Veterinaria Italiana | ISTITUTO ZOOPROFILATTICO CAPORALE (original) (raw)

Papers by Veterinaria Italiana

Objective To validate the Italian translation of the Glasgow Composite Measure Pain Scale – Short... more Objective To validate the Italian translation of the Glasgow Composite Measure Pain Scale
– Short Form (ICMPS‑SF) in order to assess acute pain in dogs. The original English‑version
of the scale (the Glasgow Composite Measure Pain Scale – Short Form ‑ CMPS‑SF) was
translated into Italian according to a standard protocol to ensure linguistic and cultural
validity. Nine Italian veterinary surgeons then recorded pain scores in dogs undergoing
orthopaedic or soft tissue surgery using the ICMPS‑SF at 2, 6, and 24 hours post‑extubation.
Construct validity was demonstrated using hypothesis testing. A total of 95 dogs were
recruited into the study. Thirty‑seven dogs underwent orthopaedic procedures and 58 dogs
underwent soft tissue procedures. Twenty‑three, 45, and 27 procedures were classified as
mild, moderate, and severe, respectively. Statistically significant differences in the median
pain scores were demonstrated between orthopaedic and soft tissue cases as well as among
mild, moderate, and severe cases. Median pain scores decreased with time and changes were
statistically significant. The ICMPS‑SF demonstrated construct validity similar to the original
English‑language scale, resulting in a valid and reliable instrument for the assessment of
acute pain in dogs by Italian veterinarians.

Zika Virus (ZIKV) is a RNA virus belonging to the genus Flavivirus, family Flaviviridae. This vir... more Zika Virus (ZIKV) is a RNA virus belonging to the genus Flavivirus, family Flaviviridae. This virus
is transmitted through bite of Aedes mosquitoes, in particular Ae. aegypti. On February 1st 2016,
the World Health Organization (WHO) has declared ZIKV a Public Health Emergency of
International Concern. Successively, considering the establishment of Ae. albopictus, WHO
has classified Italy as having a moderate likelihood of local transmission of ZIKV, preceded
in Europe only by France. For this reason an entomological surveillance plan was been
activated in Sardinia in 2016. BG Sentinel Mosquito Traps have been positioned in 29 sites,
comprising urban areas and points of entry, as ports and airports. Mosquitoes were collected
fortnightly from April to December. A total of 3,089 mosquitoes were collected belonging
to 10 species. The most numerous species have been Cx. pipiens s.l. and Ae. albopictus. All
mosquitoes sampled have been assayed by real time reverse transcriptase PCR for detection
of ZIKV RNA. A total of 584 pool have been analyzed and have been reported no evidence
of ZIKV. A permanent entomological surveillance should be implemented principally in the
urban areas and points of entry, as ports and airports, because Ae. albopictus, susceptible to
ZIKV, is established in Sardinia and also know the recent introduction of invasive mosquitoes
species Ae. koericus and Ae. japonicus in Italy.

Canine leishmaniasis (CanL) is a systemic zoonotic disease caused by the protozoan Leishmania, an... more Canine leishmaniasis (CanL) is a systemic zoonotic disease caused by the protozoan Leishmania,
an intracellular macrophage parasite, transmitted by the bite of phlebotomine sandflies. In dogs,
the clinical disease is mostly characterised by symptoms associated with viscerocutaneous
lesions such as lymphadenopathy, splenomegaly, skin lesions, and renal and ocular disease
caused by the deposition of immune complexes. The parasite may provoke mucosal lesions
which cause atypical clinical signs. The aim of this study is to describe an atypical nostril mass
in a dog infected by Leishmania. Clinical examination did not show any systemic clinical signs,
while haematological, biochemical, and urinary parameters demonstrated a mild disease
stage. Diagnosis was confirmed through the isolation of cultured live parasites by biopsy. The
dog was treated with a combination of miltefosine and allopurinol, showing full remission of
clinical symptoms after 2 months. The authors outline the importance of considering CanL in
the differential diagnosis of mucous and tumour-like lesions.

The objective of this study was to determine the effect of the porcine circovirus type 2 (PCV2) v... more The objective of this study was to determine the effect of the porcine circovirus type 2 (PCV2)
vaccination on the levels of viremia, the number of viremic-positive pigs, and production
performance [i.e. nursery mortality, post-weaning mortality, and average daily weight
gain (ADWG)] under field conditions. There were 140 farrow-to-finish pig herds involved in
this study. The vaccination of piglets was implemented in 82 of the 140 herds. In each herd
blood samples were collected from sows and pigs in different age category. In addition, a
questionnaire regarding the production performance was provided for each herd. Results
demonstrate that the vaccination of piglets prevented the development of viremia in 23.2% of
herds. Significant decreases in the levels of PCV2 DNA in serum and in the number of viremic
pigs were also noted. These results indicate that the vaccination of piglets against PCV2 is a
useful tool in controlling the PCV2 infection in herds with a high risk of a wide range of viral
and bacterial agents, poor management strategies, and a low level of biosecurity practices.

Contagious agalactia (CA) is a serious disease of small ruminants that occurs in many countries, ... more Contagious agalactia (CA) is a serious disease of small ruminants that occurs in many countries, and is usually characterized by mastitis, arthritis, keratoconjunctivitis, pleuropneumonia, and septicemia. Mycoplasma agalactiae (Ma) is the main causative agent in sheep and goats but other pathogens including Mycoplasma mycoides subsp. capri (Mmc, which incorporates the former M. mycoides subsp. mycoides Large Colony type), Mycoplasma capricolum subsp. capricolum (Mcc), and Mycoplasma putrefaciens (Mp) might be involved. They are all usually associated with infections in goats and may cause similar clinical signs. A total of 116 sheep and 16 goats suffering from the acute form of the disease were included in this study. They were recruited following a number of outbreaks suspected to be CA in the Ardebil province of Iran. Milk, lachrymal or synovial fluid were collected exclusively from the affected animals in order to identify the pathogen involved. Of the 132 collected samples, 33 (25%) were positive for Mycoplasma species by culture in PPLO broth and agar. The polymerase chain reaction followed by denaturing gradient gel electrophoresis (PCR/DGGE) method identified 18 (12 sheep and 6 goats) of the 33 Mycoplasma positive samples with mixed Mycoplasma population. In particular, 25 Ma (47.2%), 23 Mp (43.4%), 4 Mcc (7.5%), and 1 Mmc (1.9%) were identified. This confirms that the several Mycoplasma species rather than the Ma only are in circulation, and are able to cause CA in sheep and goats in Iran. This is the first report on the isolation and identification of Mp, Mmc and Mcc in infected small ruminant flocks in Iran.

In this study 252 poultry samples comprised of poultry meat (n = 228) and poultry eggs (n = 24) w... more In this study 252 poultry samples comprised of poultry meat (n = 228) and poultry eggs
(n = 24) were screened for the isolation of Escherichia coli (E. coli). A total of 62 E. coli isolates
were recovered from poultry meat. The E. coli isolates belonged to different serogroups
based on ‘O’ serotyping of the isolates viz O29 (10.8%), O8 (7.7%), O40 (6.15%), O2 (4.61%),
O60 (3.08%), O106 (3.08%), 42 (1.54%), O 87 (1.54%), and 01 serotypes of O1, O7, O30, O45,
O59, O66, O105, O1116, O136, O141, O147, O148, O166, and O172. Sixteen (24.62%) of the
isolates were UT (untypable) and 6 (9.23 %) were rough types. Molecular characterisation of
the isolates was performed, targeting stx1 and stx2 virulence gene fragment. Out of 62 E. coli
isolates, 10 (16.12%) were carrying virulence gene stx2, whereas none of the isolate was
carrying stx1 gene. The E. coli isolates showed wide variation in resistance pattern against
the antimicrobial agents that we used (9-90%). Among E. coli isolates, maximum resistance
was observed against cefuroxime (89.1%) and penicillin (89.4%), followed by ampicillin
(80.43%), vancomycin (74.1%), co-trimoxazole (73.1%), cephalothin (60.8%), ceftriaxone
(28.2%), tetracycline (17.4%), gentamicin (13%), amikacin (13.04%), ofloxacin (13%), and
ciprofloxacin (6.5%). A high degree of susceptibility was observed against amikacin (84.7%)
and ciprofloxacin (76%) followed by gentamicin (71.73) and ofloxacin (60.86%). High multiple
antibiotic resistances were observed and a total of 34 resistance patterns were identified.

The virulence gene profile of 26 rabbit enteropathogenic Escherichia coli strains, isolated from ... more The virulence gene profile of 26 rabbit enteropathogenic Escherichia coli strains, isolated from 17 colibacillosis outbreaks located in two regions of Northern Italy, was determined using an Echerichia coli virulence DNA microarray. All strains were classified according to their determined biotype, sero-and phylo-group. The distribution of virulence genes encoding for the Locus of enterocyte effacement (LEE), LEE type III secretion system (T3SS), non-LEE T3SS translocated proteins and adherence factors was also determined. All strains but one belonged to phylogroups A and B1. A prevalent association between the O103 serogroup with the rhamnose-negative phenotype (biotype 12 or 14) was found. The most prevalent LEE profile found in tested strains was ler/cesT/espA-1/espB-3/tir-1/eae(beta)/espD-2/escN/eprJ. All strains possessed either the adhesive factor rabbit-2 (afr/2) or the plasmid Rabbit adherence locus (ral) gene and 24 of them an additional individual or combined set of colonization factors efa1/lifA, lpfA and paa genes. Finally, the combined or single presence of a set of LEE and/or non-LEE effector proteins encoding genes, namely espG, cif, map and nle family genes, attested to the genetic potential of investigated strains to induce pathologic lesions to the host. The application of microarray-based technologies in assessing the genetic profile of rabbit E. coli is a reliable, cost-effective candidate for large scale investigations in monitoring programs aimed to survey the circulation of pathogenic strains within rabbit production units, their zoonotic genetic potential and to select E. coli strains eligible for vaccinal prophylaxis in fattening rabbit production.

