Liver DNA Research Articles

Preeclampsia-induced alterations in brain and liver gene expression and DNA methylation patterns in fetal mice.

Exposure to pregnancy complications, including preeclampsia (PE), has lifelong influences on offspring's health. We have previously reported that experimental PE, induced in mice by administration of adenoviral sFlt1 at gestational day 8.5 combined with LPS at day 10.5, results in symmetrical growth restriction in female and asymmetrical growth restriction in male offspring. Here, we characterize the molecular phenotype of the fetal brain and liver with respect to gene transcription and DNA methylation at the end of gestation.In fetal brain and liver, expression and DNA methylation of several key regulatory genes is altered by PE exposure, mostly independent of fetal sex. These alterations point toward a decreased gluconeogenesis in the liver and stimulated neurogenesis in the brain, potentially affecting long-term brain and liver function. The observed sex-specific growth restriction pattern is not reflected in the molecular data, showing that PE, rather than tissue growth, drives the molecular phenotype of PE-exposed offspring.

Occult hepatitis B - the result of the host immune response interaction with different genomic expressions of the virus.

With over 40 years of history, occult hepatitis B infection (OBI) continues to remain an important and challenging public health problem. Defined as the presence of replication-competent hepatitis B virus (HBV) DNA (i.e., episomal HBV covalently closed circular DNA) in the liver and/or HBV DNA in the blood of people who test negative for hepatitis B surface antigen (HBsAg) in currently available assays, OBI is currently diagnosed using polymerase chain reaction (PCR) and real-time PCR assays. However, all efforts should be made to exclude a false negative HBsAg in order to completely follow the definition of OBI. In recent years, significant advances have been made in understanding the HBV lifecycle and the molecular mechanisms that lead to the persistence of the virus in the occult form. These factors are mainly related to the host immune system and, to a smaller proportion, to the virus. Both innate and adaptive immune responses are important in HBV infection management, and epigenetic changes driven by host mechanisms (acetylation, methylation, and microRNA implication) are added to such actions. Although greater genetic variability in the S gene of HBV isolated from OBIs was found compared with overt infection, the mechanisms of OBI are not mainly viral mutations.

Abstract 740: Obesity promotes liver pro-oxidative and DNA damage in mammary tumor of Zucker rats

Abstract Obesity is a major health problem in the US and world. Obesity is associated with the risk of cardiovascular disease, type 2 diabetes, and certain types of cancers such as breast cancer. Our laboratory reported that obesity increased mammary tumor model using obese Zucker rat model. However, the effects of obesity in the liver in regard to oxidative stress and DNA reaction is less known. In present study, we used a DMBA induced-mammary tumor model to further investigate the effects of obesity on creation of pro-oxidative environment in liver and oxidative DNA reaction. Lean and obese female rats were at 50 days of age received 7,12-dimethylbenz(α)anthracene (DMBA) and were sacrificed 20 weeks later. HPLC-ECD, HPLC-UV and LC-MS were used. Obesity capable significantly modify methylation status in liver caused higher (p < 0.001) SAM/SAH ratio (know as “methylation ratio”) and increased (p < 0.05) global DNA methylation. Also, obesity decrease GSH/GSSG ratio (known as “oxidative stress ratio”) with p < 0.0001 inducing higher oxidative stress and as results, increase presence both oxidized guanosine (8-OH-Guanosine) with p < 0.001 and oxidized 5metylCytosine (5hmCytosine) p<0.04 in liver DNA. 8-OH-Guanosine/5hmCytosine ratio was not significantly affected. Obesity caused significant oxidative stress in liver, causing oxidative DNA damage and change of DNA methylation pattern. These changes may contribute to the development of fatty liver in breast cancer models. Citation Format: Reza Hakkak, Stepan Melnyk, Soheila Korourian. Obesity promotes liver pro-oxidative and DNA damage in mammary tumor of Zucker rats [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 740.

Arsenic Exposure, Blood DNA Methylation, and Cardiovascular Disease.

