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Dietzia strains are widely distributed in the environment, presenting an opportunistic role, and some species have undetermined taxonomic characteristics. Here, we propose the existence of errors in the classification of species in this genus using comparative genomics. We performed ANI, dDDH, pangenome and genomic plasticity analyses better to elucidate the phylogenomic relationships between Dietzia strains. For this, we used 55 genomes of Dietzia downloaded from public databases that were combined with a newly sequenced. Sequence analysis of a phylogenetic tree based on genome similarity comparisons and dDDH, ANI analyses supported grouping different Dietzia species into four distinct groups. The pangenome analysis corroborated the classification of these groups, supporting the idea that some species of Dietzia could be reassigned in a possible classification into three distinct species, each containing less variability than that found within the global pangenome of all strains. Additionally, analysis of genomic plasticity based on groups containing Dietzia strains found differences in the presence and absence of symbiotic Islands and pathogenic islands related to their isolation site. We propose that the comparison of pangenome subsets together with phylogenomic approaches can be used as an alternative for the classification and differentiation of new species of the genus Dietzia.

Tailoring the cell organelles and thus changing cell homeostatic behavior has permitted the discovery of fascinating metabolic features enabling enhanced viability, differentiation, or quenching inflammation. Recently, photobiomodulation (PBM) has been accredited as an effective cell manipulation technique with promising therapeutic potential. In this prospective, in vitro results revealed that 808-nm laser light emitted by a hand-piece with a flat-top profile at an irradiation set up of 60 J/cm2 (1 W, 1 W/cm2; 60 s, continuous wave) regulates bone marrow stromal cell (BMSC) differentiation toward osteogenesis. Considering the importance of actin cytoskeleton reorganization, which controls a range of cell metabolic activities, comprising shape change, proliferation and differentiation, the aim of the current work is to assess whether PBM therapy, using a flat-top hand-piece at higher-fluence irradiation on BMSCs, is able to switch photon signals into the stimulation of biochemical/differentiating pathways involving key activators that regulate de novo actin polymerization. Namely, for the first time, we unearthed the role of the flat-top hand-piece at higher-fluence irradiation on cytoskeletal characteristics of BMSCs. These novel findings meet the needs of novel therapeutically protocols provided by laser treatment and the manipulation of BMSCs as anti-inflammatory, osteo-inductive platforms.

This in vitro study was carried out to evaluate the potential antibacterial properties of canine non-transfusional hemo-components. Therapeutic formulations commonly used for regenerative medicine purposes (platelet-rich plasma, platelet gel, platelet lysate, fibrin glue), considering both leukocyte-rich and leukocyte-poor formulations, but also platelet-poor plasma and activating substances (thrombin, calcium gluconate), were tested to detect elements with potential antimicrobial properties. The antibacterial effect was tested on different bacterial strains (Staphylococcus aureus subspecies aureus, Staphylococcus cohnii subspecies cohnii, Escherichia coli, Pseudomonas aeruginosa, and Klebsiella pneumoniae subspecies pneumoniae) isolated from canine wounds and classified as susceptible, multidrug-, extensively, and pandrug-resistant bacteria toward a known panel of human and veterinary antibiotics. The evaluation was carried out by agar gel diffusion method (Kirby–Bauer) and micro-inhibition in broth using microplates and spectrophotometer reading. The study findings confirmed the hypothesized antibacterial properties of canine non-transfusional hemo-components. A more effective bacteriostatic effect was found against Gram-negative bacteria, drug-resistant too. The presence of leukocytes or platelets does not appear to be essential for the antibacterial effect. Further studies should be conducted to evaluate the exact mechanism of action of the antimicrobial activity. However, non-transfusional hemo-components could be a useful natural aid in controlling bacterial infections in dogs.

