PrPC Governs Susceptibility to Prion Strains in Bank Vole, While Other Host Factors Modulate Strain Features.

Bank vole is a rodent species that shows differential susceptibility to the experimental transmission of different prion strains. In this work, the transmission features of a panel of diverse prions with distinct origins were assayed both in bank vole expressing methionine at codon 109 (Bv109M) and in transgenic mice expressing physiological levels of bank vole PrPC (the BvPrP-Tg407 mouse line). This work is the first systematic comparison of the transmission features of a collection of prion isolates, representing a panel of diverse prion strains, in a transgenic-mouse model and in its natural counterpart. The results showed very similar transmission properties in both the natural species and the transgenic-mouse model, demonstrating the key role of the PrP amino acid sequence in prion transmission susceptibility. However, differences in the PrPSc types propagated by Bv109M and BvPrP-Tg407 suggest that host factors other than PrPC modulate prion strain features. IMPORTANCE: The differential susceptibility of bank voles to prion strains can be modeled in transgenic mice, suggesting that this selective susceptibility is controlled by the vole PrP sequence alone rather than by other species-specific factors. Differences in the phenotypes observed after prion transmissions in bank voles and in the transgenic mice suggest that host factors other than the PrPC sequence may affect the selection of the substrain replicating in the animal model.

Strengthening preparedness against global health threats: A paradigm shift based on One Health approaches.

The implementation of preparedness strategies to prevent and mitigate the impact of global health threats poses several challenges. It should promptly identify cross-cutting drivers of pandemic threats, assess context-specific risks, engage multiple stakeholders, and translate complex data from multiple sources into accessible information for action. This requires a coordinated, multidisciplinary and multisectoral effort engaging systems that, most of the time, work in isolation. The One Health (OH) approach promotes the collaboration and communication among different disciplines and sectors, and could be applied across the preparedness phases at national and international level. We discuss here gaps and needs in preparedness strategies, which can benefit from the OH approach, and a set of actionable recommendations, as shared with the G20-2021 with a dedicated Policy Brief. The discussion adds to the current debate about OH operationalization and promotes a paradigm shift towards coordinated prevention and preparedness strategies for early assessment and management of global health threats.

Evidence of exposure to SARS-CoV-2 in cats and dogs from households in Italy.

SARS-CoV-2 originated in animals and is now easily transmitted between people. Sporadic detection of natural cases in animals alongside successful experimental infections of pets, such as cats, ferrets and dogs, raises questions about the susceptibility of animals under natural conditions of pet ownership. Here we report a large-scale study to assess SARS-CoV-2 infection in 817 companion animals living in northern Italy, sampled at a time of frequent human infection. No animals tested PCR positive. However, 3.4% of dogs and 3.9% of cats had measurable SARS-CoV-2 neutralizing antibody titers, with dogs from COVID-19 positive households being significantly more likely to test positive than those from COVID-19 negative households. Understanding risk factors associated with this and their potential to infect other species requires urgent investigation. ONE SENTENCE SUMMARY: SARS-CoV-2 antibodies in pets from Italy.

Evidence of exposure to SARS-CoV-2 in cats and dogs from households in Italy.

SARS-CoV-2 emerged from animals and is now easily transmitted between people. Sporadic detection of natural cases in animals alongside successful experimental infections of pets, such as cats, ferrets and dogs, raises questions about the susceptibility of animals under natural conditions of pet ownership. Here, we report a large-scale study to assess SARS-CoV-2 infection in 919 companion animals living in northern Italy, sampled at a time of frequent human infection. No animals tested PCR positive. However, 3.3% of dogs and 5.8% of cats had measurable SARS-CoV-2 neutralizing antibody titers, with dogs from COVID-19 positive households being significantly more likely to test positive than those from COVID-19 negative households. Understanding risk factors associated with this and their potential to infect other species requires urgent investigation.

Quantitative profiling of the pathological prion protein allotypes in bank voles by liquid chromatography-mass spectrometry.

The conversion of the cellular prion protein (PrP(C)) into a misfolded isoform (PrP(TSE)) that accumulates in the brain of affected individuals is the key feature of transmissible spongiform encephalopaties (TSEs). Susceptibility to TSEs is influenced by polymorphisms of the prion gene suggesting that the presence of certain amino acid residues may facilitate the pathological conversion. In this work, we describe a quantitative, fast and reliable HPLC-MS method that allowed to demonstrate that in the brain of 109(Met/Ile) heterozygous bank voles infected with the mouse adapted scrapie strain 139A, there are comparable amounts of PrP(TSE) with methionine or isoleucine in position 109, suggesting that in this TSE model the two allotypes have similar rates of accumulation. This method can be easily adapted for the quantitative determination of PrP allotypes in the brain of other natural or experimental TSE models.

