IZS Istituto Zooprofilattico Sperimentale
Recent publications
Nearly two decades ago, pathologic examination results suggested that acoustic factors, such as mid-frequency active naval military sonar (MFAS) could be the cause of acute decompression-like sickness in stranded beaked whales. Acute systemic gas embolism in these whales was reported together with enigmatic cystic liver lesions (CLL), characterized by intrahepatic encapsulated gas-filled cysts, tentatively interpreted as “gas-bubble” lesions in various other cetacean species. Here we provide a pathologic reinterpretation of CLL in odontocetes. Among 1,200 cetaceans necropsied, CLL were only observed in four striped dolphins (Stenella coeruleoalba), with a low prevalence (2%, N = 179). Together, our data strongly suggest that CLL are the result of the combination of a pre-existing or concomitant hepatic vascular disorder superimposed and exacerbated by gas bubbles, and clearly differ from acute systemic gas embolism in stranded beaked whales that is linked to MFAS. Budd-Chiari-like syndrome in dolphins is hypothesized based on the present pathologic findings. Nonetheless, further researched is warranted to determine precise etiopathogenesis(es) and contributing factors for CLL in cetaceans.
Human orthopneumovirus (HRSV) is a virus belonging to the Pneumovirus genus that causes lower respiratory tract infections (LRTI) in infants worldwide. In Tunisia, thousands of infants hospitalized for LRTI are found to be positive for HRSV but no whole genome sequences of HRSV strains circulating in this country are available thus far. In this study, five nasal swab samples collected at different time points from a three-month-old female baby with severe immunodeficiency that was hospitalized for acute bronchiolitis were investigated by next generation sequencing. The Tunisian sequences from this study originated from samples collected in 2021, belong to the ON1 genotype of HRSV-A, and are clustered with European sequences from 2019 and not from 2020 or 2021. This is most likely related to local region-specific transmission of different HRSV-A variants due to the COVID-19 related travel restrictions. Overall, this is the first report describing the whole genome sequence of HRSV from Tunisia. However, more sequence data is needed to better understand the genetic diversity and transmission dynamic of HRSV.
The genus Motacilla belongs to Motacillidae (Passeriformes), where mitochondrial features are poorly understood and phylogeny is controversial. Whole mitochondrial genome (mitogenome) data and large taxon sampling are considered to be ideal strategies to obtain this information. We generated four complete mitogenomes of M. flava , M. cinerea , M. alba and Dendronanthus indicus , and made comparative analyses of Motacilla species combined with mitogenome data from GenBank, and then reconstructed phylogenetic trees based on 37 mitochondrial genes. The mitogenomes of four mitogenome sequences exhibited the same gene order observed in most Passeriformes species. Comparative analyses were performed among all six sampled Motacilla mitogenomes. The complete mitogenomes showed A-skew and C-skew. Most protein-coding genes (PCGs) start with an ATG codon and terminate with a TAA codon. The secondary structures of RNAs were similar among Motacilla and Dendronanthus . All tRNAs except for trnS(agy) could be folded into classic clover-leaf structures. Three domains and several conserved boxes were detected. Phylogenetic analysis of 90 mitogenomes of Passeriformes using maximum likelihood (ML) and Bayesian inference (BI) revealed that Motacilla was a monophyletic group. Among Motacilla species, M. flava and M. tschutschensis showed closer relationships, and M. cinerea was located in a basal position within Motacilla . These data provide important information for better understanding the mitogenomic characteristics and phylogeny of Motacilla .
Bluetongue virus (BTV) is the etiologic agent of bluetongue, a WOAH (founded as Office International des Épizooties, OIE)-notifiable economically important disease of ruminants. It is transmitted by its biological vector Culicoides biting midge and 24 different “classical” serotypes have been reported to date. In recent years, several putative novel BTV serotypes, often referred to as “atypical” BTVs, have been documented. These are characterized by unusual biological characteristics, most notably avirulence and vector-independent transmission. Here, we describe the recurrence of such an atypical virus strain BTV-X ITL2021 detected in goats six years after its first discovery in Sardinia, Italy. Combined serological and genome analysis results clearly suggest that the two strains belong to the same BTV serotype. However, unlike the 2015 strain, BTV-X ITL2021 was successfully isolated in BSR cell-culture allowing further serological characterization. Lastly, seropositivity for BTV-X ITL2021 was detected by virus-neutralization in approximately 74% of animals tested, suggesting that this atypical BTV serotype has been circulating undetected in asymptomatic animals for years.
