AHEAD OF PRINT

The aim of the study was to determine the genetic diversity of Echinococcus multilocularis in pigs in highly endemic areas in Poland, as well as to attempt to confirm the occurrence and geographical distribution of haplotypes characteristic for these areas, which were previously described on the basis of examination of adult tapeworms isolated from foxes.

Twenty samples of E. multilocularis larval forms were obtained from pigs’ livers in four provinces of Poland. Genetic analyses were conducted on sequences of two mitochondrial genes: cox1 and nad2.

Seven haplotypes were found for the cox1 gene (OQ874673–OQ874679) and four haplotypes for nad2 (OQ884981–OQ884984). They corresponded to the haplotypes described earlier in foxes in Poland (some of them differing only in one nucleotide). The analysis showed the presence of the Asian-like haplotype in both the cox1 and nad2 genes. The remaining haplotypes were grouped in the European clade. The geographical distribution of haplotypes identified in the pig samples was noticed to bear a similarity to the distribution of haplotypes previously isolated from foxes in the same regions.

The characteristic geographical distribution of E. multilocularis haplotypes in Central Europe (including the presence of the Asian-like haplotype) previously described in the population of definitive hosts (foxes) has now been confirmed by the analysis of samples from non-specific intermediate hosts (pigs).

Milk from cows, goats and sheep was analysed in terms of content of fourteen perfluoroalkyl substances (PFASs).

Altogether, 73 milk samples from cows (n = 38), goats (n = 20) and sheep (n = 15) were collected from various regions of Poland. Concentrations of analytes were determined using liquid chromatography–tandem mass spectrometry (LC-MS/MS).

The lower-bound sum of four PFAS (∑4 PFASs) concentrations (perfluorooctanesulfonic acid (PFOS), perfluorooctanoic acid, perfluorononanoic acid and perfluorohexanesulfonic acid) were highest in sheep’s (0.0055 μg/kg), lower in goat’s (0.0046 μg/kg), and lowest in cow’s milk (0.0008 μg/kg). Goat’s and sheep’s milk was statistically significantly more contaminated than cow’s milk. None of the samples exceeded the indicative values set by Commission Recommendation (EU) 2022/1431, and even the maximum detected concentrations were an order of magnitude lower. The most frequently detected was linear PFOS, which was found in 33%, 76% and 93% of cow’s, goat’s and sheep’s milk samples, respectively. Based on mean upper-bound ∑4 PFAS concentrations and average milk consumption, the estimated intake of ∑4 PFASs ranged from 0.153 to 0.266 ng/kg body weight (b.w.) for children and from 0.050 to 0.88 ng/kg b.w. for adults, which indicates that exposure is very low and is merely <7% of the tolerable weekly intake (TWI) for children and <2% of the TWI for adults.

Regardless of the milk type, the intake of PFASs via consumption of Polish milk does not contribute significantly to the overall PFAS intake of either adults or children.

Gastrointestinal nematodes pose a threat to animal health and affect farmers by negatively impacting farm management.

The study was conducted on a sheep farm with suspected reductions in the efficacies of anthelmintics. Efficacy was determined using in vivo faecal egg count reduction, in vitro egg hatch (EHT) and larval development (LDT) tests. In the first phase, 60 sheep were equally split into six groups. Group 1 received the recommended dose of albendazole (ALB), group 2 received the same after fasting for 24 h, group 3 received the dose divided into two halves at 6 h intervals, group 4 received a double dose of ALB, and group 5 received the recommended dose of ivermectin (IVM). Group 6 served as a control. The second phase of the experiment had two groups: one treated with levamisole (LEV) and a control group. Faecal samples were collected from all sheep.

No reduction of egg output was observed in the groups treated with single, double, or divided doses of ALB, but one of 13.7–16.9% was noted in the fasting group. Efficacy in the IVM group ranged from 31.50 to 39.97%. The mean concentrations sufficient to prevent 50% of the eggs from hatching in the in vitro EHT and the mean concentrations in which the development of larvae to the L3 stage was inhibited by 50% in the LDT exceeded established thresholds for benzimidazoles and IVM. Haemonchus contortus was the only species identified after treatment. The LDT did not indicate the presence of resistance to LEV. All animals treated with LEV were negative for eggs 10 d after treatment.

Resistance to ALB and IVM in Haemonchus contortus was confirmed. Alternative approaches to improve the efficacies of benzimidazole did not sufficiently increase the efficacy, but LEV was an efficient anthelmintic treatment.

Eosinophils represent the most active cells in mammals that show protective and assistive activity in the host immune defence against helminth parasites. These cells are also responsible for the reduction of allergic and inflammatory reactions. The eosinophils play a key role in allergic reactions by secretion of different chemical molecules leading to swelling, lesions and granuloma onset.

The study was carried out on 30 cats with inflammatory skin lesions. The cats ranged in age from seven months to 13 years, and had an average age of three years. The research methodology included information on the disease, dermatological conclusions, concomitant disorders, medical and laboratory data and the treatment strategy.

