KASP (KBioscience Competitive Allele Specific PCR) and Amplifluor (Amplification with fluorescence) SNP markers are two prominent technologies based upon a shared identical Allele-specific PCR platform. Amplifluor-like SNP and KASP analysis was carried out using published and own design of Universal probes (UPs) and Gene-specific primers (GSPs). Advantages of the Amplifluor-like system over KASP include the significantly lower costs and much greater flexibility in the adjustment and development of ‘self-designed’ dual fluorescently-labelled UPs and regular GSPs. The presented results include optimisation of ‘tail’ length in UPs and GSPs, protocol adjustment, and the use of various fluorophores in different qPCR instruments. The compatibility of the KASP Master-mix in both original and Amplifluor-like systems has been demonstrated in the presented results, proving their similar principles. Results of SNP scoring with rare alleles in addition to more frequently occurring alleles are shown. The Amplifluor-like system produces SNP genotyping results with a level of sensitivity and accuracy comparable to KASP but at a significantly cheaper cost and with much greater flexibility for UPs with self-designed GSPs.

Background and Aim: Trichinellosis is caused by a species of roundworm called Trichinella and is an invasive disease causing severe medical, veterinary, and socioeconomic problems worldwide. More than 100 mammalian species are Trichinella hosts. Among domestic animals, pigs and dogs are prone to trichinellosis. An essential aspect of controlling the spread of infection is to identify the number and level of infections in wild carnivores in the country. However, the number, habitats, and movements of wild animal Trichinella hosts in Kazakhstan have not been reported yet. This study aimed to monitor the wild animal habitat nearby the settlements for tracking the trichinellosis speading among carnivores. Materials and Methods: Wild carnivorous animals were captured in seven regions of the Republic of Kazakhstan. The carcasses of corsacs, wolves, foxes, wild boars, and badgers were studied. Muscle tissue samples from spontaneously infected wild animals were collected. The digestion method in “GASTROS-2M” was used to isolate Trichinella spp. from animal muscles. The species of the parasite was determined by a polymerase chain reaction for 5S spacer of Trichinella ribosomal DNA with subsequent sequencing by Senger. Statistical analysis methods were performed for average value in Microsoft Excel 2010. Results: The results of the research showed that among 155 animals wolves (20.4%) and foxes (26.7%) were the most infected with invasive Trichinella larvae. The invasion intensity was 503.6% in foxes and 289.7% in wolves. However, badgers (164%), wild boars (0%), and corsacs (0%) presented lower invasion levels. Using specific primers, larvae samples were identified as Trichinella nativa. Conclusion: The results of monitoring revealed the spread of trichinosis among wild animals: wolves, foxes, badgers. The Karaganda, Kostanay, Western Kazakhstan, and Akmola regions had the largest distribution of wild animals infected with trichinellosis. In total, 20% of the 155 studied animals were infected. The greatest invasion intensity was typical for wolves, foxes and badgers. It is necessary to monitor the spread of trichinellosis among wild carnivores to control the epidemiological situation and reduce the level of spontaneous infection among animals. Regular monitoring of habitats and carnivores must be conducted within the country and in the border areas. Keywords: distribution, intensity of infection, trichinellosis, wild carnivores.

Background and Aim: Trichinellosis remains a dangerous disease for humans and animals, which can lead to a lethal outcome. The study of specific body reactions in response to invasion by different types of Trichinella can help in the early diagnosis of the disease. This study aimed to investigate the hematological, biochemical, and serological characteristics of rabbits experimentally infected with trichinellosis, as well as the possibility of using changes in these parameters at various disease stages for early hematological, biochemical, and serological diagnosis of trichinellosis. Materials and Methods: Three groups of rabbits were orally infected with Trichinella nativa and Trichinella spiralis derived from encysted T. spirtalis larvae in pork muscle samples. The first and second groups were infected with T. nativa and T. spiralis, respectively, while the third group served as control by receiving a physiological solution. An ADVIA 2120i automatic hematology analyzer with a blood smear staining module was used to determine the hematological parameters of rabbits. Antigens were used in an enzyme-linked immunosorbent assay (ELISA) to detect antibodies in the sera of infected rabbits that were supernatants containing excretory-secretory antigens (ES-Ag) and somatic antigen (S-Ag). Results: The detection of biochemical responses to the invasion of T. nativa and T. spiralis isolates was detected and hematological parameters were featured in two cases. Trichinella nativa increased the number of erythrocytes, neutrophils, eosinophils, monocytes, basophils, and thrombocytes on day 7 in rabbits. Creatine kinase (CK) is regarded as the most important indicator for the early detection of parasite invasion. Blood biochemistry showed no active response to T. spiralis infection. However, counts of erythrocytes, neutrophils, lymphocytes, and CK rose significantly. In both color indicators, the number of thrombocytes decreased. Enzyme-linked immunosorbent assay with ES-Ag and S-Ag of these isolates demonstrated the ability to detect antibodies as early as 7 days after infection, with a significant increase in the marker up to 70 days. Conclusion: On the 7th day after infection, blood tests of infected animals revealed CK-N-acetyl-cysteine (18.2%) and neutrophils (43%) when infected with T. nativa and neutrophils (26.7%) and lymphocytes (20%) when infected with T. spiralis. These indicators may serve as specific parameters for the early detection of Trichinella spp. invasion.

