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Gram-positive catalase-negative cocci (GPCNCs) are significant components of the genital microbiota in sheep and goats. However, characterizing them can be difficult due to overlapping culture features and the limited information on their susceptibility to antibiotics. In this study, 97 foreskin and 13 vaginal swabs were investigated using a culturomic approach. Of 110 animals, 76 (69.09 %) hosted GPCNCs, including strains from Streptococcaceae (37, 33.64 %), Aerococcaceae (30, 27.27 %), Enterococcaceae (6, 5.45 %) and other minor species. With increasing antimicrobial resistance rates in livestock, surveillance programs are globally required, so we conducted a pilot study on GPCNCs isolated from the genital mucosa surfaces of sheep and goats using the minimal inhibitory concentration assay (MIC). Due to gaps in interpretative standard breakpoints, normalized resistance interpretation was used for setting epidemiological susceptibility cut-off values (CO WTs ). Of 57 suitable strains, the majority (80.71 %) showed high CO WTs with decrease susceptibility to at least one antimicrobial class, with 22.81 % displaying multiresistant profiles. Of interest, combined resistances to beta-lactams, macrolides, lincosamides, and tetracyclines were detected in strains of Streptococcus plurianimalium. Further combinations, including resistance to beta-lactams, pleuromutilins, aminoglycosides, and lincosamides, were also recorded in both Streptococcus uberis and Enterococcus spp. strains. Being beta-lactams, macrolides, and tetracyclines the most used antibiotics in livestock worldwide, our results highlight the need for their prudent use. Collectively, our findings highlight that small ruminant genital microbiota can serve as reservoirs for opportunistic severe pathogens, often zoonotic, carrying multidrug resistances, thus standing for high risks for both animals and humans., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Several strategies have been in place in food animal production to reduce the unnecessary use of antimicrobial agents. Beyond the monitoring of their use, the evaluation of the effect of these strategies on the occurrence and types of antimicrobial resistance (AMR) associated genes is crucial to untangle the potential emergence and spread of AMR to humans through the food chain. In the present study, the occurrence of these genes was evaluated in commensal Escherichia coli isolated from broiler carcasses "produced without the use of antibiotics" in 3 antibiotic-free (AB-free) farms in Italy in 2019. Sequenced data were analyzed along with publicly available genomes of E. coli collected in Italy from the broiler food chain from previous years (2017-2018). The genetic relationships among all 93 genomes were assessed on de novo assemblies by in silico MLST and SNP calling. Moreover, the resistomes of all genomes were investigated. According to SNP calling, genomes were gathered in three clades. Clade A encompassed, among others, ST117, ST8070, and ST1011 genomes. ST10 belonged to clade B, whereas Clade C included ST58, ST297, ST1101, and ST23 among others. Regarding the occurrence of AMR genes, a statistically significant lower occurrence of these genes in the genomes of this study in comparison to the public genomes was observed considering the whole group of genes as well as genes specifically conferring resistance to aminoglycosides, β-lactams, phenicols, trimethoprim, and lincosamides. Moreover, significant reductions were observed by comparing the whole group of AMR associated mutations, as well as those specifically for fluoroquinolones and fosfomycin resistance. Although the identification of 3° generation cephalosporin resistance associated genes in AB-free E. coli is a concern, this study provides a first indication of the impact of a more prudent use of antimicrobial agents on the occurrence of AMR genes in Italian broiler production chain. More studies are needed in next years on a higher number of genomes to confirm this preliminary observation., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Objectives: In December 2020, Italy began a national immunization campaign using the BNT162b2 coronavirus disease 2019 (COVID-19) mRNA vaccine, prioritizing healthcare workers (HCWs). Immune serum from vaccinated subjects seems (largely) to retain titres