Background and Aim: The study of Echinococcus infection among farm animals in Kazakhstan was carried out to monitor the invasion among livestock and map the data obtained. Unfortunately, there are only partial data on the study of echinococcosis among wild carnivores in Kazakhstan, which makes it difficult to conduct a comparative analysis of the epidemiological situation among wild animals. The present study aimed to estimate the genetic diversity of Echinococcus spp. (Leuckart, 1863) in Kazakhstan based on sequence analysis of cytochrome c oxidase subunit 1 (cox1) and dehydrogenase subunit 1 (nad1) of worms isolated from wild carnivorous animals wolf (Canis lupus), red fox (Vulpes vulpes) and corsac (Vulpes corsac). Materials and Methods: DNA from parasite tissue was used as a template for the amplification of the two mitochondrial genes cox1 and nad1. Sequencing was performed according to the manual for the Seq Studio Genetic Analyzer. The multiple alignments of obtained sequences were performed using the ClustalW algorithm in Mega (v.11) software. Alignments were exported as a Nexus extension and used as input for TCS v1.21 for the identification of haplotypes. The phylogenetic analysis was constructed according to the neighbor-joining method using Mega (v.11) software. Results: Analysis of the extensiveness of echinococcosis invasion showed that 6.3% were wolves, 18.2% were corsacs, and 85% were foxes. In total, 159 adults of Echinococcus spp. from the three species of animals in different parts of Kazakhstan were analyzed, and 17 individual biological samples were successfully sequenced. Sequence analysis of cox1 and nad1 genes revealed two types of echinococcosis – Echinococcus granulosus in red foxes and wolves, and Echinococcus multilocularis in corsacs. Sequencing of a portion of the mitochondrial genome made it possible to determine seven haplotypes of the pathogen in the studied samples of E. granulosus. Molecular analysis of cox1 and nad1 genes of E. multilocularis revealed three new haplotypes, which have significant variability compared with other studied Asian haplotypes. Conclusion: This study made it possible to fill the gaps in understanding the localization of the foci of the spread of the echinococcosis pathogen among the main wild carnivores and to determine the species reservoir of the pathogen in the greater territory of 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. Keywords: Echinococcus, epidemiology, helminth prevalence, Kazakhstan.

Apples are widely consumed by people all over the world due to their taste and nutritional value. However, apple trees are prone to various environmental stresses, including fungal diseases. Among them, Cytospora canker (or Valsa canker) can cause dieback of branches and twigs. Although Kazakhstan is well known as an origin of apples, very little is known about canker diseases that spread across all commercial orchards. Therefore, an accurate identification of the causal agents of those diseases is needed for further application of informed disease management strategies. In this study, eleven isolates belonging to four Cytospora species were isolated from multiple cultivars, grown in six different orchards within the Almaty region, Kazakhstan. As a result of a multilocus phylogenetic analysis using ITS, LSU and tef1-α marker genes and morphological characterization, these isolates were described as Cytospora parasitica, Cytospora sorbina, Cytospora pruinopsis and Cytospora chrysosperma. Moreover, a pathogenicity test was conducted on detached twigs, and it demonstrated that two of these fungi were highly virulent. Overall, this paper is a first report of the causal agents of apple canker disease in Kazakhstan and could be a trigger for conducting future studies to better understand the disease epidemiology, as well as build management strategies.

