Comparison of Oral Microbial Composition and Determinants Encoding Antimicrobial Resistance in Dogs and Their Owners.

Consolidated studies on animal, human, and environmental health have become very important for understanding emerging zoonotic diseases and the spread of antimicrobial resistance (AMR). The aim of this study was to analyse the oral microbiomes of healthy dogs and their owners, including determinants of AMR. Shotgun metagenomic sequencing detected 299 bacterial species in pets and their owners, from which 70 species were carried by dogs and 229 species by humans. Results demonstrated a unique microbial composition of dogs and their owners. At an order level, Bacteroidales were the most prevalent oral microbiota of dogs with significantly lower prevalence in their owners where Actinomycetales and Lactobacillales predominated. Porphyromonas and Corynebacterium were the most prevalent genera in dogs, whereas Streptococcus and Actinomyces were in animal owners. The resistances to macrolides, tetracyclines, lincosamides and Cfx family A class broad-spectrum ß-lactamase were detected in both animal and human microbiomes. Resistance determinants to amphenicols, aminoglycosides, sulphonamides, and quaternary ammonium compounds were detected exceptionally in dogs. In conclusion, the study demonstrated different bacterial composition in oral microbiomes of healthy dogs without clinical signs of periodontal disease and their owners. Due to the low numbers of the samples tested, further investigations with an increased number of samples should be performed.

Effect of low frequency oscillations during milking on udder temperature and welfare of dairy cows.

The study aimed to investigate the effect of low-frequency oscillations on the cow udder, milk parameters, and animal welfare during the automated milking process. The study's objective was to investigate the impact of low-frequency oscillations on the udder and teats' blood circulation by creating a mathematical model of mammary glands, using milkers and vibrators to analyze the theoretical dynamics of oscillations. The mechanical vibration device developed and tested in the study was mounted on a DeLaval automatic milking machine, which excited the udder with low-frequency oscillations, allowing the analysis of input parameters (temperature, oscillation amplitude) and using feedback data, changing the device parameters such as vibration frequency and duration. The experimental study was performed using an artificial cow's udder model with and without milk and a DeLaval milking machine, exciting the model with low-frequency harmonic oscillations (frequency range 15-60 Hz, vibration amplitude 2-5 mm). The investigation in vitro applying low-frequency of the vibration system's first-order frequencies in lateral (X) direction showed the low-frequency values of 23.5-26.5 Hz (effective frequency of the simulation analysis was 25.0 Hz). The tested values of the first-order frequency of the vibration system in the vertical (Y) direction were 37.5-41.5 Hz (effective frequency of the simulation analysis was 41.0 Hz), with higher amplitude and lower vibration damping. During in vivo experiments, while milking, the vibrator was inducing mechanical milking-similar vibrations in the udder. The vibrations were spreading to the entire udder and caused physiotherapeutic effects such as activated physiological processes and increased udder base temperature by 0.57°C (p < 0.001), thus increasing blood flow in the udder. Used low-frequency vibrations did not significantly affect milk yield, milk composition, milk quality indicators, and animal welfare. The investigation results showed that applying low-frequency vibration on a cow udder during automatic milking is a non-invasive, efficient method to stimulate blood circulation in the udder and improve teat and udder health without changing milk quality and production. Further studies will be carried out in the following research phase on clinical and subclinical mastitis cows.

microRNAs Are Abundant and Stable in Platelet-Rich Fibrin and Other Autologous Blood Products of Canines.

Various microRNAs (miRNAs) present in autologous blood products of canines have not been studied recently. We aimed to elucidate the existence of miRNAs in platelet-rich fibrin (PRF) and the stability of canine autologous blood products under various storage conditions. Total RNAs were isolated from PRF and other autologous blood products following newly adapted protocols used in commercial kits for plasma and tissue samples. Quantitative real-time polymerase chain reaction analysis (qPCR) was used to detect miRNAs in autologous blood products. The miR-16, miR-21, miR-155, and miR-146a were abundant in PRF and other autologous blood products of canines. Furthermore, we found they could maintain stability under protracted freezing temperatures of -30 °C for at least one month. Our findings revealed that PRF might be a stable resource for various canine miRNAs.

Peripheral mononuclear cells composition in platelet-rich fibrin in canines with chronic conditions.