Background In free-ranging reptile populations, bacterial, fungal, viral and parasitic pathogens may affect hosts through impairment in movements, thermoregulation, reproduction, survival, and population dynamics. The speckled dwarf tortoise (Chersobius [Homopus] signatus) is a threatened species that is mostly restricted to the Succulent Karoo biome in South Africa, and little information on pathogens of this species is available yet. We derived baseline parameters for five males and five females that were captured to genetically enhance a conservation breeding program in Europe. Upon collection of the tortoises, ticks were removed and identified. Immediately upon arrival in Europe, ocular, nasal, oral and cloacal swabs were taken for viral, bacteriological and mycological examinations. Fecal samples were collected before and 1 month after fenbendazole treatment, and analyzed for parasites. A panel of PCR, aiming to detect herpesviruses, adenoviruses and iridoviruses, was carried out. Results Samples were negative for viruses, while bacteriological examination yielded detectable growth in 82.5% of the swabs with a mean load of 16 × 107 ± 61 × 108 colony forming units (CFU) per swab, representing 34 bacterial species. Cloacal and oral swabs yielded higher detectable growth loads than nasal and ocular swabs, but no differences between sexes were observed. Fungi and yeasts (mean load 5 × 103 ± 13 × 103 CFU/swab) were detected in 25% of the swabs. All pre-treatment fecal samples were positive for oxyurid eggs, ranging from 200 to 2400 eggs per gram of feces, whereas after the treatment a significantly reduced egg count (90–100% reduction) was found in seven out of 10 individuals. One remaining individual showed 29% reduction, and two others had increased egg counts. In five tortoises, Nycthocterus spp. and coccidian oocysts were also identified. Soft ticks were identified as Ornithodoros savignyi. Conclusions Our baseline data from clinically healthy individuals will help future studies to interpret prevalences of microorganisms in speckled dwarf tortoise populations. The study population did not appear immediately threatened by current parasite presence.

The breeding of meat rabbits is an important sector in the livestock industry in Italy. The focus of this study was to describe the antibiotic resistance profile distribution among the Methicillin-sensitive Staphylococcus aureus isolated in a rabbit farm. From 400 animals of different ages and three farm workers, 96 randomly selected strains isolated from various anatomical sites and lesions were analysed. According to spa typing and the resistance profiles towards veterinary and human antibiotics, 26 pathotypes were identified. The highest resistance was observed against Tetracyclines (92.3%) and Macrolides (80.8%), while almost all were susceptible to Penicillins, according to the limited use of &beta, lactams on the farm. In total, 92.3% of pathotypes were multidrug resistant (MDRs). Two MDR pathotypes belonging to the t2802 spa type were isolated from both farmers and rabbits. Age categories harboured significantly different pathotypes (p = 0.019), while no association was found between pathotypes and lesions (p = 0.128) or sampling sites (p = 0.491). The antibiotic resistance was observed to increase with the time spent in the farm environment (age category). The selective pressure exerted by antibiotic use acted by giving advantage to more resistant strains rather than by lowering susceptibility to various drug categories within strains.

A study in an intensive Italian rabbit farm was carried out to assess the effect of age category and anatomical site on detection of S. aureus and to describe the diversity of spa-types within farm, including workers. On 400 rabbits of different age, 2066 samples from the ear, nose, axilla, groin, perineum and lesions were analyzed and 595 S. aureus were isolated. In total, 284 (71.0%) were colonized by S. aureus in at least one anatomical site and 35 animals (8.8%) had lesions. S. aureus prevalence was higher in adults than other age categories. Ear (29.4%) and nose (27.2%) were the most frequently colonized sites. The presence of lesions due to S. aureus was strongly associated with the colonization of at least one body site and the risk of lesions was proportionate to the number of sites colonized by S. aureus. In addition, a genotypic characterization was performed for 96 S. aureus strains randomly selected. All isolates resulted as low-virulence methicillin-sensitive-Staphylococcus aureus (LV-MSSA). Five different clonal lineages were found: spa-t2802, the most frequently detected (53.1%), also in human samples, spa-t491 (38.5%), spa-t094, t605, and spa-t2036. Strains t094, t491 and t2036 have not been isolated in Italy before.

Infectious causes of myositis are reported relatively uncommonly in horses. Among them, bacterial causes include Streptococcus equi subsp. zooepidemicus, Actinobacillus equuli, Fusobacterium spp. Staphylococcus spp, and Corynebacterium pseudotuberculosis. Infection can be spread to muscles via haematogenous or extension from skin lesions. Parasitic myositis has also been documented. In this report, a 12 year-old Italian Quarter Horse mare presented with diffuse subcutaneous nodules and masses ranging from 2 × 3 to 5 × 20 cm in size, and adherent to subcutis and muscles that were first macroscopically and cytologically diagnosed as pyogranulomas. Subsequently, histological, molecular, bacteriological, and biochemical investigations were performed. All the data obtained allowed to diagnose a severe and diffuse multibacterial granulomatous myositis caused by Corynebacterium pseudotuberculosis and Corynebacterium amycolatum. Following the therapy and an initial disappearance of most of the lesions together with a general improvement of the mare, the clinical condition deteriorated, and new nodules appeared. Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) and PCR techniques revealed the presence of bacteria as Glutamicibacter creatinolyticus and Dietzia spp. To the authors’ knowledge, this case report represents the first description of multibacterial granulomatous myositis due to Corynebacterium pseudotuberculosis, Corynebacterium amycolatum, Glutamicibacter creatinolyticus, and Dietzia spp. in a horse reared in Italy.