Advances in scrapie research.

The bovine spongiform encephalopathy (BSE) crisis clearly demonstrated the need to keep animal transmissible spongiform encephalopathies (TSE) under control in order to protect animal and human health. Scrapie is the most widespread TSE of livestock in the world. For this reason, health authorities in different countries have elaborated plans that aim towards scrapie eradication. The unusual nature of the scrapie agent and the fragmented status of scientific knowledge about it, along with the limitations of currently available diagnostic tools, make it unlikely that the objective of eradication will be achieved in the near future. Scientific research is focused on acquiring the knowledge that will improve the efficiency of these efforts.

Identification of the pathological prion protein allotypes in scrapie-infected heterozygous bank voles (Clethrionomys glareolus) by high-performance liquid chromatography-mass spectrometry.

Cerebral formation of the pathological isoform of the prion protein (PrP) is a crucial molecular event in prion diseases. The bank vole (Clethrionomys glareolus) is a rodent species highly susceptible to natural scrapie. The PrP gene of bank vole is polymorphic (Met/Ile) at codon 109. Here we show that homozygous 109Met/Met voles have incubation times shorter than heterozygous 109Met/Ile voles after experimental challenge with three different scrapie isolates. An HPLC-MS/MS method was optimized and applied to investigate whether in heterozygous animals both PrP allotypes are able to undergo pathological conversion. The results demonstrate that both allotypes of the prion protein participate to pathological deposition.

Detection and characterization of Shiga toxin-producing Escherichia coli in feral pigeons.

Escherichia coli strains producing a variant of Shiga toxin 2 (Stx2), designated Stx2f, have been recently described in the stools of feral pigeons. During 1997-1998, 649 pigeons were trapped and examined in three different squares of Rome. Stool samples were collected from each bird and enrichment cultures were examined for the presence of Stx by the vero cell assay. Stx-producing E. coli (STEC) were isolated from the positive cultures and characterized by serotyping and PCR analysis of stx and other virulence-related genes. Stx was detected in 10.8% of the stool enrichment cultures. The percentage of positive birds did not differ significantly for the three flocks considered and the season of sample collection. Conversely, STEC carriage was significantly more frequent in young than in adult birds (17.9 versus 8.2%). None of the birds examined showed signs of disease. STEC strains were isolated from 30 of 42 Stx-positive cultures examined. All the strains produced Stx2f, and most of them possessed genes encoding for intimin and the cytolethal distending toxin (CLDT). Six serogroups were identified, but most of the isolates belonged to O45, O18ab, and O75. Molecular typing indicated that most of the isolates within a flock were clonally-related. This work confirms that pigeons represent a natural reservoir of STEC strains characterized by the production of the toxin variant Stx2f, and by the frequent presence of eae and cldt genes. Further work is needed to clarify whether these STEC may represent a cause of avian disease or even a potential health hazard for humans.

Amyloid, amyloid-inducers, cytokines and heavy metals in scrapie and other human and animal subacute spongiform encephalopathies: some hypotheses.

A number of hypotheses regarding the pathogenesis of scrapie and other human and animal spontaneous and experimental subacute spongiform encephalopathies (SSE) are presented here. In particular, it is speculated that a PrPsc 27-30-induced suppression of host's defense system is responsible, through the existence of the different and synergistically operating mechanisms, for the absence of any documented inflammatory or immunologic response during SSE. This could be therapeutically counterparted by the utilization of cytokines (TNF, IL-1, etc) or cytokine-inducers, provided that synthesis and secretion of the above inflammation mediators on behalf of reticulo-endothelial cells were strongly depressed by PrP 27-30, as suggested. Finally, some hypotheses are also made in relation to the blood-brain-barrier's integrity in susceptible hosts. Similarly to Alzheimer's disease (AD) in man, this could play a role also in SSE pathogenesis, facilitating cerebral localization of metals such as aluminum, lead, or silica, which have already been shown to enhance amyloid fibrils' polymerization and deposition within human brain tissue.

Histological, histochemical and ultrastructural features of hyaline inclusions in hepatocytes of striped dolphins (Stenella coeruleoalba).