of neutralizing antibodies, even against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) VOC 202012/01-lineage B.1.1.7. Here, we describe an outbreak of SARS-CoV-2 lineage B.1.1.7 infection in three HCWs in a hospital setting; two of the HCWs were fully vaccinated (i.e. had received two doses)., Methods: Two physicians and one nurse working on the same shift on 20th February 2021 were involved in the outbreak. Real-time PCR, antigen tests, and serological tests for the IgG anti-spike protein of SARS-CoV-2 were performed, along with whole-genome sequencing (WGS)., Results: SARS-CoV-2 infection was confirmed in all three HCWs; all presented with mild symptoms of COVID-19. The two physicians were fully vaccinated with BNT162b2 vaccine, with the second dose administered 1 month before symptom onset. Both had high titres of IgG anti-spike antibodies at the time of diagnosis. WGS confirmed that all virus strains were VOC 202012/01-lineage B.1.1.7, suggesting a common source of exposure. Epidemiological investigation revealed that the suspected source was a SARS-CoV-2-positive patient who required endotracheal intubation due to severe COVID-19. All procedures were carried out using a full suite of personal protective equipment (PPE)., Conclusions: This mini-outbreak highlights some important issues about the efficacy of vaccines against transmission of SARS-CoV-2 variants, the high risk of exposure among HCWs, and the need for optimized implementation of PPE in hospitals. The wide circulation of VOC 202012/01 in Europe and Italy highlights the need to improve surveillance and genetic sequencing., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Vector-borne diseases (VBDs) of domestic and wild carnivores are of major public health concern both in industrialized and developing countries, especially in poor socioeconomic settings. War-torn areas specifically suffer from absence of veterinary surveillance of VBDs, resulting in lack of scientific knowledge on this topic. To investigate occurence and prevalence of several vector-borne pathogens (VBPs) in some carnivore species from Iraq, blood samples (n = 397) were obtained from 190 canids [97 stray dogs (Canis familiaris), 55 jackals (Canis aureus) and 38 red foxes (Vulpes vulpes)] and 207 stray cats (Felis catus) collected during a feral animal control and zoonotic disease surveillance program in several United States military bases in Iraq. The presence of Babesia spp., Hepatozoon spp., Ehrlichia spp., Anaplasma spp., Dirofilaria spp. and Leishmania spp. DNA was molecularly investigated. Out of 397 animals tested, 176 (44.3%; 95% CI: 39.5-49.2%) were positive for at least one pathogen with the highest prevalence in foxes (73.7%; 95% CI: 58-85%), followed by jackals (54.5%; 95% CI: 41.5-67%), dogs (38.1%; 29.1-48.1%) and cats (39.1%; 95% CI: 32.7-45.9%). Up to five pathogens were diagnosed in dogs. Hepatozoon canis was the most prevalent VBP in jackals (49.1%; 95% CI: 36.4-61.9%), foxes (47.3%; 95% CI: 32.5-62.7%) and dogs (33%; 95% CI: 24.4-42.8%), whereas Hepatozoon felis was the only species detected in cats (39.1%; 95% CI: 32.7-45.9%). A species of Babesia related to but different from Babesia lengau and designated as Babesia sp. MML was detected in six foxes (15.8%; 95% CI: 7.4-30.4%) and in one jackal (1.8%; 95% CI: 0.3-9.6%). This finding suggested the existence of a new species in the genus Babesia as inferred by molecular and phylogenetical analysis. Further, Babesia vulpes was identified only in two foxes (5.3%; 95% CI: 1.5-17.3%). All samples were negative for Leishmania spp. and Ehrlichia spp. Co-infection with H. canis and Babesia spp. was the most prevalent (5/176, 2.8%, i.e., 4 foxes and 1 jackal), followed by H. canis and Dirofilaria immitis (1/176, 1.3%, i.e., in 1 jackal), H. canis and Dirofilaria repens or Acanthocheilonema reconditum (1/176, 1.3%, i.e., in one dog, each). Data presented fill gaps into knowledge of VBPs in dogs, cats and wild canids in Iraq, indicating that different pathogens circulate amongst animal populations living in the same areas, possibly sharing the same tick vectors. Large-scale surveys are urgently needed to further assess VBPs distribution in Iraq and establish preventative strategies in domestic animals to minimize the risk of infection for animals and humans., (Copyright © 2019 Elsevier B.V. All rights reserved.)

