Bacterial Periodontitis in Horses: An Epidemiological Study in Southern Italy.

Equine periodontal disease (EPD) is a painful oral inflammatory syndrome characterized by multifactorial pathogenesis. Although it is well known that bacterial proliferation and consequent gingivitis are caused by the decomposition process of food residues, in hypsodont species, the pathogenetic role and the different bacterial species involved in the progression of EPD must be fully clarified. This study aimed to investigate the association of bacteria, including the complex red bacteria (RCB), with EPD, and to evaluate possible EPD risk factors. Bacterial species, including Treponema denticola, Tannerella forsythia, Porphyromonas gingivalis (belonging to the RCB), Fusobacterium nucleatum, Veilonella parvula, and Prevotella intermedia, were investigated in 125 oral swabs from healthy and EPD-affected horses using real-time multiplex PCR. Subsequently, possible risk factors (i.e., age, gender, and breed of the animals and type of feed used) were evaluated using univariate and multivariate analyses. Tannerella spp. and Treponema spp. were detected in a significantly higher proportion of horses affected by EPD than in healthy animals, although pathogens belonging to RCB were detected in low number of horses. At the same time, none of the investigated pathogens was significantly associated with a particular stage of disease severity. Horses aged older than 20 years were at higher risk of EPD. The high rate of coinfection, statistically associated with EPD, supports the hypothesis that EPD is a complex syndrome characterized by the possible simultaneous involvement of several pathogens and an increased risk depending on the animal's age. Constant oral hygiene is the best prevention to prevent and treat the disease, especially in old animals.

Antibacterial and Biofilm Production Inhibition Activity of Thymus vulgaris L. Essential Oil against Salmonella spp. Isolates from Reptiles.

Salmonellosis is an infectious disease affecting both animals and humans. Antimicrobial resistant (AMR) and biofilm-producing Salmonella spp., frequently detected in reptiles (who can then act as asymptomatic carriers for warm-blooded animals), have developed resistance to biocides; this represents a warning for the emergence of biocide/antimicrobial cross-resistance. The aim of this study was to evaluate the efficacy of Thymus vulgaris L. essential oil (TEO) in inhibiting bacterial growth and biofilm production of Salmonella spp., which had been isolated from wild reptiles housed in a Zoo in Italy. The resistance profile against different classes of antibiotics showed that all the isolates were susceptible to the tested antibiotics, despite the presence of several AMR genes. All the isolates were also tested with aqueous solutions of TEO at different dilutions (5% to 0.039%). Interestingly, TEO proved effective both in inhibiting bacterial growth at low dilutions, with MIC and MBC values ranging between 0.078% and 0.312%, and in inhibiting biofilm production, with values ranging from 0.039% to 0.156%. TEO demonstrated effective bioactivity against the biofilm producer Salmonella spp., proving to be a valid disinfectant for the prevention of salmonellosis from reptiles, a possible source of infection for humans exposed to the reptiles' environment.

Evaluation of antibacterial oxygen/ozone mixture in vitro activity on bacteria isolated from cervico-vaginal mucus of cows with acute metritis.

