Occurrence of Escherichia coli virulence genes in feces of wild birds from Central Italy

Objective: To investigate the potential role of wild birds as fecal spreaders of enteropathogenic, enterohemorrhagic and Shiga-toxins producing Escherichia coli (E. coli), enteropathogenic E. coli, enterohemorrhagic E. coli and Shiga toxin-producing E. coli strains. Methods: Fecal samples collected from 121 wild birds of different orders and species were submitted to molecular analyses. In particular, eaeA encoding intimin, hlyA encoding for hemolysin, stx1 and stx2 genes encoding Shiga-toxins 1 and 2, respectively, were investigated. Results: Overall, 21(17.35%) fecal samples resulted positive for at least one of the investigated genes. In detail, 12(9.91%) samples were positive for eaeA, 10(8.26%) for stx1, 4(3.31%) for hylA and 1(0.83%) for stx2. An owl (Athene noctua) positive for the four investigated genes suggesting that it harbored a STEC strain. However, virulence genes characterizing EPEC, and EHEC strains were mainly found among seagulls, waterfowl and feral pigeons. Conclusions: Seagulls, waterfowl and feral pigeons, which frequently reach and contaminate rural, urban and peri-urban areas with their droppings, may be important sources of E. coli infection for other animals and humans

Morphological study of the iris musculature in diurnal and nocturnal raptors

In literature it is established that the iris musculature consists of striate muscle fibers in birds while in mammals it consists of smooth muscles. Some authors report the presence of smooth muscle tissue also in the iris of some species of birds. In the present study we report on the iris muscle tissues (type of tissue, direction and mean diameter of muscle fibers or cells) in five species of Accipitriformes (diurnal raptors) and four species of Strigiformes (nocturnal raptors) because they show different way of life depending of their predatory behavior. This morphological study was carried out from raptors died or euthanized at the Wild Life Rescue Centre of Sea and Water birds in Livorno (Italy). From histological examination of iris serial radial sections we find both striated and smooth musculature even if with marked differences among analyzed species, not directly correlated with diurnal or nocturnal lifestyle. Striated fibers are always present, mainly with cross direction, throughout the iris stroma, while the histological differences concern the smooth cells. Indeed, harrier and sparrow hawk (Accipitriformes) and great horned owl and little owl (Strigiformes) show a compact layer of cross smooth muscle cells throughout the iris stroma. In the other species analyzed smooth muscle cells are slightly detectable as scattered or not detectable. Since the cross smooth muscle tissue allows to maintain a myotic state for extended periods of time, our results might be correlated more to the predatory behavior than the taxonomic order

Landscape and Climatic Variations Shaped Secondary Contacts amid Barn Owls of the Western Palearctic

The combined actions of climatic variations and landscape barriers shape the history of natural populations. When organisms follow their shifting niches, obstacles in the landscape can lead to the splitting of populations, on which evolution will then act independently. When two such populations are reunited, secondary contact occurs in a broad range of admixture patterns, from narrow hybrid zones to the complete dissolution of lineages. A previous study suggested that barn owls colonized the Western Palearctic after the last glaciation in a ring-like fashion around the Mediterranean Sea, and conjectured an admixture zone in the Balkans. Here, we take advantage of whole-genome sequences of 94 individuals across the Western Palearctic to reveal the complex history of the species in the region using observational and modeling approaches. Even though our results confirm that two distinct lineages colonized the region, one in Europe and one in the Levant, they suggest that it predates the last glaciation and identify a secondary contact zone between the two in Anatolia. We also show that barn owls recolonized Europe after the glaciation from two distinct glacial refugia: a previously identified western one in Iberia and a new eastern one in Italy. Both glacial lineages now communicate via eastern Europe, in a wide and permeable contact zone. This complex history of populations enlightens the taxonomy of Tyto alba in the region, highlights the key role played by mountain ranges and large water bodies as barriers and illustrates the power of population genomics in uncovering intricate demographic patterns

Boat anchoring on Posidonia oceanica beds in a marine protected area (Italy, western Mediterranean): effect of anchor types in different anchoring stages