This study investigated for the first time the decontamination efficacy of a probiotic-based cleaning product containing Bacillus subtilis, Bacillus pumilus, and Bacillus megaterium spores on fresh and reused broiler litters during 3 rearing cycles of 6 wk each. Moreover, the impact of reused litters treated with the cleaning product on the chicken caeca microbiota was assessed at the end of the rearing cycles in comparison to untreated litter. The Bacillus spores provided with the cleaning treatment were able to successfully colonize the reused poultry litters, decreasing the mean counts of total aerobic bacteria, Enterobacteriaceae, and coagulase positive Staphylococci. The decrease of Enterobacteriaceae, mainly represented by the genus Escherichia, was also observed in the caeca of broilers reared on reused litters treated with the cleaning product. Moreover, the treatment retained the caeca content of Ruminococcaceae and Faecalibacterium as well as the level of biodiversity among the bacteria genera colonizing the caeca of animals reared on reused litter. Overall, the results of this study highlight a positive effect of the probiotic-based cleaning strategy on the microbial decontamination of reused litters and on broiler caeca stability, thereby enhancing animal health and prevention of poultry diseases., (© 2019 Poultry Science Association Inc.)

Methicillin-resistant Staphylococcus aureus (MRSA) is an important medical issue, since it causes serious and sometimes fatal infections in humans. Intensively reared swine may serve as reservoirs for MRSA that can infect swine workers, and also consumers (via contaminated meat). In this study, MRSA strains were isolated from 55 of the 85 (64.7%) intensive pig farms surveyed, and prevalence was greater on pig fattening farms than on breeding farms. In addition, we included in the study 63 foreign pigs imported for slaughter. Overall, the prevalence of MRSA in the 418 sampled swine was 59.1%; 12 genotypes were identified among the isolates; ST398 (96.4%) was most prevalent, followed by ST97 (2%), ST9 (0.8%) and ST1 (0.8%). MRSA isolates were also detected in 26 (17.3%) of the 150 operators included in the study; the genotypes detected were ST398 (85%), ST9 (7.6%), ST5 (3.8%) and ST1 (3.8%). All the strains were pvl negative and pia positive. Both swine and human strains displayed a multi-resistance pattern, and almost all were resistant to tetracycline. The results obtained in this study confirm the high prevalence of MRSA in swine reared and slaughtered in Italy, and underline the public health risk linked to the spread of antimicrobial-resistant Staphylococcus aureus among intensively reared pigs., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Antimicrobial resistance in bacteria represents one of the most important challenges for public health worldwide. Human infections from antimicrobial-resistant bacteria can be transmitted from person to person, via the environment (especially in the hospital environment), or via handling or eating contaminated foods. Colistin is well known as a last-resort antibiotic for the treatment of human infections; a recent study performed in the People's Republic of China has revealed that colistin resistance is also conferred by the plasmid-mediated mcr-1 gene in Escherichia coli . After that discovery, further plasmid-mediated, colistin resistance genes have been detected. However, to date, only reports on E. coli carrying the mcr-1 gene ( E. coli mcr-1 + ) in foodstuff are available. E. coli mcr-1 + has been isolated from food of animal origin and vegetables; this discovery has opened a debate among food safety experts. This review aims to provide a critical overview of the currently available scientific literature on the presence of the plasmid-mediated, colistin resistance gene E. coli mcr-1 in foodstuffs, focusing on the main implications and future perspectives for food safety.