Large game animals play an important role as carriers and transmitters of O157:H7 and non‑O157:H7... more Large game animals play an important role as carriers and transmitters of O157:H7 and non‑O157:H7 Shiga toxin‑producing Escherichia coli (STEC) in nature. Fresh meat obtained from game animals has been identified as an important source of food‑borne STEC infections. The aim of this study was to evaluate the incidence of the top 6 non‑O157 STEC strains (serogroups O26, O45, O103, O111, O121, and O145) in Namibian game meat based on testing for stx, eae, and O‑group‑specific genes. Meat samples from gemsboks (Oryx gazella) (n = 75), springboks (Antidorcas marsupialis) (n = 41), greater kudus (Tragelaphus strepsiceros) (n = 5), and wildebeests (Connochaetes taurinus) (n = 5) were collected from 2 Namibian abattoirs and tested for STEC using real‑time PCR techniques. Both Shiga toxin (stx) and intimin (eae) virulence genes were detected in 94 out of 126 samples (74.6%). Five of the top 6 STEC serogroup‑specific genes were also detected in samples that were positive for both the stx and eae genes. The results of this study show a high incidence of non‑O157 STEC O‑group genes in Namibian game meat, which suggests that further scrutiny and testing may be necessary to avoid foodborne outbreaks.

According to European Union (EU) regulations, the serological tests for the eradication of bovine... more According to European Union (EU) regulations, the serological tests for the eradication of bovine and ovine brucellosis are the Rose Bengal Test, Complement Fixation Test, and i‑ELISA. These methods, also recommended by the World Organization for Animal Health (OIE) for international trades, have limitations related to the use of suspensions of smooth Brucellae or LPS extracts. Limitations include false‑positive serological reactions to brucellosis, which in turn impedes accurate diagnosis in some herds. False positive reactions should be considered carefully during the final stages of an eradication programme and for surveillance purposes in brucellosis‑free areas. In this study, we produced specific sera through the experimental infection of sheep with Y. enterocolitica O:9 and E. coli O157:H7. These are the most important cross‑reactive bacteria with Brucella. We then evaluated the antibody response of groups of sheep that had been immunised towards homologous antigens and official antigens for brucellosis, in order to identify a differential diagnostic protocol to distinguish cross‑reaction in Brucella‑infected animals. Cross‑reactivity in serological tests for brucellosis: a comparison of immune response of Escherichia coli O157:H7 and Yersinia enterocolitica O:9 vs Brucella spp.

The present case study concerns a case of predation of 4 individuals of captive pink flamingo in ... more The present case study concerns a case of predation of 4 individuals of captive pink flamingo in Emilia Romagna Region, Northeastern Italy. The pink flamingo (Phoenicopterus roseus) is a species included in the Red List of Threatened Species established by the International Union for Conservation of Nature (IUCN) which lists species in danger of extinction. During the Winter of 2013, 4 flamingos (2 in the Comacchio area, and 2 from Argenta and Codigoro oases – Ferrara province) were found dead some of them headless, with their bodies severely bitten. At first, a fox (Vulpes vulpes) was suspected to be the predator responsible for the killing and the birds were taken to the laboratory for further investigations. The investigations included: field observations, study of the predator behaviour, necropsy examinations, assessment of the intercanine distance, and genetic analysis on the predator's traces. The intercanine distance indicated that the predator could not have been a fox. The analysis of salivary DNA samples enabled us to establish that the predator was in fact a dog. This case highlights the importance of co‑operation among the various branches of forensic sciences and the great usefulness of the roles filled by other veterinary forensic experts involved in solving crime. Veterinary forensic sciences to solve a fatal case of predation on flamingos (Phoenicopterus roseus)

Reproductive disorders are responsible for significant economic losses in canary aviculture due t... more Reproductive disorders are responsible for significant economic losses in canary aviculture due to embryo and newborn chick mortality. Most of the time, deaths are caused by bacterial pathogens, however little published data exist about the prevalence of bacterial isolates that are identified during diagnostic protocols. This study reports on data collected from previous investigations carried out on cloacal swabs (n 456), unhatched eggs (n 52), and dead newborn chicks (n 68) collected from canary aviaries with a history of reproductive disorders. Of the examined samples, 41% were positive for the presence of pathogenic or potentially pathogenic bacteria, with particular regard to Gram negative species during bacteriological investigations. The most prevalent microorganisms were Gram‑negative (55%). A predominance of Klebsiella, Escherichia, and Pantoea genera was observed. These are usually associated to pathological conditions in pet birds. Among Gram‑positive bacteria, Bacillus spp. and Staphylococcus spp. were most prevalent. The antimicrobial susceptibility testing carried out on bacterial isolates showed a multiple resistance, especially against amoxycillin, erythromycin, spiramycin, tiamulin, and tylosin. This study represents a first attempt to provide an update on microbial causes of embryonic and neonatal mortality in canary aviaries in Italy; in addition, it provides further understandings about the efficacy of antimicrobial therapy. Reproductive disorders in domestic canaries (Serinus canarius domesticus): a retrospective study on bacterial isolates and their antimicrobial resistance in Italy from 2009 to 2012

In this study we examined macroscopic and microscopic lesions in the mitral valves of 50 regularl... more In this study we examined macroscopic and microscopic lesions in the mitral valves of 50 regularly slaughtered horses in Italy. Macroscopically the results were minor. Microscopic lesions to the valve were found in both young and elderly subjects, confirming that there is no correlation between age and onset of lesions. We set out to evaluate whether the lesions correspond to valve disorder during the life of the animal.

Pteropine Orthoreoviruses (PRVs) are fusogenic bat‑borne orthoreoviruses that cause flu‑like uppe... more Pteropine Orthoreoviruses (PRVs) are fusogenic bat‑borne orthoreoviruses that cause flu‑like upper respiratory tract infections in humans. The presence of this group of viruses in bats and humans has been well documented in areas where their biological reservoirs – fruit bats (family Pteropodidae) – live densely. In the present study, a serum neutralization (SN) assay to detect neutralizing antibodies against PRV Indonesia/2010 isolate was set up and used to assess the seroprevalence of this virus in Italian domestic animals. The new developed assay was able of detecting PRV neutralizing antibodies in the hyper‑immune polyclonal serum produced in rabbits (titer of 1:160). The negative serum was negative at all tested dilutions. No cross‑reactions have been evidenced neither against reference MRVs nor against their respective hyper‑immune sera. Eight hundred and fifty‑three serum samples collected from 524 bovines, 271 small ruminants, and 58 horses (all used as sentinel animals in the Bluetongue and West Nile disease National surveillance program) were also tested with the new developed SN assay. According to the results of this survey, neither PRV nor PRV cross‑reacting viruses antibodies have been demonstrated in Italian domestic animals. However, the new developed SN assay could be a very valuable diagnostic tool to detect infection in animals and humans.

The healing of wounds located on the equine distal limbs can often be impaired, which can result ... more The healing of wounds located on the equine distal limbs can often be impaired, which can result in exuberant granulation tissue and its associated aesthetic alteration and functional failures. Although a number of therapies have been developed and assessed, the treatment of these wounds still presents challenges. This report describes the treatment of exuberant granulation tissue in a horse using an intra‑lesional injection of 4% formaldehyde solution. The successful outcome of this treatment suggests that further investigations are required in order to better evaluate its efficacy, as well as the incidence and the severity of any adverse reaction.

Noroviruses and hepatitis A virus are the pathogens most frequently involved in non‑bacterial gas... more Noroviruses and hepatitis A virus are the pathogens most frequently involved in non‑bacterial gastroenteritis and hepatitis worldwide. They are mainly transmitted via the faecal–oral route, direct person‑to‑person contact or through the consumption of contaminated water and foods. In food virology, detection methods of these viruses are currently based on real‑time reverse transcription‑polymerase chain reaction (RT‑PCR). A crucial step in this process is the acid nucleic extraction, since its performance can negatively influence viral detection and thus give false negative results. The aim of this study is to evaluate the efficiency of 2 automated extraction systems, MagMAX Express and EZ1 Advanced XL, in recovering hepatitis A virus and norovirus RNA from mussels. In the present study, we used mengovirus as a process control to assess the efficiency of the extraction process. Samples were tested for mengovirus, hepatitis A, and norovirus by real time one‑step RT‑PCR assay. Our data indicates the MagMax Express is the better system to extract hepatitis A and norovirus RNA from mussels since its extraction efficiency was higher (p < 0.05) than EZ1 Advanced XL.

In the last decades, adverse food reactions have increased considerably in dogs and cats. In this... more In the last decades, adverse food reactions have increased considerably in dogs and cats. In this study we report on the possible onset of food intolerances symptoms, including otitis, diarrhoea, generalised anxiety, and dermatitis in a cohort of 8 dogs consuming commercial diets. All dogs received an organic chicken‑based diet for 15 days. We performed analysis of blood biochemical parameters, kibble composition, and oxytetracycline (OTC) serum concentration before and after 15 days of organic chicken‑based diet supplementation. We hypothesised that a chronic intake of contaminated food enhanced by the presence of nanoparticle aggregates might be at the base of the onset of pharmacologic or idiopathic food intolerances. At the end of the evaluation period, an overall significant reduction of otitis, diarrhoea, generalised anxiety, and dermatitis was observed. Biochemical analyses indicate a significant increase in the alkaline phosphatase, from 41 to 52.5 U/L, after 15 days (**p < 0.01), while a significant decrease in Gamma‑glutamyl transferase and urea, from 9.37 to 6.25 U/L and from 32.13 ± 8.72 to 22.13 ± 7.8 mg/dL, respectively, was observed (*p < 0.05). A significant decrease, from 0.22 to 0.02 μg/mL, in mean OTC serum concentration was also observed (**p < 0.01). Composition analysis revealed the presence of OTC, calcium, aluminium, silicon, and phosphorous nanoparticle aggregates. Further research on a wider sample size would help to confirm the hypothesis proposed here.

This study aimed to isolate, define the genetic profile, assess potential pathogenicity and evalu... more This study aimed to isolate, define the genetic profile, assess potential pathogenicity and evaluate the seasonal distribution of Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus strains isolated from the Vibrata river (Abruzzo Region, Italy) during a monitoring period of one year. Detection was performed according to ISO/TS 21872-1-2:2007. Species identification and characterisation were achieved using molecular methods. Vibrio spp. were detected in 50% (23) of the water samples. In particular, V. cholerae, V. parahaemolyticus, and V. vulnificus were isolated in 18 (39.1%), 4 (8.7%), and 2 (4.3%) samples, respectively. All V. parahaemolyticus strains were tdh gene negative, 75% were positive for trh gene. In 30 V. cholerae isolates, the polymerase chain reaction (PCR) assay for detecting virulence and regulatory genes (ctxA, toxR, tcpA, ompU, hlyA, tcpI, zot, and stn/sto) revealed 6 genotypes associated to different levels of pathogenicity. Pulsed-field gel electrophoresis (PFGE) characterisation of the V. cholerae strains identified 13 different pulsotypes. A greater degree of similarity was shown for strains isolated in the same period of the year. Results of our study reveal a potential health risk associated with the waters of the Vibrata river, which are used for irrigation and next to the swimming areas of Abruzzo coastline.