Ozone is an oxidating gas showing a strong microbicidal activity on bacteria, fungi, viruses and protozoa. The aim of this study was to test the in vitro bacteriocidal action of an Ozone/Oxygen gas mixture on bacteria isolated from the cervico-vaginal mucus of cows affected by acute metritis. A pilot study was initially carried out on reference strains (Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923 and Mycoplasma bovigenitalium ATCC 19852) that were tested with three different treatments: a control positive baseline group (B-group) was incubated without gas treatment, a control positive oxygen group (O2-group) was treated with pure oxygen 100%, and the treated group (T-group) was exposed to a gaseous constant flow of an Ozone/Oxygen mixture, at 50, 35, 20 µg Ozone/ml and for 5, 3 and 1 min for every different Ozone concentration. In both positive control groups, the number of colony forming units (CFU) per ml was higher than 300 CFU/ml (E. coli and S. aureus) and higher than 30 CFU/ml for M. bovigenitalium, after incubation. The T-groups showed a minimal bacterial growth equal to or lower than 1 CFU/ml per plate. Based on the results of the pilot study, a second phase was performed on bacteria isolated from the cervico-vaginal mucus (Klebsiella pneumoniae, Enterobacter agglomerans, E. coli, Proteus mirabilis and M. bovigenitalium) using the lower concentration of 20 µg/ml of Ozone for the minimum exposure time of 1 min. The E. coli and S. aureus reference strains and the clinical isolates (K. pneumoniae, E. agglomerans, E. coli, P. mirabilis) were incubated at 37 °C for 48 h and the colonies were manually counted at 24 h and 48 h following inoculation. The cultures of M. bovigenitalium (both ATCC and clinical isolate) were incubated in a jar with modified atmosphere conditions with 5% CO2 at 37 °C for 4-7 days and colony counting was performed. The second phase showed a low number of CFUs (equal to or less than 7 CFU/ml) for the clinical isolates K. pneumoniae, E. agglomerans, E. coli and P. mirabilis, and, of note, for M. bovigenitalium, both ATCC and clinical isolate, the growth was completely inhibited. Ozone was demonstrated to have a bacteriocidal activity. This study encourages further research of in vivo application of low doses of gaseous Ozone for the treatment of metritis in cows by using minimal exposure times.

Alveolar osteitis associated with methicillin-resistant Staphylococcus epidermidis.

A critical point in dentistry is the empiric prescription of broad-spectrum antibiotics that could increase the levels of antimicrobial resistance. Alveolar osteitis is one of the most common post-op- erative complications in which antibiotic use is controversial. A 35-year-old female, with pain in the right mandibular region and treated with cefixime, was diagnosed with cracked tooth syndrome and pulpitis. The tooth was extracted and a massive purulent bleeding drainage was observed. Irrigation of the socket and a new therapy with azithromycin were done. Bacteriological analysis, a specific mecA gene PCR for the methicillin resistance, and the antimicrobial susceptibility test were per- formed on the bacterial isolate. A Staphylococcus epidermidis isolate was methicillin-resistant and showed resistance to erythromycin, azithromycin, clarithromycin, and sulfamethoxazole + trimeth- oprim. After 7 days, intraoral examination showed a complete resolution. The aim of this report is to suggest that systemic antibiotics may provide insufficient efficacy during alveolar osteitis, especially when caused by a multidrug-resistant organism.

New approaches for risk assessment and management of bovine protothecosis.

Protothecosis is a potential zoonosis related to bovine mastitis. In several countries, a higher incidence of protothecal bovine mastitis that is being recorded and the resistance of Prototheca species to various factors (chlorine, high temperatures, antimicrobial and antiseptic treatments, pH variations), make it difficult to control its spread among farms. The authors aim to describe the infection caused by microalgae, focusing on the problems within cattle farms and proposing new approaches to farm management, based on Regulation (EU) No 2016/429 on transmissible animal diseases. This new flexible approach, based on risk analysis, is a further tool in protecting against Prototheca species. The list of transmissible animal diseases under Regulation (EU) No 2016/429 includes those caused by microorganisms resistant to antimicrobials, which can have important implications for human and animal health, feed and food safety. This approach would involve a series of changes to the rules used for Official Controls (Regulation (EU) No 2017/625) moving from the concept of the food chain to that of the agri-food chain.

Antimicrobial Activity of Essential Oils Evaluated In Vitro against Escherichia coli and Staphylococcus aureus.

