers that anneal to regions of the mitochondrial cytochrome b gene. The presence of DNA from one or more species not declared on the label was identified in all 21 diets: cow (Bos taurus), pig (Sus scrofa), sheep (Ovis sp.), goat (Capra hircus) and bison (Bison bison). Twenty diets were positive for the declared protein source and one diet was negative for the declared species. Cat (Felis catus), dog (Canis sp.), horse (Equus sp.), mouse (Mus musculus) and rat (Rattus norvegicus) DNA was not identified in any samples. The presence of undeclared mammal species in OTC canine dry diets marketed as having limited or single protein source ingredients may complicate AFR diagnosis and treatment. However, PCR can detect a miniscule amount of DNA which might not be clinically significant, because the amount needed to elicit a response is unknown. Quantification of the contamination was not determined in this study, precluding discrimination of intentional adulteration from unavoidable cross‐contamination.

Leaf blight caused by Alternaria alternata (Fr.) Keissler is a severe disease of Gloriosa, causing severe yield loss in every part of Gloriosa growing areas of Tamil Nadu. The foliar pathogen was isolated from disease-infected leaves and proved its pathogenicity. Further, the molecular analysis of the pathogen using 18S rDNA confirmed the pathogen as Alternaria alternata. Attempts were made to explore the biocontrol agents and fungicides for the management of leaf blight incidence under field conditions. Four field trials conducted from 2014 to 2018 revealed that foliar application of talc-based formulation of Bacillus subtilis significantly reduced the leaf blight incidence and increased the seed yield under field conditions. Besides, prophylactic application of these biocontrol agents has also increased the plant growth parameters like plant height, number of flowers/plant, no. of pods/plant and number of seeds/pod. Similarly, foliar application of chlorothalonil (0.1%) and mancozeb (0.2%) was also credited to managing the leaf blight disease under field conditions. [ABSTRACT FROM AUTHOR]

This study aimed to prepare the sugar industry for the possible introduction of genetically modified (GM) sugarcane and derived retail sugar products and to address several potential public concerns regarding the characteristics and safety of these products. GM sugarcane lines with integrated Cry1Ab and EPSPS foreign genes were used for GM sugar production. Traditional PCR, real-time fluorescent quantitative PCR (RT-qPCR), and enzyme-linked immunosorbent assay (ELISA) were performed in analyzing leaves, stems, and other derived materials during sugar production, such as fibers, clarified juices, filter mud, syrups, molasses, and final GM sugar product. The toxicity of GM sugar was examined with a feeding bioassay using Helicoverpa armigera larvae. PCR and RT-qPCR results showed that the leaves, stems, fibers, juices, syrups, filter mud, molasses, and white granulated sugar from GM sugarcane can be distinguished from those derived from non-GM sugarcane. The RT-qPCR detection method using short amplified product primers was more accurate than the traditional PCR method. Molecular analysis results indicated that trace amounts of DNA residues remain in GM sugar, and thus it can be accurately characterized using molecular analysis methods. ELISA results showed that only the leaves, stems, fibers, and juices sampled from the GM sugarcane differed from those derived from the non-GM sugarcane, indicating that filter mud, syrup, molasses, and white sugar did not contain detectable Cry1Ab and EPSPS proteins. Toxicity analysis showed that the GM sugar was not toxic to the H. armigera larvae. The final results showed that the GM sugar had no active proteins despite containing trace amounts of DNA residues. This finding will help to pave the way for the commercialization of GM sugarcane and production of GM sugar.

The genetic polymorphism of Iris pumila L., a rare ornamental species involved in hybridization, was studied by PCR analysis using three types of primers: those based on microsatellite sequences (ISSR), MGE sequences (IRAP and iPBS), and abiotic stress response genes (LP-PCR). High levels of intraspecific and intrapopulation genetic polymorphism were revealed, which were comparable to those in other species of the Iris genus. The main indicators of genetic polymorphism of five I. pumila populations from the territory of Ukraine were the proportion of polymorphic loci (P) of 26.5–68.5%, the Shannon index (S) of 0.105–0.285, and genetic diversity (He) of 0.069–0.190. The dependence of the variability level on the population size was direct in the ISSR analysis and inverse according to the other two markers. The direct relationship between genetic and geographic distances between populations was found only using ISSR markers. The highest level of genetic polymorphism was detected by LP-PCR markers, while the population identification of all individual plants was possible only using ISSR markers. The tested system of PCR markers can be used to monitor the state of the gene pool and investigate the genetic structure of populations and migration processes. [ABSTRACT FROM AUTHOR]