Widely spread in nature, Yersinia enterocolitica (YE) is a foodborne pathogen of major health and... more Widely spread in nature, Yersinia enterocolitica (YE) is a foodborne pathogen of major health
and economic significance in developed countries. The aim of this study is to analyse YE strains
isolated from 400 slaughtered pigs from the Abruzzo region, Italy, using biochemical tests
and a multiplex polymerase chain reaction PCR detecting 6 chromosomal genes (ystA, irp2,
16s, ail, inv, hemR) and one plasmid‑borne virulence gene (yadA). Antimicrobial susceptibility
was evaluated and pulsed‑field gel electrophoresis (PFGE) was also performed in order to
assess phylogenetic diversity. In total, 56 samples of porcine tonsils (14%) were found to be
positive for the presence of pathogenic YE. All YE belonged to the pathogenic bioserotype
4/O:3. All YE samples were positive for the chromosomal virulence genes ystA, ail, and inv,
whereas results for the presence of yadA and hemR were variable. This study found that YE
isolates were resistant to ampicillin (100%), streptomycin (26.79%), sulfisoxazole (19.65%),
tetracycline (16.08%), nalidixic acid (14.30%), and chloramphenicol (10.72%). The strains
characterised by PFGE showed a high similarity. This study demonstrates the usefulness of
multiplex polymerase chain reaction (PCR) compared with conventional phenotypic assays
for the identification of pathogenic YE isolates and the limitations of PFGE for the molecular
typing of YE bioserotype 4/O:3.

In the South Mediterranean and Middle East region, interactions between humans, animals, and the ... more In the South Mediterranean and Middle East region, interactions between humans, animals, and the surrounding environment are frequently close. This fact is mainly manifested in traditional farming settings (by sedentary, semi‑sedentary, and nomadic communities) as well as where livestock intensification has been introduced. A combination of complex factors in these settings (e.g. challenges in country infrastructures and cross‑sectoral collaboration/ coordination, traditional habits, poor social information, etc.) contribute to the emergence and, occasionally, to the endemic pattern of zoonoses. The phenomenal growth of international travel and trade, population displacement, and unhygienic settlements has increased the speed and ease with which pathogens and vectors can cross continents and cause outbreaks and epidemics. Programmes for the prevention and control of zoonoses have been implemented in several countries in this region; however, the expected results have not always been realised. The conflicts and civil unrest affecting certain countries in this region during the last decade, together with the mass displacement of people seeking refuge, have resulted in serious epidemiological and social impacts. Zoonoses and related food‑borne diseases are, indeed, a worldwide challenge, whose prevention and control mainly depend on the actions of national authorities. Once peace has been established in this region, authorities need to address the burden of these diseases through resource mobilisation, the implementation of international agencies' technical guidance, and inter‑country collaboration.

Bluetongue (BT) and African horse sickness (AHS) are considered the most important orbiviral dise... more Bluetongue (BT) and African horse sickness (AHS) are considered the most important orbiviral diseases in Southern Africans countries. The general endemic status makes these diseases challenging to be quantified in terms of their economic impact. Using country reported data from BT and AHS outbreaks and cases, as well as international trade data, the economic impact of BT and AHS is evaluated on local, regional, and global scales. Local scale impact in the Southern African region is underestimated as shown by the underreporting of BT and AHS. Exceptions occur during epidemic cycles of the diseases and when the diseases impact regional animal movement and global trade, as in the case of AHS in South Africa. While BT is not directly implicated as a significant non-tariff barrier for regional movement, there are unspecified clauses in import permits which refer to the 'OIE listed diseases' and the freedom thereof includes endemic diseases like BT. African horse sickness has a much more tangible regional and global economic impact because of movement restrictions within AHS control zones in South Africa and through international movement of horses from this country.

Epizootic haemorrhagic disease (EHD) is the most important infectious disease of white-tailed dee... more Epizootic haemorrhagic disease (EHD) is the most important infectious disease of white-tailed deer (WTD), however little is known about the role of inflammatory mediators in the pathogenesis. We characterized the expression of tumor necrosis factor-alpha (TNF-α) ex vivo in tissues of WTD experimentally or naturally infected with EHD virus serotype 2 and in WTD peripheral blood mononuclear cells (PBMC) infected with EHD virus serotype 2 in vitro. Circulating levels of TNF-α were evaluated in serum from experimentally infected deer via cytotoxicity assay. The expression of TNF-α in tissues was evaluated via immunohistochemistry (IHC) in both experimentally and naturally infected deer. Semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) was used to assess the level of TNF-α mRNA in tissues from experimentally infected deer and WTD's PBMC. Circulating levels of TNF-α were not increased in infected animals and TNF-α was not detected in tissues of infected deer. Increased transcription of TNF-α was detected neither in infected WTD nor in the PBMC. Tumor necrosis factor-alpha may not play a significant role in the pathogenesis of EHD virus infection in WTD.

Epizootic haemorrhagic disease virus (EHDV) replicates in endothelium and it has been shown that ... more Epizootic haemorrhagic disease virus (EHDV) replicates in endothelium and it has been shown that EHDV serotype 2 (Ibaraki) is able to cause cell death by apoptosis in cow pulmonary artery endothelial cells. However, the underlying mechanism has not been established. For some viruses, such as influenza, a p53 dependent mechanism has been demonstrated in viral induced apoptosis. In this study, we investigate the involvement of p53 in the induction of apoptosis in a US isolate of EHDV serotype 2 in cow endothelium. We inoculated cow carotid artery endothelial cell cultures with live and inactivated EHDV‑2 isolated from a white‑tailed deer (Odocoileus virginianus). Using in situ nick end‑labeling (TUNEL), caspase‑3 (cleaved) immunohistochemistry (IHC), flow cytometry and annexin staining we documented the development of apoptosis and its direct relation to viral replication. p53 gene regulation and protein expression were assessed by reverse transcription polymerase chain reaction and IHC, respectively, in infected cells. We show that p53 mRNA was not upregulated and protein expression was not significantly increased. No increase of p53 mRNA or protein expression was observed in cells that stained positive for EHDV antigen. Our results indicate that EHDV induces apoptosis through a p53 independent mechanism.

Viruses of the Simbu serogroup cause lesions to foetuses that are seen at birth and that correlat... more Viruses of the Simbu serogroup cause lesions to foetuses that are seen at birth and that correlate with the stage of pregnancy at which the dam first contracts the virus. The Simbu serogroup comprises arboviruses known to cause outbreaks of abnormal parturitions in domestic ruminants; these abnormalities include abortion, stillbirth, and congenitally deformed neonates. Simbu serogroup members include: Akabane virus (AKAV), Aino virus, Cache Valley virus, and Schmallenberg virus. Lately, dairy herds calf malformations have been observed in Europe, where there have been reports of clinical manifestations such as diarrhoea, fever, and reduced milk yield in adult lactating cows. The Israeli dairy cattle industry has experienced 2 major episodes of abnormal parturitions that resulted from 2 arboviral Simbu serogroup episodes, which occurred 35 years apart. A wave of apparently newly introduced AKAV was noted from the beginning of January 2012. Investigations carried out throughout the period of late Summer 2011 to early Winter 2012, associated the Israeli AKAV strain with central nervous system manifestations in lactating cows. A lack of clinical/epidemiological 'uniformity' among the AKAV infections was noted during these investigations. Here we describe and discuss the clinical and spatial distribution differences found among the 3 above-mentioned outbreaks. Comparable features in the clinical presentation, spatial distribution, and target-animal issues relating to Akabane disease are discussed.

Bluetongue (BT), an arthropod‑borne viral disease of ruminants, affects sheep most sev... more Bluetongue (BT), an arthropod‑borne viral disease of ruminants, affects sheep most severely than other domestic animals. Bluetongue virus serotype 24 (BTV‑24) is one of 26 known Bluetongue virus (BTV) serotypes. In this article, we present data of phylogenetic
analysis of 9 viral genes (Seg1, Seg2, Seg3, Seg4, Seg5, Seg6, Seg8, Seg9, and Seg10) from 8 Israeli BTV‑24 isolates and relate the genotype of the BTV‑24 isolates to their phenotype with regard to clinical manifestations. The high level of genetic identity (> 99.6%) between
Seg2, Seg4 and Seg5 in all 8 BTV‑24 isolates indicated that these segments shared the same viral ancestor. Phylogenetic analysis of Seg1, Seg3, Seg5, Seg8, Seg9, and Seg10 revealed that the Israeli BTV‑24 strains comprised 4 variants. Five of the viruses revealed high identity among all 9 segments, and represented variant 1. A second variant (BTV24/3027/6/10), isolated in 2010, showed significant variation from variant 1 in 3 gene segments (VP‑1,
VP‑3, and NS‑3 genes). A third variant (BTV24/3027/1/10) showed significant variation from variant 1 in 6 segments (VP‑1, VP‑3, VP‑6 and NS‑1, NS‑2 and NS‑3 genes), while a fourth variant (BTV24/2214/1/10) showed significant variation from variant 1 in 4 segments (VP‑1, NS‑1, NS‑2 and NS‑3 genes). These marked differences in sequence identity indicate that a high level of genetic reassortment is occurring between co‑circulating BTV strains in Israel.

In January 2013, an outbreak of Bluetongue (BT) affecting a Lacaune sheep flock occurred in Vasso... more In January 2013, an outbreak of Bluetongue (BT) affecting a Lacaune sheep flock occurred in Vassouras, Rio de Janeiro state, Brazil. From March to August 2013, blood samples collection and clinical examination were performed monthly, in order to monitor the epidemiological profile of Bluetongue virus (BTV) circulation and clinical disease in the flock. Agar gel immunodiffusion (AGID) and reverse transcription polymerase chain reaction (RT‑PCR) targeting BTV segment 10 were used as diagnostic assays. Additionally, insect trapping was conducted in the farm from May to July 2013. The flock serological prevalence to BTV was 80% since the first month of monitoring, with a variation in the serological rate depending on the sheep age categories. The number of susceptible lambs increased with time, probably due to the decrease of passive immunity. Viral RNA was detected in blood samples, demonstrating viral circulation, prolonged viraemia, and potential source for virus transmission in the region, even in a dry and cool season. The presence of Culicoides pusillus and Culicoides insignis was confirmed in the farm. The emergence of this outbreak in a Brazilian endemic area for BTV emphasises an urgent need of animal surveillance and BTV epidemiological studies.