The spread of extended-spectrum ß-lactamase-producing Escherichia coli and methicillin-resistant Staphylococcus aureus has caused a reduction in antibiotic effectiveness and an increase in mortality rates. Essential oils (EOs), known for their therapeutic efficacy, can be configured as novel broad-spectrum biocides. Accordingly, the bacteriostatic-bactericidal activity of Citrus Lemon (LEO), Pinus Sylvestris (PEO), Foeniculum Vulgaris (FEO), Ocimum Basilicum (BEO), Melissa Officinalis (MEO), Thymus Vulgaris (TEO), and Zingiber Officinalis Rosc. (GEO), at concentrations ranging from 1.25 to 40% (v/v), were tested in vitro against different E. coli and S. aureus strains using minimal inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs). The chemical compositions of the EOs were analyzed using GC/MS. The major components of all seven tested oils were limonene, α-pinene, anethole, estragole, citral, thymol, and zingiberene, respectively. We found that the bacteriostatic-bactericidal activity of the EOs was related to their chemotypes and concentrations, as well as the strain of the bacteria. A dose-effect correlation was found when testing GEO against S. aureus strains, whilst FEO was found to have no activity regardless of concentration. PEO, MEO, and BEO were found to have bactericidal effect with a MIC and MBC of 1.25% (v/v) against S. aureus strains, and LEO was found to have values of 1.25% (v/v) and 5% (v/v) against ATCC and clinical isolate, respectively. Interestingly, the antimicrobial activity of TEO was not related to oil concentration and the complete inhibition of growth across all E. coli and S. aureus was observed. Although preliminary, our data demonstrate the efficacy of EOs and pave the way for further investigations on their potential synergistic use with traditional drugs in the human and veterinary fields.

Prevalence of bacteriuria in cats with neurogenic bladder.

Urinary tract infections are defined as the adherence, multiplication, and persistence of an infectious agent within the urogenital system, causing an associated inflammatory response and clinical signs; instead, the presence of bacteria in urine as determined by positive bacterial culture (PUC) from a properly collected urine specimen, in the absence of clinical signs, is defined subclinical bacteriuria. Limited information on the prevalence of PUC in spinal cord injury cats affected by neurogenic bladder (NB) is available. On contrary, in NB dogs and humans the prevalence of bacteriuria is well documented. Moreover, while in humans information about bacteriemia associated with NB is already available, this aspect has never been studied in NB cats. The aim of this prospective study was to determine the prevalence of PUC in cats with NB, compared to animals affected by chronic kidney disease (CKD) and healthy cats. Furthermore, the prevalence of bacteriemia in cats with NB was evaluated. Fifty-one cats met the inclusion criteria: 12 cats were affected by NB, 22 had CKD and 17 were healthy. The prevalence of PUC was 58.33% and 18% in NB and CKD cat populations, respectively. All blood cultures were negative. The incomplete bladder emptying and the decreased resistance in the bladder wall could be considered predisposing elements to PUC in the NB feline population. The results of this study highlight, for the first time, an high prevalence of PUC in cats affected by NB, which was not found to be associated with bacteriemia.

Multidrug resistant Enterococcus faecium isolate from cholangitis/cholecystitis in a dog.

Mucocele and cholecystitis were diagnosed in a 10 year-old Shetland-sheepdog presenting aspecific clinical signs. Blood examinations and ultrasonography investigation were performed before to surgical approach, which allowed to collect biopsies and samples for bacteriological analyses. In addition, the patient was subjected to cholecystectomy. A multidrug resistant Enterococcus faecium was isolated from the gallbladder specimens. On the basis of antimicrobial susceptibility test, nitrofurantoin was used. The correct bacteriological diagnosis is necessary to set up effective therapy, influencing the patient's prognosis and improving the recovery time.

African Swine Fever-How to Unravel Fake News in Veterinary Medicine.