Epigenetic dysregulation has been proposed as a key mechanism for arsenic-related cardiovascular disease (CVD). We evaluated differentially methylated positions (DMPs) as potential mediators on the association between arsenic and CVD. Blood DNA methylation was measured in 2321 participants (mean age 56.2, 58.6% women) of the Strong Heart Study, a prospective cohort of American Indians. Urinary arsenic species were measured using high-performance liquid chromatography coupled to inductively coupled plasma mass spectrometry. We identified DMPs that are potential mediators between arsenic and CVD. In a cross-species analysis, we compared those DMPs with differential liver DNA methylation following early-life arsenic exposure in the apoE knockout (apoE-/-) mouse model of atherosclerosis. A total of 20 and 13 DMPs were potential mediators for CVD incidence and mortality, respectively, several of them annotated to genes related to diabetes. Eleven of these DMPs were similarly associated with incident CVD in 3 diverse prospective cohorts (Framingham Heart Study, Women's Health Initiative, and Multi-Ethnic Study of Atherosclerosis). In the mouse model, differentially methylated regions in 20 of those genes and DMPs in 10 genes were associated with arsenic. Differential DNA methylation might be part of the biological link between arsenic and CVD. The gene functions suggest that diabetes might represent a relevant mechanism for arsenic-related cardiovascular risk in populations with a high burden of diabetes.

Hepatic Impact of Different Concentrations of Hibiscus rosa Zinc Oxide Nanoparticles on Rats.

Nanomaterial, especially zinc oxide nanoparticles, has entered the manufacture of many materials used in daily lives. The current study aimed to assess the impact of three concentrations of hibiscus rosa zinc oxide nanoparticles (HrZnONPs) and hibiscus rosa extract (Hre) on the liver tissue and DNA fragmentation of liver cells. A total of 35 adult male Wistar rats were grouped as follows: The first group which was the control (n=7) did not receive any treatment. The remaining 28 animals were randomly assigned to four groups. Group 1 (n=7) were subcutaneously injected with 100mg\kg BW of Hibiscus rosa extract for 60 days; the rats in group 2 were subcutaneouslyinjected with 25 mg\kg BW of HrZnONPs for 60 days; rats in group 3 were subcutaneouslyinjected with 75mg\kg BW of HrZnONPs for 60 days; rats in group 4 were subcutaneously injected with 100mg\kg BW of HrZnONPs for 60 days. The liver biomarkers, aspartate aminotransferase (AST), alkaline phosphatase (ALP), and alanine aminotransferase (ALT) have been assessed in serum at zero time, after one month, and after two months of the experiment. At the end of the experiment, all animals were euthanized, the liver was dissected, the specimen underwent a pathohistological investigation, and the percentage of DNA fragmentation was evaluated. The results pointed out that the rats which were treated with HrZnONPs at concentrations of 75 and 100 mg\kg B.W. demonstrated a salient elevation in serum AST, ALT, or ALP activity, a modulation in hepatic tissue architecture, and an elevated percentage of high DNA damage, as compared to those treated with HrZnONPs at a concentration of 25 mg\kg B.W. On the other hand, the recorded data indicated that the administration of Hre has some ameliorative effects on AST, ALP, and ALT levels, histological cross-section, and the value of comet assay for liver cells due to the role of Hre antioxidant. In conclusion, the results of the current study demonstrated that high doses of HrZnONPs had exerted more adverse effects, compared to low doses. Moreover, the findings confirmed the ameliorative impact of Hre on liver biomarkers, a histological cross-section of the liver, and DNA damage.