Bacteria of the genusGlutamicibacterare considered ubiquitous because they can be found in soil, water and air. They have already been isolated from different habitats, including different types of soil, clinical samples, cheese and plants. Glutamicibacter creatinolyticus is a Gram-positive bacterium important to various biotechnological processes, however, as a pathogen it is associated to urinary tract infections and bacteremia. Recently,Glutamicibacter creatinolyticusLGCM 259 was isolated from a mare, which displayed several diffuse subcutaneous nodules with heavy vascularization. In this study, sequencing, genomic analysis ofG. creatinolyticusLGCM 259 and comparative analyseswere performedamong 4representatives of different members of genusfromdifferent habitats, available in the NCBI database. The LGCM 259 strain's genome carries important factors of bacterial virulence that are essential in cell viability, virulence, and pathogenicity. Genomic islands were predicted for 4 members of genusGlutamicibacter,showing ahigh number of GEIs,which may reflect a high interspecific diversity and a possible adaptive mechanism responsible for the survival of each species in its specific niche. Furthermore,G. creatinolyticusLGCM 259 sharessyntenicregions, albeit with a considerable loss of genes, in relation to the other species. In addition,G. creatinolyticusLGCM 259 presentsresistancegenes to 6 differentclasses ofantibiotics and heavy metals, such as: copper, arsenic, chromium and cobalt-zinc-cadmium.Comparative genomicsanalysescouldcontribute to the identification of mobile genetic elements particular to the speciesG. creatinolyticuscompared to other members of genus. The presence of specific regions inG. creatinolyticuscould be indicative of their rolesin host adaptation, virulence, and the characterization ofastrain that affects animals.

Pestiviruses are widespread in the world among ungulates and infect both domestic and wild animals causing severe economic losses in livestock. Bovine Viral Diarrhea Virus type 1 (BVDV-1), now re-designated as Pestivirus A, causes diseases mainly in cattle, while few data are available about infection in wild ruminants and about the role of these animals in viral maintenance and spread. In order to investigate BVDV-1 infection in domestic and wild ruminants, especially at the wildlife/livestock interface, bulk tank milk from dairy cattle and sheep and spleen from red deer, roe deer and fallow deer were analysed. Furthermore, faecal samples from Apennine chamois and from wild deer were evaluated as a suitable sample for detecting and genotyping pestiviruses. BVDV-1 RNA was found in all animal species tested but not sheep. Genotyping based on partial 5'UTR and Npro sequences detected BVDV-1a in samples from Apennine chamois, red deer, roe deer and pasture-raised cattle, while BVDV-1c was found in a faecal sample from Apennine chamois and in a spleen sample from roe deer. For the first time BVDV-1 RNA was found and genotyped from faecal samples of wild ruminants and of cattle. BVDV-1a detection in Apennine chamois, red deer, roe deer and pasture-raised cattle suggests that the eventuality of viral transmission at the wildlife/livestock interface should be carefully evaluated. BVDV subgenotype 1c was found for the first time in roe deer and Apennine chamois in Central Italy, therefore the epidemiological role of these animals and the viral ecology should be further investigated.

Streptococcus equi subsp. zooepidemicus (SEZ) is an opportunistic and zoonotic pathogen of horses. In this study, genetic intraspecies variability of SEZ obtained mainly from respiratory and genital samples of horses was investigated by analysis of the 16S–23S rRNA intergenic spacer region (ISR) and of the 16S rRNA gene. 16S–23S ISR rRNA type A1 was predominant, although a high rate of multiple products (30.5%) was obtained. Phylogenetic analysis of the 16S rRNA gene detected three genogroups (I, II and III). 16S rRNA variable regions V1 and V2 are the most important regions for evaluating SEZ intraspecies variability, but at least V1-V5 regions should be considered to avoid mistakes. Analysis of all 16S rRNA sequences available in databases assigned human SEZ to groups I and III but not to group II. These results show a high genetic variability in SEZ collected from different specimens of horses from various regions of Italy.

