ABSTRACT
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.
ABSTRACT
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.
ABSTRACT
[This corrects the article on p. 1736 in vol. 7, PMID: 27917186.].
ABSTRACT
BACKGROUND: 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. METHODS: Amplifluor-like SNP and KASP analysis was carried out using published and own design of Universal probes (UPs) and Gene-specific primers (GSPs). RESULTS: 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. CONCLUSIONS: 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.
Subject(s)
Genotyping Techniques/methods , Plants/genetics , Polymorphism, Single Nucleotide/genetics , Alleles , Genetic Markers/genetics , Nucleic Acid Amplification Techniques/methods , Reproducibility of ResultsABSTRACT
A panel of 89 local commercial cultivars of bread wheat was tested in field trials in the dry conditions of Northern Kazakhstan. Two distinct groups of cultivars (six cultivars in each group), which had the highest and the lowest grain yield under drought were selected for further experiments. A dehydration test conducted on detached leaves indicated a strong association between rates of water loss in plants from the first group with highest grain yield production in the dry environment relative to the second group. Modern high-throughput Amplifluor Single Nucleotide Polymorphism (SNP) technology was applied to study allelic variations in a series of drought-responsive genes using 19 SNP markers. Genotyping of an SNP in the TaDREB5 (DREB2-type) gene using the Amplifluor SNP marker KATU48 revealed clear allele distribution across the entire panel of wheat accessions, and distinguished between the two groups of cultivars with high and low yield under drought. Significant differences in expression levels of TaDREB5 were revealed by qRT-PCR. Most wheat plants from the first group of cultivars with high grain yield showed slight up-regulation in the TaDREB5 transcript in dehydrated leaves. In contrast, expression of TaDREB5 in plants from the second group of cultivars with low grain yield was significantly down-regulated. It was found that SNPs did not alter the amino acid sequence of TaDREB5 protein. Thus, a possible explanation is that alternative splicing and up-stream regulation of TaDREB5 may be affected by SNP, but these hypotheses require additional analysis (and will be the focus of future studies).