Bluetongue (BT) is one of the most economically important transboundary animal diseases. In recen... more Bluetongue (BT) is one of the most economically important transboundary animal diseases. In recent years, it has been considered a disease related to climate change. A study was undertaken in 2013 in Nepal to measure the prevalence of Bluetongue virus (BTV) infection among domestic ruminants inhabiting the 3 agro-climatic zones with altitudes ranging from 150 to 2,400 metres above sea level. Twelve clusters representing the 3 altitudes were selected. The presence of antibodies against BTV was demonstrated in serum samples of sheep, goats, cattle, buffaloes, yaks/chauries, and chyangra goats (Himalayan goat) of Nepal. For this purpose, a total of 2,084 sera were collected from a population of 202 sheep, 739 goats, 590 cattle, 379 buffaloes, 105 yaks/chauries, and 69 chyangra goats between February 2013 and January 2014. The presence of antibodies against BTV was investigated using competitive enzyme-linked immunosorbent assay (c-ELISA). Of the 2,084 collected sera, 45.20% were positive for BTV antibodies. Species-wise prevalence was 17.82%, 47.50%, 53.05%, 58.05%, 7.62%, and 20.29% in sheep, goats, cattle, buffaloes, yak, and chyangra goats, respectively. Contrary to the general belief, maximum numbers of seropositive cases were recorded in buffaloes followed by cattle, goats, chyangra goats, sheep, and yak/chauries. The samples collected in the post-monsoon period (July-August is the monsoon period) show a seroprevalence higher than the pre-monsoon samples. This study shows the seroprevalence of BT in domestic ruminant population of Nepal at all altitudes. The highest prevalence has been reported in the plains of Terai followed by gradual decline in the mid-hills, and in the high mountains. Furthermore, detection of antibodies against BTV in both small and large ruminants (chyangra goats and yak/chauries) dwelling in high altitudes in the absence of BT vaccination is suggesting vector movement to the highlands as a consequence of warmer climate. These findings suggest that the climatic conditions, even at the higher elevation, are suitable for the survival of biting midges responsible for the transmission of BTV.

Bluetongue (BT) is a non-contagious arthropod-borne viral disease of domestic and wild ruminants.... more Bluetongue (BT) is a non-contagious arthropod-borne viral disease of domestic and wild ruminants. It is endemic to India and clinical outbreaks of disease have been reported mainly in sheep, although BT is often asymptomatic in other ruminant species. In the present serological survey, a total of 576 serum samples, comprising of 416 cattle and 160 sheep, covering different agro-climatic zones of Rajasthan, Uttar Pradesh, and Karnataka states, were screened for the presence of Bluetongue virus (BTV) specific antibodies using competitive enzyme-linked immunosorbent assay (c-ELISA). Overall 73.08% (304/416) of the cattle and 53.30% (87/160) of the sheep serum samples were positive for BTV antibodies. The prevalence of BTV antibodies in cattle in different agro-climatic zones ranged between 60-80% in Rajasthan and 66-70% in Uttar Pradesh. During the study, a nested polymerase chain reaction (PCR) based on the BTV NS1 gene (genome segment 5) was optimized for detection of BTV's nucleic acid from a cell adapted strain of BTV-23, and field derived clinical blood samples. In the present study, 19/70 of cattle and 9/30 of sheep blood samples tested positive for BTV RNA by the nested PCR, which amplified specific products of 274 bp and 101 bp sizes, respectively. From this study, it can be concluded that cattle showed higher percentage of sero-positivity in comparison to sheep. The improved sero-surveillance system for BTV in endemic areas will be of great help to understand the epidemiology of BTV and to formulate effective control and preventive strategies.

High sheep population density, congenial climatic conditions for Culicoides propagation, and sus... more High sheep population density, congenial climatic conditions for
Culicoides propagation, and susceptible sheep breeds may be contributing to the higher incidence of Bluetongue (BT) in Southern states of India. Sheep farming in this part of the country is nomadic in nature and BT is one of the major infectious diseases inflicting huge losses. Andhra Pradesh is one of the Southern states with high sheep population in India. Although isolation studies in this
region were started in 1993, concerted efforts only began in 2002. More than 50 isolates were obtained in the last decade, and 7 Bluetongue virus (BTV) serotypes (1, 2, 9, 10, 12, 16 and 21) were isolated. Among them, BTV-10, BTV-12, and BTV-21 were reported for the first time from India and the genome analysis of these viruses revealed that BTV-10 and BTV-12 have high sequence identity with the modified live virus (MLV) vaccines used in USA and
South Africa, respectively. At the same time, BTV-21 has probably originated from Southeast Asia. Furthermore, some of the BTV isolated from Europe have high sequence identity with viruses isolated from Andhra Pradesh indicating common ancestry. The analysis of different
isolates involved in outbreaks revealed that more than 1 BTV serotype is involved and that mixed infections with different serotypes is not uncommon. In a limited study conducted during 2005-2009, it was observed that most of the sheep seroconverted to more than
1 serotype, which further supports circulation of multiple serotypes and mixed infections in Andhra Pradesh. Based on the virus isolation data, in this study it was observed that a few serotypes dominate for 3-4 years followed by domination of others. Continuous monitoring
of circulating serotypes is essential to understand the distribution and spread BTV in endemic areas and for devising suitable control measures.

Tamil Nadu is located in the South-Eastern part of Indian peninsula, between 8.087° ... more Tamil Nadu is located in the South-Eastern part of Indian peninsula, between 8.087° and 13.09°N and 76.50° and 80.27°E. Bluetongue (BT) was first reported in this region in sheep
during 1982 with regular occurrence thereafter. In 1989-1990, 1997-1998 and 2005-2006, there was wide spread occurrence of BT resulting in huge mortality of sheep. The present study had the goal of isolating the BTV from outbreaks in sheep occurred in Tamil Naadu
between 2003-2011 and comparing the VP2 gene sequences of the BTV isolates involved in such outbreaks. Serotypes 1, 2, 16, and 23 of the Bluetongue virus (BTV) have been isolated from sheep during BT outbreaks. BTV-16 has also been isolated in goats and cattle in the
region; BTV-2 isolated in Tamil Nadu has homology with BTV-2 isolated in Africa; whereas the BTV-23 isolated in this area has homology with BTV-23 from South East Asia, indicating that both Eastern and Western topotypes of BTV are circulating in ruminant population in Tamil Nadu.

Bluetongue virus (BTV) was sporadically isolated over a four year period (2010-2014) ... more Bluetongue virus (BTV) was sporadically isolated over a four year period (2010-2014) from several alpaca carcasses that were presented for necropsy at the Western Cape Provincial Veterinary Laboratory, South Africa. Typically, the affected animals had a history of acute dyspnoea and progressive weakness before death. Consistent hydrothorax and severe lung oedema in all lead to a preliminary diagnosis of Bluetongue, despite the absence of
ulceration and hyperaemia of the oral mucosa which is characteristic of this viral infection in sheep. The diagnosis was confirmed by virus isolation in embryonated eggs and subsequent sequencing of the extracted RNA. Assembled sequences were subjected to Blast analysis and two of the isolates could be verified as BTV 3. These cases, originating from the Western Cape Province of South Africa, represents the first official report of BTV infection in alpacas in Africa and demonstrates the susceptibility of the species to this disease when maintained in BTV endemic areas.

The seasonal abundance of Culicoides midges, the vector of Bluetongue and African horse sickness ... more The seasonal abundance of Culicoides midges, the vector of Bluetongue and African horse sickness viruses (BTV/AHSV) and the presence of viruses in midges were determined in 3 geographic areas in South Africa. In the Onderstepoort area, more than 500,000 Culicoides midges belonging to 27 species were collected. Eighteen midge species were collected throughout Winter and the presence of AHSV and BTV RNA in midges was detected using real time reverse transcription quantitative polymerase chain reaction. The nucleic acid of AHSV was found in 12 pools out of total pools of 35 Culicoides. Twenty-five Culicoides species were detected in the Mnisi area. The RNA of BTV was detected in 75.9% of the midge pools collected during Winter and 51.2% of those collected during Autumn. Antibodies for BTV were detected in 95% of cattle sampled using a competitive enzyme-linked immunosorbent assay (cELISA). The dominant species in these 2 areas was Culicoides imicola. Eight Culicoides species were collected in Namaqualand. Culicoides imicola represented the 0.9% and Culicoides bolitinos the 1.5% of total catches, respectively. Antibodies for AHSV were detected in 4.4% of 874 equines tested using an indirect ELISA. Results showed that transmission of AHSV and BTV can carry on throughout Winter and the outbreak may begin as soon as Culicoides populations reach a certain critical level.

Insecticidal sugar baits for mosquitoes and houseflies have proven efficacy to reduce insect popu... more Insecticidal sugar baits for mosquitoes and houseflies have proven efficacy to reduce insect populations and consequently, disease transmission rates. The new insecticidal sugar trap (IST) is designed specifically for controlling biting midge disease vector populations around livestock and near larval habitats. The trap operates by combining light-emitting diode (LED) technology with insecticidal sugar baits. The positive photo attraction of Culicoides elicited by the LEDs, draws the insects to the insecticidal sugar bait, which can be made from various commercial insecticide formulations (pyrethroids, neonicotinoids, etc.) or naturally derived formulations (boric acid, garlic oil, etc.) lethal to Culicoides. Insecticidal sugar trap advantages include: customizable LED lights, they can be used with several different oral insecticides that have different modes of action to help combat the evolution of pesticide resistance, screening on the trap reduces non-target insect feeding (for example bees and butterflies), targets males and females of the species because both must feed on sugar, and low energy LEDs and a solar panel reduce trap maintenance to refilling sugar baits, rather than replacing batteries. This article discusses key components of an IST, which increase the traps effectiveness for biting midge control.

In most instances, vaccination is accepted to be the most effective method of preventing Culicoid... more In most instances, vaccination is accepted to be the most effective method of preventing Culicoides-borne arbovirus transmission, as it has proven to be successful in large-scale campaigns. Under certain scenarios, however, vaccines require time to be developed and deployed or are not used due to financial, logistical or trade constraints. In the absence of vaccines, animal movement restrictions and techniques to reduce either the number of Culicoides biting livestock or their subsequent survival are the only responses available to prevent or reduce arbovirus transmission and spread. This review evaluates the progress made during the past 10 years in the development of Culicoides control techniques for this purpose and assesses their potential impact in reducing arbovirus transmission. In addition, the future prospects and challenges facing Culicoides control are examined and suggestions are made as to research directions and opportunities.