In total, 30 cats were diagnosed with eosinophilic granuloma complex. The distribution of lesions was 87.1% in the skin and 12.9% at the skin–mucosal junction. The lesions increased and decreased with the seasons of spring and summer, and the onset of the disease usually coincided with exposure to fleas.

Eosinophilic granuloma complex in cats is a serious pathology and frequently requires lifelong treatment, so it is important to diagnose it quickly and accurately to ensure optimal treatment of affected animals.

Enterovirus E (EV-E) is a common viral pathogen endemic in cattle worldwide. Little is known, however, about its potential interactions with bovine immune cells.

The EV-E-permissiveness of bovine peripheral blood mononuclear cells (PBMCs) was evaluated. The infectious titres of extracellular virus were measured and the intracellular viral RNA levels were determined by reverse transcription quantitative PCR after cell inoculation. The effects of EV-E on cell viability and proliferative response were investigated with a methyl thiazolyl tetrazolium bromide reduction assay, the percentages of main lymphocyte subsets and oxidative burst activity of blood phagocytes were determined with flow cytometry, and pro-inflammatory cytokine secretion was measured with an ELISA.

Enterovirus E productively infected bovine PBMCs. The highest infectious dose of EV-E decreased cell viability and T-cell proliferation. All of the tested doses of virus inhibited the proliferation of high responding to lipopolysaccharide B cells and stimulated the secretion of interleukin 1β, interleukin 6 and tumour necrosis factor α pro-inflammatory cytokines.

Interactions of EV-E with bovine immune cells may indicate potential evasion mechanisms of the virus. There is also a risk that an infection with this virus can predispose the organism to secondary infections, especially bacterial ones.

In the light of the problem of antibiotic resistance, the use of combined alternative therapies in combatting bacteria-related disorders has gained popularity. Bacteriophages are one element implemented in new combination therapy. Stevia rebaudiana is known to have antimicrobial activity and regarded as potentially having a synergistic effect with bacteriophages. Therefore, possible interactions of lytic bacteriophages (MS2, T4 and Phi6) with acetone and methanol S. rebaudiana extracts (SRa and SRm) in the bacterial environment were examined.

The interactions were tested using a microdilution method, phage-extract co-incubation assay, static interaction (synography) and dynamic growth profile experiments in a bioreactor.

The interactions of the tested factors in a static environment differed from those in a dynamic environment. Dynamic conditions altered the effect of the extracts in a concentration-dependent manner. How different the effect of the SRa extract was to that of the SRm extract on bacterial growth in a dynamic environment depended on the species of the phage and bacterial host. The greatest differences were observed for E. coli strains and their phages, whereas Pseudomonas syringae and the Phi6 phage reacted very similarly to both extracts. Differences also emerged for the same extract in different E. coli strains and their phages.

Every extract type should be tested on a case-by-case basis and experiment outcomes should not be generalised before gathering data. Moreover, many varied experiments should be performed, especially when examining such multifactorial mixtures. The tested mixtures could potentially be used in multidrug-resistant bacterial infection treatments.

Testin is a protein involved in cell mobility, adhesion and colony formation. In rats, testin presence has been reported in the testes, and its possible role in spermatogenesis has been suggested. Studies in humans also suggest a possible role of testin as a cancer suppressor protein. In the dog, which represents both an important pet species and a good animal model for studying biological and pathological testicular processes, the presence of testin has never been reported.

In the present study, the expression of testin in foetal, prepubertal, adult and aged canine testes was investigated. Testes from 5 adult and 3 aged dogs, from 2 one-month-old puppies and from 2 foetuses miscarried at the end of pregnancy were immunohistochemically examined with a commercial antibody against testin.

Testin was intensely expressed in Sertoli cells in every testis examined. Spermatids were also positive for testin in mature dogs and in the testicular areas of the aged ones which were not atrophic. Weak expression of testin was also detected in all testes examined.

The present study, the first demonstrating the presence of testin in canine testes, provides the basis for further dog–human comparative research and for studies on the role of this protein in canine physiology, reproduction and testicular pathologies.

MicroRNAs (miRNAs), a class of noncoding small RNAs, have been recognised as potential biomarkers of mammary gland conditions, including bovine mastitis diagnosis. The aim of this study was to quantify selected miRNAs in the milk of mastitic cows.

Milk samples (n = 90) were collected from healthy and mastitic dairy cows originating from local dairy cattle farms located in the west of Poland. MicroRNAs of the miR-21a, miR-92a, miR-146a and miR-383 species were quantified using the highly sensitive droplet digital PCR method. Direct measurement of somatic cell count (SCC) was performed using a cell counter. Cows were divided into three groups: those with an SCC below 200,000/mL were designated Low (n = 25), those with an SCC between 200,000 and 999,999 were Medium (n = 34), and those with an SCC of 1,000,000 or higher were High (n = 31). Microbiological analyses were performed using standard culture testing.