Seagrasses worldwide are noted for suffering from mechanical damage caused by boat anchoring. This is particularly so in sites highly frequented by boaters (marine protected areas or coastal urbanised areas). In the last decades, different strategies have been put into practice to reduce such impacts on seagrasses (i.e. by anchoring bans or by deploying boat moorings), More recently, in consideration that few marine protected area (MPA) management bodies or local administrations have the resources to enforce their anchorage regulations, the self-regulatory approach based on education and information of boaters has been preferred in several cases. At present, however, very little is known on the correct anchoring practices to ensure the safeguarding of seagrasses. The aim of the present study was to experimentally quantify in the field the damage caused to Posidonia oceanica shoot density by anchoring. A multifactorial experiment was designed to test whether the damage is dependent on (1) different anchor types (Hall, Danforth and Folding grapnel), (2) the use of a chain vs. a rope, (3) the three anchoring stages (anchor fall, dragging/lock-in and weighing), and finally (4) whether the pattern is consistent among different locations of the meadow. As expected, the three anchor types employed in the present study differed in the levels of damage inflicted on the P. oceanica meadows of the Ustica Island MPA. In particular, the use of the Hall type anchor seems to be preferable to minimise this impact in comparison with the other two anchor types. Moreover, the effect on the meadow of the three anchor types is greatly dependent on the anchoring stage. These results confirm that the weighing stage is the critical stage of the anchoring process. The number of damaged shoots of P oceanica was not affected by the presence of the chain. These patterns were consistent between locations. In the long term, even anchoring on P. oceanica by small boats using low-impact anchors may potentially have detrimental consequences. For this reason, we suggest that in vulnerable sites, it is preferable to implement an educational program based on information of boaters on correct anchoring practices and anchor typology to use, rather than adopting strong restrictions to boat anchoring or deploying mooring buoys. Although the use of these management strategies is still recommended in the case of anchorage frequented by bigger vessels using heavier anchors and chains. (C) 2003 Elsevier B.V. All rights reserved

Effect of topical ophthalmic instillation of rocuronium bromide on the intraocular pressure of kestrels (Falco tinnunculus) and little owls (Athene noctuae)

OBJECTIVE: To assess the effect of rocuronium bromide-induced mydriasis on the intraocular pressure (IOP) of kestrels (Falco tinnunculus) and little owls (Athene noctuae). ANIMALS: 13 adult kestrels and 13 adult little owls. PROCEDURES: All birds were ophthalmologically normal. During the first of 2 treatment periods, a 1% rocuronium bromide solution was topically instilled in both eyes of all birds at a dose of 0.12 mg (12 μL) for kestrels and 0.20 mg (20 μL) for little owls. No ophthalmic treatments were administered during the second (control) treatment period, which was conducted 1 week after the first. During both treatment periods, rebound tonometry was used to measure IOP before rocuronium bromide instillation or at the beginning of the control period (baseline) and at predetermined times after baseline or until the pupillary light reflex returned to normal. All IOP measurements were obtained between 8 am and 5 pm. RESULTS: The mean IOP did not differ significantly from baseline for either species during the control treatment period. During the rocuronium bromide treatment period, complete mydriasis was achieved in all birds. The mean IOP was significantly decreased from baseline and from the corresponding mean IOP for the control period beginning 60 and 30 minutes after drug instillation for kestrels and owls, respectively, and reached its nadir at 60 minutes after drug instillation for both species. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that topical instillation of rocuronium bromide in the eyes successfully induced mydriasis and decreased the IOP of common kestrels and little owls

Effetto midriatico del rocuronio bromide applicato topicamente nel gheppio (Falco Tinniculus)