Correlation Investigation between Pyrrole-DNA and Pyrrole-Protein Adducts in Male ICR Mice Exposed to Retrorsine, a Hepatotoxic Pyrrolizidine Alkaloid

Pyrrolizidine alkaloids (PAs) have been found in over 6000 plants worldwide and represent the most common hepatotoxic phytotoxins. Catalyzed by hepatic cytochrome P450 enzymes, PAs are metabolized into reactive pyrrolic metabolites, which can alkylate cellular proteins and DNA to form pyrrole-protein adducts and pyrrole-DNA adducts, leading to cytotoxicity, genotoxicity, and tumorigenicity. To date, the correlation between these PA-derived pyrrole-protein and pyrrole-DNA adducts has not been well investigated. Retrorsine is a representative hepatotoxic and carcinogenic PA. In the present study, the correlations among the PA-derived liver DNA adducts, liver protein adducts, and serum protein adducts in retrorsine-treated mice under different dosage regimens were studied. The results showed positive correlations among these adducts, in which serum pyrrole-protein adducts were more accessible and present in higher abundance, and thus could be used as a suitable surrogate biomarker for pyrrole-DNA adducts to indicate the genetic or carcinogenic risk posed by retrorsine.

IN SITU HYBRIDIZATION AND VIRUS CHARACTERIZATION OF SKUNK ADENOVIRUS IN NORTH AMERICAN WILDLIFE REVEALS MULTISYSTEMIC INFECTIONS IN A BROAD RANGE OF HOSTS.

Skunk adenovirus-1 (SkAdV-1) has been reported infecting several North American wildlife species; however, lesions associated with disease have not yet been completely characterized, particularly in porcupines. We describe and characterize the tissue distribution and lesions associated with SkAdV-1 infection in 24 wildlife diagnostic cases submitted between 2015 and 2020, including 16 North American porcupines (Erethizon dorsatum), three striped skunks (Mephitis mephitis), and five raccoons (Procyon lotor), which constitute a new host species. The most common lesion in all species was severe necrotizing bronchopneumonia with (n=12) or without (n=10) interstitial involvement. Intranuclear inclusion bodies were common in respiratory epithelium (n=21) and less often in renal tubular (n=6) and biliary epithelium (n=1). Several cases (n=4) had secondary bacterial infections, including Bordetella bronchiseptica, Pasteurella multocida, and Streptococcus zooepidemicus. In situ hybridization in porcupine (n=6), raccoon (n=1), and skunk (n=1) revealed SkAdV-1 DNA in multiple tissue types, including lung, trachea, turbinates, liver, kidney, lymph node, and brain, and multiple cell types including epithelial, endothelial, and mesothelial cells. These findings were consistent across species. Comparison of viral genomes from a porcupine and a raccoon with that originally isolated from a skunk demonstrated DNA point mutations affecting several viral genes, including the fiber protein gene. Our findings show the spectrum of disease associated with SkAdV-1 infection in a broad host range of wildlife species.

Lactoferrin modulates increased liver DNA damage by reducing serum cytokine, hepcidin and iron levels in lipopolysaccharide-induced endotoxemia in rats

Lactoferrin modulates increased liver DNA damage by reducing serum cytokine, hepcidin and iron levels in lipopolysaccharide-induced endotoxemia in rats

Fertaric acid amends bisphenol A-induced toxicity, DNA breakdown, and histopathological changes in the liver, kidney, and testis.

BACKGROUNDBisphenol A (BPA) is present in many plastic products and food packaging. On the other hand, fertaric acid (FA) is a hydroxycinnamic acid. AIMTo investigate the effect of FA on BPA-related liver, kidney, and testis toxicity, DNA breakdown, and histopathology in male rats.METHODSThirty male albino rats were divided into five equal groups (6 rats/group): Control, paraffin oil, FA-, BPA-, and FA + BPA-treated groups. The control and paraffin oil groups were administered orally with 1 mL distilled water and 1 mL paraffin oil, respectively. The FA-, BPA-, and FA+ BPA-treated groups were administered orally with FA (45 mg/kg, bw) dissolved in 1 mL distilled water, BPA (4 mg/kg, bw) dissolved in 1 mL paraffin oil, and FA (45 mg/kg, bw) followed by BPA (4 mg/kg, bw), respectively. All these treatments were given once a day for 6 wk. RESULTSBPA induced a significant decrease in serum alkaline phosphatase, acid phosphatase, sodium, potassium and chloride, testosterone, dehydroepiandrosterone sulfate, glucose-6-phosphate dehydrogenase, 3β-hydroxysteroid dehydrogenase, and testis protein levels but a highly significant increase in serum aspartate aminotransferase, alanine aminotransferase, γ-glutamyl transpeptidase, lactate dehydrogenase, bilirubin, urea, creatinine, uric acid, luteinizing hormone, follicle stimulating hormone, sex hormone binding globulin, blood urea nitrogen, and testis cholesterol levels. Also, FA inhibited the degradation of liver, kidney, and testis DNA content. Oral administration of FA to BPA-treated rats restored all the above parameters to normal levels. CONCLUSIONFA ameliorates BPA-induced liver, kidney, and testis toxicity, DNA breakdown, and histopathological changes.

