Publications by authors named "Giorgia Borriello"

37 Publications

SIT42 Infection in an Abused Dog in Southern Italy.

Front Vet Sci 2021 22;8:653360. Epub 2021 Jun 22.

Department of Animal Health, Unit of Virology, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Naples, Italy.

A case of infection is described in a dead adult male dog in Southern Italy. The carcass was found by the Health Authority in a gypsy encampment. It was admitted to our forensic veterinary medicine unit, with a suspicion of cruelty to the animal. Necropsy showed beating and traumatism signs, and mistreating was confirmed. Gross lesions included multiple nodular hepatic lesions, hemorrhagic enteritis with enlarged mesenteric lymph nodes, body cavity effusions, and an adrenal neoplasm. Bacteriological and molecular analyses were carried out on the liver lesions that enabled to identify SIT42 (LAM9). Drug-resistance patterns were evaluated by screening mutations on the and genes that showed susceptibility to both rifampin and isoniazid, respectively. Very few studies report canine tuberculosis, and little is known about the disease in Italy. To the authors' knowledge, this is the first report of SIT42 infection in a dog in Italy.
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http://dx.doi.org/10.3389/fvets.2021.653360DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8258403PMC
June 2021

Long-chain polyphosphates impair SARS-CoV-2 infection and replication.

Sci Signal 2021 07 6;14(690). Epub 2021 Jul 6.

CEINGE Biotecnologie Avanzate, Naples 80145, Italy.

Inorganic polyphosphates (polyPs) are linear polymers composed of repeated phosphate (PO ) units linked together by multiple high-energy phosphoanhydride bonds. In addition to being a source of energy, polyPs have cytoprotective and antiviral activities. Here, we investigated the antiviral activities of long-chain polyPs against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. In molecular docking analyses, polyPs interacted with several conserved amino acid residues in angiotensin-converting enzyme 2 (ACE2), the host receptor that facilitates virus entry, and in viral RNA-dependent RNA polymerase (RdRp). ELISA and limited proteolysis assays using nano- LC-MS/MS mapped polyP120 binding to ACE2, and site-directed mutagenesis confirmed interactions between ACE2 and SARS-CoV-2 RdRp and identified the specific amino acid residues involved. PolyP120 enhanced the proteasomal degradation of both ACE2 and RdRp, thus impairing replication of the British B.1.1.7 SARS-CoV-2 variant. We thus tested polyPs for functional interactions with the virus in SARS-CoV-2-infected Vero E6 and Caco2 cells and in primary human nasal epithelial cells. Delivery of a nebulized form of polyP120 reduced the amounts of viral positive-sense genomic and subgenomic RNAs, of RNA transcripts encoding proinflammatory cytokines, and of viral structural proteins, thereby presenting SARS-CoV-2 infection in cells in vitro.
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http://dx.doi.org/10.1126/scisignal.abe5040DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8432949PMC
July 2021

Different Non-Structural Carbohydrates/Crude Proteins (NCS/CP) Ratios in Diet Shape the Gastrointestinal Microbiota of Water Buffalo.

Vet Sci 2021 May 31;8(6). Epub 2021 May 31.

Department of Veterinary Medicine, University of Naples "Federico II", 80137 Naples, Italy.

The microbiota of the gastrointestinal tract (GIT) are crucial for host health and production efficiency in ruminants. Its microbial composition can be influenced by several endogenous and exogenous factors. In the beef and dairy industry, the possibility to manipulate gut microbiota by diet and management can have important health and economic implications. The aims of this study were to characterize the different GIT site microbiota in water buffalo and evaluate the influence of diet on GIT microbiota in this animal species. We characterized and compared the microbiota of the rumen, large intestine and feces of water buffaloes fed two different diets with different non-structural carbohydrates/crude proteins (NSC/CP) ratios. Our results indicated that Bacteroidetes, Firmicutes and Proteobacteria were the most abundant phyla in all the GIT sites, with significant differences in microbiota composition between body sites both within and between groups. This result was particularly evident in the large intestine, where beta diversity analysis displayed clear clustering of samples depending on the diet. Moreover, we found a difference in diet digestibility linked to microbiota modification at the GIT level conditioned by NSC/CP levels. Diet strongly influences GIT microbiota and can therefore modulate specific GIT microorganisms able to affect the health status and performance efficiency of adult animals.
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http://dx.doi.org/10.3390/vetsci8060096DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8229247PMC
May 2021

SERS Biosensor Based on Engineered 2D-Aperiodic Nanostructure for In-Situ Detection of Viable Bacterium in Complex Matrix.

Nanomaterials (Basel) 2021 Mar 31;11(4). Epub 2021 Mar 31.

Institute of Applied Sciences and Intelligent Systems "E. Caianiello" of CNR, 80078 Pozzuoli, Italy.

is a foodborne pathogen globally affecting both the economy and healthcare. Surface Enhanced Raman Spectroscopy (SERS) nano-biosensing can be a promising strategy for its detection. We combined high-performance quasi-crystal patterned nanocavities for Raman enhancement with the use of covalently immobilized Tbilisi bacteriophages as high-performing bio-receptors. We coupled our efficient SERS nano-biosensor to a Raman system to develop an on-field phage-based bio-sensing platform capable of monitoring the target bacteria. The developed biosensor allowed us to identify in milk by our portable SERS device. Upon bacterial capture from samples (10 cells), a signal related to the pathogen recognition was observed, proving the concrete applicability of our system for on-site and in-food detection.
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http://dx.doi.org/10.3390/nano11040886DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8067257PMC
March 2021

First Detection of in a Buffalo Aborted Foetus in Campania Region (Southern Italy).