Scopo del lavoro. Lo scopo del presente lavoro è stato quello di valutare il potere midriatico di un bloccante neuromuscolare (BNM), il rocuronio bromide, applicato topicamente in entrambi gli occhi di rapaci diurni. Materiali e metodi. Nello studio sono stati inseriti dieci gheppi (Falco Tinnunculus) non affetti da patologie oftalmiche, appartenenti a entrambi i sessi, con peso tra 200 e 250 g. I soggetti sono stati sottoposti a una visita oculistica completa durante la quale la midriasi è stata ottenuta utilizzando un BNM, il rocuronio bromide (Esmeron® 10 mg/ml) applicato topicamente senza diluizione, con una pipetta. Gli uccelli hanno ricevuto una singola dose di 0.12 mg in ciascun occhio (dose totale: 0.24 mg/uccello). Durante la procedura la terza palpebra è stata trattenuta con un retrattore palpebrale per un minuto al fine di prevenirne il movimento e quindi evitare la rapida eliminazione del farmaco dalla superficie oculare. Il diametro pupillare e il riflesso pupillare diretto sono stati valutati con una fonte di luce standard prima della somministrazione del farmaco (Tbase), dopo 10 minuti e poi ogni 20 minuti fino a un tempo massimo di 290 minuti. Le variazioni del diametro pupillare sono state misurate con un gauge pupillare con approssimazione a 0.5 mm mentre il riflesso pupillare è stato valutato con una scala di 3 punti (2 normale, 1 diminuito, 0 assente). Sono stati monitorati eventuali effetti collaterali locali e sistemici. Le differenze tra i valori registrati a Tbase e tutti gli altri tempi, per ogni occhio, sono state analizzate con ANOVA ad una via per dati ripetuti con Dunnett come test Post Hoc, mentre le differenze tra i due occhi sono stati esaminate statisticamente con ANOVA a una via con test di Tukey come Post Hoc, considerando significativi valori di P < 0.05 (Graph Pad Prism4®). Risultati. La midriasi massima è stata ottenuta a T90 ed è risultata di 6.30 ± 0.42 mm per l’occhio destro e 6.35 ± 0.41 mm per l’occhio sinistro. Una differenza statistica è stata rilevata tra Tbase e tutti gli altri intervalli di valutazione per ciò che riguarda il diametro pupillare e tra Tbase e T10 fino a T110 per il riflesso fotomotore. Non sono state evidenziate differenze tra occhio destro e sinistro sia per il diametro pupillare che per il riflesso fotomotore nei vari tempi di osservazione. Nei 20 occhi trattati il grado di dilatazione pupillare ottenuto ha consentito una completa valutazione del fondo oculare che in 6/10 soggetti era già possibile a T30. Non sono stati evidenziati effetti collaterali locali e/o sistemici in nessuno degli uccelli trattati. Conclusioni. I risultati del presente lavoro suggeriscono che una singola dose di 0.12 mg di rocuronio bromide applicata topicamente in ciascun occhio di rapaci diurni consenta di ottenere una midriasi bilaterale in assenza di effetti collaterali locali o sistemici. Il rocuronio quindi sembra essere, alla dose impiegata, un BNM di sicuro impiego per indurre la midriasi nei gheppi. In passato l’efficacia midriatica di altri BNM è stata valutata in rapaci diurni della stessa specie oggetto di questo studio, con risultati variabili1. Infatti il pancuronio bromide determina una midriasi transitoria e inconsistente mentre l’alcuronio cloride è un ottimo midriatico ma causa effetti collaterali sistemici anche gravi. Soltanto il vecuronio bromide è considerato efficace in assenza di effetti collaterali, ma per determinare midriasi necessita di somministrazioni ripetute (almeno 3 ogni 15 minuti)1. È importante sottolineare che nella maggior parte delle specie aviarie nelle quali i BNM sono stati impiegati topicamente a scopo midriatico, la somministrazione è sempre stata monolaterale poiché probabilmente la dose cumulativa necessaria per indurre la dilatazione pupillare in entrambi gli occhi poteva essere pericolosa e determinare gravi effetti collaterali1-3. In uno studio recente condotto su rapaci notturni, il rocuronio si è dimostrato un efficace midriatico dopo una singola somministrazione topica determinando una dilatazione pupillare sovrapponibile a quella ottenuta con 2 somministrazioni ripetute4. Per tale motivo gli autori del presente lavoro hanno impiegato nei gheppi una singola somministrazione di farmaco ma su entrambi gli occhi, ottenendo una midriasi bilaterale contestuale. In conclusione, l’applicazione topica di rocuronio induce una buona midriasi nei gheppi e poiché non necessita di somministrazioni ripetute nel singolo occhio per ottenere la dilatazione pupillare, può essere impiegato bilateralmente offrendo notevoli vantaggi di carattere pratico

Occurrence of Escherichia coli virulence genes in feces of wild birds from Central Italy