Within Northern California, Culicoides sonorensis is the major vector of Bluetongue virus (BTV) a... more Within Northern California, Culicoides sonorensis is the major vector of Bluetongue virus (BTV) and annual infection of livestock is distinctly seasonal (typically July‑November). Our recent studies compare the population dynamics of C. sonorensis midges with occurrence of BTV infection of C. sonorensis and sentinel dairy cattle throughout both the seasonal and interseasonal ('overwintering') periods of BTV activity. Spring emergence and seasonal abundance of adult C. sonorensis on the sampled farms coincided with rising vernal temperature. Intensive surveillance confirmed widespread infection of both sentinel cattle and vector midges during the August‑November period of seasonal BTV transmission. Bluetongue virus infection of parous female midges captured in traps set during daylight hours was also detected during the interseasonal period of virus activity, whereas there was no concurrent active infection of sentinel cattle during the overwintering period. The finding of BTV‑infected vector midges during mid‑Winter suggests that BTV can overwinter in Northern California by infection of long‑lived female C. sonorensis midges that were infected during the prior seasonal period of virus transmission and which, then, entered a quiescence in the fall (Autumn) and re‑emerged sporadically during the overwintering period. Notably, vertical transmission of BTV was not detected among progeny of midges infected in the laboratory nor in field‑collected larvae. In addition to defining the mechanism of BTV over‑wintering in a temperate region, the studies reviewed in this article also provide precise documentation of temporal changes in the annual abundance, dispersal and dynamics of BTV infection of Culicoides midges. Collectively these findings are critical to the creation of accurate predictive models of BTV infection in livestock and to development of sound abatement strategies.

As Bluetongue virus (BTV) spread in Italy following its first incursion in 2000, it soon became a... more As Bluetongue virus (BTV) spread in Italy following its first incursion in 2000, it soon became apparent that, besides Culicoides imicola, additional species of the subgenus Avaritia were involved as vectors, namely one or more of the species that belong to the so-called ‘Culicoides obsoletus group’, which comprises C. dewulfi, C. chiopterus, C. obsoletus sensu stricto, C. scoticus and C. montanus; the three last named species are considered generally as
forming the Obsoletus complex. This study presents the findings made over the last decade and more, within the Italian entomological surveillance program for Bluetongue. It describes the integrated morphological and molecular approach used to identify the species of the ‘C. obsoletus group’, maps in detail their relative abundances and geographic ranges in Italy, clarifies the hitherto unknown comparative seasonal abundances of C. obsoletus s.s.
and C. scoticus in a site in Central Italy, and provides further details on the potential vector status of five species of the ‘C. obsoletus group’, with emphasis on C. obsoletus s.s., C. scoticus and
C. montanus. Unlike the situation in Northern Europe, Culicoides dewulfi
and C. chiopterus are uncommon to rare in Italy. In contrast, the Obsoletus complex occurs abundantly throughout Italy, with
C. obsoletus s.s. being the most prevalent and ecologically adaptive of the three species making up the complex. A longitudinal study conducted at a site in Central Italy revealed that: (i) species of the Obsoletus complex prefer horses to sheep; (ii) their parity rates range from 10% (March) to 56% (November); (iii) throughout the year C. scoticus is consistently more abundant than C. obsoletus s.s.
; (iv) abundances in both, C. obsoletus s.s. and C. scoticus,
peak in May-June, with the peak of the latter species being more evident. Bluetongue virus was first isolated from wild caught midges of the Obsoletus complex in 2002. Thereafter,
pools of selected parous midges collected across Italy, and during multiple outbreaks of BT, have been found consistently PCR-positive for the virus. More recently, viral RNA has been detected in field specimens of C. dewulfi, C. obsoletus s.s., C. scoticus and C. montanus.

Light trapping for biting midges was performed on cattle farms in 3 provinces of We... more Light trapping for biting midges was performed on cattle farms in 3 provinces of Western
Russia in 2013. A total of 9,272, 2,457, and 10,245
Culicoides
midges were collected from farms
in Smolenskaya, Pskovskaya, and Vladimirskaya provinces, respectively. More than 99% of
collected midges belonged to either the Obsoletus or Pulicaris complexes. Species belonging to the Obsoletus complex were the most abundant in Pskovskaya, whereas species from the Pulicaris complex dominated the catches in Vladimirskaya and Smolenskaya provinces. The seasonal duration of adult activity in all 3 regions was approximately 4.5 months.

In 2012, six years after the previous epidemic, Bluetongue virus serotype 1 (BTV-1) re-emerged i... more In 2012, six years after the previous epidemic, Bluetongue virus serotype 1 (BTV-1) re-emerged
in Sardinia causing a limited number of outbreaks. Due to impossibility of implementing a
vaccination campaign, the BTV-1 then spread all over the island in 2013 with about 7,000
outbreaks and, in September 2013, the virus reached Central Italy, with a limited number
of outbreaks located along the Tyrrhenian coast. The surveillance system in place in Italy
detected viral circulation during the following winter, when a few seroconversions were
notified. Starting from mid July 2014, a huge number of outbreaks were reported and the
disease spread toward inland territories, affecting Umbria, Abruzzo and Marche. In 2014,
BTV-1 affected areas where
Culicoides
species belonging to the Obsoletus and Pulicaris
complexes were identified as main vectors. The analysis of temperature and rainfall in Central
Italy revealed a significant warmer winter (2013-2014) and a cooler and rainy summer season
(2014). These climatic aspects might have certainly favored the overwintering of the virus in
local vector or host populations in the Tyrrhenian coast, and, then, the spread of the virus to
the rest of Central Italy. However, the heavy circulation of BTV-1 and the severity of clinical
outbreaks recorded leave a number of ‘open questions’ that are currently under investigations.

Bluetongue, Schmallenberg and African horse sickness viruses are transmitted by vectors... more Bluetongue, Schmallenberg and African horse sickness viruses are transmitted by vectors belonging to the genus Culicoides (Diptera: Ceratopogonidae). Among this genus, species of the Obsoletus complex play a crucial role in Europe. In Italy the complex includes three species: Culicoides obsoletus sensu strictu, Culicoides scoticus and Culicoides montanus. These three sibling species were identified in 80 collection sites across Italy through a multiplex PCR test. Sixty-four sites were classified with a predominant species through a Bayesian approach. Environmental, topographic and climatic factors together with landscape metrics were investigated in each species group using the non-parametric Mann-Whitney test.
Culicoides obsoletus s.s. resulted the most widely distributed species across the peninsula (51 sites). This species was collected in many eco-climatic conditions, at a wide range of temperature and
altitudes, mainly in anthropogenic landscape. Conversely C. scoticus
collection sites were dominated by natural vegetated areas or forest, at medium higher altitudes, preferably in a wilder and more pristine environment (predominant species in 8 sites). Culicoides montanus was a rarer species, statistically predominant in only 3 sites located in Southern Italy, characterized by temperatures higher than the other species. These results, together with other on-
going researches on the vector competence of the species belonging to the Obsoletus complex, and with investigation of larval and breeding habitats, could greatly improve epidemiological knowledge of possible spreading Culicoides-borne viruses.

Nowadays, leptospirosis is a re‑emerging widespread infectious disease often underestimate worldw... more Nowadays, leptospirosis is a re‑emerging widespread infectious disease often
underestimate worldwide. The National Reference Centre for Leptospirosis (NRCL), at
the Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna, Brescia
(Italy), with the cooperation of all the other Istituti Zooprofilattici Sperimentali (IIZZSS),
evaluated the distribution of such important zoonosis in Italy. Serological data obtained
between 2010‑2011 by each laboratory were collected by the NRCL and discussed. Serum
samples collected from 43,935 animal specimens were analysed by the Microscopic
Agglutination Test (MAT), using a panel of 8 serogroups as antigens (Australis, Ballum,
Canicola, Grippotyphosa, Icterohaemorrhagiae, Pomona, Sejroe, Tarassovi). A MAT cut‑off
of 1:100 was used to identify the serological positivities, 6,279 sera showed positive titers.
Bovine (46.9%), swine (27.5%), ovine and goat (7.4%), dog (6.9%), and wild boar (4.5%)
samples were delivered to the Laboratories more frequently than equine and other species
sera. Data analysis showed that the most common serogroups in Italy are: Australis present
in dogs, wild boars, horses, hares, swine, foxes, and rodents; Sejroe detected in cattle, sheep,
goats, and buffaloes; Icterohaemorrhagiae present in dogs, goats, and foxes; Pomona
detected in swine, cattle, and wild species; Grippotyphosa reported in hares.

Multilocus enzyme electrophoresis is the tool most frequently used to classify Leishmania spp., a... more Multilocus enzyme electrophoresis is the tool most frequently used to classify Leishmania spp., although it is time consuming and, sometimes, a not enough discriminative method. In the present study a kinetoplast DNA polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to characterize 16 zymodeme MON-1 Leishmania infantum strains: 15 were from dogs housed in public kennels of 7 geographical areas in the Emilia-Romagna region, Northern Italy, 1 was the L. infantum reference strain MHOM/TN/1980/IPT1. Six enzymatic patterns were observed. Kinetoplast DNA RFLP-PCR confirmed to have a good discriminatory power within the same zymodeme and proved to be useful for comparing few strains or discriminating between relapse and reinfection in the same host. We therefore recommend it use for discriminating between relapse and reinfection in the same host rather than supporting large-scale epidemiological studies.