This study aimed to prepare the sugar industry for the possible introduction of genetically modified (GM) sugarcane and derived retail sugar products and to address several potential public concerns regarding the characteristics and safety of these products. GM sugarcane lines with integrated Cry1Ab and EPSPS foreign genes were used for GM sugar production. Traditional PCR, real-time fluorescent quantitative PCR (RT-qPCR), and enzyme-linked immunosorbent assay (ELISA) were performed in analyzing leaves, stems, and other derived materials during sugar production, such as fibers, clarified juices, filter mud, syrups, molasses, and final GM sugar product. The toxicity of GM sugar was examined with a feeding bioassay using Helicoverpa armigera larvae. PCR and RT-qPCR results showed that the leaves, stems, fibers, juices, syrups, filter mud, molasses, and white granulated sugar from GM sugarcane can be distinguished from those derived from non-GM sugarcane. The RT-qPCR detection method using short amplified product primers was more accurate than the traditional PCR method. Molecular analysis results indicated that trace amounts of DNA residues remain in GM sugar, and thus it can be accurately characterized using molecular analysis methods. ELISA results showed that only the leaves, stems, fibers, and juices sampled from the GM sugarcane differed from those derived from the non-GM sugarcane, indicating that filter mud, syrup, molasses, and white sugar did not contain detectable Cry1Ab and EPSPS proteins. Toxicity analysis showed that the GM sugar was not toxic to the H. armigera larvae. The final results showed that the GM sugar had no active proteins despite containing trace amounts of DNA residues. This finding will help to pave the way for the commercialization of GM sugarcane and production of GM sugar. [ABSTRACT FROM AUTHOR]

The objective of this study was to investigate the genetic diversity of hexaploid wheat varieties of Uzbekistan breeding using simple sequence repeat (SSR) markers. These varieties are adapted to local conditions, and can be considered as the most important supplier of genetic resources for cultivation in Uzbekistan and other countries. Microsatellite markers are now most widely used and effective classes of DNA markers for genotyping, certification and classification of plant varieties. In this paper, genotyping results of 32 hexaploid wheat domestic varieties using 144 microsatellite primer pairs are presented. Microsatellite primer pairs were chosen from literature data and 36 primer pairs (from 144) gave polymorphic well-reproducible PCR-fragments. The individual SSR spectra differing in number of amplicons were obtained for each variety. A total number of 141 alleles for 36 microsatellite loci were detected. The number of alleles per locus ranged from 2 to 6, the mean number of alleles per locus (Na) was 3 alleles. For the studied genotypes group the effective number of alleles (ne) characterizing the loci by the allele frequency, varied from 1.7 to 4.8, the mean number of alleles per locus was 2.8. The expected heterozygosity (He) ranged from 0 to 0.792, averaging 0.626, in studied wheat population. The amplified fragment sizes ranged from 93 to 552 bp. The polymorphic index content (PIC) ranged from 0 to 0.758. A dendrogram was constructed using the alleles set of microsatellite loci, reflecting the phylogenetic differences of the studied hexaploid wheat varieties. It showed that Uzbekistan breeding varieties are divided into two main clusters, which may be evidence of their common origin. A genetic formula has been developed for each Uzbek wheat variety. It can be used for identification, certification of these varieties, as well as for the selection of parental pairs in the wheat breeding programs.

