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Biological risk assessment in occupational settings currently is based on either qualitative or semiquantitative analysis. In this study, a quantitative microbial risk assessment (QMRA) has been applied to estimate the human adenovirus (HAdV) health risk due to bioaerosol exposure in a wastewater treatment plant (WWTP). A stochastic QMRA model was developed considering HAdV as the index pathogen, using its concentrations in different areas and published dose–response relationship for inhalation. A sensitivity analysis was employed to examine the impact of input parameters on health risk. The QMRA estimated a higher average risk in sewage influent and biological oxidation tanks (15.64% and 12.73% for an exposure of 3 min). Sensitivity analysis indicated HAdV concentration as a predominant factor in the estimated risk. QMRA results were used to calculate the exposure limits considering four different risk levels (one illness case per 100, 1.000, 10.000, and 100.000 workers): for 3 min exposures, we obtained 565, 170, 54, and 6 GC/m3 of HAdV. We also calculated the maximum time of exposure for each level for different areas. Our findings can be useful to better define the effectiveness of control measures, which would thus reduce the virus concentration or the exposure time.

Metabarcoding studies using environmental DNA (eDNA) and high‐throughput sequencing (HTS) are rapidly becoming an important tool for assessing and monitoring marine biodiversity, detecting invasive species, and supporting basic ecological research. Several barcode loci targeting teleost fish and elasmobranchs have previously been developed, but to date primer sets focusing on other marine megafauna, such as marine mammals, have received less attention. Similarly, there have been few attempts to identify potentially “universal” barcode loci which may be informative across multiple marine vertebrate orders. Here we describe the design and validation of two new sets of primers targeting hypervariable regions of the vertebrate mitochondrial 12S and 16S rRNA genes, which have conserved priming sites across virtually all cetaceans, pinnipeds, elasmobranchs, boney fish, sea turtles, and birds, and amplify fragments with consistently high levels of taxonomically diagnostic sequence variation. “In silico” validation using the OBITOOLS software showed our new barcode loci outperformed most existing vertebrate barcode loci for taxon detection and resolution. We also evaluated sequence diversity and taxonomic resolution of the new barcode loci in 680 complete marine mammal mitochondrial genomes demonstrating that they are effective at resolving amplicons for most taxa to the species level. Finally, we evaluated the performance of the primer sets with eDNA samples from aquarium communities with known species composition. These new primers will potentially allow surveys of complete marine vertebrate communities in single HTS metabarcoding assessments, simplifying workflows, reducing costs, and increasing accessibility to a wider range of investigators. Several barcode loci targeting teleost fish and elasmobranchs have previously been developed, but to date primer sets focusing on other marine megafauna, such as marine mammals, have received less attention. Similarly, there have been few attempts to identify potentially “universal” barcode loci which may be informative across multiple marine vertebrate orders. Here we describe the design and validation of two new sets of primers targeting hypervariable regions of the vertebrate mitochondrial 12S and 16S rRNA genes, which have conserved priming sites across virtually all cetaceans, pinnipeds, elasmobranchs, boney fish, sea turtles, and birds, and amplify fragments with consistently high levels of taxonomically diagnostic sequence variation. These new primers will potentially allow surveys of complete marine vertebrate communities in single HTS metabarcoding assessments, simplifying workflows, reducing costs, and increasing accessibility to a wider range of investigators.

Marine environmental DNA (eDNA) is an important tool for biodiversity research and monitoring but challenges remain in scaling surveys over large spatial areas, and increasing the frequency of sampling in remote locations at reasonable cost. Here we demonstrate the feasibility of sampling from commercial vessels (Mediterranean ferries) while underway, as a strategy to facilitate replicable, systematic marine eDNA surveys in locations that would normally be challenging and expensive for researchers to access. Sixteen eDNA samples were collected from four fixed sampling stations, and in response to four cetacean sightings, across three cruises undertaken along the 300 km ferry route between Livorno (Tuscany) and Golfo Aranci (Sardinia) in the Ligurian/Tyrrhenian Seas, June-July 2018. Using 12SrDNA and 16SrDNA metabarcoding markers, we recovered diverse marine vertebrate Molecular Operational Taxonomic Units (MOTUs) from teleost fish, elasmobranchs, and cetaceans. We detected sample heterogeneity consistent with previously known variation in species occurrences, including putative species spawning peaks associated with specific sea surface temperature ranges, and increased night time abundance of bathypelagic species known to undertake diel migrations through the water column. We suggest commercial vessel based marine eDNA sampling using the global shipping network has potential to facilitate broad-scale biodiversity monitoring in the world’s oceans.