Babesiosis due to Babesia bigemina is a relevant tick-borne disease, affecting cattle worldwide. ... more Babesiosis due to Babesia bigemina is a relevant tick-borne disease, affecting cattle worldwide.
Many surface proteins of the pathogen including the Apical Membrane Antigen 1 (AMA-1) -
have been analysed for vaccine and diagnostic purposes. This study focused on B. bigemina
AMA-1 and on its use for the assessment of diagnostic tests. After bioinformatic analyses,
AMA-1 codifying region was amplified and cloned into an expression vector used to induce
protein synthesis in Escherichia coli cells. AMA-1 was purified by affinity chromatography
and used to set up the best condition for an ELISA protocol. Bovine field sera positive to B.
bigemina were used to evaluate the presence of anti-AMA-1 antibodies. In order to verify
the assay specificity, sera positive to Babesia bovis or to the piroplasm Theileria annulata
were also included. Significant differences were obtained between sera negative to both B.
bigemina and B. bovis and samples positive to B. bigemina, to B. bovis or to both pathogens.
No significant reaction was observed with T. annulata positive sera. The results showed that
AMA-1 protein is suitable to be used as antigen in diagnostic assays for babesiosis diagnosis
in cattle, as it does not show any cross reaction with anti-T. annulata antibodies

Small breed dogs are 12 times more likely to develop medial patellar luxation (MPL) than large br... more Small breed dogs are 12 times more likely to develop medial patellar luxation (MPL) than large breed dogs and breed predisposition has been reported. Many surgical techniques are available for correction of patellar luxation in dogs. However, recent studies reported an 8% incidence of reluxation when traditional techniques are used. The relatively high frequency of major complications and patellar reluxation may be partially caused by inadequate appreciation of the underlying skeletal deformity and subsequent incorrect selection and application of traditional techniques. The aims of this study were to report the normal values of the anatomic and mechanical joint angles of the femur and tibia in small breed dogs and to compare these data to a population of small breed dogs affected by different degrees of MPL. Normal values of the anatomic and mechanical angles of the femur are similar to the ones reported in literature in Pomeranian dogs. Normal values of the anatomic and mechanical angles of the tibia have been described for the first time. Significant differences were found between normal population and dogs affected by grade 4 MPL in relation to anatomical Lateral Distal Femoral Angle (aLDFA), mechanical Medial Proximal Tibial Angle (mMPTA), and mechanical Caudal Proximal Tibial Angle (mCaPTA).

Given the limited information regarding the anatomy of the thoracic limb in European avian specie... more Given the limited information regarding the anatomy of the thoracic limb in European avian species, we decided to investigate the related muscles in the grey heron (Ardea cinerea), in the eurasian buzzard (Buteo buteo), and in the common kestrel (Falco tinnunculus). Therefore we performed a stratigraphic dissection of the wing in 3 subjects. The pars major and minor of the musculus deltoideus, despite being roughly in line with those reported by other authors in other species, displayed unique features. Concerning the pars propatagialis of the musculus deltoideus, from what was observed in the grey heron, we believe this structure can contribute to maintain the propatagial tension. In this way vibrations of this structure, which could cause diminished lift, are avoided. Moreover the peculiarity evidenced in the distal insertion of the common kestrel could influence the control of the pronation-supination of the wing during hovering. With respect to the musculus flexor carpi ulnaris, we believe the presence of a sesamoid-like structure at the base tendon, found in the grey heron and in the eurasian buzzard, may help complete the articular surfaces of the elbow. This study shows interesting data on species not previously examined and provides a possible functional correlation between the peculiarity observed and the kind of flight of each species.

Progressive ethmoid hematoma is frequently reported among the pathologies of the upper airways in... more Progressive ethmoid hematoma is frequently reported among the pathologies of the upper airways in horses. While several hypotheses have been proposed such as repeated trauma, inflammatory processes and/or chronic infections, congenital phenomena, complication of hemangiomas, and neoplastic processes, the aetiology of this condition is yet unknown. Clinical symptoms are non‑specific and includes haemorrhagic nasal discharge, respiratory noise not related to physical exercise and, in most severe cases, phenomena of coughing. The purpose of this case report is to contribute to a correct diagnostic approach to this particular pathology of upper respiratory tract in horses. Magnetic Resonance allows for a very detailed imaging of the development of the disease and suggests appropriate therapeutic choices.

Riassunto Il termine «macellazione religiosa» comunemente fa riferimento alla pratica di macellaz... more Riassunto Il termine «macellazione religiosa» comunemente fa riferimento alla pratica di macellazione senza stordimento, in accordo con i precetti sanciti delle religioni ebraica e musulmana. Scopo della presente ricerca è fornire una panoramica generale della situazione italiana in materia di macellazione religiosa di bovini, ovi‑caprini e avicoli non preventivamente storditi nel 2012. Il lavoro è stato suddiviso in due fasi: la prima fase prevede la raccolta di informazioni generali sullo stabilimento mediante la compilazione di un questionario da parte dei mattatoi italiani abilitati alla macellazione rituale. La seconda fase, invece, consiste nel campionamento sul posto di animali macellati ritualmente senza preventivo stordimento in 5 mattatoi selezionati. Tutte le informazioni riguardanti la gestione, le tecniche di contenimento e le informazioni sulle pratiche effettuate sono state raccolte e analizzate tenendo particolarmente in considerazione la tematica del benessere animale. Pratiche di macellazione religiosa in Italia Summary The term 'religious slaughter' commonly refers to the practice of killing animals without stunning, according to the precepts of Jewish and Muslim religions. The aim of this paper is to assess the situation concerning ritual slaughtering in not‑stun bovines, small ruminants, and poultry in Italy in 2012. The study was divided into 2 phases. During the first phase, preliminary data about all slaughterhouses authorized for ritual slaughter in Italy in 2012 are collected through the compilation of a questionnaire sent to each plants. The second step involved a sampling of not‑stun animals religiously slaughtered in 5 selected plants. Authors collected and compiled all informations about management, restrain system and rite taking into account in particular animal welfare.

The myostatin (MSTN) gene encodes a protein known to be a negative regulator of muscle mass in ma... more The myostatin (MSTN) gene encodes a protein known to be a negative regulator of muscle mass in mammalian species. Different polymorphisms of the horse (Equus caballus) MSTN gene have been identified, including single nucleotide polymorphisms and a short interspersed nuclear element (SINE) insertion of 227 bp within the promoter of the gene. The SINE insertion has been associated with performance traits in Thoroughbred racehorses and it was proposed as a predictor of optimum racing distance. The aims of this study were to perform in silico analysis to identify putative gains or abrogation of transcription-factor binding sites (TFBSs) generated by the SINE allele of the promoter and to analyse the frequency of the SINE insertion in horses used for racing (gallop and trot) and other purposes. The SINE insertion was genotyped in 227 horses from 10 breeds belonging to different morphological types (brachimorphic, mesomorphic, meso-dolichomorphic and dolichomorphic). The presence of the insertion was confirmed in the Quarter Horse (SINE allele frequency of 0.81) and in the Thoroughbred (0.51), whereas the SINE allele did not segregate in any of the other analysed breeds. As the SINE MSTN gene polymorphism may be population or breed specific, it is not a useful marker for association studies in all breeds.

Small breed dogs are 12 times more likely to develop medial patellar luxation (MPL) than large br... more Small breed dogs are 12 times more likely to develop medial patellar luxation (MPL) than
large breed dogs and breed predisposition has been reported. Many surgical techniques are
available for correction of patellar luxation in dogs. However, recent studies reported an 8%
incidence of reluxation when traditional techniques are used. The relatively high frequency
of major complications and patellar reluxation may be partially caused by inadequate
appreciation of the underlying skeletal deformity and subsequent incorrect selection and
application of traditional techniques. The aims of this study were to report the normal values
of the anatomic and mechanical joint angles of the femur and tibia in small breed dogs and
to compare these data to a population of small breed dogs affected by different degrees
of MPL. Normal values of the anatomic and mechanical angles of the femur are similar to
the ones reported in literature in Pomeranian dogs. Normal values of the anatomic and
mechanical angles of the tibia have been described for the first time. Significant differences
were found between normal population and dogs affected by grade 4 MPL in relation to
anatomical Lateral Distal Femoral Angle (aLDFA), mechanical Medial Proximal Tibial Angle
(mMPTA), and mechanical Caudal Proximal Tibial Angle (mCaPTA).

Given the limited information regarding the anatomy of the thoracic limb in European avian specie... more Given the limited information regarding the anatomy of the thoracic limb in European avian
species, we decided to investigate the related muscles in the grey heron (Ardea cinerea), in
the eurasian buzzard (Buteo buteo), and in the common kestrel (Falco tinnunculus). Therefore
we performed a stratigraphic dissection of the wing in 3 subjects. The pars major and minor
of the musculus deltoideus, despite being roughly in line with those reported by other
authors in other species, displayed unique features. Concerning the pars propatagialis of the
musculus deltoideus, from what was observed in the grey heron, we believe this structure can
contribute to maintain the propatagial tension. In this way vibrations of this structure, which
could cause diminished lift, are avoided. Moreover the peculiarity evidenced in the distal
insertion of the common kestrel could influence the control of the pronation‑supination of
the wing during hovering. With respect to the musculus flexor carpi ulnaris, we believe the
presence of a sesamoid‑like structure at the base tendon, found in the grey heron and in the
eurasian buzzard, may help complete the articular surfaces of the elbow. This study shows
interesting data on species not previously examined and provides a possible functional
correlation between the peculiarity observed and the kind of flight of each species.

Cross-sectoral assessment of health risks arising or existing at the human-animal interface is cr... more Cross-sectoral assessment of health risks arising or existing at the human-animal interface is crucial to identifying and implementing effective national disease control measures. This requires availability of information from 4 functional information 'streams' – epidemiological, laboratory, animal, and human health. The Food and Agriculture Organization of the United Nations (FAO)/ World Organisation for Animal Health (OIE)/ World Health Organization (WHO) Four-Way Linking (4WL) project promotes the establishing of a national-level joint framework for data sharing, risk assessment, and risk communication, in order to both improve communications within and among governmental public health and animal health influenza laboratories, epidemiology offices, national partners, with the aim of strengthening the national capacity to detect, report and assess risks arising from emerging influenza viruses. The project is currently being implemented in countries where H5N1 avian influenza is endemic and where human cases have been reported. The project is comprised of two main activities at country level: a 'review mission', which is the project launch in the country and has the objective to assess the existing situation; and a 'scenario based workshop', with the scope to bring together key national partners and build relationships among people working in the 4 information streams and to improve understanding of national strengths and gaps. During the workshop the delegates engaged in interactive sessions on basic risk assessment and devoted to specify the needs and roles of the 4 different streams. The participants work through a mock influenza outbreak scenario, which practically illustrates how risk assessment and communication of an emergency at the animal-human interface is more effective when there is linking of the 4 streams, collaboration, communication, and coordinated action. In 2010, Egypt was the first country where the project was successfully implemented, followed by Vietnam and Indonesia.

