Protection by Herpes Simplex Virus Glycoprotein D against Fas-mediated Apoptosis ROLE OF NUCLEAR FACTOR κB

Bovine postpartum uterine disease, metritis, affects about 40% of animals and is widely considered to have a bacterial aetiology. Although the gamma-herpesvirus bovine herpesvirus 4 (BoHV-4) has been isolated from several outbreaks of metritis or abortion, the role of viruses in endometrial pathology and the mechanisms of viral infection of uterine cells are often ignored. The objectives of the present study were to explore the interaction, tropism and outcomes of BoHV-4 challenge of endometrial stromal and epithelial cells. Endometrial stromal and epithelial cells were purified and infected with a recombinant BoHV-4 carrying an enhanced green fluorescent protein (EGFP) expression cassette to monitor the establishment of infection. BoHV-4 efficiently infected both stromal and epithelial cells, causing a strong non-apoptotic cytopathic effect, associated with robust viral replication. The crucial step for the BoHV-4 endometriotropism appeared to be after viral entry as there was enhanced transactivation of the BoHV-4 immediate early 2 gene promoter following transient transfection into the endometrial cells. Infection with BoHV-4 increased cyclooxygenase 2 protein expression and prostaglandin estradiol secretion in endometrial stromal cells, but not epithelial cells. Bovine macrophages are persistently infected with BoHV-4, and co-culture with endometrial stromal cells reactivated BoHV-4 replication in the persistently infected macrophages, suggesting a symbiotic relationship between the cells and virus. In conclusion, the present study provides evidence of cellular and molecular mechanisms, supporting the concept that BoHV-4 is a pathogen associated with uterine disease.

https://rep.bioscientifica.com/downloadpdf/journals/rep/134/1/1340183.pdf

Bovine herpesvirus 4 is tropic for bovine endometrial cells and modulates endocrine function

The idea of using oncolytic viruses for the treatment of cancers was proposed a century ago. During the last two decades, viruses able to replicate specifically in cancer cells and to induce their lysis were identified and were genetically modified to improve their viro-oncolytic properties. More recently, a new approach consisting of inducing selective apoptosis in cancer cells through viral infection has been proposed; this approach has been called viro-oncoapoptosis. In the present study, we report the property of bovine herpesvirus-4 (BoHV-4) to induce, in vitro and in vivo, apoptosis of some human carcinomas. This conclusion relies on the following observations: (a) In vitro, BoHV-4 infection induced apoptosis of A549 and OVCAR carcinoma cell lines in a time- and dose-dependent manner. (b) Apoptosis was induced by the expression of an immediate-early or an early BoHV-4 gene, but did not require viral replication. (c) Cell treatment with caspase inhibitors showed that apoptosis induced by BoHV-4 relied mainly on caspase-10 activation. (d) Infection of cocultures of A549 or OVCAR cells mixed with human 293 cells (in which BoHV-4 does not induce apoptosis) showed that BoHV-4 specifically eradicated A549 or OVCAR cancer cells from the cocultures. (e) Finally, in vivo experiments done with nude mice showed that BoHV-4 intratumoral injections reduced drastically the growth of preestablished A549 xenografts. Taken together, these results suggest that BoHV-4 may have potential as a viro-oncoapoptotic agent for the treatment of some human carcinomas. Moreover, further identification of BoHV-4 proapoptotic gene(s) and the cellular pathways targeted by this or these gene(s) could lead to the design of new cancer therapeutic strategies.

https://cancerres.aacrjournals.org/content/canres/65/20/9463.full.pdf

Bovine Herpesvirus 4 Induces Apoptosis of Human Carcinoma Cell Lines In vitro and In vivo

Today a growing number of studies are focusing on antibiotic resistance in wildlife. This is due to the potential role of wild animals as reservoirs and spreaders of pathogenic and resistant bacteria. This study focused on isolating and identifying Escherichia coli from the feces of wild animals living in the Apuan Alps Regional Park (Tuscany, Italy) and evaluating some of their antibiotic resistance and pathogenicity traits. Eighty-five fecal samples from different species were studied. Seventy-one E. coli were identified by matrix assisted laser desorption ionization-time of flight mass spectrometry analysis, subjected to antibiograms and polymerase chain reaction for the detection of antibiotic resistance genes and pathogenicity factors. The highest resistance rates were found against cephalothin (39.4%) and ampicillin (33.8%), followed by amoxicillin/clavulanic acid (15.5%), streptomycin (12.7%), and tetracycline (5.6%). Regarding resistance genes, 39.4% of the isolates were negative for all tested genes. The remaining isolates were positive for blaCMY-2, sul2, strA-strB and aadA1, tet(B), and tet(A), encoding resistance to beta-lactams, trimethoprim/sulfamethoxazole, streptomycin, and tetracycline, respectively. With regard to virulence factors, 63.4% of the isolates were negative for all genes; 21.1% carried astA alone, which is associated with different pathotypes, 9.9% carried both escV and eaeA (aEPEC); single isolates (1.4%) harbored escV (aEPEC), escV associated with astA and eaeA (aEPEC), astA with stx2 and hlyA (EHEC) or astA and stx1, stx2, and hlyA (EHEC). These results show that wildlife from nonanthropized environments can be a reservoir for antibiotic-resistant microorganisms and suggest the need for a deeper knowledge on their origin and diffusion mechanisms through different ecological niches.

Antibiotic Susceptibility and Virulence Factors in Escherichia coli from Sympatric Wildlife of the Apuan Alps Regional Park (Tuscany, Italy).