Platelet-rich fibrin (PRF) is a hot research topic because of its regenerative effect in humans. However, data reporting about its application in companion animals is lacking. The study aimed to supplement currently available data on PRF cell composition in canine patients by isolating peripheral blood mononuclear cells (PBMC), namely T cells, matured B cells, monocytes and macrophages, and adapting current protocols of cell flow cytometry for PRF analysis. The canine patient population was divided into three subgroups: animals with periodontitis only, animals with neoplasia and periodontitis, and healthy controls. Individual clinical parameters of the patients and evaluation of the wound healing quality were included in the research. In the present study, canine PRF cell composition was analyzed for the first-time using cell flow cytometry protocol. A higher proportion of PBMC cells related to wound healing (CD3+, CD3+ CD4+ CD8-, CD14+) were found in the PRF of control, periodontitis and neoplasia groups compared to the respective blood samples, which implies a positive outcome associated with clinical PRF usage in canine patients. Proportions of monocytes and macrophages were higher in PRF samples compared to the blood of healthy patients and periodontitis-affected patients. However, inflammatory and neoplastic processes do not affect the distribution of PBMC.

Effect of Electromagnetic Field Exposure on Mouse Brain Morphological and Histopathological Profiling.

INTRODUCTION: Mobile phones (MP) and other electronic and communication devices that are used daily expose users to electromagnetic fields (EMF) and contribute to an increasing incidence of neurological disorders. Brain tissue is the closest organ to the MP as it operates, thus the influence of MP radiation on brain tissue is of particular concern, although research is still inconclusive. The present study investigated the possible effect of an EMF (1,350-1,375 megahertz (MHz)) from an MP on morphological and histopathological profiles in the mouse brain. MATERIAL AND METHODS: Healthy BALB/c mice were assigned to three equal groups (a control and two experimental groups, n = 10 each). Experimental mice were exposed to EMFs continuously for 72 h, those of experimental group I to a 1,350 MHz field at a specific absorption rate (SAR) of 4.0 W/kg, and group II to a 1,375 MHz field EMF at an SAR of 4.0 W/kg. Brain segmentation and histopathological analysis were applied to detect changes in the morphometric parameters of the brain lobes and identify pathological lesions, respectively. RESULTS: Histopathology results revealed shrinkage of pyramidal neurons, presence of mild perivascular and perineural oedema, and some vacuolation of neurons and glial cells derived from mouse great hemispheres. The lesions also included reduction of Purkinje cells, vacuolisation of neurons and glial cells, and interstitial oedema in the cerebellum. CONCLUSION: MP distance of 3 cm from the cage may induce appreciable morphological changes in mouse brain structures; therefore, more comprehensive research is essential for assessment of safe distance. These pronounced effects may interfere with the results of laboratory tests on murine experimental models in veterinary or biomedical research.

Evaluation of automatic milking system variables in dairy cows with different levels of lactation stage and reproduction status.

In this study, we hypothesized that differences of automatic milking systems (AMS) variables in dairy cows during estrus and through diverse stages of lactation can be suggested as alternative indicators to support the pregnancy in dairy farms using automatic milking systems. The key objectives were: (1) to determine the variation of automatic milking system indicators during lactation and to estimate the relationship with reproduction status in dairy cows; (2) to test the hypothesis that milking traits of cows can be influenced by estrus and conceiving, and can be used as a predictor of the likelihood of reproductive success in dairy herds. Estrus synchronization was performed in 368 healthy Lithuanian Black and White cows. All cows (n = 368) were synchronized and inseminated for the first time on the 91st day in milk (DIM). Cows not pregnant (17.39%) were synchronized and inseminated again at 132 DIM. After the first insemination pregnant (n = 304) cows were identified as group 1, after the second insemination pregnant (n = 58) cows - as group 2. Overall, 12 01 713 records of udder quarters in cows from 5 to 305 DIM were evaluated. The results revealed the reduction in milk yield during estrus 11.05% on 91 DIM and 13.89% on 132 DIM (P < 0.001) and an increment in milk flow traits in cows after 91 DIM (P < 0.05), also a slight decline in milk flow traits on 132 DIM. Furthermore, milking frequency (MF) of cows decreased significantly (P < 0.001) after conceiving. The interval between milkings (MI) increased (40.30%) during estrus of cows in group 1 (P < 0.001), and thereafter gradually increased, however in group 2 there was a temporary increment (6.06%) on the 91 DIM and steady rise (42.13%) on 132 DIM was noticed. The results highlight that changes in AMS indicators of cows may be considered as an additional tool for improvement of reproductive management in dairy herds, but further research-based studies are necessary before practical application.

Correction to: Prevalence and phylogenetic analysis of hepatitis E virus in pigs, wild boars, roe deer, red deer and moose in Lithuania.

Following publication of the original article [1], we have been notified of a typing error in the "Discussion" section.