A Polysaccharide From Eupolyphaga sinensis Walker With Anti-HBV Activities In Vitro and In Vivo

Hepatitis B virus (HBV) infection remains a major global threat to human health worldwide. Recently, the Chinese medicines with antiviral properties and low toxicity have been a concern. In our previous study, Eupolyphaga sinensis Walker polysaccharide (ESPS) has been isolated and characterized, while its antiviral effect on HBV remained unclear. The anti-HBV activity of ESPS and its regulatory pathway were investigated in vitro and in vivo. The results showed that ESPS significantly inhibited the production of HBsAg, HBeAg, and HBV DNA in the supernatants of HepG2.2.15 in a dose-dependent manner; HBV RNA and core protein expression were also decreased by ESPS. The in vivo studies using HBV transgenic mice further revealed that ESPS (20 and 40 mg/kg/2 days) significantly reduced the levels HBsAg, HBeAg, and HBV DNA in the serum, as well as HBV DNA and HBV RNA in mice liver. In addition, ESPS activated the Toll-like receptor 4 (TLR4) pathway; elevated levels of IFN-β, TNF-α, and IL-6 in the serum were observed, indicating that the anti-HBV effect of ESPS was achieved by potentiating innate immunity function. In conclusion, our study shows that ESPS is a potential anti-HBV ingredient and is of great value in the development of new anti-HBV drugs.

Occult Hepatitis B virus (HBV) infection and challenges for hepatitis elimination: A literature review.

Occult hepatitis B infection (OBI) is characterized by the detection of hepatitis B virus (HBV) DNA in serum or liver but negativity for hepatitis B surface antigen. OBI, which is thought to be maintained by host, immunological, viral and/or epigenetic factors, is one of the most challenging clinical features in the study of viral hepatitis. Currently, there is no validated detection test for OBI. It is believed that OBI is widely distributed throughout the world, with a higher prevalence in populations at high-risk HBV, but the detailed worldwide prevalence patterns are unknown. We conducted a survey of recently published studies on OBI rates across all continents. High prevalence rates of OBI are observed in some specific groups, including patients with hepatitis C virus, human immunodeficiency virus co-infection or hepatocellular carcinoma. In 2016, the World Health Organization adopted strategies to eliminate viral hepatitis by 2030, but the difficulties in detecting and treating OBI currently challenge this goal. Subjects with OBI can transmit HBV, and episodes of reactivation can occur. Further studies to understanding the mechanisms that drive the development of OBI are needed and can contribute to efforts at eliminating viral hepatitis.

Potential Toxic Effects of Exposure to Titanium Silicon Oxide Nanoparticles in Male Rats.