The distinction between lower respiratory tract infections caused by Rhodococcus equi and those caused by other pathogens is difficult. The aim of this retrospective study was to describe cytological findings in bronchoalveolar lavage fluid (BALF) of foals with pneumonia caused by R. equi or other bacteria. Nineteen foals aged from 3 weeks to 6 months with evidence of respiratory disease between 2015 and 2016 were selected from the database of the Veterinary Teaching Hospital “Mario Modenato” of the University of Pisa. Eight foals of 19 (42.1%) had R. equi pneumonia, whereas 11 of 19 (57.9%) had another bacterial pneumonia. R. equi–positive foals had statistically significant higher total nucleated cell count (P = .02) and neutrophils percentage (P = .002) than R. equi–negative ones. Macrophages proportion (P = .01) was lower in R. equi–positive than in R. equi–negative foals. Bronchoalveolar lavage is a quite easy procedure that can be performed in the field with minimal equipment required. Here we reported significant differences in the cellular composition of BALF that can be used to differentiate foals with R. equi bronchopneumonia from those with other bacterial pneumonias, whereas waiting for culture results.

In order to determine the association between the lactation stage with serum and milk ELISA results, 697 ewes reared in 17 Paratuberculosis infected dairy farms, were sampled: 10 flocks at early/late and 7 flocks at peak lactation period. Prevalences of 7.75% (n = 697) and 5.31% (n = 697) were observed analyzing serum and milk samples, respectively, and the difference was not significant (P = 0.065). A higher number of Mycobacterium avium subsp. paratuberculosis (MAP) seropositive sheep were found in the larger flocks than in the smaller ones (P = 0.04), when sampled in early/late lactation. Moreover, the milk ELISA revealed that 7.8% and 3.7% of the ewes were positive to MAP antibodies at early/late and peak of lactation, respectively, and the difference was highly significant (P = 0.021). Considering the importance of early diagnosis, milk ELISA could be a valid alternative tool in early/late lactation stage. Further studies, investigating the effect of MAP positivity on production parameters of ovine dairy breeds, are needed.

The aims of this work were to evaluate the potential role of enrofloxacin in controlling the severity of the clinical mastitis in sheep caused by Staphylococcus aureus; to improve cure rates and to minimize the related effects of the disease on the mammary glands. This study was conducted in commercial dairy flocks where there was on going intensive monitoring of subclinical mastitis by Somatic Cell Count (SCC) and bacteriology. Two groups of animals were selected from these flocks. Group A (n = 34 animals) and Group B (n = 39 animals) were treated with 2.5 mg/kg bw and 5 mg/kg bw, respectively of enrofloxacin (Baytril® 5% injectable solution, Bayer, Italy) for three consecutive days (2 doses per day). The effectiveness of the enrofloxacin in curing the S. aureus-induced clinical mastitis was monitored through SCC, rectal temperature, and by systemic and local mammary gland reactions from the 1st to the 14th day post treatment. The presence of S. aureus in milk samples was confirmed by bacteriological examination and PCR before and after treatment. Bacteriological cure was 39% in Group A and 82% in Group B. Both doses significantly reduced SCC (P

EHV1 and EHV4 are the most important herpesviruses in horses. Repeated cases of abortion in mares regularly vaccinated, prompted us to investigate the immune response after vaccination with the same inactivated vaccine, but with three different protocols. Eighteen mares were chosen and randomly divided in three study groups (G1-G2-G3) and a control group (Ctrl). For serologic and PCR investigations nasal swabs, sera and blood were collected. The protocol used in G3 (4 doses) increased the titer recorded by ELISA and seroneutralization (SN). Poor agreement and no correlation were observed in titer values between ELISA and SN and between SN and PCR. A very weak positive correlation between ELISA and PCR was obtained. Seven out of 18 nasal swabs were positive by PCR; none showed viremia and no abortion occurred, regardless of vaccination status and despite active circulation of EHV-1 in the farm at the time of the study. The study was conducted in field conditions, in a susceptible population with a known history of infection and abortion, and among the three protocols, the one proposed in the G1 was the least efficient while the one proposed for the G3, seems to have induced a higher antibody titer in both SN and ELISA.