Front Vet Sci 2020 10;7:571654. Epub 2021 Feb 10.

Unit of Virology, Department of Animal Health, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Naples, Italy.

(LM) is the causative agent of listeriosis in both animals and humans, representing one of the most severe food-borne diseases in humans. Out of 13 serotypes, only three (i.e., 1/2a, 1/2b, and 4b) are responsible for 95% of human outbreaks of listeriosis. Ruminants have been hypothesised to represent the main natural reservoir for this pathogen and to be involved in the transmission of to humans. During pregnancy, listeriosis in ruminants cause various reproductive disorders as well as abortion. However, little is known about abortion due to LM in water buffaloes (). In this study, we report for the first time the detection of LM in a water buffalo foetus in the region of Campania, Italy. Complete necropsy was performed, and samples and swabs from the abomasum, kidneys, liver, lungs, and spleen were collected. Microbiological and molecular analyses were carried out to detect bacterial, viral, and protozoarian abortive pathogens. The results revealed the presence of LM in the liver, lungs, and abomasum, and no other agent was detected. Isolation was confirmed by biochemical and molecular tests. Molecular serotype characterisation was performed, and serogroup IVb was identified. In conclusion, because of the zoonotic implications of our findings, this report highlights the importance of including LM in the diagnostic panel in cases of bubaline abortion.
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http://dx.doi.org/10.3389/fvets.2020.571654DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7902923PMC
February 2021

A type II variant of Toxoplasma gondii infects the Eurasian otter (Lutra lutra) in southern Italy.

Transbound Emerg Dis 2021 Feb 1. Epub 2021 Feb 1.

Department of Animal Health, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Portici, Italy.

Toxoplasmosis, caused by the protozoan parasite Toxoplasma gondii, is a widespread zoonosis capable to affect a wide range of warm-blooded vertebrates. In the past two decades, T. gondii emerged as a significant aquatic pathogen with some pathogenic atypical genotypes isolated and characterized from stranded marine mammals. In contrast, no information is available for mammals in freshwater environment. Although otters are considered highly susceptible to T. gondii infection, to date molecular evidence of T. gondii in Eurasian otter (Lutra lutra) does not exist. We report the first molecular evidence of T. gondii in a free-ranging Eurasian otter from southern Italy and characterized the present strain as a genotype type II variant, with all loci type II except PK1 (locus sequence corresponding to type II variant B), B1 (locus sequence corresponding to type II/X A) and C29-2 (locus with SNPs). Our results indicate circulation of a type II variant in freshwater environment which suggests potential risk of transmission to animals and humans. The finding of a potential pathogenic strain is of great concern for future conservation programmes of the critically endangered Eurasian otter in southern Italy.
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http://dx.doi.org/10.1111/tbed.14012DOI Listing
February 2021

Cerumen microbial community shifts between healthy and otitis affected dogs.

PLoS One 2020 25;15(11):e0241447. Epub 2020 Nov 25.

Istituto Zooprofilattico Sperimentale del Mezzogiorno, Portici, Italy.

Otitis externa is a common multifactorial disease in dogs, characterized by broad and complex modifications of the ear microbiota. The goal of our study was to describe the ear cerumen microbiota of healthy dogs, within the same animal and between different animals, and to compare the cerumen microbiota of otitis affected dogs with that of healthy animals. The present study included 26 healthy dogs, 16 animals affected by bilateral otitis externa and 4 animals affected by monolateral otitis externa. For each animal cerumen samples from the right and left ear were separately collected with sterile swabs, and processed for DNA extraction and PCR amplification of the 16S rRNA gene. Amplicon libraries were sequenced using an Ion Torrent Personal Genome Machine (PGM), and taxonomical assignment and clustering were performed using QIIME 2 software. Our results indicate that the bacterial community of the cerumen in healthy dogs was characterized by extensive variability, with the most abundant phyla represented by Proteobacteria, Actinobacteria, Firmicutes, Bacteroidetes and Fusobacteria. The analysis of both alpha and beta diversity between pairs of left and right ear samples from the same dog within the group of affected animals displayed higher differences than between paired samples across healthy dogs. Moreover we observed reduced bacterial richness in the affected group as compared with controls and increased variability in population structure within otitis affected animals, often associated with the proliferation of a single bacterial taxon over the others. Moreover, Staphylococcus and Pseudomonas resulted to be the bacterial genera responsible for most distances between the two groups, in association with differences in the bacterial community structure. The cerumen microbiota in healthy dogs exhibits a complex bacterial population which undergoes significant modifications in otitis affected animals.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0241447PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7688138PMC
December 2020

Fatal Interstitial Pneumonia Associated with Bovine Coronavirus in Cows from Southern Italy.

Viruses 2020 11 19;12(11). Epub 2020 Nov 19.

Unit of Virology, Department of Animal Health, Experimental Zooprophylactic Institute of Southern Italy, Via Salute 2, 80055 Portici, Italy.