Methicillin-resistant Staphylococcus aureus (MRSA) is a pathogen present in the hospital environment (HA-MRSA), in the community (CA-MRSA) and in livestock, including pigs (LA-MRSA). MRSA may enter the human food chain during slaughtering and may infect humans coming into direct contact with pigs or pork products. This study aimed to determine the prevalence and characteristics of MRSA isolated from pigs and workers at industrial abattoirs in southern Italy. A total of 215 pig nasal swabs were screened for the presence of MRSA using PCR. An MRSA isolate was detected from each mecA/nuc PCR-positive sample and characterized by spa-typing, Multi-Locus Sequence Typing, SCC-mec and Panton-Valentine Leukocidin (PVL), and also tested for the production of staphylococcal enterotoxins (SEs). Eighty-one MRSA isolates (37.6%) were obtained from the 215 pig nasal swabs; 37 of these isolates were further characterized, and showed 18 different spa-types and 8 different STs. The most frequently recovered STs were ST398 (CC398-t034, t011, t899, t1939 - 43.2%) followed by ST8 (CC8-t008, t064, t2953, t5270 - 24.3%) and ST1 (CC1-t127, t174, t2207 - 10.8%). Nine MRSA isolates were obtained from the 113 human swabs; the isolates showed 5 different spa-types and 5 different STs, including the novel ST2794 (t159). The most representative STs recovered were ST1 (CC1-t127) and ST398 (CC398-t034) (33.3%). None of the MRSA isolates showed the ability to produce SEs and PVL and all resulted resistant to two or more classes of antimicrobials. This study shows the great genetic diversity of MRSA strains in slaughtered pigs and in abattoir employees in Italy, and clearly demonstrates the need for improved hygiene standards to reduce the risk of occupational and food-borne infection linked to the handling/consumption of raw pork containing MRSA., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Natural enemies of ticks include the parasitoid wasp Ixodiphagus hookeri (Hymenoptera: Encyrtidae). The aim of this preliminary study was to investigate the occurrence of I. hookeri DNA in a community of ticks (Ixodes ricinus, Dermacentor marginatus, Hyalomma marginatum, Haemaphysalis inermis and Rhipicephalus turanicus). From May 2010 to March 2012, ticks were collected monthly by dragging and flagging, identified, and 481 adults and 305 nymphs screened molecularly for infection with I. hookeri. Of the samples tested (n=786), 3.1% (n=25) were positive for I. hookeri DNA, 7.2% (n=22) in nymphs and 0.6% (n=3) in adults. I. hookeri DNA was only detected in I. ricinus. This study shows that I. hookeri infests I. ricinus in southern Italy, with nymphs being the main developmental stage affected by this wasp., (Copyright © 2015 Elsevier GmbH. All rights reserved.)

This study reports a combined analysis of mitochondrial and ribosomal DNA target regions of phlebotomine sand flies (Diptera: Psychodidae) from the Mediterranean region. A ∼900 bp long fragment of the mitochondrial DNA encompassing regions within cytb and nd1 gene and the complete ITS2 ribosomal region (∼500 bp) were sequenced and characterized for Phlebotomus perniciosus, Phlebotomus perfiliewi, Phlebotomus neglectus, Phlebotomus papatasi, and Sergentomyia minuta, captured in two sites of southern Italy. From one to eight mitochondrial haplotypes and from one to three ITS2 sequence types were found for the examined specimens according to the different sand fly species. The mean interspecific difference in the mitochondrial sequences was of 16.1%, with an overall intraspecific nucleotide variation from 0.1 to 2.8%. A higher interspecific difference (mean 25.1%) was recorded for the ITS2 sequence, with an overall intraspecific nucleotide variation up to 4.9%. The sequence types alignment of ITS2 region showed that all phlebotomine specimens possessed a split 5.8S rRNA, consisting of a mature 5.8S rRNA and a 2S rRNA separated by a short transcribed spacer. Phylogenetic analysis of the Phlebotomus spp. sequences, herein determined and of those available in GenBank™ were concordant in clustering P. neglectus, P. perfiliewi and P. papatasi with the same species collected from different geographic areas of the Mediterranean basin in four main clades for mtDNA and ITS2, respectively. This study demonstrates the utility of multilocus sequencing, provides a dataset for the molecular identification of the most prevalent phlebotomine sand flies in southern Europe and defines the phylogenetic relationships among species examined., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Hypoderma spp. larvae cause internal myiasis in domestic and wild animals characterized by subcutaneous warbles. Their differentiation is usually performed at species level based on the morphology of third stage larvae. The recent release of the whole mtDNA of Hypoderma lineatum and the retrieval of a 102 bp noncoding region occurring between tRNA(Ser) and NADH dehydrogenase subunit 1 (nd1) genes represented the foundation for this study. The noncoding region and the two flanking mitochondrial genes (i.e., tRNA(Ser) and nd1) of the most diffused Hypoderma spp. (i.e., Hypoderma actaeon, Hypoderma bovis, Hypoderma diana, H. lineatum, Hypodermasinense