Many striped dolphins (Stenella coeruleoalba) which died in the recent morbillivirus epizootic in the Mediterranean Sea had hyaline inclusions in hepatocytes. We investigated the histological, histochemical and ultrastructural features of these inclusions in two affected dolphins. Histochemical tests indicated that they contained glycoprotein but not lipid. Ultrastructurally, they consisted of granular, moderately electron-dense material, bounded by a membrane. A central or eccentric core of highly electron-dense material was usually apparent. The inclusions were probably of lysosomal origin.

Squamous cell carcinoma of the skin in a Père David's deer (Elaphurus davidianus).

A solitary mass overlying the right carpal joint region's skin in an 18-year-old female Père David's deer (Elaphurus davidianus) was surgically excised and histologically diagnosed as a squamous cell carcinoma (SCC). The tumor was locally infiltrative, showing rather few "horn pearls" and many mitotic figures. This is the first report of a cutaneous SCC in the Père David's deer.

Embryonic and neonatal mortality from salmonellosis in captive bred raptors.

In a captive breeding center near Rome (Italy), cases of embryonic and neonatal death were recorded during the breeding seasons in the European eagle owl (Bubo bubo), peregrine falcon (Falco peregrinus), buzzard (Buteo buteo), and lanner falcon. (Falco biarmicus). Salmonella havana and S. virchow were isolated. Three pulli, clinically infected with S. havana, were successfully treated with enrofloxacin. From two groups of healthy 3- to 4-wk-old eagle owls, Salmonella sp. group 61 (61:r:-) and S. havana were collected. A strain of S. paratyphi B was detected in a pharyngeal swab and a fecal sample from an adult female goshawk (Accipiter gentilis), affected with pharyngeal trichomoniasis. A S. hadar strain was collected from a healthy 1-yr-old female eagle owl and S. livingstone was isolated from a 1-mo-old female peregrine, dead of an acute respiratory syndrome. Lesions of fibrinous polyserositis and multivisceral congestion were observed. From frozen 1-day-old chicks, on which adult and young raptors were fed, S. havana and S. livingstone isolates with similar biochemical and drug susceptibility patterns to those isolated from raptors were identified. A surveillance program on infectious diseases reduced embryonic and neonatal death rates in the following breeding seasons.

Post mortem investigations on cetaceans found stranded on the coasts of Italy between 1990 and 1993.

Detailed pathological and virological examinations were carried out on 25 cetaceans found stranded between 1990 and 1993 on the coasts of six Italian regions (Latium, Tuscany, Apulia, Abruzzo, Veneto and Sicily). There were 16 striped dolphins (Stenella coeruleoalba), three bottlenosed dolphins (Tursiops truncatus), three Risso's dolphins (Grampus griseus), one rough-toothed dolphin (Steno bredanensis), one fin whale pup (Balaenoptera physalus), and one minke whale (Balaenoptera acutorostrata). Apart from parasitic diseases (44 per cent), the most frequently detected lesions were pneumonia (68 per cent), enteritis (44 per cent), non-purulent hepatitis (40 per cent), interstitial nephritis (32 per cent) and encephalitis (32 per cent). Morbilivirus infection was diagnosed by immunocytochemistry in four striped dolphins, two stranded on the coasts of Latium in 1991 and two on the coasts of Tuscany in 1993. Despite the presence of lesions consistent with morbilliviral pneumonia in two other striped dolphins stranded on the coast of Apulia in 1991, no morbillivirus antigen was demonstrated in the tissues of these animals. Anticanine distemper virus antibodies were detected in the serum of the adult minke whale found stranded on the coast of Tuscany in 1993. However, no viruses were isolated from the tissues of any of the 25 cetaceans.

Clinicopathological findings in sheep from Sardinia showing neurological signs of disease.

Histopathological and bacteriological examinations were performed on 178 brains from Sardinian sheep which were showing neurological signs. The sheep represented the total number of sheep with neurological syndromes submitted for diagnostic investigations over a three-year period in Sardinia. Scrapie was detected in 57 cases, cerebrocortical necrosis in 25, intoxication by a typical Mediterranean plant (Cistus species) was suspected in 25, coenurosis was detected in 11 cases, Listeria monocytogenes in eight cases and focal symmetrical encephalomalacia in six cases. Non-suppurative inflammatory changes were observed in three of the brains and suppurative changes were noted in two. Lesions restricted to the spinal cord were found in three cases. In the remaining 38 cases there were no significant neuropathological changes.