The level of miR-383 was very low and this miRNA was excluded from analysis. The miR-92a was used to normalise miR-21a and miR-146a expression levels. The obtained results of expression of miR-21a and miR-146a correlated with somatic cell number (R = 0.53 and 0.79, respectively).

These results show that ddPCR is a useful method for quantifying miRNAs in raw cow milk. It seems that miR-146a is a promising marker for bovine mastitis, although further studies are needed to select a panel of miRNAs that can be used in mastitis monitoring in Poland.

Strains of Leptospira interrogans belonging to two very closely related serovars, Icterohaemorrhagiae and Copenhageni, have been associated with disease in mammalian species and are the most frequently reported agents of human leptospirosis. They are considered the most pathogenic serovars and represent more than half of the leptospires encountered in severe human infections.

Nineteen such isolates from the United Kingdom – human, domestic and wildlife species – were typed using three monoclonal antibodies (F12 C3, F70 C14 and F70 C24) in an attempt to elucidate their epidemiology. They were further examined by restriction endonuclease analysis (REA), multiple-locus variable-number tandem repeat analysis (MLVA) and lic12008 gene sequence analysis.

Monoclonal antibody F12 C3, which is highly specific for Icterohaemorrhagiae and Copenhageni, confirmed that all the strains belonged to these two serovars. Sixteen strains were identified as Copenhageni and three as Icterohaemorrhagiae serovar. Only one restriction pattern type was identified, thus confirming that REA is not able to discriminate between the Icterohaemorrhagiae and Copenhageni serovars. Variable-number tandem-repeat analysis found three loci with differences in the repeat number, indicating genetic diversity between British isolates. Sequences of the lic12008 gene showed that all isolates identified as the Icterohaemorrhagiae serotype have a single base insertion, in contrast to the same sequences of the Copenhageni serotype.

Copenhageni is the predominant serovar in the Icterohaemorrhagiae serogroup isolated in British Isles. There is a genetic diversity of MLVA patterns of the isolates but no genetic tool used in the study was able to determine serovars.

Maedi-visna virus and caprine arthritis encephalitis virus are two closely related lentiviruses which cause multisystemic, progressive and persistent infection in goats and sheep. Because these viruses frequently cross the species barrier, they are considered to be one genetic group called small-ruminant lentiviruses (SRLV). They have in vivo tropism mainly for monocytes and macrophages and organ tropism with unknown mechanisms. Typical clinical signs are pneumonia in sheep, arthritis in goats, and mastitis in both species. Infection with SRLV cannot currently be treated or prevented, and control programmes are the only approaches to avoiding its spread. These programmes rely mainly on annual serological testing and elimination of positive animals. However, the high genetic and antigenic variability of SRLV complicate their early and definitive diagnosis. The objective of this review is to summarise the current knowledge of SRLV genetic variation and its implications for tropism, the development of diagnostic tests and vaccines and the effectiveness of control and eradication programmes.

Subject literature was selected from the PubMed and the Google Scholar databases.

The high genetic diversity of SRLV affects the performance of diagnostic tools and therefore control programmes. For the early and definitive diagnosis of SRLV infection, a combination of serological and molecular tests is suggested. Testing by PCR can also be considered for sub-yearling animals. There are still significant gaps in our knowledge of the epidemiology, immunology and biology of SRLV and their impact on animal production and welfare.

This information may aid selection of the most effective SRLV spread reduction measures.

The molecular contamination of an animal facility was investigated during and after an infection with highly pathogenic African swine fever virus (ASFV) among domestic pigs. The investigation evaluated the risk of indirect transmission of the disease and indicated points that may facilitate cleaning and disinfection processes.

Six domestic pigs were infected oronasally with the highly pathogenic Georgia 2007 strain. Environmental samples from the floors, walls, rubber floor mats, feeders, drinkers, high-efficiency particulate-absorbing filter covers and doors were collected 7 days post infection (dpi), 7 days later and 24 h after disinfection of the facility. The samples were investigated by real-time PCR and in vitro assays to find genetic traces of ASFV and infectious virus.

Typical clinical outcomes for ASF (i.e. fever, apathy, recumbency and bloody diarrhoea) were observed, and all animals died or required euthanasia before or at 9 dpi. No infectious virus was found in environmental samples at the sampling time points. Genetic traces of ASFV were found in all locations except the doors. The initial virus load was calculated using real-time PCR threshold cycle values and was the highest at the drain. A statistically significant decrease of virus load over time was found on non-porous surfaces mechanically cleaned by water (the floor and drain).

The gathered data confirmed different routes of virus excretion (oral and nasal, faeces and urine, and aerosol) and showed virus locations and different initial concentrations in the animal facility. Maintaining the facility with mechanical cleaning and using personal protection (gloves) and hand disinfection may efficiently minimise the risk of further virus spread. Together with the results of previously published studies, the present investigations’ failure to isolate infectious virus may suggest that if stable environmental conditions are assured, the time needed before the introduction of new herds into previously ASF-affected farm facilities could be shortened and in this way the economic losses caused by the disease outbreak mitigated.