Beginning in the 1960s, restoration of the European bison (Bison bonasus) in Ukraine has resulted in seven herds with approximately 350 total individuals currently. For the first time, we describe characteristics of habitats used by these seven European bison herds located in forest, forest-steppe, and mountain zones of Ukraine. During the growing season across all ecological zones and herds, we observed variation in bison use of woodland and open habitats associated with temporal variation in landscape scale habitat characteristics. During winter across all ecological zones and herds, variation in habitat use declined dramatically with supplementary feeding that incentivized sedentary association with woodlands. Our descriptive field observations are intended to improve understanding of European bison landscape ecology and serve as a basis for additional quantitative investigations.
Clinical and subclinical ketosis (SCK) in dairy cows occurs during the lactation period frequently in many herds, causing a reduction in milk yield and alterations in milk quality with significant economic losses for farmers. SCK is defined as a preclinical stage of ketosis characterized by an elevated ketone body level without clinical signs. Often many cows develop an elevated ketone body level during the first weeks of lactation even though it never goes up to a critical point causing clinical signs. This study aimed to evaluate the prevalence of SCK in Sicily and assess the effect of a treatment with propylene glycol (PG) to control the SCK, thus, reducing the negative effect on milk quality yield. This cross-sectional study was carried out on 22 farms located south-east of Sicily and 1,588 cows in lactation. A total of 3,989 individual milk samples were collected from calving to 80 subsequently days to check the β-hydroxybutyrate (BHB) values in order to establish the SCK status by the Fourier Transform Infrared Spectroscopy. Moreover, the contents of fat, protein, lactose, casein, urea, somatic cell count and acetone were evaluated to identify a correlation between SCK and milk quality. A total of 1,100 cows showed BHB values higher than 0.10 mmol/L. These cows were considered SCK positive, were separated from the rest of the herd, and treated with PG (400 g/head per day), all SCK cows were treated with PG and cows without SCK were not treated. The results showed a prevalence of 41.5% of SCK-positive cows during the first 9 days of lactation. The comparison among the cure rate of treated cows shows that the treatment was most effective in the first 7 days of lactation (76.5% of treated cows) than in the following days. PG positively influenced the milk quality parameters, except for the fat proportion. Moreover, the animals treated with PG showed also an increase in milk yield, supporting the economical sustainability of treatment.
When alien species enter new environments, they face new challenges. They encounter new predators, parasites, or pathogenic bacteria, which leads to the rapid evolution of invasive species to adapt to new habitats, where immune-related genes play a significant role. Lithobates catesbeianus is one of the worst invasive species in the world; it rapidly spread worldwide in the 19th century and invaded China in the 1950s. We predicted the possible transmission routes of bullfrogs in China through microsatellite loci and clustered the populations at different points. We studied the adaptive selection and drift reshaping of the genetic structure of an immune genetic major histocompatibility complex (MHC) in invasive bullfrogs in 23 Chinese invasive populations (13 island and 10 mainland sites) and two American populations (Kansas and California). The Chinese bullfrog populations were divided into 6 clusters by microsatellite structure, and the MHC diversity in each cluster was different. The genetic diversity of both microsatellite and MHC genes decreased due to the bottleneck effect and the rate of diversity loss at functional and neutral loci, and there was no significant difference. We found that two MHC alleles were endemic in Chinese populations, which corresponded to distinct functional supertypes. We also analyzed the impact of environmental factors and island effects on the diversity of MHC genes. These results indicate that rapid evolution plays an important role in maintaining functionally important MHC variation during the bullfrog invasion process and provide evidence that low genetic diversity can result in successful invasion.