[Study and management of zoonoses transmitted by wild animals]. / Approccio allo studio e alla gestione delle zoonosi trasmesse dagli animali selvatici.

In the present work the criteria to approach the study, the prophylaxis and the control of wildlife-related zoonoses are discussed, along with the role of veterinarians as responsible for wildlife and public health management. More in detail, the ecological method in approaching the study of wildlife infections is discussed, along with the relationships existing among hosts, parasites and the environment. Finally, the role of man as a host of zoonotic agents is discussed, and the basic principles of prophylaxis and control of infections caused by such agents are also considered.

Bovine spongiform encephalopathy: an overview.

Bovine spongiform encephalopathy (BSE), a novel disease of cattle first described in 1986, has now reached epidemic proportion in Great Britain with about 500 cases a week. The clinical, epidemiological, and pathological characteristics of BSE are described. Moreover, the etiopathogenetic mechanisms of spongiform encephalopathies are reviewed. Legislative measures to prevent the spread of BSE in Italy and in other EEC countries and to minimize the theoretical risk to man are reported.

Animal transmissible spongiform encephalopathies and genetics.

The genotype of the host plays a crucial role in the pathogenesis of transmissible spongiform encephalopathies (TSEs). In this respect, the most important factor is represented by the gene of the prion protein (PrP). The present work summarizes the currently available knowledge on the genetic basis of TSEs focusing, in particular, on sheep scrapie. Interest in this disease has grown markedly following the discovery of bovine spongiform encephalopathy, both for scientific and health reasons. In Italy, specific research grants from the Ministry of Health and the National Research Council (CNR), together with cooperation between the Istituto Superiore di Sanità and the Istituti Zooprofilattici Sperimentali, have allowed us to study the PrP genotype and to investigate the genetic susceptibility to scrapie in the most important Italian sheep breeds, with special reference to Sarda, Comisana and Massese. The PrP genotype in relation to scrapie susceptibility was also studied in goats of Ionica breed.

Neurochemistry of myenteric plexus neurons of bank vole (Myodes glareolus) ileum.

The neurochemistry of enteric neurons differs among species of small laboratory rodents (guinea-pig, mouse, rat). In this study we characterized the phenotype of ileal myenteric plexus (MP) neuronal cells and fibers of the bank vole (Myodes glareolus), a common rodent living in Europe and in Northern Asia which is also employed in prion experimental transmission studies. Six neuronal markers were tested: choline acetyltransferase (ChAT), neuronal nitric oxide synthase (nNOS), calbindin (CALB), calcitonin gene-related peptide (CGRP) and substance P (SP), along with HuC/D as a pan-neuronal marker. Neurons expressing ChAT- and nNOS-immunoreactivity (IR) were 36 ± 12% and 24 ± 5%, respectively. Those expressing CGRP-, SP- and CALB-IR were 3 ± 3%, 21 ± 5% and 6 ± 2%, respectively. Therefore, bank vole MPs differ consistently from murine MPs in neurons expressing CGRP-, SP- and CALB-IR. These data may contribute to define the prion susceptibility of neuron cell populations residing within ileal MPs from bank voles, along with their morpho-functional alterations following oral experimental prion challenge.

Co-existence of classical scrapie and Nor98 in a sheep from an Italian outbreak.

Nor98 is an atypical scrapie strain characterized by a molecular pattern and brain distribution of the pathological prion protein (PrP(Sc)) different from classical scrapie. In Italy, 69 atypical cases have been identified so far and all were characterized as Nor98 strain. In this paper we report an unusual case in a sheep which showed immunohistochemical and molecular features of PrP(Sc) different from the other atypical cases. The sheep was from an outbreak where the index and the other four cases were affected by classical scrapie. Histopathological, immunohistochemical and Western blot analyses on the brain of the unusual case revealed the simultaneous presence of pathological features characteristic of Nor98 and classical scrapie. Interestingly, the prevalent disease phenotype in the brainstem was classical scrapie-like, while in the cerebral cortex and cerebellum the Nor98 phenotype was dominant. The sub-mandibular lymph node was positive and showed a PrP(Sc) molecular pattern referable to classical scrapie. The PrP genotype was AL(141)RQ/AF(141)RQ. Taken together, the occurrence of classical scrapie in the outbreak, the PrP genotype, the involvement of different cellular targets in the brain and the pathological and molecular PrP(Sc) features observed suggest that this unusual case may result from the co-existence of Nor98 and classical scrapie.