Objective: To investigate the potential role of wild birds as fecal spreaders of enteropathogenic, enterohemorrhagic and Shiga-toxins producing Escherichia coli (E. coli), enteropathogenic E. coli, enterohemorrhagic E. coli and Shiga toxin-producing E. coli strains. Methods: Fecal samples collected from 121 wild birds of different orders and species were submitted to molecular analyses. In particular, eaeA encoding intimin, hlyA encoding for hemolysin, stx1 and stx2 genes encoding Shiga-toxins 1 and 2, respectively, were investigated. Results: Overall, 21(17.35%) fecal samples resulted positive for at least one of the investigated genes. In detail, 12(9.91%) samples were positive for eaeA, 10(8.26%) for stx1, 4(3.31%) for hylA and 1(0.83%) for stx2. An owl (Athene noctua) positive for the four investigated genes suggesting that it harbored a STEC strain. However, virulence genes characterizing EPEC, and EHEC strains were mainly found among seagulls, waterfowl and feral pigeons. Conclusions: Seagulls, waterfowl and feral pigeons, which frequently reach and contaminate rural, urban and peri-urban areas with their droppings, may be important sources of E. coli infection for other animals and humans

Serological and Molecular Investigation on <i>Toxoplasma gondii</i> Infection in Wild Birds

Toxoplasma gondii is an obligate apicomplexan zoonotic parasite that infects humans and other animals and is responsible for toxoplasmosis. This parasite causes one of the most common parasitic infections in humans worldwide. Toxoplasmosis meets the requirements for a One Health Disease due to its ability to affect the health of human beings as well as domestic and free ranging animals. Integrating human, domestic animal, and wildlife data could better assess the risk and devise methods of control. A first step of such an approach would be the knowledge of the prevalence of parasitosis in humans and animals in selected areas. Therefore, the aim of the present study was to evaluate the occurrence of Toxoplasma infection in 216 free ranging birds belonging to different genera/species by serology and molecular techniques. Twenty-five out of 216 animals (11.6%) were positive to the immunofluorescence antibody test (IFAT) with antibody titers ranging from 1/20 to 1/320, and 19 of them (8.8%) also showed a positive PCR for Toxoplasma DNA. The results confirmed the widespread occurrence of Toxoplasma infection in wild birds and serological data were corroborated by molecular results in birds that also had low antibody titers. The knowledge of the wide occurrence of the parasite in game and wild birds should enhance the accurate estimation of the risks in handling, managing, and eating these species with regard to domestic carnivores as well as the impact of viscera and offal in the environment

Antimicrobial-Resistant Enterococcus spp. in Wild Avifauna from Central Italy

Bacteria of the genus Enterococcus are opportunistic pathogens, part of the normal intestinal microflora of animals, able to acquire and transfer antimicrobial resistance genes. The aim of this study was to evaluate the possible role of wild avifauna as a source of antimicrobial-resistant enterococci. To assess this purpose, 103 Enterococcus spp. strains were isolated from the feces of wild birds of different species; they were tested for antimicrobial resistance against 21 molecules, vancomycin resistance, and high-level aminoglycosides resistance (HLAR). Furthermore, genes responsible for vancomycin, tetracycline, and HLAR were searched. E. faecium was the most frequently detected species (60.20% of isolates), followed by E. faecalis (34.95% of isolates). Overall, 99.02% of the isolated enterococci were classified as multidrug-resistant, with 19.41% extensively drug-resistant, and 2.91% possible pan drug-resistant strains. Most of the isolates were susceptible to amoxicillin/clavulanic acid (77.67%) and ampicillin (75.73%), with only 5.83% of isolates showing an ampicillin MIC &ge; 64 mg/L. HLAR was detected in 35.92% of isolates, mainly associated with the genes ant(6)-Ia and aac(6&prime;)-Ie-aph(2&Prime;)-Ia. Few strains (4.85%) were resistant to vancomycin, and the genes vanA and vanB were not detected. A percentage of 54.37% of isolates showed resistance to tetracycline; tet(M) was the most frequently detected gene in these strains. Wild birds may contribute to the spreading of antimicrobial-resistant enterococci, which can affect other animals and humans. Constant monitoring is essential to face up to the evolving antimicrobial resistance issue, and monitoring programs should include wild avifauna, too