and Hypoderma tarandi) were analysed. Interspecific variations in amplicon size (from 20 to 102 bp in H. tarandi and H. lineatum, respectively) and nucleotide sequences allowed a genetic discrimination of the examined species providing information instrumental to a rapid molecular identification of Hypoderma spp., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Standardized tools for typing Listeria monocytogenes isolates are required in epidemiological surveys investigating food-borne disease outbreaks and in the food-processing environment to identify the sources of contamination and routes by which the organisms are spread. In this survey Amplified Fragment Length Polymorphism (AFLP) and Multi-Locus Sequence Typing (MLST) have been used to study 103 L. monocytogenes isolates from food and environmental sources. A total of 62 AFLP types and 66 MLST Sequence Types were identified. AFLP and MLST produced similar results in terms of discriminating power. The Discrimination Index calculated for the two techniques was 0.976 for AFLP and 0.972 for MLST. These values were appreciably higher compared to serotyping (0.739). A good congruence was observed between AFLP and MLST. The present study demonstrated that AFLP and MLST subtyping are suitable tools for studying the epidemiology of L. monocytogenes. The great advantage of MLST over AFLP and other molecular typing methods based on fragment fingerprinting lies in the unambiguity of sequence data while AFLP is less costly and highly processive. In conclusion the two methods can be perfectly integrated for high-resolution genotyping of L. monocytogenes.

In this study, 19 chickens were experimentally infected by Leishmania infantum and tissue samples, collected at different times, were cultured and subjected to conventional PCR and/or real time PCR (qPCR) to assess their susceptibility to infection. In addition, 121 serum samples from rural chickens (n=73) and backyard birds (n=48) were tested for anti-L. infantum antibodies by indirect immunofluorescence test. All the 19 animals showed to be molecularly positive at least at one tissue sample. In particular, 26 tissue samples from the experimentally infected chickens were positive on conventional PCR and/or qPCR but no clinical signs or seroconversion were detected and all tissue cultures were negative. Accordingly, all serum samples from rural chickens were negative whereas four (8.4%) from game birds (three Anser anser and one Phasianus colchicus) were positive. These results indicate that chickens are not suitable hosts for L. infantum under experimental condition. The occurrence of anti-L. infantum antibodies in domestic gooses (A. anser) and in a pheasant (P. colchicus) points out their possible role in the epidemiology of visceral leishmaniasis.

Escherichia coli 026 is known as a verocytotoxin-producing E. coli (VTEC) organism that causes severe foodborne diseases such as hemorrhagic colitis and hemolytic uremic syndrome. Although cattle are the most important reservoir of VTEC, only a few reports on the role of water buffalo (Bubalus bubalis) as a reservoir of VTEC and on the presence of these organisms in their milk are available. However, in Southern Italy, where water buffalo are intensively reared, an outbreak of hemolytic uremic syndrome due to E. coli 026 has recently been reported, in which the consumption of typical dairy products was considered to be a common risk factor. The aims of this work were to assess the prevalence of E. coli O26 in raw water buffalo milk, to characterize the virulence gene profiles of the isolates, and to evaluate their phenotypic antimicrobial resistance pattern. Of 160 analyzed samples, 1 (0.6%) tested positive for E. coli O26, and the isolate showed the stx1+/stx2+/eae-/hlyA+ genotypic profile. The strain showed resistance against glycopeptides, macrolides, and penicillins. The presence of VTEC organisms in raw water buffalo milk could be considered to be a potential threat to consumers; however, the strict adherence to the processes used in the preparation of the most common buffalo dairy products could strongly mitigate the foodborne risk. To our knowledge, this article reports the first isolation and characterization of E. coli O26 VTEC in raw water buffalo milk.

The study of animal growth is a longstanding crucial topic of theoretical biology. In this paper we introduce a new class of stochastic growth models that enjoy two crucial properties: the growth path of an individual is monotonically increasing and the mean length at time t follows the classic von Bertalanffy model. Besides the theoretical development, the models are also tested against a large set of length-at-age data collected on Atlantic herring (Clupea harengus): the mean lengths and variances of the cohorts were directly estimated by least squares. The results show that the use of subordinators can lead to models enjoying interesting properties, in particular able to catch some specific features often observed in fish growth data. The use of subordinators seems to allow for an increased fidelity in the description of fish growth, whilst still conforming to the general parameters of the traditional von Bertalanffy equation.