Small Ruminant Lentiviruses (SRLVs) are widespread in many countries and cause economically relevant, slow, and persistent diseases in sheep and goats. Monitoring the genetic diversity of SRLVs is useful to improve the diagnostic tools used in the eradication programs. In this study, SRLVs detected in Spanish Assaf sheep with different grades of lymphoproliferative mastitis were sequenced. Genetic characterization showed that most samples belonged to type A and were closer to Spanish SRLV isolates previously classified as A2/A3. Four samples belonged to subtype B2 and showed higher homology with Italian B2 strains than with Spanish B2 isolates. Amino acid sequences of immuno-dominant epitopes in the gag region were very conserved while more alterations were found in the LTR sequences. No significant correlations were found between grades of mastitis and alterations in the sequences although samples with similar histological features were phylogenetically closer to each other. Broader genetic characterization surveys in samples with different grades of SRLV-lesions are required for evaluating potential correlations between SRLV sequences and the severity of diseases.

During a serological survey, 157 out of 681 unvaccinated buffaloes resulted seropositive for bovine alphaherpesvirus 1 (BoHV1) glycoprotein B (gB) and seronegative for BoHV1 glycoprotein E (gE). These serological results were generally expected in animals vaccinated with a BoHV1 gE-deleted vaccine but not in unvaccinated animals. Seroneutralization tests on 36 selected sera detected neutralizing antibody titers more than three times higher for BuHV1 than for BoHV1. In order to investigate the virus, one of these buffaloes was injected with dexamethasone, and from nasal and vaginal swabs collected at different time points, a ruminant herpesvirus was isolated, characterized and also detected by PCR. Restriction enzyme analysis, sequencing and phylogenic analysis of gB and gD genes showed that the virus was genetically similar but not identical to BuHV1 strain b6. Intranasal inoculation of the virus in a healthy seronegative buffalo resulted in a mild and transient upper respiratory disease; the virus was isolated from clinical specimens and DNA was detected by PCR in nasal and vaginal swabs up to 9 days after infection. Further investigations should be aimed at sequencing the whole viral genome and at evaluating the host-range of this virus. Specific tests are needed to discriminate infections by different ruminant herpesviruses and to improve eradication programs of infectious bovine rhinotracheitis/infectious pustular vulvovaginitis in cattle.

The literature has supported the concept of mesenchymal stromal cells (MSCs) in bone regeneration as one of the most important applications in oro-maxillofacial reconstructions. However, the fate of the transplanted cells and their effects on the clinical outcome is still uncertain. Photobiomodulation (PBM) plays an important role in the acceleration of tissue regeneration and potential repair. The aim of this in vitro study is to evaluate the effectiveness of PBM with 808 nm diode laser therapy, using a flat-top hand-piece delivery system at a higher-fluence (64 J/cm2) irradiation (1 W, continuous-wave) on bone marrow stromal cells (BMSCs). The BMSCs of 3 old female Balb-c mice were analyzed. The cells were divided into two groups: irradiated group and control group. In the former the cells were irradiated every 24 h during 0 day (T0), 5 (T1), 10 (T2), and 15 (T3) days, whereas the control group was non-irradiated. The results have shown that the 64 J/cm2 laser irradiation has increased the Runt-related transcription factor 2 (Runx2). Runx2 is the most important early marker of osteoblast differentiation. The higher-fluence suppressed the synthesis of adipogenic transcription factor (PPARγ), the pivotal transcription factor in adipogenic differentiation. Also, the osteogenic markers such as Osterix (Osx) and alkaline phosphatase (ALP) were upregulated with an increase in the matrix mineralization. Furthermore, western blotting data demonstrated that the laser therapy has induced a statistically valid increase in the synthesis of transforming growth factor β1 (TGF-β1) but had no effects on the tumor necrosis factor α (TNFα) production. The data has statistically validated the down-regulation of the important pro-inflammatory cytokines such as interleukin IL-6, and IL-17 after 808 nm PBM exposition. An increase in anti-inflammatory cytokines such as IL-1rα and IL-10 was observed. These in vitro studies provide for first time the initial proof that the PBM of the 808 nm diode laser therapy with flat-top hand-piece delivery system at a higher-fluence irradiation of 64 J/cm2 (1 W/cm2) can modulate BMSCs differentiation in enhancing osteogenesis.