Thuja is one of the ornamental plants used for landscaping parks and health resorts. The plant is distinguished by a pyramidal and conical crown shape and the presence of many thin branches with scale-shaped needles, green all year round. In addition, this plant has a number of antimicrobial properties, which affects the popularity of the plant in landscaping the health resort territory (Bakht et al. 2020, Chindyaeva et al. 2020). In January 2020, symptomatic Thuja plants were observed in Southern Kazakhstan. Symptoms included distortion of the crown. External examination of the trees revelaed the presence of white fluffy mycelium on Thuja branches. The branches acquired a yellow color with a necrotic lesion developing below the affected area. Samples of infected branches from different Thuja trees (n = 13) were collected. The infected branches were cut into small pieces (5 × 5 mm), washed in 70% ethanol for 30 min, and then rinsed three times with sterile distilled water. Later, these pieces were placed on Sabouraud's medium (Laboratorios Conda S.A., Spain) and incubated at 28°C for 7 days. Yellow-green colonies grew from the pieces of wood. The colonies had a light gray-whitish aerial mycelium. Conidia (n = 35) were pale to dark brown in color, irregular and ellipsoid to ovoid conical in shape. The size of the conidia varied from 5 to 25 µm × 6 to 12 µm (n = 40) with longitudinal and transverse septations. These morphological characters were previously described and corresponded to the Alternaria alternata (Simmons et al. 2007). Genomic DNA was extracted from mycelium using the liquid nitrogen and phenol-chloroform extraction method (Butler 2012). A 568 bp product of the Alt a1 gene and 472 bp product of the calmodulin protein-coding gene was amplified using following primer pairs Alt-for/Alt-rev (Hong et al. 2005) and CALDF1/CALDR1, respectively (Lawrence et al. 2013) (Integrated DNA Technologies, Inc., Coralville, IA, USA). The PCR reaction was done in a SimpliAmp thermal cycler (Applied biosystems, Waltham, MA, USA) under the following conditions: initial denaturation at 94 °C for 1 min, 35 cycles at 94°C for 30 s to denature, 57°C for 1 min for annealing, and 72°C for 1 min for extension. A final extension step at 72°C for 10 min was also included. The sequencing was done using BigDye® Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems) and the sequence was deposited in GenBank with accession no. OL542696 calmodulin). These sequences were compared with other sequences in the GenBank by using the BLAST analysis (MZ222274.1 and MN473132.1). The phylogenetic analysis was carried out with MEGA 6 software (The Pennsylvania State University, University Park, PA, USA). To confirm the pathogenicity, 10 thuja branches from healthy trees from another area without visible pathologies were inoculated with a suspension of conidia (100 conidia/ml; obtained from 2-week-old cultures). Control samples were inoculated with sterile distilled water. The inoculated branches were placed in sterile plastic containers to maintain high humidity and incubated for 10 days at 28°C. After 7 days, irregular shaped lesions and fungal growth was observed at the site of inoculation. The affected area gradually increased in size with simioar symptomatology to that described above. Re-isolation of the pathogen and identification based on morphological features and sequencing confirmed the presence of the A. alternata pathogen. To our knowledge, this is the first report of A. alternata causing branches of thuja in Kazakhstan. Thuja is a rare plant species for this region; the cost and care are expensive. This case will allow timely diagnosis of the disease caused by Alternaria spp. in the future. It is necessary to develop preventive measures and a protocol for the treatment of thuja from a fungal infection. Bakht, J., 2020. Antibacterial activity of the crude extracts from medicinally important Thuja occidentalis. Pak J Pharm Sci. 33(2): 627-630. PMID: 32276908. Butler, J.M., 2012. Chapter 2 - DNA extraction methods. pp. 29-47 in Butler JM (Ed) Advanced topics in forensic DNA typing: Methodology. San Diego, Academic Press. doi: 10.1016/C2011-0-04189-3. Chindyaeva., L.N., et al. 2020. Comparative assessment of the phytoncidity of woody plants in the selection of species for landscaping: the possibility of use in sanatorium-and-spa practice. Vopr Kurortol Fizioter Lech Fiz Kult. 97(4): 44-51. Russian. doi: 10.17116/kurort20209704144. Hong, S.G. et al. 2005. Alt a1 allergen homologs from Alternaria and related taxa: analysis of phylogenetic content and secondary structure. Fungal Genet Biol 42:119-129. doi:10.1016/j.fgb.2004.10.009 Lawrence, D.P. et al. 2013. The sections of Alternaria: formalizing species-group concepts. Mycologia 105: 530-546. DOI: 10.3852/12-249. Simmons, E. G., 2007. Alternaria: An Identification Manual. CBS, Fungal Biodiversity Center, Utrecht, Netherlands.

Cysticercosis in wild and domestic ungulates, caused by the larval metacestode stages of Taenia hydatigena and Taenia multiceps (formerly known as Cysticercus tenuicollis and Coenurus cerebralis, respectively), is a widespread parasitic disease and poses a significant concern worldwide, particularly in endemic regions. Although Taenia species have been extensively studied globally, their epidemiology and genetic diversity in Kazakhstan remain poorly understood. In this study, wild (roe deer, red deer, moose) and domestic (cattle, sheep) ungulates, serving as intermediate hosts for Taenia spp., were examined for cysticerci in muscle tissues and internal organs. Phylogenetic analysis and pairwise nucleotide variation assessments of the cox1 and nad1 genes were conducted. An overall prevalence of 5.2% was recorded among 1370 ruminant carcasses (cattle = 773, sheep = 563, roe deer = 25, moose = 9), with infection rates of 0.6% in cattle, 1.1% in sheep, 8.0% in roe deer, and 11.1% in moose. Cattle, sheep, and moose were infected with T. hydatigena, while roe deer were infected with T. multiceps. DNA sequence analysis of all isolates revealed four nad1 gene haplotypes for T. hydatigena, with Hap_3 being the most common (10 isolates). Phylogenetic analysis showed that T. multiceps isolates from roe deer clustered within the clade defined by the reference sequences for this species. This study provides important baseline data on the prevalence and genetic variation in T. hydatigena and T. multiceps in Kazakhstan and lays the groundwork for future research on the epidemiology and population genetics of Taenia species in the region.

The increasing reliance on imported fish products in Kazakhstan raises concerns about the presence of fish-borne parasitic infections, particularly zoonotic helminths that pose risks to public health. This study aimed to assess the diversity and prevalence of helminths in commercially imported marine fish using both traditional and molecular diagnostic methods. A total of 670 specimens representing 17 fish species were collected from retail markets in Astana, Almaty, and Karaganda. Macroscopic inspection and muscle compression techniques were used to detect larval parasites, followed by DNA extraction and PCR amplification targeting the ITS-2, 5.8S, 18S rRNA, and mitochondrial COX gene regions. Sequencing and phylogenetic analysis confirmed the presence of cestodes (Eubothrium crassum, Hepatoxylon trichiuri, Nybelinia surmenicola), acanthocephalans (Echinorhynchus gadi), and nematodes, with a predominance of zoonotic species from the Anisakidae family, including Anisakis simplex, A. pegreffii, Pseudoterranova decipiens, and Contracaecum osculatum. The highest levels of infection were detected in Atka mackerel (97.1%), herring (96.0%), mackerel (92.0%), and blue whiting (88.1%), while the lowest rates were recorded in smelt (6.8%), flounder (10.2%), and haddock (16.0%). This is the first molecular-based survey of fish helminths in Kazakhstan and highlights the need to integrate genetic screening into food safety control systems to better protect consumers and improve parasite monitoring of imported seafood.