Prevalence and phylogenetic analysis of hepatitis E virus in pigs, wild boars, roe deer, red deer and moose in Lithuania.

BACKGROUND: Hepatitis E virus (HEV) is one of the major causes of acute viral hepatitis worldwide. In Europe, food-borne zoonotic transmission of HEV genotype 3 has been associated with domestic pigs and wild boar. Controversial data are available on the circulation of the virus in animals that are used for human consumption, and to date, no gold standard has yet been defined for the diagnosis of HEV-associated hepatitis. To investigate the current HEV infection status in Lithuanian pigs and wild ungulates, the presence of viral RNA was analyzed by nested reverse transcription polymerase chain reaction (RT-nPCR) in randomly selected samples, and the viral RNA was subsequently genotyped. RESULTS: In total, 32.98 and 22.55% of the domestic pig samples were HEV-positive using RT-nPCR targeting the ORF1 and ORF2 fragments, respectively. Among ungulates, 25.94% of the wild boar samples, 22.58% of the roe deer samples, 6.67% of the red deer samples and 7.69% of the moose samples were positive for HEV RNA using primers targeting the ORF1 fragment. Using primers targeting the ORF2 fragment of the HEV genome, viral RNA was only detected in 17.03% of the wild boar samples and 12.90% of the roe deer samples. Phylogenetic analysis based on a 348-nucleotide-long region of the HEV ORF2 showed that all obtained sequences detected in Lithuanian domestic pigs and wildlife belonged to genotype 3. In this study, the sequences identified from pigs, wild boars and roe deer clustered within the 3i subtype reference sequences from the GenBank database. The sequences obtained from pig farms located in two different counties of Lithuania were of the HEV 3f subtype. The wild boar sequences clustered within subtypes 3i and 3h, clearly indicating that wild boars can harbor additional subtypes of HEV. For the first time, the ORF2 nucleotide sequences obtained from roe deer proved that HEV subtype 3i can be found in a novel host. CONCLUSION: The results of the viral prevalence and phylogenetic analyses clearly demonstrated viral infection in Lithuanian pigs and wild ungulates, thus highlighting a significant concern for zoonotic virus transmission through both the food chain and direct contact with animals. Unexpected HEV genotype 3 subtype diversity in Lithuania and neighboring countries revealed that further studies are necessary to understand the mode of HEV transmission between animals and humans in the Baltic States region.

Detection and molecular characterization of porcine reproductive and respiratory syndrome virus in Lithuanian wild boar populations.

BACKGROUND: Porcine reproductive and respiratory syndrome virus (PRRSV) is recognized worldwide as an important and economically devastating pathogen in pig production. Although PRRSV is widespread in domestic swine, there is a lack of information regarding PRRSV infection in European wild boars (Sus scrofa). Currently available information does not provide conclusive evidence that wild boars are a reservoir of PRRSV. Nevertheless, wild boars may be likely to become infected by domestic swine through occasional direct or indirect contact. Furthermore, wild boars can act as a reservoir for infectious diseases of domestic pigs. Therefore, the objectives of the present study were to determine the virus prevalence and further explore the epidemiology and diversity of PRRSV strains present in Lithuanian wild boars over a 5-year period. A total of 1597 tissue and serum samples from wild boars inhabiting 44 districts and ten counties in Lithuania were analysed using conventional nested reverse transcription polymerase chain reaction (RT-PCR) and real-time Taqman RT-PCR for the detection of PRRSV-specific open reading frame (ORF) 1 and 6 sequences. RESULTS: PRRSV was highly prevalent in Lithuanian wild boar populations, with an average rate of 18.66 % using conventional RT-PCR and 19.54 % using real-time RT-PCR. PRRSV was detected in 36.71 and 41.77 % of 237 hunting grounds tested by conventional RT-nPCR and real-time RT-PCR, respectively. No statistically significant differences in PRRSV prevalence were observed by geographic area in the ten Lithuanian counties. Animals infected with PRRSV were identified in all age groups; however, significantly higher prevalence rates were identified in subadult and adult wild boars than in juveniles up to 12 months old. No positive results were obtained using conventional PCR with Type 2 specific primers. Phylogenetic analysis of the partial ORF5 region revealed that ten wild boars harboured virus sequences belonging to genetic subtypes 3 and 4 and may therefore pose a serious threat to Lithuanian pig farms in which only subtype two strains are circulating. CONCLUSIONS: The results of virus prevalence and phylogenetic analyses strongly support the role of wild boars as a possible natural reservoir for PRRSV in Lithuania.