The objective of the study was to review current literature to determine whether the topical application of platelet-rich plasma (PRP) promotes healing in experimentally-induced full-thickness skin wounds in animals. The hypothesis was that the adjunct of PRP has a positive effect on wound healing. An electronic search was carried out on the following databases: Web of Science, Cochrane Library, PubMed, Research Gate, Cochrane Wounds Group, Veterinary Information Network. No publication date nor language restrictions were applied. Randomised and not randomised controlled clinical trials comparing PRP with placebo or with other treatments were included. The reduction of open wound area in PRP-treated (test) wounds compared to control wounds was the primary outcome. Secondary outcomes were healing time and number of healed cases in test group compared to control. The following effect sizes were calculated: the Hedges' g for continuous variables; the odds ratio for binary data. Eighteen controlled clinical trials were included in the qualitative and quantitative synthesis, with a total of 661 wounds. All studies were published in the period 2007-2016. Eight studies were carried out on rodent/lagomorph mammals and 10 on non-rodent/lagomorph mammals. In all included studies, control wounds underwent placebo or were left untreated. The PRP group showed a better healing performance than the control group in each outcome. The effect size was statistically significant considering the primary outcome and the overall aggregation of the three outcomes. The effect size, although in favour of the treatment with PRP, was not significant considering the healing time and the number of healings. The overall heterogeneity was mild or moderate. Five studies reported a high risk of selection bias. The publication bias was always mild or absent. The results support the hypothesis of the positive effects of the PRP when compared to control groups in the treatment of experimentally-induced full-thickness skin wounds in animals. PRP can therefore be considered an effective adjunctive therapy in stimulating second intention healing of acute wounds in healthy animals.

Necropsies were conducted on a female blue-fronted Amazon (Amazona aestiva) and a female yellow-headed Amazon (Amazona oratrix) that died after depression, ruffled feathers, diarrhea, and biliverdin in the urine. Gross and microscopic examinations revealed multifocal necrosis in the liver, spleen, lungs, kidneys, intestines, and heart caused by acute bacteremia. Yersinia pseudotuberculosis, serogroup O:1a, was isolated by culturing from the visceral lesions in the liver, intestines, and spleen. Virulence gene analysis showed the presence of the inv gene and the complete pathogenicity island: IS100, psn, yptE, irp1, irp2 ybtP-ybtQ, ybtX-ybtS, and int asnT-Int. Histopathologic findings and chemical analysis also demonstrated hepatic hemosiderosis. As has been demonstrated in other species, hemosiderosis may predispose Amazona spp. to systemic infection with Y. pseudotuberculosis after enteric disease.

The literature has supported the concept of mesenchymal stromal cells (MSCs) in bone regeneration as one of the most important applications in oro-maxillofacial reconstructions. However, the fate of the transplanted cells and their effects on the clinical outcome is still uncertain. Photobiomodulation (PBM) plays an important role in the acceleration of tissue regeneration and potential repair. The aim of this in vitro study is to evaluate the effectiveness of PBM with 808 nm diode laser therapy, using a flat-top hand-piece delivery system at a higher-fluence (64 J/cm2) irradiation (1 W, continuous-wave) on bone marrow stromal cells (BMSCs). The BMSCs of 3 old female Balb-c mice were analyzed. The cells were divided into two groups: irradiated group and control group. In the former the cells were irradiated every 24 h during 0 day (T0), 5 (T1), 10 (T2), and 15 (T3) days, whereas the control group was non-irradiated. The results have shown that the 64 J/cm2 laser irradiation has increased the Runt-related transcription factor 2 (Runx2). Runx2 is the most important early marker of osteoblast differentiation. The higher-fluence suppressed the synthesis of adipogenic transcription factor (PPARγ), the pivotal transcription factor in adipogenic differentiation. Also, the osteogenic markers such as Osterix (Osx) and alkaline phosphatase (ALP) were upregulated with an increase in the matrix mineralization. Furthermore, western blotting data demonstrated that the laser therapy has induced a statistically valid increase in the synthesis of transforming growth factor β1 (TGF-β1) but had no effects on the tumor necrosis factor α (TNFα) production. The data has statistically validated the down-regulation of the important pro-inflammatory cytokines such as interleukin IL-6, and IL-17 after 808 nm PBM exposition. An increase in anti-inflammatory cytokines such as IL-1rα and IL-10 was observed. These in vitro studies provide for first time the initial proof that the PBM of the 808 nm diode laser therapy with flat-top hand-piece delivery system at a higher-fluence irradiation of 64 J/cm2 (1 W/cm2) can modulate BMSCs differentiation in enhancing osteogenesis.

Streptococcus dysgalactiae subsp. equisimilis (SDSE) can be severely pathogenic in humans and is increasingly isolated from horses with respiratory, reproductive or other diseases, although it is often considered a commensal bacterium. Here a PCR protocol is described for identifying SDSE recovered from humans. A multiplex PCR targeting the 16S rRNA and the streptokinase precursor gene has been optimized for differentiating between SDSE strains isolated from humans and those isolated from horses. Previously, the sequence of the streptokinase precursor gene of SDSE recovered from horses has been found in two human cases of pneumonia in Japan. Although further evaluation is required, the findings of this study suggest that SDSE strains are host-specific and this multiplex PCR protocol can be useful in further epidemiological studies and for investigating the zoonotic potential of SDSE.