Brucella ceti, a zoonotic pathogen of major concern to cetacean health and conservation, is responsible for severe meningo-encephalitic/myelitic lesions in striped dolphins (Stenella coeruleoalba), often leading to their stranding and death. This study investigated, for the first time, the cellular prion protein (PrPc) expression in the brain tissue from B. ceti-infected, neurobrucellosis-affected striped dolphins. Seven B. ceti-infected, neurobrucellosis-affected striped dolphins, found stranded along the Italian coastline (6) and in the Canary Islands (1), were investigated, along with five B. ceti-uninfected striped dolphins from the coast of Italy, carrying no brain lesions, which served as negative controls. Western Blot (WB) and immunohistochemistry (IHC) with an anti-PrP murine monoclonal antibody were carried out on the brain parenchyma of these dolphins. While PrPc IHC yielded inconclusive results, a clear-cut PrPc expression of different intensity was found by means of WB analyses in the brain tissue of all the seven herein investigated, B. ceti-infected and neurobrucellosis-affected cetacean specimens, with two dolphins stranded along the Italian coastline and one dolphin beached in Canary Islands also exhibiting a statistically significant increase in cerebral PrPc expression as compared to the five Brucella spp.-negative control specimens. The significantly increased PrPc expression found in three out of seven B. ceti-infected, neurobrucellosis-affected striped dolphins does not allow us to draw any firm conclusion(s) about the putative role of PrPc as a host cell receptor for B. ceti. Should this be the case, an upregulation of PrPc mRNA in the brain tissue of neurobrucellosis-affected striped dolphins could be hypothesized during the different stages of B. ceti infection, as previously shown in murine bone marrow cells challenged with Escherichia coli. Noteworthy, the inflammatory infiltrates seen in the brain and in the cervico-thoracic spinal cord segments from the herein investigated, B. ceti-infected and neurobrucellosis-affected striped dolphins were densely populated by macrophage/histiocyte cells, often harboring Brucella spp. antigen in their cytoplasm, similarly to what was reported in macrophages from mice experimentally challenged with B. abortus. Notwithstanding the above, much more work is needed in order to properly assess the role of PrPc, if any, as a host cell receptor for B. ceti in striped dolphins.
Background: Tamias Sibiricus is the only member of genus Tamias, which is a very important and vigorous seed distributors, and also a vital food for their predators. No information is known about the strict diet, Gut microbiota structure and metabolism profile of chipmunk and how it divers seasonally. Analysis of above mentioned factors, and flexibility toward seasonal shift is crucial to define its growth rates, health, survivorship, and population stability. Results: In this study, we explored the diet, gut microbiota composition, and chipmunk metabolism. Additionally, the influence of different seasons was also investigated by using next-generation sequencing. Results revealed that seasons had a strong effect on a diet : as streptophyte accounted for 37% in spring, which was lowest in summer (34.3%) and autumn (31.4%). Further, Ascomycota was observed 43.8% in spring which reduced to 36.6% in summer and lowest (31.3%) in autumn. Whereas, Nematoda showd maximum abundance from spring (15.8%) to summer (20.6%) and autumn (24.1%). These results are signifying the insectivorous nature of the chipmunk in summer and autumn. While herbivorous and fungivorous nature in spring. The DNA analysis revealed that chipmunk mainly feeds on fungi, including Aspergillus and Penicillium genus. Like diet composition, microbiome also exhibited highly significant dissimilarity (p<0.001, R=0.235) between spring/autumn and spring/summer seasons. Proteobacteria (35.45%), Firmicutes (26.7%) and Bacteroidetes (23.59%) were shown to be the better discriminators as they contributed most to cause differences between seasons. Moreover, PICRUSt was exposed that the assimilation of nutritions were also variated seasonally. The abundance of carbohydrate, lipid, nucleotide, xenobiotics, energy, terpenoids and polyketides metabolism were highest in spring than other seasons. Conclusions: Our study illustrates that seasonal reconstruction in chipmunk diet have a significant role in shaping temporal variations in gut microbial community structure and metabolism profile.