Colibacillosis, caused by E. coli, is responsible for economic losses in the poultry industry due to mortality, decreased production, and the cost of antibiotic treatments. Prevention of colibacillosis is based on improved biosecurity measures and the use of the vaccine performed with O78 E. coli strains, which is responsible for most cases of colibacillosis. Recently, there has been increased interest in other infection control methods, such as the use of natural compounds. The aim of this study was to evaluate the antimicrobial efficacy of cinnamon essential oil (CEO) against E. coli strains isolated from poultry. The MIC50 and MIC90 of CEO were determined by testing 117 strains belonging to serogroups O78, O2, O128, O139, isolated from laying hens (91 strains), broilers (10 strains), and turkeys (16 strains). The bacterial strains were tested at cell densities of 108 and 106 CFU/mL. At the cell density of 108 CFU/mL, MIC50 and MIC90 were 0.4 and 0.5 µL/mL for most of the tested strains, while they corresponded to 0.5 µL/mL for all strains isolated from broilers and for strains belonging to serogroup O139. At the cell density of 106 CFU/mL, MIC50 and MIC90 were 0.3 and 0.4 µL/mL, regardless of bird species of origin and for strains belonging to serogroups O78 and O2. In addition, a concentration of 0.04 µL/mL of CEO corresponded both to MIC50 and MIC90 for strains belonging to serogroups O139 and O128. Based on these results, cinnamon essential oil showed an effective antibacterial activity against E. coli strains from poultry and could find field application for the prevention of colibacillosis.

Climate change, excessive exploitation of agricultural land which reduces natural habitats, wildlife shooting, and the use of pesticides all cause difficulties for wildlife, with considerable numbers of animals being brought to wildlife rescue centres. Although the efforts of staff involved in wildlife management at these centres usually focus on therapeutic treatments to reintroduce them into the wild, the monitoring of pathogens that may be transmitted to humans is of relevance. Campylobacter (C.) jejuni and C. coli are frequently carried by animals without inducing clinical signs and are responsible for enteric disorders and more rarely extra-intestinal disease in humans. Farm species and poultry, in particular, are the main reservoirs of C. jejuni and C. coli, but wild animals may also be carriers. The aim of this paper was to investigate the presence of C. jejuni and C. coli in wild birds housed at a wildlife rescue centre and to evaluate the sensitivity of the detected strains to antibiotics. Campylobacter was found in 52 out of 209 (24.88%) birds from 33 different species. C. jejuni was more prevalent, while C. coli was only detected in three Long-eared Owls (Asio otus). The incidence of the infection was particularly high (72.22%) among omnivorous species. Infection rates were higher in birds housed indoors (57.14%) than outdoors (31.74%). Moreover, Campylobacter was not detected in species whose mean temperature body is below 40 °C or higher than 42.2 °C. The most common antibiotic resistance in the tested strains was against trimethoprim/sulfamethoxazole, ciprofloxacin and enrofloxacin. In addition, multi-drug resistance was also found. The results highlight the need to increase biosecurity measures at rescue centres so as to reduce health-related risks to workers involved in wildlife management.

The number of pets such as dogs, cats, rabbits, and parrots has increased in European families. Social benefits to owners such as decreasing feelings of loneliness and anxiety are provided by pets which are also used in Animal-assisted Therapy (AAT). Nevertheless, human-animal interactions are also associated with health problems including allergies, asthma, and zoonosis. Rabbits may carry potential pathogens for humans. One of the most common bacteria that colonizes the oro-pharynx and the upper respiratory tract of rabbits is Pasteurella (P.) multocida. Transmission of the infection to humans results from scratches, licks, and bites but it also can occur from the inhalation of air particles containing the microorganism. Immunocompromised people or persons with pulmonary disorders are particularly susceptible to the infection. Infected rabbits may carry P. multocida with or without clinical signs. In this paper, the sensitivity to antibiotics and the invasiveness ability of P. multocida identified in a farm of pet rabbits affected by severe pasteurellosis were investigated. The strain was P. multocida belonging to capsular type A which is the type most often detected in humans. The identified strain was susceptible to the tested antibiotics, but it appeared equipped with several virulence genes which are responsible for fimbriae production, adhesion processes to host cells, enzyme production, and are involved in iron acquisition processes. These findings are of particular interest because rabbits recovered from pasteurellosis very often become carriers of the bacteria. Therefore, we suggest considering P. multocida screening in the routine medical checks of rabbits, especially if they are meant to be companion animals for children and elder people, given that the transmission of the pathogen cannot be excluded.