An outbreak of winter dysentery, complicated by severe respiratory syndrome, occurred in January 2020 in a high production dairy cow herd located in a hilly area of the Calabria region. Of the 52 animals belonging to the farm, 5 (9.6%) died with severe respiratory distress, death occurring 3-4 days after the appearance of the respiratory signs (caught and gasping breath). Microbiological analysis revealed absence of pathogenic bacteria whilst Real-time PCR identified the presence of RNA from Bovine Coronavirus (BCoV) in several organs: lungs, small intestine (jejunum), mediastinal lymph nodes, liver and placenta. BCoV was therefore hypothesized to play a role in the lethal pulmonary infection. Like the other CoVs, BCoV is able to cause different syndromes. Its role in calf diarrhea and in mild respiratory disease is well known: we report instead the involvement of this virus in a severe and fatal respiratory disorder, with symptoms and disease evolution resembling those of Severe Acute Respiratory Syndromes (SARS).
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http://dx.doi.org/10.3390/v12111331DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7699522PMC
November 2020

Genotyping of Toxoplasma gondii in wild boar (Sus scrofa) in southern Italy: Epidemiological survey and associated risk for consumers.

Zoonoses Public Health 2020 11 4;67(7):805-813. Epub 2020 Sep 4.

Istituto Zooprofilattico Sperimentale del Mezzogiorno, Portici, Italy.

Toxoplasma gondii is a widespread protozoan parasite (phylum Apicomplexa), which causes a zoonotic parasitic disease, known as toxoplasmosis. The aim of this study was to evaluate the occurrence and genotypes of T. gondii in wild boars of southern Italy and thus to assess the risk of infection for consumers. The boars were inspected during the hunting season within the regional project 'Wild Boar Emergency Plan in Campania', and molecular analyses were performed on 338 boars analysing a total number of 884 matrices (263 brains, 310 hearts and 311 masseter muscles). Toxoplasma gondii was detected in 134 out of 338 boars (39.6%). No significant statistical difference between genders was found (χ  = 0.15 p = .70). The prevalence was 47.1%, 39.3% and 39.2% in piglets, yearlings and adults, respectively (χ  = 0.41; p = .81). The highest prevalence of T. gondii was found in masseter muscles (74/311, 23.8%), followed by the heart (70/310, 22.6%) and brain (58/263, 22.0%), respectively. Microsatellite (MS) analysis of 11 samples revealed eleven T. gondii genotypes (nine atypical, one belonging to type II one to type III). Most of the genotypes found were thus atypical and may be virulent in humans. Hierarchical clustering analysis showed the presence of three distinct clusters, with the majority of atypical genotypes in the GII-GIII cluster. The high prevalence of infection in masseters highlights the potential risk for public health, considering that this muscle is commonly used to prepare raw meat products ('guanciale' and sausages), which may be a source of T. gondii infection in humans. Wild boars may act as an interface role between wildlife, livestock and humans. Our data highlight the urgent need to minimize the risk of infection for animals and humans by setting up a surveillance programme and preventive strategies in a One Health approach to wildlife species.
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http://dx.doi.org/10.1111/zph.12762DOI Listing
November 2020

Identification of Altered miRNAs in Cerumen of Dogs Affected by Otitis Externa.

Front Immunol 2020 29;11:914. Epub 2020 May 29.

Dipartimento di Medicina Veterinaria, Università degli Studi di Milano, Milan, Italy.

Otitis externa is one of the most common diseases in dogs. It is associated with bacteria and yeast, which are regarded as secondary causes. Cerumen is a biological substance playing an important role in the protection of ear skin. The involvement of cerumen in immune defense is poorly understood. MicroRNAs can modulate the host immune response and can provide promising biomarkers for several inflammatory and infectious disorder diagnosis. The aims of this study were to profile the cerumen miRNA signature associated with otitis externa in dogs, integrate miRNAs to their target genes related to immune functions, and investigate their potential use as biomarkers. Cerumen was collected from healthy and otitis affected dogs and the expression of miRNAs was profiled by Next Generation Sequencing; the validation of the altered miRNAs was performed using RT-qPCR. The potential ability of miRNAs to modulate immune-related genes was investigated using bioinformatics tools. The results pointed out that 32 miRNAs, of which 14 were up- and 18 down-regulated, were differentially expressed in healthy vs. otitis-affected dogs. These results were verified by RT-qPCR. To assess the diagnostic value of miRNAs, ROC analysis was carried out, highlighting that 4 miRNAs are potential biomarkers to discriminate otitis-affected dogs. Bioinformatics showed that cerumen miRNAs may be involved in the modulation of host immune response. In conclusion, we have demonstrated for the first time that miRNAs can be efficiently extracted and quantified from cerumen, that their profile changes between healthy and otitis affected dogs, and that they may serve as potential biomarkers. Further studies are necessary to confirm their diagnostic value and to investigate their interaction with immune-related genes.
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http://dx.doi.org/10.3389/fimmu.2020.00914DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7273745PMC
April 2021

Antibiotic Activity of a -Produced Diketopiperazine against .

J Fungi (Basel) 2020 Jun 10;6(2). Epub 2020 Jun 10.

Dipartimento di Medicina Veterinaria e Produzioni Animali-Università degli Studi di Napoli Federico II, Via Federico Delpino, 1, 80137 Napoli, Italy.