The genetic diversity of Brucella melitensis and Brucella abortus strains isolated in 199 cattle ... more The genetic diversity of Brucella melitensis and Brucella abortus strains isolated in 199 cattle and sheep from 156 brucellosis outbreaks which occurred in 8 regions of Southern Italy in 2011, was determined using a Multiple-Locus Variable Number of Tandem Repeats Analysis (MLVA‑16) approach. The existence of possible genetic clusters was verified through a hierarchical cluster analysis based on 'single link', which is closely related to the minimum spanning tree. The Hamming weighted distance matrix was adopted in the analysis. All calculations were performed using R and the additional libraries phangorn and Cluster. For a number of clusters, ranging from 2 to 15, the average silhouette width was calculated. The number of clusters adopted was identified according to the maximum average silhouette width. For B. abortus and B. melitensis, 6 and 11 genetic clusters were identified, respectively. Three out of 6 B. abortus clusters included the 96.7% of all B. abortus isolates. Clusters were clearly geographically separated, and this highlighted the known epidemiological links among them. Brucella melitensis genotypes resulted more heterogeneous; the 3 more representative genetic clusters included 79.7% of all B. melitensis isolates. A clear geographical clusterization of genotypes is recognizable only for 1 cluster, whereas the others are more widespread across Southern Italy. The genetic characterization of Brucella strains isolated from animals may be a useful tool to better understand the epidemiology and dissemination patterns of this pathogen through host populations.

Feline morbillivirus was detected in urine samples of a 15 year old cat suffering from severe nep... more Feline morbillivirus was detected in urine samples of a 15 year old cat suffering from severe nephropathy. Viral RNA was not detected in blood and faecal samples and also the most common pathogens associated to cat kidney failure were not found. This report describes the first evidence of feline morbillivirus in Europe.

In the last decades, adverse food reactions have increased considerably in dogs and cats. In this... more In the last decades, adverse food reactions have increased considerably in dogs and cats. In
this study we report on the possible onset of food intolerances symptoms, including otitis,
diarrhoea, generalised anxiety, and dermatitis in a cohort of 8 dogs consuming commercial
diets. All dogs received an organic chicken‑based diet for 15 days. We performed analysis
of blood biochemical parameters, kibble composition, and oxytetracycline (OTC) serum
concentration before and after 15 days of organic chicken‑based diet supplementation.
We hypothesised that a chronic intake of contaminated food enhanced by the presence of
nanoparticle aggregates might be at the base of the onset of pharmacologic or idiopathic
food intolerances. At the end of the evaluation period, an overall significant reduction of
otitis, diarrhoea, generalised anxiety, and dermatitis was observed. Biochemical analyses
indicate a significant increase in the alkaline phosphatase, from 41 to 52.5 U/L, after 15 days
(**p < 0.01), while a significant decrease in Gamma‑glutamyl transferase and urea, from
9.37 to 6.25 U/L and from 32.13 ± 8.72 to 22.13 ± 7.8 mg/dL, respectively, was observed
(*p < 0.05). A significant decrease, from 0.22 to 0.02 μg/mL, in mean OTC serum concentration
was also observed (**p < 0.01). Composition analysis revealed the presence of OTC, calcium,
aluminium, silicon, and phosphorous nanoparticle aggregates. Further research on a wider
sample size would help to confirm the hypothesis proposed here.

Pteropine Orthoreoviruses (PRVs) are fusogenic bat-borne orthoreoviruses that cause flu-like uppe... more Pteropine Orthoreoviruses (PRVs) are fusogenic bat-borne orthoreoviruses that cause flu-like
upper respiratory tract infections in humans. The presence of this group of viruses in bats
and humans has been well documented in areas where their biological reservoirs – fruit bats
(family Pteropodidae) – live densely. In the present study, a serum neutralization (SN) assay
to detect neutralizing antibodies against PRV Indonesia/2010 isolate was set up and used to
assess the seroprevalence of this virus in Italian domestic animals. The new developed assay
was able of detecting PRV neutralizing antibodies in the hyper-immune polyclonal serum
produced in rabbits (titer of 1:160). The negative serum was negative at all tested dilutions.
No cross-reactions have been evidenced neither against reference MRVs nor against their
respective hyper-immune sera. Eight hundred and fifty-three serum samples collected
from 524 bovines, 271 small ruminants, and 58 horses (all used as sentinel animals in the
Bluetongue and West Nile disease National surveillance program) were also tested with the
new developed SN assay. According to the results of this survey, neither PRV nor PRV crossreacting
viruses antibodies have been demonstrated in Italian domestic animals. However,
the new developed SN assay could be a very valuable diagnostic tool to detect infection in
animals and humans.

Highly pathogenic avian influenza (HPAI) virus H5N1 spread throughout Nigeria between 2006 and 20... more Highly pathogenic avian influenza (HPAI) virus H5N1 spread throughout Nigeria between
2006 and 2007. Bird samples collected across the country were submitted through the
free‑of‑charge (FOC) program to the National Veterinary Research Institute, Vom (NVRI‑Vom)
laboratory. The present article describes the spatial distributions and evaluated clustering of
the FOC submissions from poultry farms at the global, local, and focal levels between 2006
and 2007 epidemic in Nigeria. Spatial statistics evaluating clustering of the FOC submissions
were implemented using the Moran’s I test, the purely spatial cluster analysis with the
SaTScan Poisson model, and the Bithell's linear score test. A significant global clustering of
the FOC submissions was observed. Significant local clusters of submissions were observed
in the North‑East, North‑Central, and South‑West zones. There was significant decline in
FOC submissions with increasing distance from NVRI‑Vom. These results indicated that
the geographic area of influence of the FOC submission program in Nigeria was limited to
regions closer to the diagnostic laboratory. This work provides a detailed insight into the
surveillance activities during the HPAI outbreaks in Nigeria, and should assist policy‑makers
and field veterinarians to improve the effectiveness of national eradication plans in the face
of any outbreak of animal diseases.

Anaplasma and Ehrlichia spp. are tick‑transmitted bacteria of clinical relevance in European dogs... more Anaplasma and Ehrlichia spp. are tick‑transmitted bacteria of clinical relevance in European dogs. The diagnosis of infection is often difficult due to the wide spectrum of disease caused by them. During infection, reduction in platelet count is considered the most common haematological abnormality, frequently representing the sole alteration in asymptomatic dogs. In this study, the presence of bacteria belonging to the genera Anaplasma and Ehrlichia was investigated in Northern Italy in blood samples from 159 thrombocytopenic dogs using a polymerase chain reaction (PCR) assay amplifying a portion of the heat shock gene (groEL). Obtained amplicons were sequenced and analysed. Two dogs were positive for A. phagocytophilum, while A. platys and E. canis were not detected. None of the PCR‑positive dogs were diagnosed at the time of hospital admission, even in the presence of clinical signs and clinicopathological abnormalities potentially related to A. phagocytophilum infection. Nucleotide sequence analysis showed that the 2 detected strains belonged to the cluster Europe 1 and were different from each other. This study confirms the presence of A. phagocytophilum infections in dogs of Northern Italy, causing clinical signs and laboratory abnormalities that could not be properly diagnosed and treated.

Epizootic haemorrhagic disease virus (EHDV) was detected for the first time in Tunisia and in oth... more Epizootic haemorrhagic disease virus (EHDV) was detected for the first time in Tunisia and in
other Northern African countries in 2006. The objective of the present study was to investigate
whether EHDV circulated in Tunisian livestock before and after the officially‑reported outbreak
of 2006. Thus, serum samples from cattle and dromedaries collected in different time periods
(before and after 2006) and from different regions of Tunisia were screened for the presence
of EHDV antibodies. Serological investigations conducted on cattle and dromedary sera
collected in 2000 and 2001 demonstrated no virus circulation on these dates. However, viral
circulation was evidenced in 2012 and 2013, although no EHDV cases were officially reported
in these years. Serum‑neutralization assessed on few ELISA positive samples, confirmed the
presence of antibodies against EHDV serotype 6, which was the serotype involved in the
EHDV outbreak in the Maghreb region in 2006.

In this study a mink showing hard pad disease like symptoms was euthanised. Heart blood and vario... more In this study a mink showing hard pad disease like symptoms was euthanised. Heart blood and various tissue samples collected during necroscopy and tested by specific RT-PCR were found positive for CDV. H and F gene segments of the CDV strain was also partially sequenced using the appropriate primers, and subsequently the sequences were analysed and compared with same gene fragment sequence of other CDV isolates from different countries. The results of the phylogenetic analysis showed that the Turkish-Mink distemper strain is closely related to European CDV strains of lineage 1. Additionally, the distemper antigen was also detected when the tissue samples were examined by histology or immunohistochemistry.

The study described in this paper had the goal to validate the Italian version of the UNESP‑Botuc... more The study described in this paper had the goal to validate the Italian version of the UNESP‑Botucatu multidimensional composite pain scale (UNESP‑Botucatu MCPS) to assess postoperative pain in cats using video analysis and psycometric testing. The English version of the scale was translated into Italian. Thirty videos of the perioperative period of ovariohysterectomy surgery were analysed by 5 Italian observers with the aim to determine the pain score using the Italian version of the scale and to verify the need for analgesic treatment for each cat. Obtained scores were submitted to psycometric validity, responsiveness, and reliability tests. Of the 3 domains identified by factor analysis, the internal consistency was excellent for 'Psychomotor changes' and 'Protection of the painful area and vocal expressions of pain' , while 'Physiological variables' showed moderate internal consistency. Significant changes in pain scores in response to surgery and analgesics confirmed content and construct validity. The agreement between the 'gold standard' and the blinded observers supported the criterion validity. Inter‑ and intra‑rater reliability ranged from good to very good for all scale items. The cut‑off point for rescue analgesia was > 7. The study concluded that the Italian version of the UNESP‑Botucatu MCPS is a valid and reliable instrument for assessing postoperative pain in cats. The cut‑off point for rescue analgesia provides an additional tool for guiding analgesic therapy.