In recent years, fake scientific news has spread much faster through the Internet and social media within the so-called "infodemic". African Swine Fever (ASF) is a perfect case study to prove how fake news can undermine the public health response, even in the veterinary field. ASF is a highly contagious infective disease affecting exclusively domestic and wild pigs such as wild boars. ASF can cause social damage and economic losses both directly (due to the high mortality rate) and indirectly (due to international sanctions). Although ASF is not a threat to human health, since 2018 newspapers have often reported false or misleading news, ranging from misinterpreted findings/data to fake or alarmistic news. In some cases, fake news was spread, such as the use of snipers at the border of nations to kill wild boars, or those reports concerning possible risks to human health. In order to provide real and fact-based news on epidemics, some organizations have created easy-to-read infographic and iconographic materials, available on their websites, to help the readers identifying the fake news. Indeed, it is crucial that governments and scientific organizations work against fear and anxiety, using simple and clear communication.

Bayesian estimation of prevalence of Johne's disease in dairy herds in Southern Italy.

Johne's disease (JD) is a chronic granulomatous disease caused by Mycobacterium avium subsp. paratuberculosis (MAP) causing important losses on dairy farms. In Italy, voluntary programs to control MAP infection in dairy cattle are implemented in the Northern part of the country, where several studies have been carried out. Conversely, the disease status has not been fully investigated in the Southern regions. The aims of this study were to (i) determine the herd-level true prevalence (HTP) and (ii) the conditional within herd animal-level prevalence (CWHP) of JD in selected dairy cattle herds in Southern Italy. Serum samples were taken from 27 farms and analysed using a commercial ELISA test. A Bayesian model was fitted to the data. The estimated posterior mean of HPT was 0.46 (89 % CI 0.25-0.67), while the mean CWHP was 0.03 (89 % CI: 0.012-0.045). The results presented in this study call for designing and implementing an effective JD control program at national level.

Basic knowledge and misconceptions on antibiotic use: a comparative survey between Veterinary College and High School students in Bari (Italy).

Misconceptions about the use and effectiveness of antibiotics contribute to the persistence of antimicrobial resistance. The aim of this study was to gather information on appropriate use of antibiotics in students from the Veterinary Medicine College (G1, n = 119) and from High School (G2, n = 220), from Bari (Italy) through a questionnaire. The response rate was 89% in G1 and 89.5% in G2. Fifty­five % of college students and 79% of high­school students had taken antibiotics in the last 12 months. Unsurprisingly, high­school students had more misconceptions about antibiotics than G1. The majority of misconceptions stated that i) antibiotics kill viruses (OR 8.4, CI 4.8­14.7, p < 0.001); ii) they are active against cold and flu (OR 4.6, CI 2.6­8.1, p < 0.001); iii) it is possible to purchase antibiotics without a medical prescription (OR 7.3, CI 4.3­12.5, p < 0.001). Information campaigns among young people are urgently needed to reduce misuse and to improve knowledge on antibiotic.

Characterization of oral and cloacal microbial communities of wild and rehabilitated loggerhead sea turtles (Caretta caretta).

BACKGROUND: Microbial communities of wild animals are being increasingly investigated to provide information about the hosts' biology and promote conservation. Loggerhead sea turtles (Caretta caretta) are a keystone species in marine ecosystems and are considered vulnerable in the IUCN Red List, which led to growing efforts in sea turtle conservation by rescue centers around the world. Understanding the microbial communities of sea turtles in the wild and how affected they are by captivity, is one of the stepping stones in improving the conservation efforts. Describing oral and cloacal microbiota of wild animals could shed light on the previously unknown aspects of sea turtle holobiont biology, ecology, and contribute to best practices for husbandry conditions. RESULTS: We describe the oral and cloacal microbiota of Mediterranean loggerhead sea turtles by 16S rRNA gene sequencing to compare the microbial communities of wild versus turtles in, or after, rehabilitation at the Adriatic Sea rescue centers and clinics. Our results show that the oral microbiota is more sensitive to environmental shifts than the cloacal microbiota, and that it does retain a portion of microbial taxa regardless of the shift from the wild and into rehabilitation. Additionally, Proteobacteria and Bacteroidetes dominated oral and cloacal microbiota, while Kiritimatiellaeota were abundant in cloacal samples. Unclassified reads were abundant in the aforementioned groups, which indicates high incidence of yet undiscovered bacteria of the marine reptile microbial communities. CONCLUSIONS: We provide the first insights into the oral microbial communities of wild and rehabilitated loggerhead sea turtles, and establish a framework for quick and non-invasive sampling of oral and cloacal microbial communities, useful for the expansion of the sample collection in wild loggerhead sea turtles. Finally, our investigation of effects of captivity on the gut-associated microbial community provides a baseline for studying the impact of husbandry conditions on turtles' health and survival upon their return to the wild.