Recently, nano titanium silicon oxide (TiSiO4 NPs) has been used in different fields and industries. Very few toxicological data exist for TiSiO4 NPs. In the present study, the potential adverse effects of oral exposure to a single dose of TiSiO4 NPs ≤ 50 nm (250 mg/kg b.w.) in adult male rats were investigated through the assessment of biomarkers for serum biochemical parameters, liver DNA damage, and histopathological examination and determination of Si and Ti in the exposed rat tissues. The results revealed that there were no significant changes in serum total protein, albumin, and triglycerides content, while total cholesterol level was significantly increased 7 days after exposure. TiSiO4 NPs significantly increased superoxide dismutase (SOD), glutathione peroxidase (GPx), acetylcholine esterase (AChE), lactate dehydrogenase (LDH) activity, luteinizing hormone (LH), and follicle-stimulating hormone (FSH) levels in the exposed rat serum, whereas alanine aminotransferase (ALT), aspartate aminotransferase (AST) activity, urea level, immunoglobulins (IgG and IgM) concentrations, progesterone, and testosterone levels were significantly decreased. The liver comet assay indices were significantly increased after 7 days post-exposure. Moreover, histopathological changes and the accumulation of Si and Ti in liver, kidney, spleen, and lung tissues of treated rats were recorded.

Alobar Holoprosencephaly in an Aborted American Quarter Horse Fetus

Holoprosencephaly is a central nervous system malformation, characterized by incomplete or total lack of division of prosencephalon hemispheres, which is commonly accompanied by craniofacial malformations. A 9-month-gestation aborted American Quarter Horse fetus was submitted for postmortem examination. The fetus lacked haircoat and had severe facial malformations including marked shortening/absence of the maxillary, incisive and nasal bones, bilateral anophthalmia, and pre-maxillary agenesis. The prosencephalon was small and nearly spherical, represented by a single lobe, with no visible separation between cerebral hemispheres. The olfactory bulbs, piriform lobes, and the optic chiasm were absent. At cross sectioning of the prosencephalon, the inner structures of the brain were completely absent, and replaced by a monoventricle lined by the remaining compressed cortex, and the thalami were fused. Since mutations in the sonic hedgehog (SHH) gene have been associated with human holoprosencephaly, the three coding SHH exons were sequenced using liver DNA of the aborted foal. The obtained SHH sequence was similar to the Equus caballus SHH mRNA sequence deposited in Genbank (XM_023640069.1); therefore, no polymorphism in the coding region of this gene justifying the phenotype was observed. This is the first report of alobar holoprosencephaly in horses.

Lack of germline transmission in male mice following a single intravenous administration of AAV5-hFVIII-SQ gene therapy

Valoctocogene roxaparvovec (AAV5-hFVIII-SQ) is an adeno-associated virus serotype five gene therapy under investigation for the treatment of hemophilia A. Herein, we assessed the potential for germline transmission of AAV5-hFVIII-SQ in mice. Male B6.129S6-Rag2tm1Fwa N12 mice received a single intravenous dose of vehicle or 6 × 1013 vg/kg AAV5-hFVIII-SQ. Vehicle and AAV5-hFVIII-SQ-treated mice were mated with naïve females 4 days after dosing, when the concentration of vector genomes was expected to be at its peak in semen, and 37 days after dosing, when a full spermatogenesis cycle was estimated to be complete. Quantitative PCR was used to evaluate the presence of transgene DNA in liver and testes from F0 males dosed with AAV5-hFVIII-SQ and liver tissue of F1 offspring. Transgene DNA was detected in liver and testes of all F0 males dosed with AAV5-hFVIII-SQ, confirming successful transduction. Importantly, no transgene DNA was detected in any tested F1 offspring derived from F0 males dosed with AAV5-hFVIII-SQ. Using a novel 2-stage statistical model that takes into account the number of males dosed with AAV5-hFVIII-SQ and the number of offspring sired by these males, we estimate that the risk of germline transmission is <5% with a 99.2% confidence level.