Development of conservation biological control programs requires the identification of sources that contribute to predator colonization of crops. Macrolophus pygmaeus (Rambur) (Hemiptera: Miridae) is an efficient polyphagous predator used in biological control programs in vegetable crops in Europe. We have developed a marking method based on spraying with a solution of the brine shrimp Artemia spp. (Anostraca: Artemiidae) cysts, followed by a PCR detection of Artemia DNA to monitor M. pygmaeus dispersal from banker plants to tomato crops. Experiments conducted in climatic chambers show that the topical application of this marking solution on M. pygmaeus does not significantly reduce adult longevity and that it is detected up to 6 d after the application. When this Artemia solution was applied on Calendula officinalis L. banker plants harboring M. pygmaeus and maintained outdoors, Artemia DNA was still detected on 62% of the insects after 6 d. The conducted field applications in commercial greenhouses have confirmed the usefulness of this method to monitor M. pygmaeus dispersal from banker plants to a newly planted tomato crop. This method can be used to assess arthropod movement, being an interesting molecular approach for further improving future pest management strategies. [ABSTRACT FROM AUTHOR]

The goal of this study was a molecular genetic passportization of the initial breeding material of sugar beet (male sterile lines, synanthous pollinators, and their hybrids). The following SSR primers were used: Unigene 24 552, Unigene 2305, Unigene 17 623, Unigene 14 805, and Unigene 62 524. The length range of the obtained DNA fragments varied within 100–3000 bp, and up to 11 polymorphic bands per genotype were amplified. The maximum Polymorphic Information Content (PIC) value was revealed for the Unigene 17 623 (PIC = 0.88), Unigene 2305 (PIC = 0.84), and Unigene 14 805 (PIC = 0.85) loci that provided a possibility to differentiate sugar beet breeding material. Using five of the tested SSR markers, the molecular genetic passportization of 26 genotypes of valuable breeding samples of this crop was carried out that allowed their identification and certification for further use in a marker-assisted selection. The calculated genetic distances between male sterile forms and synanthous pollinators varied from 2.236 to 4.796. Parent samples located at considerable genetic distances from each other were recommended for use in creating heterotic hybrids. [ABSTRACT FROM AUTHOR]

Using allele-specific primers and hybridological analysis, the allelic composition of the VRN and PPD loci was determined in common wheat lines derived from the Bezostaya 1 (Bez1) cultivar. In lines of the Bez1 cultivar carrying different dominant alleles of the VRN genes and their combinations, the duration of certain developmental phases was examined. It was demonstrated that, in lines with the combination of two dominant alleles of the VRN-1 locus (Bez1Vrn-A1aVrn-B1a and Bez1Vrn-A1aVrn-B1c), the duration of the "tillering–first node" and "shoots–heading" periods was statistically significantly decreased compared to the initial isogenic lines (i:Bez1Vrn-A1a, i:Bez1Vrn-B1a, and i:Bez1Vrn-B1c). In addition, the presence of two dominant alleles led to the reduction in the time span of the organogenesis stages, as shown by studying the dynamics of shoot apex size and morphology in common wheat lines of the Bez1 cultivar. The productivity analysis in the lines of the Bez1 cultivar showed that the i:Bez1Vrn-B1c line was characterized by highest productivity among isogenic lines, while the Bez1Vrn-A1a Vrn-B1c line was more productive than the Bez1Vrn-A1a Vrn-B1a line. [ABSTRACT FROM AUTHOR]

Context: Dental tissue remains are the toughest, and chemically, the most stable tissue in the body. Its high resilience in the events of fire and bacterial decomposition makes them vital for DNA analysis by PCR method.Aims: Determination of sex of children through molecular analysis of pulp tissue of exfoliated deciduous teeth stored in different media and analyzed after a different time period.Settings and Design: Sixty samples of deciduous teeth were divided into three groups. Group IA and Group IIA were stored in natural soil and wet clay for 1 month, respectively. Group IB and Group IIB were stored in natural soil and wet clay for 6 months, respectively. Group III was analyzed immediately after extraction.Methods and Material: Sex determination was carried out in five steps: Pulp tissue removal, DNA isolation, DNA quantification, PCR amplification, Sex determination. X and Y specific chromosomes from each sample were amplified and compared.Statistical Analysis Used: Kruskal-Wallis test, Dunn's test, and Wilcoxon signed rank test.Results: Group III revealed the highest amount of DNA quantified. Amount of DNA quantified after 6 months of storage in natural soil and wet clay decreased in both the groups with the samples stored in wet clay showing a maximum decrease. Results of the PCR analysis also showed 100% accuracy rate in the samples of Group III.Conclusions: Sex determination from pulp tissue depends a lot on the quality and quantity of DNA extracted. Sex could be effectively determined among the samples evaluated immediately after extraction. This ability decreases as the storage condition changes and the time period increases. Samples stored in wet clay were found to show the least sex identification ability than dry soil. [ABSTRACT FROM AUTHOR]