A diketopiperazine has been purified from a culture filtrate of the endophytic fungus , isolated from healthy tissues of strawberry plants in a survey of microbes as sources of anti-bacterial metabolites. Its structure has been determined by nuclear magnetic resonance (NMR) and liquid chromatography-mass spectrometry (LC-MS) analyses and was found to be identical to (L-Pro-L-Phe) purified from species of other fungal genera. This secondary metabolite has been selected following bioguided-assay fractionation against two strains of , the causal agent of bovine gastroenteritis. The diketopiperazine (L-Pro-L-Phe), isolated for the first time from species, showed minimum inhibitory concentration (MIC) values of 71.3 and 78.6 μg/mL against the two strains. This finding may be significant in limiting the use of synthetic antibiotics in animal husbandry and reducing the emergence of bacterial multidrug resistance. Further in vivo experiments of diketopiperazines are important for the future application of these metabolites.
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http://dx.doi.org/10.3390/jof6020083DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7344678PMC
June 2020

Different Impacts of MucR Binding to the and Promoters on Gene Expression in 2308.

Biomolecules 2020 05 19;10(5). Epub 2020 May 19.

Department of Environmental, Biological and Pharmaceutical Sciences and Technologies, University of Campania "Luigi Vanvitelli", via Vivaldi - 43, 81100 Caserta, Italy.

The protein MucR from Brucella abortus has been described as a transcriptional regulator of many virulence genes. It is a member of the Ros/MucR family comprising proteins that control the expression of genes important for the successful interaction of α-proteobacteria with their eukaryotic hosts. Despite clear evidence of the role of MucR in repressing virulence genes, no study has been carried out so far demonstrating the direct interaction of this protein with the promoter of its target gene encoding a LuxR-like regulator repressing genes. In this study, we show for the first time the ability of MucR to bind the promoter of in electrophoretic mobility shift assays demonstrating a direct role of MucR in repressing this gene. Furthermore, we demonstrate that MucR can bind the gene promoter. Analyses by RT-qPCR showed no significant differences in the expression level of genes in CC092 lacking MucR compared to the wild-type strain, indicating that MucR binding to the promoter has little impact on gene expression in 2308. The MucR modality to bind the two promoters analyzed supports our previous hypothesis that this is a histone-like protein never found before in .
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http://dx.doi.org/10.3390/biom10050788DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7277663PMC
May 2020

Parasite Load and STRs Genotyping of Isolates From Mediterranean Mussels () in Southern Italy.

Front Microbiol 2020 5;11:355. Epub 2020 Mar 5.

Istituto Zooprofilattico Sperimentale del Mezzogiorno, Portici, Italy.

Toxoplasmosis is a zoonotic food-borne disease caused by , a land-derived protozoan parasite that infects a broad range of terrestrial and aquatic hosts. may reach coastal waters via contaminated freshwater runoff and its oocysts may enter into the marine food web. Marine invertebrates as mussels being filter feeders are exposed and may concentrate oocysts representing a potential source of infection for animals and humans. The present works investigated the prevalence, parasite burden and genotypes of in the Mediterranean mussels () from southern Italy. We sampled a total of 382 individual Mediterranean mussels from May to August 2018 from seven production sites in the Gulf of Naples (Campania region). An additional sample including 27 farmed Mediterranean mussels was obtained in February 2018 from a mollusk depuration plant in Corigliano Calabro (Calabria region). DNA was detected in 43 out of 409 (10.5%) Mediterranean mussels from seven out of eight sampling sites. The number of copies/g in the digestive gland ranged from 0.14 to 1.18. Fragment analysis of Short Tandem Repeats (STRs) at 5 microsatellite loci was performed from 10 PCR positive samples revealing the presence of five distinct genotypes including one corresponding to type I and four atypical genotypes. These findings suggest potential implications of epidemiological importance for human and animal health because both type I and atypical genotypes could be highly pathogenic.
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http://dx.doi.org/10.3389/fmicb.2020.00355DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7066981PMC
March 2020

Screen Printed Based Impedimetric Immunosensor for Rapid Detection of in Drinking Water.

Sensors (Basel) 2020 Jan 3;20(1). Epub 2020 Jan 3.

Department of Physics, Politecnico di Milano, Piazza Leonardo da Vinci, 32, 20133 Milano, Italy.

The development of a simple and low cost electrochemical impedance immunosensor based on screen printed gold electrode for rapid detection of in water is reported. The immunosensor is fabricated by immobilizing anti- antibodies onto a gold surface in a covalent way by the photochemical immobilization technique, a simple procedure able to bind antibodies upright onto gold surfaces. Impedance spectra are recorded in 0.01 M phosphate buffer solution (PBS) containing 10 mM Fe(CN)/Fe(CN) as redox probe. The Nyquist plots can be modelled with a modified Randles circuit, identifying the charge transfer resistance as the relevant parameter after the immobilization of antibodies, the blocking with BSA and the binding of . The introduction of a standard amplification procedure leads to a significant enhancement of the impedance increase, which allows one to measure in drinking water with a limit of detection of 3 × 10 CFU mL while preserving the rapidity of the method that requires only 1 h to provide a "yes/no" response. Additionally, by applying the Langmuir adsorption model, we are able to describe the change of in terms of the "effective" electrode, which is modified by the detection of the analyte whose microscopic conducting properties can be quantified.
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http://dx.doi.org/10.3390/s20010274DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6982893PMC
January 2020

Molecular Detection of spp. (Apicomplexa: Piroplasma) in Free-Ranging Canids and Mustelids From Southern Italy.