Feline herpesvirus type 1 (FHV-1) is a common causative agent of domestic cats’ rhinotracheitis in domestic cats, and it increasingly threatens wild felids worldwide. The endangered snow leopard (Panthera uncia) belongs to the family Felidae, and it is the top predator on the Tibetan Plateau. Here we report the identification and isolation of FHV-1 from three dead captive snow leopards that presented with sneezing and rhinorrhea. To explore the relationship between FHV-1 and their deaths, organs and nasal swabs were collected for histopathology, viral isolation and sequence analysis. The results revealed that all three snow leopards were infected with FHV-1. The first animal died primarily of cerebral infarction and secondary non-suppurative meningoencephalitis that was probably caused by FHV-1. The second animal died mainly of renal failure accompanied by interstitial pneumonia caused by FHV-1. The cause of death for the third animal was likely related to the concurrent reactivation of a latent FHV-1 infection. The gD and gE gene sequence alignment of the isolated FHV-1 isolate strain revealed that the virus likely originated from a domestic cat. It was found that FHV-1 infection can cause different lesions in snow leopards than in domestic cats and is associated with high risk of disease in wild felids. This suggests that there should be increased focus on protecting wild felids against FHV-1 infections originating from domestic cats.
Background Interstitial lung disease is a heterogeneous group of conditions characterized by severe radiographic changes and clinicopathological findings. However, in the vast majority of cases, the cause remains unknown. Case description In the present study, we reported the clinical case of a 3 years old female Bull Terrier presented in October 2020 to the Advanced Diagnostic Imaging Department of the Turin Veterinary Teaching Hospital with a progressive pulmonary illness characterized by dyspnea, exercise intolerance, and a diffuse and severe pulmonary interstitial pattern at imaging investigations. Considering the clinical findings, the dog was included in a serological survey for Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection in companion animals, showing positive results. Due to the further clinical worsening, the owners opted for euthanasia. At necroscopy, dog showed severe and chronic bronchopneumonia compatible with a Canine Idiopathic Pulmonary Fibrosis and with serological features linked to a SARS-CoV-2 infection. Conclusions The comparison of these lesions with those reported in humans affected by Coronavirus Disease 2019 (COVID-19) supports the hypothesis that these findings may be attributable to the post-acute sequelae of SARS-CoV-2 infection in a dog with breed predisposition to Canine Idiopathic Pulmonary Fibrosis (CIPF), although direct evidence of SARS-CoV-2 by molecular or antigenic approaches remained unsolved.
When we flip a coin, we expect a 50 percent chance of it landing on heads. But what happens when an insect jumps? In this study, we report the self-righting strategy of the flea beetle, Altica cirsicola , which lives in a dense maze of leaves. How can the flea beetle coordinate landing on surfaces of such a range of orientation and position? In this experimental study, we film the take-off, flight, and landing of flea beetles on a configurable angled platform. Dead flea beetles tossed through the air have a 46% chance of landing on their legs, similar to a fair coin. However, live beetles land successfully 85% of the time, suggesting the importance of active self-righting strategies to accommodate the different orientations of the landing platform. We observe that beetles may extend their wings to slow down and reduce impact as well as perform numerous rotations. These behaviors may inspire jumping robots that can land softly.
Animal hair is a useful biomonitoring tool for assessing the occurrence of trace elements in ecosystems. Essential (chromium, copper, iron, manganese, nickel, and zinc) and nonessential (aluminum, arsenic, cadmium, mercury, lead, palladium, platinum, rubidium, tin, and vanadium) elements were investigated in hair of badger (Meles meles), wild boar (Sus scrofa), marmot (Marmota marmota), wolf (Canis lupus), fox (Vulpes vulpes) and deer (Cervus elaphus) from Northwestern Italy. Badger was found to be the highest bioaccumulator of metals, while wolf, fox and deer recorded the lowest values. Essential elements contribute in higher degree to the sum of metals for all species except for wild boar. Results have shown that animals with omnivorous diet such as badger, marmot and wild boar have metal content (especially Al, As, Cr, Cu, Fe, Ni, V) higher than carnivores (wolf and fox) and herbivores (deer) and could represent an effective sentinel of environmental exposure to metals.