Antimicrobial Resistance in Loggerhead Sea Turtles (Caretta caretta): A Comparison between Clinical and Commensal Bacterial Isolates.

Gram negative organisms are frequently isolated from Caretta caretta turtles, which can act as reservoir species for resistant microorganisms in the aquatic environment. C. caretta, which have no history of treatment with antimicrobials, are useful sentinel species for resistant microbes. In this culture-based study, commensal bacteria isolated from oral and cloacal samples of 98 healthy C. caretta were compared to clinical isolates from the wounds of 102 injured animals, in order to investigate the presence of AMR bacteria in free-living loggerheads from the Adriatic Sea. A total of 410 isolates were cultured. Escherichia coli and genera such as Serratia, Moraxella, Kluyvera, Salmonella were isolated only in healthy animals, while Acinetobacter, Enterobacter, Klebsiella and Morganella were isolated only from the wounds of the injured animals. When tested for susceptibility to ampicillin, amoxicillin + clavulanic acid, ceftazidime, cefuroxime, gentamicin, doxycycline, ciprofloxacin and enrofloxacin, the clinical isolates showed highly significant differences in AMR rates vs. commensal isolates for all the drugs tested, except for doxycycline. The detection of high AMR rates in loggerheads is of clinical and microbiological significance since it impacts both the choice of a proper antibiotic therapy and the implementation of conservation programs.

Next-Generation Sequencing Analysis of Root Canal Microbiota Associated with a Severe Endodontic-Periodontal Lesion.

A patient with an unusual endo-periodontal lesion, without coronal decay or damage, likely caused by a deep periodontal lesion with subsequent endodontic bacterial migration, required medical care. Next-generation sequencing (NGS) was used to assess the endodontic microbiota in vestibular and palatal canals after tooth extraction, evidencing a predominant population (Fusobacterium nucleatum) in one endodontic canal, and a mixed bacterial population with six major populations almost equally distributed in the other endodontic canal (F. nucleatum, Porphyromonas gingivalis, P. endodontis, Parvimonas, Peptostreptococcus stomatis, Prevotella multiformis). These data could suggest different, separated ecologic niches in the same endodontic system, with potentially different pathogenicity levels, clinical manifestations and prognoses for every single canal of the same tooth.

Antibiotics or No Antibiotics, That Is the Question: An Update on Efficient and Effective Use of Antibiotics in Dental Practice.

The antimicrobial resistance (AMR) phenomenon is an emerging global problem and is induced by overuse and misuse of antibiotics in medical practice. In total, 10% of antibiotic prescriptions are from dentists, usually to manage oro-dental pains and avoid postsurgical complications. Recent research and clinical evaluations highlight new therapeutical approaches with a reduction in dosages and number of antibiotic prescriptions and recommend focusing on an accurate diagnosis and improvement of oral health before dental treatments and in patients' daily lives. In this article, the most common clinical and operative situations in dental practice, such as endodontics, management of acute alveolar abscesses, extractive oral surgery, parodontology and implantology, are recognized and summarized, suggesting possible guidelines to reduce antibiotic prescription and consumption, maintaining high success rates and low complications rates. Additionally, the categories of patients requiring antibiotic administration for pre-existing conditions are recapitulated. To reduce AMR threat, it is important to establish protocols for treatment with antibiotics, to be used only in specific situations. Recent reviews demonstrate that, in dentistry, it is possible to minimize the use of antibiotics, thoroughly assessing patient's conditions and type of intervention, thus improving their efficacy and reducing the adverse effects and enhancing the modern concept of personalized medicine.