Front Vet Sci 2019 23;6:269. Epub 2019 Aug 23.

Istituto Zooprofilattico Sperimentale del Mezzogiorno, Portici, Italy.

Babesiosis is an emerging tick-borne disease caused by apicomplexan parasites with widespread geographical distribution and various wildlife species as reservoir hosts. The aims of this study were to investigate the prevalence and assess the role of free-ranging canids and mustelids in the maintenance of spp. in southern Italy. PCR analysis of splenic samples targeting the 18S rRNA gene revealed the presence of spp. in 36 of 82 (43.9%) red foxes () including 29 (58%) from Campania region and seven (21.8%) from Calabria region, in seven of 13 (53.8%) Eurasian badgers (), and in one of 13 (7.7%) gray wolves (). Samples from other host species including 9 Eurasian otters (), 1 stone marten (), 1 least weasel (), and 1 European polecat () tested spp. negative. Sequence analysis of the 18S rRNA gene demonstrated the presence of in the red fox and two sequence types of badger-associated spp. in the Eurasian badger. The sp. sequence detected in the gray wolf was identical to a badger-associated sp. This study shows that the number of spp. infecting free-ranging carnivores in Italy is higher than currently believed, and suggests that these hosts may play an important role in the maintenance of the sylvatic cycle of these parasites. It is the first report of badger-associated spp. in Italy and in a gray wolf.
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http://dx.doi.org/10.3389/fvets.2019.00269DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6716145PMC
August 2019

Characterization of circulating miRNA signature in water buffaloes (Bubalus bubalis) during Brucella abortus infection and evaluation as potential biomarkers for non-invasive diagnosis in vaginal fluid.

Sci Rep 2019 02 13;9(1):1945. Epub 2019 Feb 13.

Università degli Studi di Milano, Dipartimento di Medicina Veterinaria, Milano, 20133, Italy.

Brucellosis is an infectious disease caused by bacteria from the Brucella genus that can be transmitted to humans through contact with infected animals or contaminated animal products. Brucellosis also causes financial losses in animal production. Ruminants are highly susceptible to brucellosis, and the causative agent water buffaloes (Bubalus bubalis) is Brucella abortus. Circulating microRNAs (miRNAs) are cropping up as promising biomarkers for several infectious diseases. The goals of this study were to characterize the serum miRNA signature associated with brucellosis in water buffaloes and investigate the miRNAs' potential use as biomarkers in vaginal fluids. Next Generation Sequencing was used to assess miRNA expression profiles in Brucella-positive and Brucella-negative blood sera; dysregulated miRNAs in blood serum and vaginal fluids were validated using RT-qPCR. ROC curves were generated to evaluate the diagnostic value of miRNAs for Brucella. GO and KEGG pathway enrichment analyses were exploited to investigate the biological functions of dysregulated miRNAs. The results showed that 20 miRNAs were modulated, of which, 12 were upregulated and 8 were downregulated. These findings were corroborated by RT-qPCR, and ROC curves indicated that the miRNAs can serve as potential biomarkers for Brucella. GO and KEGG pathway analyses pointed out that some of these miRNAs are related to immune response and apoptosis. These results provided an overview of miRNA expression profiles and highlighted potential biomarkers for Brucella infection in water buffaloes. We also demonstrated the potential of vaginal fluids in studies involving microRNA detection. Further functional and mechanistic studies of these miRNAs may improve our understanding of the biological processes involved in Brucella infection and host immune response.
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http://dx.doi.org/10.1038/s41598-018-38365-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6374377PMC
February 2019

Complete Genome Sequencing of 10 Brucella abortus Biovar 3 Strains Isolated from Water Buffalo.

Genome Announc 2018 Apr 19;6(16). Epub 2018 Apr 19.

Istituto Zooprofilattico Sperimentale del Mezzogiorno, Portici, Italy

Brucellosis is a zoonotic disease that affects both humans and animals. Its distribution is global, concentrated in the Mediterranean area, India, Central Asia, and Latin America. Here, we present a complete genome assembly of 10 strains isolated from water buffaloes farmed in the Campania region of Italy.
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http://dx.doi.org/10.1128/genomeA.00180-18DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5908924PMC
April 2018

Complete Genome Sequencing of Eight Brucella abortus Biovar 1 Strains Isolated from Water Buffalo.

Genome Announc 2018 Mar 29;6(13). Epub 2018 Mar 29.

Istituto Zooprofilattico Sperimentale del Mezzogiorno, Portici, Italy

Brucellosis is a zoonotic disease caused by bacteria of the genus The disease is endemic in many areas, causing chronic infections responsible for reproductive disorders in infected animals. Here, we present eight complete genome assemblies of eight strains isolated from water buffaloes farmed in the Campania region.
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http://dx.doi.org/10.1128/genomeA.00179-18DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5876486PMC
March 2018

Octupolar Metastructures for a Highly Sensitive, Rapid, and Reproducible Phage-Based Detection of Bacterial Pathogens by Surface-Enhanced Raman Scattering.

ACS Sens 2017 Jul 20;2(7):947-954. Epub 2017 Jun 20.

