Molecular confirmation of the systemic toxoplasmosis in cat.

INTRODUCTION AND OBJECTIVE: Systemic toxoplasmosis with tissue-spread parasites occurring in intermediate hosts may also occur in immunocompromised cats (e.g., infected with FLV or FIV). To the best of our knowledge, no reports have been published on the detection and genotyping of T. gondii DNA in cats with extraintestinal toxoplasmosis in Poland. The article describes the case of the sudden death of 3 out of 4 cats in a cattery, and the detection and molecular characterization of T. gondii DNA detected in the tissues of one of the dead cats. MATERIAL AND METHODS: Samples of brain, lungs, heart, and liver of the cat that died suddenly were examined for the presence of T. gondii DNA (B1 gene) by nested PCR and real-time PCR. DNA positive samples were also genotyped at 12 genetic markers using multiplex multilocus nested PCR-RFLP (Mn-PCR-RFLP) and multilocus sequence typing (MLST). RESULTS: A total of 9 out of the 20 DNA samples were successfully amplified with nested and/or Real-time PCR. DNA from 3 out of 5 types of tested samples were genotyped (brain, heart and muscle). Mn-PCR-RFLP and MLST results revealed type II (and II/III at SAG1) alleles at almost all loci, except a clonal type I allele at the APICO locus. This profile corresponds to the ToxoDB#3 genotype, commonly identified amongst cats in Central Europe. CONCLUSIONS: To the best of our knowledge, this is the first study describing the genetic characteristics of T. gondii population determined in a cat in Poland. These data confirm the importance of this host as a reservoir for this pathogen, and demonstrate the genotypic variation of this parasite. Veterinarians should take into account that cats may develop disseminated toxoplasmosis, and that it is a systemic disease which may lead to the death of the cat, and to transmission of the pathogen to other domestic animals and to humans.

Parasitological contamination of arable soil in selected regions of Poland - preliminary study.

INTRODUCTION AND OBJECTIVE: The hygienic status of arable soils in most developed countries has been unknown. In the presented study, a preliminary investigation was undertaken to determine the contamination with eggs of parasitic nematodes in the soil of arable fields in Poland. The aim of the study was to determine whether such contamination is common enough to constitute a significant problem and what factors may influence it. MATERIAL AND METHODS: The study was conducted in 5 Polish provinces from autumn 2021 to spring 2022. The provinces differed significantly in terms of the area of agricultural land, agricultural suitability, type of soil, scale of cattle and pig breeding, production of manure and slurry, and the use of manures and organic fertilizers for fertilization. A total of 133 soil samples were collected. Parasitological examination of soil samples was carried out using the PN-Z-19006 method [1], with confirmed high sensitivity. RESULTS: Parasite eggs were found in a total of 67 samples, of which 56 samples contained eggs of roundworms of the genus Ascaris (an average of 3.29 eggs/100 g of soil), 23 contained eggs of whipworms (an average of 1.22 eggs/100 g), and 3 contained eggs of Toxocara (1 egg/100 g). CONCLUSIONS: Differences in the percentage of positive samples were found depending on the period in which the samples were taken. The percentage of positive samples collected in autumn (53.57%) was higher than the percentage of positive samples collected in spring (48.05%). Similarly, the average number of eggs of in positive samples collected in autumn (3.43 eggs/100 g) was higher than the average number of eggs in samples collected in spring (2.90 eggs/100 g). Differences in the percentage of positive samples were also found depending on the region of origin of the samples.

Comparison of the effectiveness of various parasitological methods in detecting nematode eggs in different types of soil.

INTRODUCTION AND OBJECTIVE: Natural fertilizers, sewage sludge, digestates, as well as organic fertilizers produced on their basis, can become a source of parasitological contamination of cultivated land. High concentration of invasive forms of parasites in the soil may pose a threat to human and animal health. Therefore, it is necessary to control the hygienic condition of fertilizers and fertilized soils with particular emphasis on parasites. The aim of the study was to compare the effectiveness of methods commonly used for parasitological examination of soil with own methods which were used to develop the standards. MATERIAL AND METHODS: The study was carried out using samples of sandy soil (SS), horticultural mix soil (HS) and peat-based substrate (PS). Each sample was spiked with 100 dyed Ascaris suum eggs and examined with the use of 6 methods: Vasilkova, Dada, Quinn, and 3 methods according to the Polish Standards (PN-19000, PN- 19005, PN-19006). For each variant, 8 repetitions were made. RESULTS: The largest number of A. suum eggs were found with PN-19006 (mean number of detected eggs was 21.25, 46.50, 23.00 for HS, SS, PS, respectively. Slightly lower results were obtained using PN-19005 - the mean number eggs was 21.25, 36.00, 16.75, respectively. On the other hand, the mean number of A. suum eggs found with the Dada method was about 2-3 times lower than with the PN-19006 - 15.75, 22.50, 6.50 for HS, SS, PS soil, respectively. Other methods were much less effective. CONCLUSIONS: PN-19006 method turned out to be the most effective in detecting A. suum eggs. This method can be used for parasitological examination of soils and can be the basis for developing a system of methods dedicated to testing different types of soils for the presence of nematode eggs.