Epizootiological monitoring of wolf helminths in Northern and Central Kazakhstan.

Background and Aim: Wolves (Canis lupus) play a role in nature, including the regulation of the number of ungulates and the use of dead animals. In addition, wolves are a natural link and carrier for the spread of many parasitic invasions. Hence, the main task in preventing the spread of parasitic invasions is to regulate the wolf population. This study aimed to monitor the endoparasitological fauna of wild wolves inhabiting Northern and Central Kazakhstan. Materials and Methods: Overall, 81 wolves were investigated for parasitic worms using the K. I. Scriabin method. Wolf intestinal materials were collected from the following six regions: North Kazakhstan, Pavlodar, Kostanay, Akmola, Ulytau, and Karaganda. The genetic diversity of the parasites was identified using a polymerase chain reaction with specific primers. After data collection, a comprehensive statistical analysis was performed. Results: Several helminth types were identified in wolves, including Echinococcus granulosus, Taenia hydatigena, Mesocestoides spp., Toxascaris leonina, Trichinella nativa, Alaria alata, and Dirofilaria repens. Based on the results of this study, young male wolves aged 1-4 years were the most vulnerable to helminthiasis. Wolves living in steppe and semi-desert regions are often exposed to helminth infections. The prevalence of T. nativa in the wolves was 20.4%. This study also revealed the presence of echinococcosis among wolf populations in Karaganda and Kostanay, with prevalence rates of 4.1% and 4.7%, respectively. The overall prevalence of tapeworms in wolves was 54.3%. Conclusion: This study highlights the significance of understanding the potential risks associated with helminth infections in wild carnivores because helminths can act as disease reservoirs and pose a threat to humans, livestock, and other wild carnivores. These results can contribute to the development of effective control and management strategies for helminth infections in wolves, which can infect humans and livestock.

Genetic diversity and haplotype analysis of cattle hydatid cyst isolates using mitochondrial markers in Kazakhstan.

Background and Aim: In Kazakhstan, the study of Echinococcus infection among farm animals is crucial to monitor the invasion among livestock and map the data obtained. Unfortunately, there are only partial data on the study of Echinococcus among cattle's in Kazakhstan, which makes it difficult to conduct a comparative analysis of the epidemiological situation among livestock animals. The present study aimed to molecularly identify the species and haplotypes of the E. granulosus complex infecting cattle in Kazakhstan and investigate their genetic variation relative to mitochondrial (mt) targets. Materials and Methods: Individual cyst isolates (n = 700) were collected from infected cattle lungs and livers after slaughter from the slaughterhouse. Total DNA was extracted from the germinal layers of the cyst from each isolate. This DNA sequenced partial mt genes of cytochrome c oxidase 1 (450 bp) and NADH dehydrogenase 1 (1200 bp). Results: We determined that all the sequences were detected as E. granulosus s.s., of which 69 (94.5%) samples belonged to G1, and only 4 (5.4%) samples belonged to the G3 genotype. After bioinformatic analysis, 38 haplotypes were identified. Conclusion: Our findings revealed that the G1 genotype of E. granulosus s.s. is the predominant cattle genotype in Kazakhstan. However, only one region showed the presence of two genotypes G1 and G3, in the sequence, which suggests that further research is needed to investigate the epidemiology of Echinococcus infection in cattle in Kazakhstan.

A Comparative Study of New Fluorescent Anthraquinone and Benzanthrone α-Aminophosphonates: Synthesis, Spectroscopy, Toxicology, X-ray Crystallography, and Microscopy of Opisthorchis felineus.

In this research, we explore the synthesis of and characterize α-aminophosphonates derived from anthraquinone and benzanthrone, focusing on their fluorescence properties and potential applications in confocal laser scanning microscopy (CLSM). The synthesized compounds exhibit notable solvatochromic behavior, emitting fluorescence from green to red across various solvents. Spectroscopic analysis, including 1H-, 13C-, and 31P-NMR, FTIR, and mass spectrometry, confirms the chemical structures. The compounds' toxicity is evaluated using etiolated wheat sprouts, revealing varying degrees of impact on growth and oxidative damage. Furthermore, the study introduces these α-aminophosphonates for CLSM imaging of the parasitic flatworm Opisthorchis felineus, demonstrating their potential in visualizing biological specimens. Additionally, an X-ray crystallographic study of an anthraquinone α-aminophosphonate provides valuable structural insights.