An outbreak of strangle-like disease involving 26 horses farmed in central Italy was investigated by clinic examination, endoscopy, cytology, bacteriology and polymerase chain reaction (PCR). At weekly interval, a total of three nasal swabs and one guttural pouches lavage fluid (GPLF) were collected, and no Streptococcus equi subsp. equi carrier was found. Some horses showed upper airways disease and endoscopic signs of pharyngeal lymphoid hyperplasia of different grade and/or abnormal endoscopic appearance of guttural pouches. Streptococcus dysgalactiae subsp. equisimilis was isolated from 14 horses while S. equi subsp. zooepidemicus was isolated from six horses. PCR confirmed the biochemical and serological identification of all isolates and was positive in 10 bacteriological negative samples. The absence of S. equi and the frequent detection of S. equisimilis and S. zooepidemicus suggest that beta-haemolytic streptococci other than S. equi could be the causative agent of strangle-like disease.

Staphylococcus aureusstrains were isolated from mastitic milk of cows with infected mammary glands. The animals were living in 12 different farms near Lublin, in Central-Eastern Poland. A biochemical identification method based on enzymatic assay was performed, followed by haemolytic and proteolytic tests. PCR-RFLP targeted on thegapgene allowed the genetic identification of strains at the species level and verified phenotypic identification results. A molecular typing method using triplex PCR was performed to recognize the genetic similarity of the analyzed strains. DNA microarray hybridization (StaphyType, Alere Technologies) was used for detection of antibiotic resistance and virulence associated markers. The results obtained indicate high genetic similarity in strains isolated from the same sites. High genetic similarities were also detected between strains isolated from cows from different farms of the same region. A slightly lower similarity was noted however, in strains from various regions indicating that the strains are herd specific and that the cow’s infections caused byS. aureuswere of a clonal character. In 21 representative isolates selected for DNA-microarray testing, only fosfomycin (fosB) and penicillin resistance markers (blaZ, blaI, blaR) were detected. The presence of genes coding for haemolysins (lukF, lukS, hlgA, hla, hld, hlb), proteases (aur, sspA, sspB, sspP), enterotoxins (entA, entD, entG, entI, entJ, entM, entN, entO, entR, entU, egc-cluster), adhesins (icaA, icaC, icaD, bbp, clfA, clfB, fib, fnbA, map, vwb) or immune evasion proteins (scn, chp, sak) was common and, with exceptions, matched triplex PCR-defined clusters.

The commercial LCx amplification assay, usually employed to detect the Myocobacterium tuberculosis complex in respiratory specimens, was evaluated by comparing the results it gave with those obtained using Lowenstein-Jensen solid medium and pathological findings on 55 lymph nodes from cattle with positive and 10 lymph nodes from cattle with negative skin tests for tuberculosis. Fifty-three cultures (51 and 2, respectively) were positive for M. bovis, while the results for the LCx assay and the histological method were positive in 48 (45, 3) and 24 (20, 4) samples, respectively. None of the samples from cattle from certified tuberculosis-free herds were positive by any of the procedures. The results obtained with the LCx assay, compared with the culture procedure, regarded as the gold standard among the diagnostic techniques, gave a specificity of 91.6% and sensitivity of 90.5%. Although the sensitivity of LCx was suboptimal, DNA of M. bovis was detected in 81.8% of the skin test-positive animals. Amplification techniques could provide a rapid and reasonably reliable tool for detecting bovine tuberculosis.

Ocular infections due to Streptococcus dysgalactiae subsp. equisimilis are rare. In the present report, three patients with a history of uncomplicated small incision cataract surgery with intraocular lens implantation developed exogenous endophthalmitis due to Streptococcus dysgalactiae subsp. equisimilis. The identification of the organisms was confirmed by PCR for a 16S rRNA sequence specific to the species S. dysgalactiae. Intravitreal treatment of cefazolin and amikacin, in addition to topical ofloxacin and tobramycin, resulted in resolution of infection in all three patients. Our reports indicate the importance of bacterial culture and molecular identification in the diagnosis of S. dysgalactiae subsp. equisimilis infection in the eye.