Circulating exosome level of indigenous fish may be a novel biomarker for the integrated ecotoxicity effect of water environment

The deficiency of effective biomarker for the toxic effects of water pollutants greatly limits the application of biological monitoring. This study aimed to investigate the possibility of circulating exosomes of indigenous fish acting as biomarker for the ecotoxicity effect of water environment. The Helong Reservoir in Guangzhou, China, was chosen as the investigating field, of which the water quality belongs to Class V (2013) (GB 3838-2002, China). The clean drinking water source of the upper reaches of the Liuxihe Reservoir was selected as the control. Indigenous fishes including Oreochromis niloticus (Nile tilapia), Labeo rohita (Rohu), Carassius auratus (Crucian carp) were sampled during the period from July 2020 to April 2021. Circulating exosomes of fish samples were isolated by using ultracentrifugation, characterized with transmission electron microscopy (TEM) and quantified by using bicinchoninic acid (BCA) assay. Oxidative stress, DNA and chromosome damage in liver, kidney, brain, gill and blood of fish samples were measured. The results showed that there were significant differences in superoxide dismutase (SOD) activity, glutathione (GSH) and malondialdehyde (MDA) contents, DNA and chromosome damage in fish samples between the Helong Reservoir and the control. Interestingly, there were also significant differences in circulating exosome levels of fish samples between them. Our data suggested that circulating exosome level of indigenous fish may be a novel biomarker for the ecotoxicity effects of water environment.

Non-Clinical Investigation of Tuberculosis Drugs: Conjugated Norbornene- Based Nanocarriers Toxic Impacts on Zebrafish

INTRODUCTION: Rifampicin conjugated (R-CP), and rifampicin -isoniazid dual conjugated (RI-CP) norbornene-derived nanocarriers are newly designed for pH stimuli-responsive delivery of tuberculosis (TB) drugs. Its biosafety level is yet to be well established. OBJECTIVES: To assess the impacts of the nanocarriers on liver cells using zebrafish animal model and human liver cell line model (HepG2). METHODS: Initially, lethal dose concentration for the norbornene-derived nanocarrier systems in zebrafish was determined. The toxic effects were analysed at the sub-lethal drug concentration by histopathological study, total GSH level, gene expression and DNA damage in zebrafish liver cells. Fish erythrocyte nuclear abnormalities were also evaluated. Cell viability and oxidative stress level (ROS generation) after exposure to the nanoconjugates was determined using HepG2 cell in the in vitro study. RESULTS: In vivo studies of both R-CP and RI-CP showed 100% mortality at 96 hours for exposure concentration &gt;100mg/l and showed toxic changes in zebrafish liver histology, GSH, and DNA damage levels. A noticeable upregulated PXR, CYP3A and cyp2p6 genes was observed in RI-CP exposure than in RIF or R-CP molecules. The in vitro study revealed a dose-dependent effect on cell viability and ROS generation for RIF, R-CP and RI-CP exposures in HepG2 cells. CONCLUSION: The current study reports that the rifampicin conjugated (R-CP) and rifampicin-isoniazid conjugated (RI-CP) norbornene derived nanocarriers exhibit enhanced toxic responses in both adult zebrafish and HepG2 cells. The pH-sensitive norbornene derived nanocarriers on conjugation with different drugs exhibited varied impacts on hepatic cells. Hence the present investigation recommends a complete metabolomics analysis and norbornene carrier-drug interaction study to be performed for each drug conjugated norbornene nanocarrier to ensure its biosafety.

Identification of new candidate biomarkers to support doxorubicin treatments in canine cancer patients