Germination is a biotechnological process that activates enzymatic reactions and alters the chemical content of grain, improving its nutritional and functional value. However, the duration of the germination process significantly affects grain composition and properties. Prolonged and uncontrolled germination can lead to undesirable changes, such as excessive enzymatic activity, microbial contamination of sprouted grains, and loss of dry matter. While short-term germination is preferable for producing various food products, including whole-grain porridge, this article analyzed the impact of short-term germination of triticale grain (for 18, 20, 22, and 24 h) on its biochemical indicators and functional-technological properties. This research established that changes in biochemical values accompany the germination of triticale grain. The most significant increase in protein content was observed after 20 h of germination (GT 20), which increased by 4.43%. In contrast, after 24 h (GT 24), a decrease of 3.27% was noted compared to the original content in the whole grain (WGT). The contents of fats, carbohydrates, and ash tended to decrease. Germination promoted an increase in the amount of essential amino acids and improved the amino acid profile, particularly during the 20 and 22 h germination intervals. At 24 h of germination, the highest increase in total phenolic compounds (28.6%) and antioxidant activity (20.8%) was recorded. B-group vitamins, thiamine, and riboflavin, were detected in the sprouts only after 22 and 24 h of germination. The highest thiamine content (0.17 mg/kg) was observed at the 22nd hour, and the highest riboflavin content (2.51 mg/kg) was observed at the 24th hour. Niacin content showed a steady increase throughout the germination period. The maximum magnesium (0.200%) and molybdenum (0.589%) contents were recorded after 24 h of germination, while the calcium content increased at all germination intervals. The functional and technological properties of sprouted triticale grain improved, while the pasting (gelatinization) properties tended to decrease. Thus, it has been established that short-term germination enhances the biochemical indicators and functional-technological properties of triticale grains, indicating their potential for use in the production of whole-grain porridge.

Drought is a primary ecological stress limiting wheat yield in water-deficient regions. Conducting targeted genetic selection of wheat cultivars can expedite the adaptation process of wheat to the climatic conditions of the region, allowing for the identification of high-yielding varieties with stable genetic traits. This study investigated the impact of the TaGW8 and TaGS3A genes, known for their contribution to wheat productivity. The effective productivity genes TaGW8-B1b/B1a and the TaGS5-3A-T genome exert a 32.8% influence on the variability of the 1000 grain weight (TGW) trait. This influence stems from both individual genes and their interactions, with at least 17.5% of TGW variability explained by the gene combinations examined in the study. Notably, the TaGS5-3A-T gene exhibits a significant positive correlation with total yield, exceeding 63%. The integration of these productivity genes, based on field phenotypic data, has resulted in an overall yield increase of selected samples by 0.8 tons/ha compared to the country's average multi-year indicator.

Background and Aim: Diagnosis of trichinellosis at the intestinal stage during larval development is the primary challenge in the early detection and treatment of trichinellosis. The use of serine protease as a diagnostic marker for serological tests has been the subject of various studies, but data on Trichinella nativa serine protease in the intestinal phase are still insufficient for a proper diagnosis. This study aimed to establish the duration of the intestinal phase for early diagnosis and to determine the level of expression of the serine protease gene in T. nativa and Trichinella spiralis larvae. Materials and Methods: We used European isolates from T. spiralis pigs and T. nativa larvae isolated from spontaneously infected wild carnivorous animals (wolf, Karaganda region) in Central Kazakhstan. Isolation of larvae from the meat of infected animals was carried out using the compressor method. For two species of Trichinella, 36 mice (in each group 18 mice) were infected with 250 larvae and euthanized by intramuscular injection of xylazine followed by an intravenous overdose of anestofol at 3, 5, 7, 14, 21, and 30 dpi (each day 3 infected mice) and one control group (3 mice). Sequencing and bioinformatics methods were used to determine the DNA and cDNA of the serine protease gene, and molecular methods (DNA extraction, reverse transcription polymerase chain reaction, and sequence) were used to measure the accumulation of serine protease transcripts in isolated larvae. Results: The results showed differences in the duration of intestinal phase between T. spiralis and T. nativa. The intestinal larvae of T. nativa were observed from 7 to 30 dpi, and the intensity of invasion increased up to 30 dpi (p < 0.001), while in the case of T. spiralis, the increase in larval growth in the intestinal phase decreased to 21 dpi, and only an increase of 1.6 ± 0.88 (p < 0.01) was detected at 30 dpi. T. nativa muscle larvae were detected at 21 dpi, compared with T. spiralis at 14 dpi. This characteristic was also reflected in the levels of serine protease transcripts in the samples. Accumulation was observed in both cases higher in the muscular stage of development, whereas the duration of the intestinal stage of T. nativa made it possible to detect serine protease at 30 dpi. Conclusion: The intestinal stage of T. nativa lasts for 30 days, indicating that the use of T. nativa serine protease is useful for the identification of intestinal infection. Furthermore, this protein can be used to identify T. spiralis and T. nativa in laboratory samples. Serine protease can be used as a marker for serological diagnosis. Within the framework of the research topic, it is important to conduct further studies on the species specificity of the obtained recombinant protein. It is necessary to focus on identifying highly specific Trichinella proteins for early disease detection. Keywords: enzyme-linked immunosorbent assay, gene expression, invasion, transmembrane serine protease, Trichinella nativa, Trichinella spiralis, trichinellosis.