Salmonella enterica Subsp. houtenae Associated with an Abscess in Young Roe Deer (Capreolus capreolus).

BACKGROUND: S. enterica subsp. houtenae has been rarely documented, and very limited genomic information is available. This report describes a rare case of primary extraintestinal salmonellosis in a young roe deer, associated with Salmonella enterica subsp. houtenae. Methods: A traditional cultural-based analysis was carried out from the contents of a neck abscess; biochemical identification and PCR assay were performed to isolate and identify the pathogen. Through whole-genome sequencing (WGS), multilocus sequence typing (MLST), core genome MLST (cgMLST), and the Salmonella pathogenicity islands (SPIs) survey, resistome and virulome genes were investigated to gain insight into the virulence and antimicrobial resistance of S. houtenae. RESULTS: Biochemical identification and PCR confirmed the presence of Salmonella spp. in the swelling. The WGS analysis identified Salmonella enterica subspecies houtenae serovar 43:z4,z23:- and ST 958. The virulence study predicted a multidrug resistance pattern with resistance shown against aminoglycosides, tetracycline, beta-lactamase, fluoroquinolones, fosfomycin, nitroimidazole, aminocoumarin, and peptide. Fifty-three antibiotic-resistant genes were identified. No plasmids were detected. CONCLUSION: This study demonstrates the importance of continuous surveillance of pathogenic salmonellae. Biomolecular analyses combined with epidemiological data can provide important information about poorly described Salmonella strains and can help to improve animal welfare.

Detection of multi-drug resistance and AmpC ß-lactamase/extended-spectrum ß-lactamase genes in bacterial isolates of loggerhead sea turtles (Caretta caretta) from the Mediterranean Sea.

Sea turtles are useful sentinels to monitor the dissemination of antimicrobial resistance (AMR) in the marine coastal ecosystems. Forty Gram negative bacteria were isolated from wounds of 52 injured Caretta caretta, living in the Mediterranean Sea. Bacteria were identified using 16S rRNA gene sequencing and tested for susceptibility to 15 antibiotics. In addition, NGS amplicon sequencing was performed to detect the presence of AmpC ß-lactamase genes (blaAmpC) and extended-spectrum ß-lactamase (ESBL) genes (blaCTX-M,blaSHV,blaTEM). Seventy-five percent of the isolates (30/40 isolates) exhibited multidrug resistance (MDR) phenotypes and 32.5% (13/40 isolates) were confirmed to be positive for at least one gene. The variants of ESBLs genes were blaCTX-M-3,blaTEM-236 and blaSHV-12. Variants of the blaAmpCß-lactamase gene i.e., blaACT-24, blaACT-2, blaACT-17, blaDHA-4 and blaCMY-37, were also detected. In addition, 4 isolates were found simultaneously harboring CTX and AmpC genes while 2 strains harbored 3 genes (blaACT-2+TEM-236+SHV-12, and blaCTX-M-3+ACT-24+TEM-236).

Bovine respiratory disease in beef calves supported long transport stress: An epidemiological study and strategies for control and prevention.