BackgroundBoth human and veterinary cancer chemotherapy are undergoing a paradigm shift from a “one size fits all” approach to more personalized, patient-oriented treatment strategies. Personalized chemotherapy is dependent on the identification and validation of biomarkers that can predict treatment outcome and/or risk of toxicity. Many cytotoxic chemotherapy agents, including doxorubicin, base their mechanism of action by interaction with DNA and disruption of normal cellular processes. We developed a high-resolution/accurate-mass liquid chromatography-mass spectrometry DNA screening approach for monitoring doxorubicin-induced DNA modifications (adducts) in vitro and in vivo. We used, for the first time, a new strategy involving the use of isotope-labeled DNA, which greatly facilitates adduct discovery. The overall goal of this work was to identify doxorubicin-DNA adducts to be used as biomarkers to predict drug efficacy for use in veterinary oncology.ResultsWe used our novel mass spectrometry approach to screen for adducts in purified DNA exposed to doxorubicin. This initial in vitro screening identified nine potential doxorubicin-DNA adduct masses, as well as an intense signal corresponding to DNA-intercalated doxorubicin. Two of the adduct masses, together with doxorubicin and its metabolite doxorubicinol, were subsequently detected in vivo in liver DNA extracted from mice exposed to doxorubicin. Finally, the presence of these adducts and analytes was explored in the DNA isolated from dogs undergoing treatment with doxorubicin. The previously identified nine DOX-DNA adducts were not detected in these preliminary three samples collected seven days post-treatment, however intercalated doxorubicin and doxorubicinol were detected.ConclusionsThis work sets the stage for future evaluation of doxorubicin-DNA adducts and doxorubicin-related molecules as candidate biomarkers to personalize chemotherapy protocols for canine cancer patients. It demonstrates our ability to combine in one method the analysis of DNA adducts and DNA-intercalated doxorubicin and doxorubicinol. The last two analytes interestingly, were persistent in samples from canine patients undergoing doxorubicin chemotherapy seven days after treatment. The presence of doxorubicin in all samples suggests a role for it as a promising biomarker for use in veterinary chemotherapy. Future studies will involve the analysis of more samples from canine cancer patients to elucidate optimal timepoints for monitoring intercalated doxorubicin and doxorubicin-DNA adducts and the correlation of these markers with therapy outcome.

Investigation on haplotypes of ixodid ticks and retrospective finding of Borrelia miyamotoi in bank vole (Myodes glareolus) in Switzerland

The current status of tick species, important tick-borne bacteria and protozoan parasites is well-documented in Switzerland. However, reports on the genetic diversity and geographical relationships of tick species in this country appear to be in part lacking or outdated. Thus, the aim of this study was to collect ticks from various host species in southern Switzerland, to compare them in a geographical context and to screen in these samples rare tick-borne pathogens hitherto not reported or having low prevalence in Switzerland.In 2019–2020 altogether 177 ixodid ticks were collected from the vegetation, as well as from humans (n = 17), dogs (n = 23), cats (n = 41), red deer (n = 8), a European rabbit and a European hedgehog at 25 locations in three cantons of south Switzerland. Tick species were identified morphologically, followed by DNA extraction and comparison of mitochondrial haplotypes with molecular-phylogenetic methods. Tick DNA extracts, as well as sixty-two rodent liver or spleen tissue DNA extracts (representing six species) available from 2005 to 2006 were screened for trypanosomes, Occidentia massiliensis and Borrelia miyamotoi.Morphologically, three tick species were identified: Ixodes ricinus (n = 170), Rhipicephalus sanguineus sensu lato (n = 6) and I. hexagonus (n = 1). In contrast to companion animals (dogs, cats) immature ticks (larvae and nymphs) predominated on humans, which was a highly significant association (P < 0.0001). Molecular comparison of the cytochrome c oxidase subunit I (cox1) gene with GenBank data established the species as R. sanguineus sensu stricto and confirmed I. hexagonus, both showing 99.8–100% sequence identity to conspecific ticks from northern Italy. Seventy-nine specimens morphologically identified as I. ricinus revealed high 16S rRNA gene haplotype diversity and represented two phylogenetic groups. Two I. ricinus haplotypes from Switzerland belonged to the same haplogroup with I. inopinatus from Spain, Germany and Austria as well as with I. ricinus reported from a broad geographical range of Europe (including Italy, the Netherlands, Poland, Latvia and Sweden). All 141 tick DNA extracts (from five R. sanguineus s.l., 135 I. ricinus and one I. hexagonus) and 62 rodent tissue DNA extracts were negative for trypanosomes and O. massiliensis. However, B. miyamotoi was identified in a bank vole (Myodes glareolus) and three ticks by sequencing.From Switzerland, this is the first report of tick haplotypes that are phylogenetically closely related to I. inopinatus. However, based on their morphology, both specimens are considered as I. ricinus. These results highlight the importance that the identification of I. inopinatus should be based on coherent morphologic and molecular properties. This is also the first report of rodent-borne B. miyamotoi in Switzerland. Taking into account the year of collection (2005), in a chronological order this might be the first indication of B. miyamotoi in any rodent species in Europe.