Institute of Applied Sciences and Intelligent Systems "E. Caianiello" of CNR , 80072 Pozzuoli, Italy.

The development of fast and ultrasensitive methods to detect bacterial pathogens at low concentrations is of high relevance for human and animal health care and diagnostics. In this context, surface-enhanced Raman scattering (SERS) offers the promise of a simplified, rapid, and high-sensitive detection of biomolecular interactions with several advantages over previous assay methodologies. In this work, we have conceived reproducible SERS nanosensors based on tailored multilayer octupolar nanostructures which can combine high enhancement factor and remarkable molecular selectivity. We show that coating novel multilayer octupolar metastructures with proper self-assembled monolayer (SAM) and immobilized phages can provide label-free analysis of pathogenic bacteria via SERS leading to a giant increase in SERS enhancement. The strong relative intensity changes of about 2100% at the maximum scattered SERS wavelength, induced by the Brucella bacterium captured, demonstrate the performance advantages of the bacteriophage sensing scheme. We performed measurements at the single-cell level thus allowing fast identification in less than an hour without any demanding sample preparation process. Our results based on designing well-controlled octupolar coupling platforms open up new opportunities toward the use of bacteriophages as recognition elements for the creation of SERS-based multifunctional biochips for rapid culture and label-free detection of bacteria.
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http://dx.doi.org/10.1021/acssensors.7b00195DOI Listing
July 2017

The Eurasian otter (Lutra lutra) as a potential host for rickettsial pathogens in southern Italy.

PLoS One 2017 7;12(3):e0173556. Epub 2017 Mar 7.

Istituto Zooprofilattico Sperimentale del Mezzogiorno, Portici (Naples), Italy.

Canine monocytic ehrlichiosis and rickettsiosis are zoonotic tick-borne diseases of canids caused by the intracellular obligate bacteria Ehrlichia canis and Rickettsia species respectively. In this study, we investigated using standard and real-time PCR and sequencing, the occurrence and molecular characterization of E. canis and Rickettsia species in the Eurasian otter (Lutra lutra) from the southern Italian population. Samples were screened by using molecular assays also for Neospora caninum, Toxoplasma gondii, Clamydophyla spp., Coxiella burnetii, Leishmania spp., Cryptosporidium spp., and Giardia spp. detection, and helminths were studied by traditional methods. Out of six carcasses tested, three were positive for E. canis and co-infection with Rickettsia sp. occurred in one of those. Sequences of the 16S rRNA E. canis gene were identical to each other but differed from most of those previously found in red foxes (Vulpes vulpes) and wolves (Canis lupus) from southern Italy. Helminths included just cystacanths of Sphaerirostris spp. from the intestine of two Eurasian otters and the nematode Angiostrongylus vasorum from the lungs of a single Eurasian otter. None of the samples was positive for the other investigated selected pathogens. This study is the first report on the evidence of infection by rickettsial pathogens in the Eurasian otter. The present result prompts some inquiries into the pathogenic role of those bacteria for the isolated sub-populations of the endangered Eurasian otter in southern Italy.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0173556PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5340359PMC
August 2017

Complete Genome Sequences of Three Siphoviridae Bacteriophages Infecting Salmonella enterica Serovar Enteritidis.

Genome Announc 2016 Nov 17;4(6). Epub 2016 Nov 17.

Istituto Zooprofilattico Sperimentale del Mezzogiorno, Portici, Italy

Three bacteriophages, 118970_sal1, 118970_sal2, and 64795_sal3, were isolated from water buffalo feces in southern Italy, exhibiting lytic activity against Salmonella enterica serovar Enteritidis. These bacteriophages belong to the Siphoviridae family and have a 60,113-bp, 123,930-bp, and 48,094-bp double-stranded DNA (dsDNA) genome containing 72, 173, and 80 coding sequences (CDSs), respectively.
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http://dx.doi.org/10.1128/genomeA.00939-16DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5114364PMC
November 2016

The tumor necrosis factor g1022G>A polymorphism is associated with resistance to tuberculosis in water buffalo (Bubalus bubalis).

Anim Genet 2017 Apr 14;48(2):250-251. Epub 2016 Oct 14.

Department of Agriculture, University of Naples Federico II, via Università 100, 80055, Portici, Napoli, Italy.

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http://dx.doi.org/10.1111/age.12512DOI Listing
April 2017

Complete Genome Sequence of a Lytic Siphoviridae Bacteriophage Infecting Several Serovars of Salmonella enterica.

Genome Announc 2016 Sep 29;4(5). Epub 2016 Sep 29.

Istituto Zooprofilattico Sperimentale del Mezzogiorno, Portici, Italy

The bacteriophage 100268_sal2 was isolated from water buffalo feces in southern Italy, exhibiting lytic activity against several subspecies of Salmonella enterica This bacteriophage belongs to the Siphoviridae family and has a 125,114-bp double-stranded DNA (ds-DNA) genome containing 188 coding sequences (CDSs).
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5043552PMC
http://dx.doi.org/10.1128/genomeA.00943-16DOI Listing
September 2016

Complete Genome Sequence of a Myoviridae Bacteriophage Infecting Salmonella enterica Serovar Typhimurium.

Genome Announc 2016 Sep 29;4(5). Epub 2016 Sep 29.

Istituto Zooprofilattico Sperimentale del Mezzogiorno, Portici, Italy.