Monitoring of infections that may be transmitted to humans by animals in wildlife rescue centres is very important in order to protect the staff engaged in rehabilitation practices. Salmonella may be a natural inhabitant of the intestinal tract of turtles, rarely causing disease. This may represent a potential risk for humans, increasing the sanitary risk for operators in wildlife rescue centres. In this paper, the occurrence of non-typhoidal Salmonella among terrestrial turtles housed in a wildlife rescue centre in Southern Italy was investigated, in order to assess the serovars more frequently carried by turtles and identify those that may represent a risk for operators involved in wildlife management. Sixty-nine adult turtles (Testudo hermanni hermanni, T. h. boettgeri, T. graeca, and T. marginata) were tested. Detection and serotyping of Salmonella strains were performed according to ISO 6579-1 and ISO/TR 6579-3:2013, respectively. The distribution of Salmonella spp. was significantly higher in T. hermanni hermanni than in other species, independent of the age and gender of the animals. Two different Salmonella species, S. enterica and S. bongori, three S. enterica subspecies (enterica, diarizonae, salamae), and five different serovars (Hermannswerder, Abony, Ferruch, Richmond, Vancouver) within the group S. enterica subspecies enterica were identified. Different combinations of Salmonella types were simultaneously found in specimens of T. h. hermanni. Most of detected Salmonella types may represent a potential risk for public health. Adopting correct animal husbandry procedures and informing on potential sanitary risks may be useful for minimising the risk of transmission of Salmonella to workers involved in wildlife management.

The poultry red mite (PRM) Dermanyssus gallinae is well known for its vectorial role for pathogens, such as Salmonella enterica ser. Gallinarum, the causative agent of fowl typhoid. Here, we ascertained the vertical transmission of S. Gallinarum across the PRM life stages, combining the Baudruche-based in vitro feeding system and a PRM-fitting DNA extraction and detection method by qPCR. Small-sized pools (4–5 specimens) of adult mites, eggs, larvae, and protonymphs, as well as single eggs, were tested for S. Gallinarum. The pathogen was detected in 89% of adult mites, 5% of single eggs, 17% of pooled eggs, 9% of larvae, and 43% of protonymphs. Additionally, the feeding rate for infected and uninfected mites was similar, while differences in ovipositing and fecundity rate were observed. The method allowed to confirm the infection of mites through the bloodmeal and to strongly suggest the transmission of S. Gallinarum across the PRM life stages. Furthermore, it allows to avoid in vivo studies and it could be useful for further investigating the vectorial role of D. gallinae or other hematophagous arthropods for infectious agents.

Avian malaria is a re-emerging threat to avian species worldwide. It is sustained by several protozoan species belonging to the genus Plasmodium, mainly Plasmodium relictum. The even wider diffusion of the disease, probably because of the increase in the areas covered by their mosquito vectors, may pose new risks for avian species lacking natural resistance (especially those from artic or sub-artic environments) or those hosted in structures like zoos and wildlife rescue centers. With that premise, this study describes the efficacy and safety of a therapeutic protocol to treat avian malaria in three snowy owls (Bubo scandiacus) hosted in a wildlife rescue center in Apulia, south of Italy, and affected by avian malaria by P. relictum. The protocol consisted of administering 10/4 mg/kg atovaquone/proguanil per os once a day for three consecutive days, repeating this seven days later. Seven days after the end of the treatment, P. relictum was not detected in the birds’ blood and no adverse effects were observed during the 60 days of monitoring after the end of the treatment. Therefore, a therapeutic regimen of 10/4 mg/kg/day may be considered safe and effective in a valuable and endangered species such as B. scandiacus.

Large numbers of wild animals are injured every year in road traffic accidents. Scant data are available for rescued wild carnivores, in particular for red foxes. Cases of foxes with head trauma were retrospectively considered for inclusion in this study. Clinical examination, modified Glasgow coma scale (MGCS), computed tomography (CT) examination, therapy, outcome, and post mortem findings of the brain were investigated. In all foxes, cranial vaults lesion occurred in single (67%) or multiple sites (33%). Midline shift and hydrocephalus were observed in this population. The mean survival was 290 (±176) days. In our study, we performed CT scans on average 260 days after fox rescue, and we speculate that persisting clinical signs could be attributed to TBI. In our study, only two foxes were alive at the time of writing. Other foxes were euthanized due to the severity of the clinical signs. CT scans help diagnose chronic lesions and their effect on prognostic judgment for animals released to wildlife environments.