A serological investigation of equine infectious anaemia (EIA), equine viral arteritis (EAV), glanders, and dourine was conducted on the sera of 346 Turkish horses using a combination of tests in series (ELISA and agar gel immunodiffusion) for EIA, virus neutralisation for EAV, and complement fixation (CF) for glanders and dourine. Forty-nine sera showed anticomplementary reactions and were not assessable in the CF test for glanders and dourine. No positive samples were detected for EIA, dourine, and glanders. Fifty-seven sera were positive for EAV. A systematic review of the distribution of these diseases in Turkey was conducted to describe their epidemiological status in the country. The serological results of this investigation confirm those of the published reports for EAV and glanders, whereas different results were reported for dourine and EIA. In fact, no previous data were found for dourine. Furthermore, all sera tested for EIA in the literature were negative, but 3 outbreaks were reported on an international official site in 2005 without details. Further studies and reports of the outbreaks are needed to better understand the real status of infections and the transmission of the diseases. The systematic review is a useful tool to improve the knowledge of public health disease in a country.

Objectives The aim was to identify conjunctival bacterial and fungal flora in clinically normal sheep. Design Prospective study. Setting Tuscany. Participants 100 eyes from 50 adult Massese female sheep were examined. The sheep included in the study were considered free of anterior ophthalmic abnormalities. Primary and secondary outcome measures Bacteria were identified by morphological assessment, Gram staining, biochemical tests. Identification of filamentous fungi was achieved at the genus level, and Aspergillus species were identified based on keys provided by other authors. Yeast colonies were highlighted, but not identified. Results Positive cultures were obtained from 100/100 eyes for bacteria, and from 86/100 eyes for fungi. A total of 14 types of bacteria and 5 types of fungi were isolated. Yeasts were isolated from 13/100 eyes. The most frequent fungal isolates were saprophytic fungi. Conclusions Conjunctival bacterial and fungal flora of clinically normal eyes were reported in sheep. The positivity obtained for conjunctival bacteria was higher compared to findings in the literature by other authors in the same species (100 per cent v 40 per cent), while our results were in line with a recent work performed on mouflons (Ovis Musimon) with a 100 per cent positivity for bacterial conjunctival fornix. In our survey, Gram-positive species were prevalent, as reported by other authors in different species. Few data are available in the literature regarding conjunctival fungal flora in healthy small ruminants. The prevalence of conjunctival fungal flora in this study was higher than findings reported in mouflons (86 per cent v 45 per cent). Differences in fungal prevalence may be due to different methods of managing herds, though further studies are required to verify this hypothesis. The similarities in bacterial and fungal isolates between sheep and mouflons suggest a genera pattern of conjunctival colonisation by bacteria and fungi.

Two horses with Rhodococcus equi infection were examined post mortem by an immunohistochemical method (peroxidase-antiperoxidase; PAP) with a monoclonal antibody (Mab 10G5) to the 15-17 kDa antigen of R. equi. One of the horses was also examined bacteriologically, R. equi being isolated in culture. Immunolabelling with this Mab was marked and widespread. On the other hand, the immunohistochemical reactivity of infected macrophages with a polyclonal antibody specific for lysozyme was slight. Thus, Mab 10G5 would appear to be a useful diagnostic reagent in R. equi infection, with or without cultural confirmation.

The polymerase chain reaction (PCR) was performed to identify reference and field strains of mycobacteria. PCR was able to identify all the reference strains of the Mycobacterium genus and sub-divide them into no Tuberculosis-no Avium complex, Tuberculosis complex and, partially, Avium complex. The primers used for the last recognised only some strains of the different types of M. avium and M. intracellulare. A number of field strains were identified as belonging to the Mycobacterium genus, but further research is required for a complete sub-division into Tuberculosis complex and Avium complex.

This study was performed to verify the safety of raw ovine milk and its role as a vehicle for Mycobacterium avium subsp. paratuberculosis (MAP) transmission. Milk samples from 697 randomly chosen adult sheep, reared in 17 dairy flocks in central Italy, were examined using Ziehl Neelsen (ZN) staining, bacteriological culture (BC), and indirect ELISA tests. Using ELISA, we identified 70 % of MAP-infected ovine dairy farms. MAP infection was confirmed in 24.3 and 21.6 % of ELISA-positive milk samples (n = 37) by ZN staining and BC, respectively. A fair (k = 0.43) and slight (k = 0.33) agreement was observed between ELISA–ZN and ELISA–BC, respectively. The presence of MAP in raw ovine milk produced in central Italy confirmed a risk of potential transmission to humans through consumption of milk and cheese manufactured from unpasteurized milk.