Background: Accurate determination of gender from the skeletal remains has a significant role in the identification process. In conditions of extreme fragmentation, information from DNA plays a vital role in establishing the gender of the person and further contributes to personal identification. Method: Sixty sound and non-carious extracted deciduous teeth were grouped into three of 20 each. Group 1 was analyzed immediately after extraction. Group 2 and Group 3 were stored at room temperature for three months and Fifteen months respectively before subjecting them to PCR analysis. The X and Y chromosomes from each sample were amplified and compared with the actual gender of the person. Shapiro-Wilk test the independent sample t-test, paired t-test and the chi-square test were used to analyze the data obtained. Result: The mean DNA volume (mg/ml) obtained immediately after extraction was significantly more than that from the teeth stored at room temperature for 15 months. The PCR analysis did not show any significant difference between group 1 and group 3 (p=0.072). Statistically significant difference was observed, when the results of group 1 were compared with that of group 3. Conclusion: Gender could be conclusively determined from the samples analyzed immediately after extraction. The accuracy of gender determination decreased as the period of storage increased. [ABSTRACT FROM AUTHOR]

Objective: The aim of our study was to determine the relation of particular genetic variants in selected genes (GSTM1, GSTT1 null genotypes; rs1695 GSTP1; rs10735781 EVI5) to the risk of multiple sclerosis (MS) development and find out the possible association with disease disability progression rate. Material and methods: Our study included 202 MS patients and 174 healthy control volunteers. MS patients were divided according to disability progression rate to three groups - slowly progressing, mid-rate progressing and rapidly progressing. All DNA samples were isolated from venous blood. Genotyping was performed by PCR-RFLP and multiplex PCR. Results: Our analysis showed that GSTT1 null genotype (OR 0.56; 95%CI 0.33 -0.95; p=0.04) and GSTM1, GSTT1 double null genotype (OR 0.32; 95%CI 0.14 - 0.74; p=0.006) are potentially protective in relation to MS. We observed similar result in GSTT1 null genotype in association with mid-rate progression (OR 0.48; 95%CI 0.24 - 0.97; p=0.05). Frequency of GSTM1 and GSTT1 double null genotype is significantly lower in subgroup of MS patients with progression rate defined as slow (OR 0.22; 95%CI 0.05 - 0.98; p=0.05) and middle (OR 0.33; 95%CI 0.11 - 0.99; p=0.045). We did not show any significant association of genetic changes rs1695 in GSTP1 and rs10735781 in EVI5 with MS or rate of disease progression. Conclusions: Genetic basis of multiple sclerosis is still not fully elucidated. Further research may clarify our results and confirm the value of studied factors for clinical practice.

Most papers on sport genetics identify differences between genotypes of athletes and a control group. It is obvious that the genetic differences should also be among sportsmen with different qualifications. Additionally, athletes’ performance depends not only on their genotypes, but also on the gene activities, which can be different during the training process in various athletes.The aim of the study was to compare genotypes of athletes with different qualifications and to analyze the change in expression of some genes responsible for the physical performance. Genotypes of 143 elite sportsmen of 18 national teams were analyzed by PCR method. A comparison of the genotypes of Masters of Sports, International Masters of Sports and Honored Masters of Sports showed that the frequencies of favorable gene variants were higher in the genotypes of more qualified athletes; it proves an appropriate genetic potential necessity for high achievements in sports. The analysis of UCP2, HIF1A and MTHFR gene expression changes in response to two-week hypoxiс training was performed on 15 skaters of high qualification. We found that average UCP2 and MTHFR mRNA levels had significantly increased after the training but the expression of the HIF1A gene had reduced. At the same time, individual athlete variability in UCP2, HIF1A and MTHFR gene expression was revealed. Genotype influence on gene expression was shown with the help of the UCP2 gene – its activity was higher in sportsmen with Val/Val than with Val/Ala or Ala/Ala genotypes. Consequently, genotyping and analysis of gene expression is very important for athlete selection and training.