The bacteriophage 118970_sal3 was isolated from water buffalo feces in southern Italy, exhibiting lytic activity against Salmonella enterica serovar Typhimurium. This bacteriophage belongs to the Myoviridae family and has a 39,464-bp double-stranded DNA (ds-DNA) genome containing 53 coding sequences (CDSs).
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5043551PMC
http://dx.doi.org/10.1128/genomeA.00940-16DOI Listing
September 2016

Molecular survey of Ehrlichia canis and Coxiella burnetii infections in wild mammals of southern Italy.

Parasitol Res 2016 Nov 17;115(11):4427-4431. Epub 2016 Aug 17.

Istituto Zooprofilattico Sperimentale del Mezzogiorno, 80055, Portici, Naples, Italy.

Ehrlichiosis and Q fever caused by the intracellular bacteria Ehrlichia canis and Coxiella burnetii, respectively, are tick-borne diseases with zoonotic potential and widespread geographical distribution. This study investigated the prevalence of both infections in wild mammals in southern Italy. Tissue samples obtained from the red fox (Vulpes vulpes), European badger (Meles meles), gray wolf (Canis lupus), beech marten (Martes foina), and crested porcupine (Hystrix cristata) were processed for molecular detection of both pathogens. E. canis was detected in 55 out of 105 (52 %) red foxes and three out of six gray wolves. Four sequence types were identified, three of which were found in the spleen and liver samples of red foxes and wolves, and one in the kidney of a red fox. None of the examined mammals was positive to C. burnetii type. This represents the first report of E. canis in free-ranging wolves worldwide, as well as the first evidence of this pathogen in red foxes in the peninsular Italy. Our results suggest that E. canis infection is common in free-ranging canids in southern Italy and that a sylvatic life cycle of this pathogen may occur.
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http://dx.doi.org/10.1007/s00436-016-5213-0DOI Listing
November 2016

Overlap of Spoilage-Associated Microbiota between Meat and the Meat Processing Environment in Small-Scale and Large-Scale Retail Distributions.

Appl Environ Microbiol 2016 07 13;82(13):4045-54. Epub 2016 Jun 13.

Department of Agricultural Sciences, Division of Microbiology, University of Naples Federico II, Portici, Italy

Unlabelled: Microbial contamination in food processing plants can play a fundamental role in food quality and safety. The aims of this study were to learn more about the possible influence of the meat processing environment on initial fresh meat contamination and to investigate the differences between small-scale retail distribution (SD) and large-scale retail distribution (LD) facilities. Samples were collected from butcheries (n = 20), including LD (n = 10) and SD (n = 10) facilities, over two sampling campaigns. Samples included fresh beef and pork cuts and swab samples from the knife, the chopping board, and the butcher's hand. The microbiota of both meat samples and environmental swabs were very complex, including more than 800 operational taxonomic units (OTUs) collapsed at the species level. The 16S rRNA sequencing analysis showed that core microbiota were shared by 80% of the samples and included Pseudomonas spp., Streptococcus spp., Brochothrix spp., Psychrobacter spp., and Acinetobacter spp. Hierarchical clustering of the samples based on the microbiota showed a certain separation between meat and environmental samples, with higher levels of Proteobacteria in meat. In particular, levels of Pseudomonas and several Enterobacteriaceae members were significantly higher in meat samples, while Brochothrix, Staphylococcus, lactic acid bacteria, and Psychrobacter prevailed in environmental swab samples. Consistent clustering was also observed when metabolic activities were considered by predictive metagenomic analysis of the samples. An increase in carbohydrate metabolism was predicted for the environmental swabs and was consistently linked to Firmicutes, while increases in pathways related to amino acid and lipid metabolism were predicted for the meat samples and were positively correlated with Proteobacteria Our results highlighted the importance of the processing environment in contributing to the initial microbial levels of meat and clearly showed that the type of retail facility (LD or SD) did not apparently affect the contamination.

Importance: The study provides an in-depth description of the microbiota of meat and meat processing environments. It highlights the importance of the environment as a contamination source of spoilage bacteria, and it shows that the size of the retail facility does not affect the level and type of contamination.
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http://dx.doi.org/10.1128/AEM.00793-16DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4907188PMC
July 2016

Identification of single nucleotide polymorphisms in Toll-like receptor candidate genes associated with tuberculosis infection in water buffalo (Bubalus bubalis).

BMC Genet 2014 Dec 14;15:139. Epub 2014 Dec 14.

Istituto Zooprofilattico Sperimentale del Mezzogiorno, Via Salute, 2, 80055, Portici, Italy.

Background: Toll-like receptors play a key role in innate immunity by recognizing pathogens and activating appropriate responses. Pathogens express several signal molecules (pathogen-associated molecular patterns, PAMPs) essential for survival and pathogenicity. Recognition of PAMPs triggers an array of anti-microbial immune responses through the induction of various inflammatory cytokines. The objective of this work was to perform a case-control study to characterize the distribution of polymorphisms in three candidate genes (toll-like receptor 2, toll-like receptor 4, toll-like receptor 9) and to test their role as potential risk factors for tuberculosis infection in water buffalo (Bubalus bubalis).