Toxascaris leonina in dogs - A nematode species of high prevalence in some regions of Eurasia.

A recent meta-analysis of studies from around the world had shown a global prevalence of Toxascaris leonina in stray dogs of about 7%. However, studies from Eurasian regions, where higher percentages of positive dogs were often found, were under-represented in this meta-analysis. This prompted the present study. Its main objective was to examine free-roaming dogs from the capital city of Kazakhstan to obtain information on the current occurrence of T. leonina egg shedding compared to Toxocara spp. Faecal samples from 500 free-roaming dogs from the city of Astana and its suburbs were collected 1-2 days after admission to the municipal animal shelter during three time periods (May to October 2019, September 2020, and April 2021). Samples were examined by a saline flotation method. Nematode egg-positive samples were subjected to a modified McMaster method to determine the number of eggs per gram of faeces (EPG). Toxascaris leonina eggs were found in 53.8% and Toxocara spp. eggs in 14.8% of the dogs examined. The prevalence of T. leonina egg shedding was significantly associated with age class but not with sex: dogs aged between 6 months and 2 years were more likely to be T. leonina egg positive than puppies. The intensity of T. leonina egg shedding was also age dependent: dogs aged between 6 months and 2 years had significantly higher EPGs compared to younger or older animals. In contrast, the prevalence of Toxocara spp. egg shedding was associated with both age class and sex: dogs older than 2 years were less likely to shed Toxocara spp. eggs than puppies, and females were less likely to be Toxocara spp. egg positive compared to males. The present results confirm those of other studies in Eurasian regions. It is therefore reasonable to assume that the global prevalence of T. leonina infection has been underestimated and should be revised accordingly.

Epizootic situation on horse helminthiases in the northern region of Kazakhstan.

The research relevance is predefined by the widespread distribution of parasitosis, including helminthiases, alongside constant depasture on the same areas of pastures, which annually causes massive re­infection of horses. In this regard, the research aims to identify the age, seasonal, and territorial dynamics of infection of horses with helminthiases in the context of the districts of the northern region of Kazakhstan. The primary research method was an experimental analysis that allowing to consider the dynamics and causes of horse helminthiases, associated with the prepatent period, the resistance of the organism, and the system of keeping and feeding horses. The study presents the results of studies of horse helminthiases on the territory of the Northern region of Kazakhstan, covering 4 regions that currently exist in 61 agricultural formations. Faecal samples of 4,395 horses were examined by scatological methods and 126 samples - by incomplete helminthological autopsy. As a result of coprolarvoscopic and coprological studies, it was found that in horses, the dominant representatives among the invasive larvae of strongylates are small and large palisade worms. In scatological studies, strongylatosis of the gastrointestinal tract was recorded in 63.9% of cases, oxyurosis in 40.8%, and parascariasis in 25.9%. The study also presents the results of post­mortem diagnostics, which was carried out during the slaughter or due to the death of the horse.

Genetic identification of Trichinella species found in wild carnivores from the territory of Kazakhstan.

Trichinellosis, also called trichinosis, is a foodborne parasitic disease caused by eating raw or undercooked meat from animals infected with Trichinella spp. larvae and affects both animals and humans. Although on the territory of Kazakhstan, the species characteristics and prevalence of this helminth were studied back in the 90s, the data have not been updated since then. Given the above, our study was aimed at identifying Trichinella spp. using parasitological and molecular genetics methods. In our work, we studied 160 samples of muscle tissue of wild animals living in the natural zones of steppes and semi-deserts. Of the animals examined, 32 were positive for Trichinella spp., including 1 lynx (Lynx lynx), 17 wolves (Canis lupus), 11 foxes (Vulpes vulpes), 1 jackal (Canis aureus) and 2 corsac foxes (Vulpes corsac). Helminths were extracted using the digestion method. DNA was extracted using a Gene Jet commercial kit (Thermo Fisher Scientific, United Kingdom). For species identification a multiplex PCR, amplification of ESV, ITS1, and ITS2 genes regions was performed. After that, uniplex PCR was performed on the 5S rDNA and ITS1 genes region for sequencing analysis. The resulting sequences were subsequently used to construct a phylogenetic tree and the studied samples were identified as Trichinella nativa and Trichinella britovi. Thus, we can conclude that there is a circulation of two species of Trichinella in Kazakhstan, highlighting that constant control and monitoring of wild animals are necessary to prevent transmission and protect the health of people.