Natural infection status of 5 pathogens in rodents in Longyou county, Zhejiang

Objective To investigate the natural infection status of 5 pathogens in rodents in Longyou County of Zhejiang province, and provide evidence for the prevention and control of related diseases. Methods Rodents were captured in the field in Longyou. The lungs of the captured rodents were frozen and fixed in cold acetone. Hanta virus was detected by direct immunofluorescence. Rodent liver DNA extraction was done for the detection of Borrelia burgdorferis, Ehrlichia chaffeensis, Phagocytophilum, and Yersinia pestis by PCR amplification. Results A total of 88 rodents in 6 species were captured in 2017, including Apodemus agrarius, Rattus acutus, Rattus commune, shrew, Rattus flavipectus and Rattus norvegicus. Among them, Apodemus agrarius was the predominant species (88.64%), followed by Rattus acutus (5.00%). In 2019, a total of 103 rodents in 6 species were captured, including Apodemus agrarius, Rattus flavipectus, Rattus acutus, Rattus norvegicus, shrew and nested rat. Apodemus agrarius was still the predominant species, accounting for 84.47% of the total captured rodents. The results of pathogen detection showed that the positive rate of Hanta virus antigen was 3.85% and 4.60% in 2017 and 2019, respectively. The positive rate of Borrelia burgdorferi nucleic acid were 1.28% and 2.30%, respectively. The positive rate of Ehrlichia chaffeensis nucleic acid was 6.41% and 5.75%, respectively. The positive rate of Phagocytophilum nucleic acid was 1.28% and 2.30%, respectively. The detection results of Yersinia pestis were negative. No related pathogens in other species were detected. Conclusion There is a large number of Apodemus agrarius in Longyou, which is a potential infection source of many important infectious diseases. Therefore, it is necessary to strengthen the surveillance for Apodemus agrarius. In addition, the investigation of related diseases should be expanded in the future to provide evidence for the prevention and control of natural focal diseases.

Mechanisms of hepatoprotective effects of white and black garlic extracts against preneoplastic lesions induced by N-nitrosodiethylamine in rats

Introduction: Garlic has many pharmacological properties such as antibacterial, anti-inflammatory, and antioxidant activities. This study aimed to investigate the possible mechanisms of the hepatoprotective effect of white garlic extract (WGE) and black garlic extract (BGE) against preneoplastic lesions induced by N-nitrosodiethylamine (NDEA) in rats. Methods: Forty-two rats were randomly distributed into six equal groups. Oral administration of WGE and BGE began 3 weeks before injection of NDEA. Group 1 was kept as the negative control, while the other groups were injected by a single intraperitoneal dose of NDEA in the 3rd week, followed by 2 subcutaneous injections/week of CCl4 till six weeks to induce preneoplastic lesions. Group 2 kept positive control and groups 3, 4, 5, and 6 were given WGE and BGE each of them at 250 and 500 mg/kg, respectively, for six weeks from the beginning. Serum liver enzymes, total protein, albumin (Alb), total bilirubin (TBil), and antioxidant enzymes were measured. Malondialdehyde (MDA), glutathione (GSH), tumor biomarkers, along with the content of 8-hydroxy-2-deoxyguanosine (8-OHdG) in liver DNA were estimated and liver histopathology was performed. Results: WGE and BGE significantly decreased the serum liver enzymes, TBil, tumor biomarkers, lipid peroxidation but increased total protein levels. The extracts significantly increased antioxidant enzymes, decreased 8-OHdG content in liver DNA, and alleviated histopathological lesions in the liver. Conclusion: The results affirm the hepatoprotective effect of WGE and BGE against NDEA-induced preneoplastic lesions in rats. This effect may be due to inhibition of lipid peroxidation, reduction of tumor biomarkers, enhancement of antioxidant enzymes, or reduction of 8-OHdG content in liver DNA.