Epilepsy is a major global issue. Epilepsy patients are treated with AED (antiepileptic drugs). Interindividual variability in drug response has been documented in several studies. The resistance to drug response may be attributed to genetic polymorphism. The current study was undertaken to investigate the CYP2C9 gene polymorphism associated with antiepileptic drug (AED) resistance in the Pakistani population. The current study included 337 individuals including 100 control subjects, 110 drug-resistant subjects, and 127 drug responders. Genomic DNA was isolated from blood, and amplification of rs1799853 (430C > T) and rs1057910 was carried out by polymerase chain reaction. Genotypes of CYP2C9 SNPs were determined by Sanger’s sequencing. Astounding results were observed in the current study that none of the well-known reported SNPs of CYP2C9 was found in our Pakistani cohorts. However, a novel missense variant (c.374G > A) was found only in drug-resistant patients of the current study. According to the in silico analysis performed by PolyPhen-2, it was observed that this nonsynonymous substitution is likely to be pathogenic. The results of our study demonstrated that rs1799853 and rs1057910 may be involved in drug resistance in the Pakistani population. However, some other variants on CYP2C9 may play a critical role in AED resistance that needs to be explored.
Epilepsy affects millions of people worldwide. Although antiepileptic drugs work for the majority of epileptic patients, these drugs do not work for some of the patients, subjecting them to drug-resistant epilepsy (DRE). Voltage-gated sodium channels act as targets for a number of antiepileptic drugs, and the genes encoding these channels can play a crucial role in developing drug-resistant epilepsy. This case-control (100 control: 101patients) study evaluated the association of sodium channel genes SCN1A and SCN2A with drug-resistant epilepsy. The cases were further accounted in two categories, drug-resistant and drug-responsive epileptic patients. The polymorphic sites rs794726754, rs1057518252, rs121918809, rs12191792, rs121917932, c.730 G > T, c.735 G > T, c.736 A > T, rs10167228, and rs2298771 of the SCN1A gene and rs17183814 of SCN2A gene were selected for mutational analysis. The DNA was isolated, amplified by PCR, and then, was run through 1% agarose gel. The sequencing was performed, and the sequences were observed through BioEdit software for any change in DNA sequence. In our study, no polymorphism was observed in the studied SNPs except for rs2298771. For rs2298771, a significant difference existed in the distribution of genotypic and allelic frequencies ( p < 0.01) among the case and control group. Furthermore, the genotypic and allelic frequencies of the two categories of cases (drug responder drug resistant) were calculated. The genotypic and allelic frequencies of drug-responsive and drug-resistant epileptic patients did not differ significantly ( p > 0.01). Our study indicated that the rs2298771 polymorphism of SCN1A may not be associated with chance of developing DRE in the Pakistani population.
Bovine besnoitiosis is a parasitic disease caused by the protozoan Besnoitia besnoiti, leading to infertility in bulls and abortions in cows. In Italy, it is considered an emerging disease, recently introduced by the importation of animals from Spain and France. In the last decade, many outbreaks have been reported and confirmed in native cattle, mostly in northern and central Italy. This study reports on an autochthonous outbreak of bovine besnoitiosis in two nearby farms located in northwestern Sicily. A 15-month-old Limousine bull born on the farm showed typical clinical signs of the chronic disease phase, such as edema of the scrotum with subsequent hyperkeratosis associated with eschars and thickening of the skin. A histopathological examination revealed the presence of Besnoitia tissue cysts containing bradyzoites in the eyes, tendons, testicles, dermis, and nictitating membrane. A serological investigation using a commercial ELISA kit revealed a high seroprevalence of the antibody anti-B. besnoiti (79.2% for the farms in this study). Clinical disease showed low prevalence (1.5%) despite the high seroprevalence of specific antibodies in the herd, confirming that bovine besnoitiosis is an emergent endemic pathogen in Sicily, but its clinical behavior still remains sporadic.