BRD is associated with infectious agents, but management and transport-stress are trigger factors. Metaphylactic administration of antimicrobial reduces colonization of respiratory tract by pathogens, but the development of antibiotic-resistance raises public health concerns leading to propose new control strategies. The study analyzed nasopharyngeal swabs of 231 imported cattle, 10% of 49 trucks, transported from France to southern Italy and, through Real-time PCR identified the prevalence of the involved pathogens speculating on strategies to reduce the impact of BRD. The samples were tested by Real-time PCR, for the detection of bovine coronavirus (BCoV), bovine respiratory syncytial virus (BRSV), bovine parainfluenza virus (BPiV), bovine adenovirus (BAdV), Mannheimia haemolytica, Pasteurella multocida, Histophilus somni, and Mycoplasma bovis. Yates-corrected chi squared, or Fisher's exact test were used to compare both animal-health status and positivity/negativity to pathogens, and the relationship between presence/absence of clinical signs and Real-time PCR-positivity. H. somni and BCoV were the most frequently identified pathogens. In BRD-diagnosed cattle, BAdV was detected in 13.8% (19/138), BRSV in 14.5% (20/138) and BPiV in 4.3% (6/138). Healthy cattle were mostly positive for H. somni (89.2%, 83/93). A statistically significant association was observed between clinical signs and positivity to M. haemolytica (p value = 0.016). Although mass-medication and vaccination are used for BRD control, it still remains a primary health problem. Our results highlight that the nasopharyngeal microbiota could be affected by transport and that strategies to enhance calf immunity for reducing BRD-risk development would be more effective if applied at farm of origin prior to loading.

Brucella melitensis B115-based ELISA to unravel false positive serologic reactions in bovine brucellosis: a field study.

BACKGROUND: Brucellosis is a zoonosis whose incidence is not declining worldwide despite the global effort to control the disease. Accurate and precise diagnosis is a crucial step in any prophylaxis program but single tests to unequivocally detect animals infected with Brucella spp. are currently unavailable. In Italy, serological diagnosis of bovine brucellosis is performed with two official tests: a rapid agglutination test (i.e., Rose Bengal Plate test, RBPT) and a complement fixation test (CFT) that detect antibodies directed mainly to the smooth lipopolysaccharide (S-LPS). Neither of the two tests is able to avoid the detection of false positive serological reactions (FPSRs) caused by bacteria sharing S-LPS components with Brucella spp. and responsible for the single reactors (SR) phenomenon. A B. melitensis R strain-based ELISA showed a good diagnostic performance in unravelling FP animals; however, since a limited number of animals were analyzed in that study, a large field study was conducted here to discriminate between Brucella-infected from FP animals, with the final aim of reducing the unnecessary slaughter of the latter. An ELISA based on a R strain of Brucella, i.e., Brucella melitensis B115, was employed to measure specific IgG responses in a collection of bovine sera (n = 648). Sera were obtained from 180 farms (either officially brucellosis-free or not brucellosis-free) recruited during an extended period of time (2014-2018) and were preliminarily assayed with the official tests by the Italian Reference Centers and then subjected to the ELISA. RESULTS: Negative sera, when subjected to the ELISA, gave O.D. values below the cutoff; SR sera, i.e. RBPT positive and CFT negative, as well as double positive (DP) sera, i.e. RBPT and CFT positive, gave O.D. values that were below the cutoff. All positive sera, i.e. from Brucella-infected animals, were RBPT positive and CFT positive (ICFTU ranging from 20 to 1280) and gave ELISA O.D. values above the cutoff. CONCLUSIONS: The B. melitensis B115-based ELISA systematically unravelled all false positive (FP) sera while confirming the diagnosis in Brucella-infected animals. Thus, the test employed in the present study may complement the official assays to avoid the costly slaughter of FP animals.

Seroprevalence of Coxiella burnetii in dairy cattle and buffalo from Southern Italy.

A cross­sectional survey was carried out in dairy cattle and buffalo herds from the Southern Italy to detect antibodies against Coxiella burnetii. From 2014 to 2018, 402 herds were monitored and 50 mL of bulk­tank milk (BTM) per farm was analyzed by indirect ELISA. Blood samples of animals from positive farms were also taken and analyzed with the same ELISA test. The overall seroprevalence was 35% [95% Confidence interval (CI):30­39] at herd level and 13% (95%CI:13­14) at animal level. Herd province seroprevalences ranged from 17% to 75%. The provinces of Matera (71%, 95%CI:38­105) and Agrigento (75%, 95%CI:51­100) showed the highest percentage of infected farms. These results describe the widespread distribution of C. burnetii in livestock from Southern Italy, highlighting the need to implement a monitoring program for Q fever.