Results: The case-control study included 184 subjects, 59 of which resulted positive to both intradermal TB test and Mycobacterium bovis isolation (cases) and 125 resulted negative to at least three consecutive intradermal TB tests. The statistical analysis indicated that two polymorphisms exhibited significant differences in allelic frequencies between cases and controls. Indeed, the TT genotype at TLR9 2340 C > T locus resulted significantly associated with susceptibility to bovine tuberculosis (P = 0.030, OR = 3.31, 95% CI = 1.05-10.40). One polymorphism resulted significantly associated with resistance to the disease, and included the CC genotype, at the TLR4 672 A > C locus (P = 0.01, OR = 0.26, 95% CI = 0.08-0.80). Haplotype reconstruction of the TLR2 gene revealed one haplotype (CTTACCAGCGGCCAGTCCC) associated with disease resistance (P = 0.04, OR = 0.51, 95% CI = 0.27-0.96), including the allelic variant associated with disease resistance.

Conclusions: The work describes novel mutations in bubaline TLR2, TLR4 and TLR9 genes and presents their association with M. bovis infection. These results will enhance our ability to determine the risk of developing the disease by improving the knowledge of the immune mechanisms involved in host response to mycobacterial infection, and will allow the creation of multiple layers of disease resistance in herds by selective breeding.
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http://dx.doi.org/10.1186/s12863-014-0139-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4278265PMC
December 2014

PCR detection of Neospora caninum in water buffalo foetal tissues.

Acta Parasitol 2014 Mar 26;59(1):1-4. Epub 2014 Feb 26.

Department of Animal Health, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Via Salute, 2, 80055, Portici, NA, Italy,

The seroprevalence of Neospora caninum was surveyed by an ELISA kit on two water buffalo herds of Southern Italy. Seropositive samples were detected in 47% and 59% of individuals, respectively, thus indicating high level of exposure to the parasite even if the possibility of vertical transmission cannot be excluded. Tissue samples collected from three aborted fetuses from the same herds were investigated for N. caninum presence by PCR assays targeting the 18S and the Nc5 DNA sequences, respectively. Both methods have shown the presence of N. caninum DNA in heart and brain. Sequencing of the Nc5 genomic DNA confirmed the presence of N. caninum in the samples; phylogenetic analysis of the obtained sequences showed high homology among the Neospora recovered from different samples. The present study suggests an important role of N. caninum as a possible abortive agent for water buffaloes.
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http://dx.doi.org/10.2478/s11686-014-0201-yDOI Listing
March 2014

Detection of Brucella abortus DNA and RNA in different stages of development of the sucking louse Haematopinus tuberculatus.

BMC Vet Res 2013 Dec 1;9:236. Epub 2013 Dec 1.

Department of Veterinary Medicine and Animal Production, Federico II University, V, Delpino 1, 80137 Naples, Italy.

Background: Brucellosis is considered the world's most widespread zoonotic infection. It causes abortion and sterility in livestock leading to serious economic losses and has even more serious medical impact in humans, since it can be a trigger to more than 500,000 infections per year worldwide. The aim of this study was to evaluate the role of Haematopinus tuberculatus, a louse that can parasitize several ruminants, as a new host of brucellosis. Louse specimens were collected from seropositive and seronegative water buffaloes and divided in 3 developmental stages: adults, nymphs and nits. All samples were separately screened for Brucella spp. DNA and RNA detection by Real Time PCR. In particular, primers and probes potentially targeting the 16S rRNA and the Brucella Cell Surface 31 kDalton Protein (bcsp31) genes were used for Real Time PCR and buffalo β actin was used as a housekeeping gene to quantify host DNA in the sample. A known amount of B. abortus purified DNA was utilized for standard curve preparation and the target DNA amount was divided by the housekeeping gene amount to obtain a normalized target value. A further molecular characterization was performed for Brucella strain typing and genotyping by the Bruce-ladder, AMOS-PCR and MLVA assays. Data were statistically analysed by ANOVA.

Results: Brucella abortus DNA and RNA were detected in all developmental stages of the louse, suggesting the presence of viable bacteria. Data obtained by MLVA characterization support this finding, since the strains present in animals and the relative parasites were not always identical, suggesting bacterial replication. Furthermore, the detection of Brucella DNA and RNA in nits samples demonstrate, for the first time, a trans-ovarial transmission of the bacterium into the louse.

Conclusions: These findings identified H. tuberculatus as a new host of brucellosis. Further studies are needed to establish the role of this louse in the epidemiology of the disease, such as vector or reservoir.
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http://dx.doi.org/10.1186/1746-6148-9-236DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4220825PMC
December 2013

Link between geographical origin and occurrence of Brucella abortus biovars in cow and water buffalo herds.

Appl Environ Microbiol 2013 Feb 26;79(3):1039-43. Epub 2012 Nov 26.

Istituto Zooprofilattico Sperimentale del Mezzogiorno, Portici, Italy.

Sixty-three Brucella isolates from water buffaloes and cattle slaughtered within the Italian national plan for brucellosis control were characterized by multiple-locus variable-number tandem repeat analysis (MLVA). Genotyping indicated a strong influence of geographic origin on the Brucella abortus biovar distribution in areas where brucellosis is endemic and highlighted the importance of rigorous management procedures aimed at avoiding inter- and intraherd spreading of pathogens.
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http://dx.doi.org/10.1128/AEM.02887-12DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3568571PMC
February 2013
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