Trichinellosis dissemination among wild carnivores in the Republic of Kazakhstan: A 10-year study.

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.

Bovine hypodermosis is highly prevalent in Kazakhstan: Results of a first serological study.

Background and Aim: Recent information on the occurrence of bovine hypodermosis in Kazakhstan is limited to the results of a few clinical studies in the northern and eastern regions. A first serological study aimed to obtain more data on its geographical distribution and to estimate the prevalence in this country. Material and Methods: Serum samples collected from 891 dairy cows on 30 dairy farms in eight Kazakh provinces during the winter season 2015/2016 were examined for antibodies to Hypoderma spp. first-stage larval antigen using a commercially available enzyme-linked immunosorbent assay (IDEXX Bovine Hypodermosis Serum Antibody Test). Results: Overall, 73.6% (95% confidence interval: 70.6%-76.5%) of the cows sampled were seropositive for Hypoderma, and antibody-positive cows were found in 28 of 30 farms and in seven of eight provinces. Conclusion: The results suggest a high prevalence of bovine hypodermosis in Kazakhstan, for which the socioeconomic changes in agriculture and village life following the country's independence are considered to be responsible.

Hematological, biochemical, and serological parameters of experimentally infected rabbits with Trichinella nativa and Trichinella spiralis for early identification of trichinellosis.

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.

Gasterophilus spp. infections in horses from northern and central Kazakhstan.

A cross-sectional survey was performed to obtain current data on the gastrointestinal myiasis of horses in the provinces of Kostanay, Akmola and Karagandy, northern and central Kazakhstan. The stomach, small intestine and rectum of 148 slaughter horses were examined for Gasterophilus spp. larvae during a 26-month study period. All horses were infected with 2nd and 3rd stage larvae (mean intensity: 803±350), and 22% of them harboured >1000 Gasterophilus spp. larvae each. Four species were identified: G. intestinalis (prevalence: 100%; mean intensity: 361±240 larvae), G. haemorrhoidalis (100%; 353±191), G. nasalis (100%; 73±36) and G. pecorum (91.2%; 18±10). Horses aged<2 years were higher infected with Gasterophilus larvae than 2-4 years old animals. Both the prevalence and extremely high intensity of Gasterophilus infections of horses in these Kazakh regions suggest respective control measurements to improve the health and performance of the animals and to increase the economic income of horse owners.

Low genetic diversity in wide-spread Eurasian liver fluke Opisthorchis felineus suggests special demographic history of this trematode species.

Opisthorchis felineus or Siberian liver fluke is a trematode parasite (Opisthorchiidae) that infects the hepato-biliary system of humans and other mammals. Despite its public health significance, this wide-spread Eurasian species is one of the most poorly studied human liver flukes and nothing is known about its population genetic structure and demographic history. In this paper, we attempt to fill this gap for the first time and to explore the genetic diversity in O. felineus populations from Eastern Europe (Ukraine, European part of Russia), Northern Asia (Siberia) and Central Asia (Northern Kazakhstan). Analysis of marker DNA fragments from O. felineus mitochondrial cytochrome c oxidase subunit 1 and 3 (cox1, cox3) and nuclear rDNA internal transcribed spacer 1 (ITS1) sequences revealed that genetic diversity is very low across the large geographic range of this species. Microevolutionary processes in populations of trematodes may well be influenced by their peculiar biology. Nevertheless, we suggest that lack of population genetics structure observed in O. felineus can be primarily explained by the Pleistocene glacial events and subsequent sudden population growth from a very limited group of founders. Rapid range expansion of O. felineus through Asian and European territories after severe bottleneck points to a high dispersal potential of this trematode species.