Background: There have been worldwide changes in the researches on Hantaviruses in the past several decades. Nevertheless, there are few bibliometric analysis studies this field. We aim to evaluate and visualize the research focuses and trends of this field using a bibliometric analysis way to help understand the developmet and future hotspots of this field.Material and Methods: Publications related to Hantavirus studies were culled from the Web of Science Core Collection to generate trend analysis. The articles and reviews were re-extracted and Countries, institutions, authors, references and keywords in this field were visually analyzed by using VOSviewer and CiteSpace.Results: A total of 4408 studies were included and the number of publications regarding Hantaviruses significantly increased yearly. 3716 research articles and reviews were retrieved to generate bibliometric analysis. These studies mainly come from 125 countries led by USA and China and 3312 institutions led by the University of Helsinki. 12529 authors were identified and Vaheri A were the most influential author. Journal of Virology was the journal with the most studies and citations. After analysis, Hemorrhagic fever with renal syndrome , Hantavirus cardiopulmonary syndrome, nephropathia epidemica and related genotypes, clinical symptoms and rodents were the most common keywords and developing areas.Conclusion: Research on Hantavirus is flourishing. Cooperation among different countries and institutions in this field must be strengthened in the future. The ecology and clinical symptoms of new genotypes, the vaccine development and factors that affect host population distribution and density are current and developing areas of study.
The fusion of the SARS-CoV-2 virus with cells, a key event in the pathogenesis of Covid-19, depends on the assembly of a six-helix fusion core (FC) formed by portions of the spike protein heptad repeats (HR) 1 and 2. Despite the critical role in regulating infectivity, its distinctive features, origin, and evolution are scarcely understood. Thus, we undertook a structure-guided positional and compositional analysis of the SARS-CoV-2 FC, in comparison with FCs of related viruses, tracing its origin and ongoing evolution. We find that clustered amino acid substitutions within HR1, distinguishing SARS-CoV-2 from SARS-CoV-1, enhance local heptad stereotypy and increase sharply the FC serine-to-glutamine (S/Q) ratio, determining a neat alternate layering of S-rich and Q-rich subdomains along the post-fusion structure. Strikingly, SARS-CoV-2 ranks among viruses with the highest FC S/Q ratio, together with highly syncytiogenic respiratory pathogens (RSV, NDV), whereas MERS-Cov, HIV, and Ebola viruses display low ratios, and this feature reflects onto S/Q segregation and H-bonding patterns. Our evolutionary analyses reveal that the SARS-CoV-2 FC occurs in other SARS-CoV-1-like Sarbecoviruses identified since 2005 in Hong Kong and adjacent regions, tracing its origin to >50 years ago with a recombination-driven spread. Finally, current mutational trends show that the FC is varying especially in the FC1 evolutionary hotspot. These findings establish a novel analytical framework illuminating the sequence/structure evolution of the SARS-CoV-2 FC, tracing its long history within Sarbecoviruses, and may help rationalize the evolution of the fusion machinery in emerging pathogens and the design of novel therapeutic fusion inhibitors.
Feline herpesvirus type 1 (FHV-1) is a common causative agent of domestic cats rhinotracheitis and gradually threatens the wild felid worldwide. The endangered snow leopard belongs to the family Felidae and is also the top predator on the Tibetan Plateau. Herein, FHV-1 was identified and isolated from three dead snow leopards with symptom of sneezing and rhinorrhea. To explore the relationship between FHV-1 and their death, histopathology and molecular biology was performed. The organs and nasal swabs were collected for examinations of histopathology, the nucleic acid of the pathogen, viral isolation, and sequence analysis. The results reveal that all three snow leopards were infected with FHV-1. The first case primarily died of old cerebral infarction and secondary non-suppurative meningoencephalitis probably caused by FHV-1. The second case mainly died of renal failure accompanied by interstitial pneumonia caused by FHV-1. The third case was doubted to be related to the reactivation of latency of FHV-1. The gD and gE gene sequence alignment of the FHV-1 isolate strain revealed that the isolated strain originated from a domestic cat. Therefore, FHV-1 infection can cause different lesions of snow leopards and shows a high risk for the wild felid. We should focus more on protecting felid against threatening of FHV-1 infection originating from domestic cats.
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190 members
Franca Rossi
  • Diagnostica Specialistica Sezione di Campobasso
Rossella Lelli
  • Department of Foreign animal diseses and Epidemiology
Lara Savini
  • Stat&GIS
Manuela Tittarelli
  • Department of Serology
Carla Ippoliti
  • Statistics&GIS
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