The diatoms test in veterinary medicine: a pilot study on cetaceans and sea turtles

Fishing activities are considered one of the most relevant threats for cetaceans and sea turtles con- servation since these animals are sometimes found dead entangled in fishing gears. Currently, postmortem diagnosis is based mainly on the presence of nets and lines on the body and the related marks and injuries evident at gross examination. A more detailed and objective evidence is needed to clarify doubts cases and the diatoms technique, used in forensic human medicine, could support drowning diagnosis also in this field. Diatoms\u2019 investigation was implemented to be applied in ma- rine vertebrate on 8 striped (Stenella coeruleoalba) and 1 bottlenose (Tursiops truncatus) dolphins and 5 sea turtles (Caretta caretta) stranded along the Italian coastlines with a likely cause of death hypothized on necropsies carried out by veterinary pathologists. Diatoms were microscopically searched in the bone marrow collected from long bones implementing protocols used in human medicine and their presence was observed in 4 cetaceans and 2 sea turtles. Despite a clear relation between diatoms\u2019 presence and amount and the likely cause of death was not proved due to the poor number of samples, the higher burden of diatoms was found in 3 animals deemed to be death for the interaction with human activity. Despite more studied are necessary to identify the possible relation between the cause of death and diatoms\u2019 findings, the present study implemented this technique to be adapted to marine animals, confirming its possible application also in veterinary forensic medi- cine

Efficient isolation on Vero.DogSLAMtag cells and full genome characterization of Dolphin Morbillivirus (DMV) by next generation sequencing

The Dolphin Morbillivirus (DMV) genome from the frst Mediterranean epidemic (1990-\u201992) is the only cetacean Morbillivirus that has been completely sequenced. Here, we report the frst application of next generation sequencing (NGS) to morbillivirus infection of aquatic mammals. A viral isolate, representative of the 2006-\u201908 Mediterranean epidemic (DMV_IZSPLV_2008), efciently grew on Vero.DogSLAMtag cells and was submitted to whole genome characterization by NGS. The fnal genome length was 15,673 nucleotides, covering 99.82% of the DMV reference genome. Comparison of DMV_IZSPLV_2008 and 1990-\u201992 DMV strain sequences revealed 157 nucleotide mutations and 47 amino acid changes. The sequence similarity was 98.7% at the full genome level. Whole-genome phylogeny suggested that the DMV strain circulating during the 2006-\u201908 epidemics emerged from the 1990-\u201992 DMV strain. Viral isolation is considered the \u201cgold standard\u201d for morbillivirus diagnostics but efcient propagation of infectious virus is difcult to achieve. The successful cell replication of this strain allowed performing NGS directly from the viral RNA, without prior PCR amplifcation. We therefore provide to the scientifc community a second DMV genome, representative of another major outbreak. Interestingly, genome comparison revealed that the neglected L gene encompasses 74% of the genetic diversity and might serve as \u201chypervariable\u201d target for strain characterization

Mediterranean Fin Whales (Balaenoptera physalus) Threatened by Dolphin MorbilliVirus

During 2011-2013, dolphin morbillivirus was molecularly identified in 4 stranded fin whales from the Mediterranean Sea. Nucleoprotein, phosphoprotein, and hemagglutinin gene sequences of the identified strain were highly homologous with those of a morbillivirus that caused a 2006-2007 epidemic in the Mediterranean. Dolphin morbillivirus represents a serious threat for fin whales

Mosquito-Borne Diseases and ‘One Health’: The Northwestern Italian Experience

In Italy, the surveillance of Mosquito-Borne Diseases (MBDs) is regulated by two national preparedness plans: (1) for West Nile and Usutu viruses, integrating human and veterinary surveillance in order to early detect viruses circulation and to quickly apply control measures aimed at reducing the risk of transmission through blood and blood components and (2) for Arbovirosis transmitted by Aedes mosquitoes, mainly Chikungunya, Dengue and Zika viruses, based on surveillance of both imported and autochthonous human cases. This chapter reports the results of the application of these National Plans in Northwestern Italy and their impact for human health. In detail, we present the coordinated activities enforced in Piemonte and Liguria Regions, as a good example of the ‘One Health approach’ to control MBDs and prevent human transmission

Detection of Invasive Borrelia burgdorferi Strains in North-Eastern Piedmont, Italy.

SummaryFollowing reports of human cases of Lyme borreliosis from the Ossola Valley, a mountainous area of Piemonte, north‐western Italy, the abundance and altitudinal distribution of ticks, and infection of these vectors with Borrelia burgdorferi sensu lato were evaluated. A total of 1662 host‐seeking Ixodes ricinus were collected by dragging from April to September 2011 at locations between 400 and 1450 m above sea level. Additional 104 I. ricinus were collected from 35 hunted wild animals (4 chamois, 8 roe deer, 23 red deer). Tick density, expressed as the number of ticks per 100 m2, resulted highly variable among different areas, ranging from 0 to 105 larvae and from 0 to 22 nymphs. A sample of 352 ticks (327 from dragging and 25 from wild animals) was screened by a PCR assay targeting a fragment of the 16S rRNA gene of B. burgdorferi s.l. Positive samples were confirmed with a PCR assay specific for the 5S‐23S rRNA intergenic spacer region and sequenced. Four genospecies were found: B. afzelii (prevalence 4.0%), B. lusitaniae (4.0%), B. garinii (1.5%) and B. valaisiana (0.3%). Phylogenetic analysis based on the ospC gene showed that most of the Borrelia strains from pathogenic genospecies had the potential for human infection and for invasion of secondary body sites

Multidisciplinary studies on a sick-leader syndrome-associated mass stranding of sperm whales (Physeter macrocephalus) along the Adriatic coast of Italy

Mass strandings of sperm whales (Physeter macrocephalus) are rare in the Mediterranean Sea. Nevertheless, in 2014 a pod of 7 specimens stranded alive along the Italian coast of the Central Adriatic Sea: 3 individuals died on the beach after a few hours due to internal damages induced by prolonged recumbency; the remaining 4 whales were refloated after great efforts. All the dead animals were genetically related females; one was pregnant. All the animals were infected by dolphin morbillivirus (DMV) and the pregnant whale was also affected by a severe nephropathy due to a large kidney stone. Other analyses ruled out other possible relevant factors related to weather conditions or human activities. The results of multidisciplinary post-mortem analyses revealed that the 7 sperm whales entered the Adriatic Sea encountering adverse weather conditions and then kept heading northward following the pregnant but sick leader of the pod, thereby reaching the stranding site. DMV infection most likely played a crucial role in impairing the health condition and orientation abilities of the whales. They did not steer back towards deeper waters, but eventually stranded along the Central Adriatic Sea coastline, a real trap for sperm whales