Wild boar are among the most widespread wild mammals in Europe. Although this species can act as a reservoir for different pathogens, data about its role as a carrier of pathogenic and antimicrobial-resistant Escherichia coli are still scarce. The aim of this study was to evaluate the occurrence of antimicrobial-resistant and pathogenic Escherichia coli in wild boar in the Tuscany region of Italy. During the hunting season of 2018-2019, E. coli was isolated from 175 of 200 animals and subjected to antimicrobial resistance tests and PCR for detection of resistance and virulence factor genes. The highest resistance rates were against cephalothin (94.3%), amoxicillin-clavulanic acid (87.4%), ampicillin (68.6%), and tetracycline (44.6%). The most detected resistance genes were blaCMY-2 (54.3%), sul1 (38.9%), sul2 (30.9%), and tetG (24.6%). Concerning genes encoding virulence factors, 55 of 175 isolates (31.4%) were negative for all tested genes. The most detected genes were hlyA (47.4%), astA (29.1%), stx2 (24.6%), eaeA (17.1%), and stx1 (11.4%). E. coli was classified as Shiga toxin-producing E. coli (STEC) (21.7%), enterohemorrhagic E. coli (EHEC) (6.3%), enteroaggregative E. coli (EAEC) (5.1%), and atypical enteropathogenic E. coli (aEPEC) (3.4%). Enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC), and typical enteropathogenic E. coli (tEPEC) were not detected. Our results show that wild boars could carry pathogenic and antimicrobial-resistant E. coli, representing a possible reservoir of domestic animal and human pathogens.

Pathotypes and Antimicrobial Susceptibility of Escherichia Coli Isolated from Wild Boar (Sus scrofa) in Tuscany.

Wild birds are considered to be reservoirs of human enteric pathogens and vectors of antimicrobial resistance dissemination in the environment.
 During their annual migration, they play a potential role in the epidemiology of human associated zoonoses. The aim of this study was to investigate the frequency of isolation and antimicrobial susceptibility profiles of microorganisms found in the cloaca of common European passerines. One hundred and twenty-one cloacal swabs were collected during a monitoring program of migratory birds in the Forest Reserve for Protection “Metaponto” (Basilicata, Italy). All samples were cultured using standard bacteriological methods and antibiotic susceptibility testing (agar disk diffusion test) of isolated strains was performed. The bacteriological analysis produced 122 strains belonging to 18 different species. The most commonly isolated species were Enterobacter cloacae and Providencia rettgeri (21 strains, 17.2%). Potentially pathogenic species including Klebsiella pneumoniae, Serratia marcescens and Pseudomonas spp. have also been identified. Isolates showed significant frequencies of antimicrobial resistance. The highest frequency of resistance was observed against amoxicillin (n = 79, 64.8%); ampicillin (n = 77, 63.1%); rifampicin (n = 75, 61.5%); amoxicillin–clavulanic acid (n = 66, 54.1%). Thirty-one strains (25.4%) showed resistance to imipenem and 8 (6.6%) to meropenem. Migratory birds play an important role in the ecology, circulation and dissemination of potentially pathogenic antimicrobial resistant organisms. They can therefore be considered sentinel species and environmental health indicators. Our results suggest that the integration of epidemiological surveillance networks during ringing campaigns of wild species can be an effective tool to study this phenomenon.

/pdf/antibiotic-resistance-assessment-in-bacteria-isolated-in-3vuiw0h2yk.pdf

Antibiotic resistance assessment in bacteria isolated in migratory Passeriformes transiting through the Metaponto territory (Basilicata, Italy)

The gamma-2-herpesvirus bovine herpesvirus 4 causes apoptotic infection in permissive cell lines.

Currently, there are no data in the literature on the topical treatment with Thymus vulgaris essential oil (EO) in bacterial skin infections of dogs. This study evaluates the effectiveness of Thymus vulgaris EO for topical use in dogs with skin 18 half-breed dogs, affected by skin infections housed in a rescue shelter were studied. The bacteria isolated from these dogs were identified by MALDI-TOF-MS. The susceptibility of the isolated microorganisms to Thymus vulgaris L. EO was estimated in vitro by bacteriological test (CLSI 2015), in comparison to some antimicrobials drugs (amoxicillin-clavulanic acid, doxycy-cline, thiamphenicol and marbofloxacin) and to Citrus bergamia EO. The dogs, divided in two groups at random, were treated topically for 7 days with Thymus vulgaris L. EO (Group 1: n. 10 animals) and Citrus bergamia Risso e Poiteau EO (Group 2: n. 8 animals) respectively. The bacteria isolated were: Staphylococcus pseudintermedius (7 samples), Staphylococcus sciuri (4), ESBL Escherichia coli (3) and Proteus mirabilis (4). In all animals treated with Thymus vulgaris EO, the clinical signs decreased rapidly within 5 days from the administration, with complete remission 7 days after the treatment. No bacterial growth was observed from skin swabs after 7 days of treatment. None of the treated animals showed local or general side effects. The use of Thymus vulgaris EO could be a possible alternative or additional treatment to antibiotics in dermatological infections, particularly in cases refractory to conventional therapy.

/pdf/effectiveness-of-thymus-vulgaris-essential-oil-in-the-3fgm4v2wtl.pdf

Bianca Maria Orlandella

Papers

Antibiotic resistance assessment in bacteria isolated in migratory Passeriformes transiting through the Metaponto territory (Basilicata, Italy)

The gamma-2-herpesvirus bovine herpesvirus 4 causes apoptotic infection in permissive cell lines.

Effectiveness of Thymus vulgaris Essential Oil in the Treatment of Skin Infections in Dogs