Canine monocytic ehrlichiosis (CME) caused by Ehrlichia canis is the most known canine tick-borne disease (TBD) spread throughout the world. Preventing tick bites is a priority to reduce the risk of TBDs and it was the aim of the present study to evaluate the efficacy of a combination of imidacloprid 10% and permethrin 50% (ImPer) (Advantix; Bayer AG, Germany) in a spot-on formulation to control CME under field conditions. On January-March 2005, 845 dogs from two kennels in southern Italy (kennels of Bari (KB)- and Ginosa (KG)), with a history of tick infestation were initially tested by serology and PCR assay for E. canis infection. Data on Leishmania infantum infection were also available from a previous study carried out on the same dog population. One hundred twenty-six dogs (14.9%) presented anti-E. canis antibodies with a relative prevalence of 15.6% (n=65 dogs in KB) and 14.2% (n=61 dogs in KG). Five hundred thirty-five animals found negative both for E. canis and L. infantum infections were enrolled in three groups (Group A--treated with ImPer once a month; Group B--treated every 2 weeks; and Group C--untreated control animals) and monitored for E. canis infection by serology and PCR in November 2005 (first follow-up) and in March 2006 (second follow-up). The E. canis infection was serologically revealed, at the first and/or second follow-up, in 26 animals from Group C in KB and KG (mean incidence density rate (IDR), 13.24%) while in none of the animals from Group A (KB and KG) and only in one animal from Group B (IDR 1.13%) in KG. The final protection efficacy of ImPer ranged from 95.57% to 100% in Groups B and A. At PCR only 15 dogs from KG were positive for Rickettsiales only at the first follow-up and at the sequence analysis two (both in Group C) revealed 100% homology with E. canis sequences while 13 with Anaplasma platys. Four out of 13 A. platys PCR-positive dogs were also seropositive for E. canis at one or both follow-ups. ImPer, by virtue of its repellent and acaricidal activity against ticks, has been shown to be efficacious to prevent E. canis infection in treated dogs living under natural conditions in endemic areas.

Dicrocoelium dendriticum (Rudolphi, 1819) and Dicrocoelium hospes (Looss, 1907) are recognised to affect the liver of domestic and wild ruminants. A third species, Dicrocoelium orientalis which was described from musk deer in the Baikal region of the former Soviet Union and re-named to Dicrocoelium chinensis (Sudarikov and Ryjikov, 1951) Tang and Tang, 1978 was isolated from other species of deer in Asian countries and from mouflon and roe deer in Europe. Scant information is available for D. chinensis, including the range of species that act as definitive and intermediate hosts. To provide morphological and molecular evidences differentiating D. chinensis versus D. dendriticum, 239 Dicrocoelium spp. specimens were collected from sheep, cattle and sika deer from different localities in Austria, Germany and Italy. Specimens were morphologically identified based on the testes orientation, overall size, and level of maximum body width and other morphometric measurements. From this sample, 10 specimens of D. chinensis and 25 of D. dendriticum from different hosts and geographical localities were characterized molecularly through sequencing of partial 18S rDNA (approximately 1400 bp) and ITS-2 (including the 5.8S and 28S flanking regions; approximately 600 bp). Interspecific differences between D. dendriticum and D. chinensis of 0.14% and 3.8% were recorded in 18S rRNA and ITS-2 sequences, respectively. Phylogenetic analyses via Bayesian inference were conducted using sequences of ITS-2 (276 bp) and partial 28S (221 bp) of the above species of Dicrocoelium together with 20 species belonging to the Xiphidiata within the Plagiorchiida available in GenBank. Both gene regions were strongly concordant in differentiating the Dicrocoeliidae, Gorgoderidae and Plagiorchiidae and were in agreement with their current classification. Morphological and molecular characterization clearly differentiate D. dendriticum and D. chinensis as two distinct digeneans infecting ruminants. The implications on the separate status of D. chinensis on the etiology, biology and diagnosis of dicrocoeliosis are discussed.