Strontium-90 (90 Sr) is a fission product, resulting from the use of uranium and plutonium in nuc... more Strontium-90 (90 Sr) is a fission product, resulting from the use of uranium and plutonium in nuclear reactors and weapons. Consequently, it may be found in the environment as a consequence of nuclear fallouts, nuclear weapon testing, and not correct waste management. When present in the environment, strontium-90 may be taken into animal body by drinking water, eating food, or breathing air. The primary health effects are bone tumors and tumors of the blood-cell forming organs, due to beta particles emitted by both 90 Sr and yttrium-90 (90 Y). Moreover, another health concern is represented by inhibition of calcification and bone deformities in animals. Actually, radiometric methods for the determination of 90 Sr in animal bones are lacking. This article describers a radiochemical method for the determination of 90 Sr in animal bones, by ultra low-level liquid scintillation counting. The method precision and trueness have been demonstrated through validation tests (CV% = 12.4%; mean recovery = 98.4%). Detection limit and decision threshold corresponding to 8 and 3 mBecquerel (Bq) kg-1 , respectively, represent another strong point of this analytical procedure. This new radiochemical method permits the selective extraction of 90 Sr, without interferences, and it is suitable for radiocontamination surveillance programs, and it is also an improvement with respect to food safety controls.

The aim of this study is to analyse and evaluate the knowledge concerning risk communication acqu... more The aim of this study is to analyse and evaluate the knowledge concerning risk communication
acquired by veterinarians during a national training course. The study analyses 694 press
releases written during the training course. Textual variables and indexes were considered to
identify the linguistic structures used by veterinarians. At first, the analysis of press releases
focused on the descriptive demographic variables, then stylistic and editorial variables were
considered, and finally a lexicographic analysis was performed on the textual variables.
The texts were found to have a moderate level of readability due to the use of scientific
and technical terms. This study shows the need to improve training activities to develop
effective risk communication, especially in terms of the language used, in order to facilitate
interactions among veterinarians, mass media, and citizens

Haemostasis is a physiological process that prevents excessive blood loss. In laboratory, the pro... more Haemostasis is a physiological process that prevents excessive blood loss. In laboratory, the prothrombin time (PT) and the activated partial thromboplastin time (aPTT) are used to examine clotting systems. However, the influence of sex on PT and aPTT values is unknown. The present work aimed to determine the values for PT and aPTT in adult sheep and to evaluate their dependence on the sex of the animal. Blood samples were collected from 40 adults (1-6 years old) of mixed breed sheep (20 males and 20 females) via jugular venepuncture conducted using vacuum tubes containing 3.8% sodium citrate as an anticoagulant. PT and aPTT were determined by visual detection of clot formation. The mean PT and aPTT values for all sheep were 7.71 ± 0.87 s and 35.7 ± 3.57 s, respectively. The aPTT values showed a significant difference (P = 0.0013) between male and female samples, while the difference in PT values was not significant (P = 0.0565). Thus, the animal sex influences the function of the plasma blood-clotting system in sheep. In contrast with table 1 data, in particular, aPTT values are significantly higher in female sheep than in males. Influence of sex on activated partial thromboplastin time (aPTT) and prothrombin time (PT) in sheep SHORT COMMUNICATION

All livestock transport within the European Union must comply with the EC Regulation 1/2005. For ... more All livestock transport within the European Union must comply with the EC Regulation 1/2005. For sheep, this law prescribes a maximum journey of 29 hours after which animals must rest in control posts (CP) for 24 hours before further transportation. However, there is no scientific evidence defining the effects of different stop duration on sheep recovery during long journeys. The aim of this study was to assess if a shorter stop could be provided without impairing ewes' welfare. Ninety-six adult ewes were divided into 4 homogenous groups. One group stayed at the farm (control) and the other 3 were transported for 29 hours (long-transport, LT), stopped at CP for different times (8 hours (S8 group); 16 hours (S16 group); 24 hours (S24 group)) and were re-transported for 6 hours (short-transport, ST). Blood and saliva were collected to assess dehydration, muscular damage, and adrenocortical stress before departure, after LT, after the stop, and after ST. The LT affected the hydration of all transported groups (i.e. higher BUN/creatinine levels than controls, p < 0.001), but basal values were restored after the ST, regardless of the stop duration. After the ST, S8 group had higher muscular damage than the other groups (p < 0.05). No differences in stress level were observed. These results suggest that, in this trial, ewe's welfare was not impaired by a stop reduction from 24 hours to 16 hours.

The efficacy of sweep nets and a CDC white light-suction trap for the sampling of Culicoides spec... more The efficacy of sweep nets and a CDC white light-suction trap for the sampling of Culicoides
species (Diptera: Ceratopogonidae) were compared on a livestock farm in Northern Spain
during the Summer of 2013. A total of 6,082 specimens representing 26 species were
collected with sweep nets in 4 areas at different heights (ground level, 1.5 m, and 3 m),
and 8,463 specimens representing 28 species with a single white light trap. Eight species -
Culicoides brunnicans, Culicoides punctatus, Culicoides obsoletus/Culicoides scoticus, Culicoides
lupicaris, Culcoides picturatus, Culicoides achrayi, and Culicoides simulator - were dominant and
accounted for 97.4% and 97.2% of the total specimens collected with both methods, sweep
nets, and light traps, respectively. The sex ratios with sweep netting and light trapping were
strongly female biased (78.4% and 97.1%, respectively). Nulliparous and parous females were
predominantly captured with both methods. A high percentage (17%) of gravid females was,
however, captured on manure at ground level while sweeping. Searches for male swarms
revealed the presence of several C. punctatus swarms consisting of 26 to 196 males and 3
swarms of C. obsoletus that ranged from 1 to 12 males in size. This study suggested that both
methods are suitable and complementary tools for Culicoides sampling.

This paper focuses on a research concerning the operational management of the pig-handling phase,... more This paper focuses on a research concerning the operational management of the pig-handling
phase, during the period of breeding and before slaughtering. Given the behaviour of these
animals during transfers, a particular tool has been designed to manage them in this phase. A
total number of 48 animals, divided in 4 groups, were moved without use of the tool (control
groups) and by using the tool described in this article. The time required by the control groups
to leave the pen ranges from 21 to 125 seconds; while, when the proposed tool was used, the
time for the movement of the animals ranged between 10 and 17 seconds. In particular, in
the groups where the tool was deployed the ‘waiting phase’ (before the first animal goes out)
lasted less than half of the time of the ‘waiting phase’ of the control group, thus showing a
minimization of the effects of the ‘panic phenomenon’ among the animals. Thus, the studied
device can be considered as valid guide technique, both for the quick exit of the first animal
and for those that follow. Once the row has been formed, the animals continue neatly to
leave the box. This study also shows that this solution can also be considered appropriate for
reducing the identified critical issues in the traditional handling. The need of only 1 worker
to move the group of pigs is important to achieve economic saving. The deployment of this
tool, thus, make possible to consider the movement of animals no longer a ‘critical stage’, but
as a routine step of the production cycle of pork’s meat.

Ticks (Acari: Ixodidae) are ectoparasites infesting livestock in every geographic area in the wor... more Ticks (Acari: Ixodidae) are ectoparasites infesting livestock in every geographic area in the world and they are vectors of several viral, bacterial, and protozoan pathogens to animals and humans worldwide. A deep knowledge of the geographical distribution of these arthropods would have a key role in the control of tick-borne diseases. Few data are available about tick presence in domestic ruminants in Lebanon. The study aimed at providing an analysis of tick presence and distribution in Lebanon. Ticks were collected from cattle, sheep, and goats farms distributed in 6 Lebanese provinces between June and September 2014. A total of 272 adult hard ticks were randomly collected from domestic ruminants (cattle, sheep, and goats) located at 37 Lebanese farms, distributed among 30 villages. Ticks belonged to 4 Ixodidae
genera: Rhipicephalus (72.4%), Haemaphysalis (11.4%), Dermacentor (8.1%), and Hyalomma
(8.1%). They included the following species: Rhipicephalus annulatus (50.7%), Rhipicephalus
turanicus (18.8%), Hyalomma anatolicum (8.1%), Haemaphylasis punctata (11.4%),
Dermacentor marginatus (8.1%), Rhipicephalus sanguineus (2.5%), and Rhipicephalus bursa
(0.4%). Rhipicephalus turanicus and H. anatolicum were found on cattle, sheep, and goats,
R. annulatus on cattle and sheep, R. sanguineus, D. marginatus and Hea. punctata on sheep
and goats, while R. bursa was collected only on sheep. Tick species involved in pathogen
transmission were found and some of the identified species were recorded in Lebanon for
the first time.

Dermestidae and Tenebrionidae are well known inhabitants of bird’s nests and poultry farms, under... more Dermestidae and Tenebrionidae are well known inhabitants of bird’s nests and poultry farms,
under favourable conditions they can be very abundant under favourable conditions. At
times, their larvae shift from a scavenging behaviour to a parasitic/predatory one, entering
nestling’s plumage and feeding on skin and feathers, and finally provoking skin damages
and blood losses. These episodes mainly involve species of the genus Dermestes, but the
tenebrionid Alphitobius diaperinus h also been reported to be responsible of similar cases.
In June 2014, a mortality of caged young pigeons occurred in a family farm in Central Italy.
Post-mortem examination of 1 of the dead nestlings revealed the presence, near the cloacal
orifice, of a triangular shaped hole of about 1 cm side, with rounded edges facing inward
and with bleeding from the cavity. Five coleoptera larvae 0.5-2 cm long were collected from
the edges of the hole. Bacteriological examination of liver, intestine, and lungs revealed
the presence of Escherichia coli in the lung samples. The 5 larvae were morphologically
identified as Dermestes bicolor (4) and Alphitobius diaperinus (1). This is the first reported
case of pigeon nestling’s mortality caused by Dermestidae and Tenebrionidae larvae acting
as parasites/predators in Italy.

Severe lung strongylosis was detected in a wild red fox (Vulpes vulpes) (1/12) from Apulia (Italy... more Severe lung strongylosis was detected in a wild red fox (Vulpes vulpes) (1/12) from Apulia (Italy). We performed routine diagnostics on 12 foxes found dead in Apulia. Eleven of them showed lesions consistent with a vehicle collision. However, the remaining fox appeared to have died from other causes. At necropsy we observed, catarrhal enteritis, fatty liver, lung congestion with some areas firm in consistence and brain haemorrhages and malacia. Histopathology revealed lung fibrosis with mononucleate cells infiltration, thrombosis a several larval nematodes spread in the parenchyma, interstitial nephritis, interstitial myocarditis, encephalitis, encephalomalacia, and a brain granuloma. The larvae recovered from the lung parenchyma were identified as the first stage larvae of Angiostrongylus vasorum. This is the first documented report of angiostrongylosis in a fox in Southern Italy.

This study reports the mosquito collections conducted in July‑August 2013 in Castiglione del Lago... more This study reports the mosquito collections conducted in July‑August 2013 in Castiglione del Lago (Umbria Region, Italy) during the entomological surveillance within the West Nile national control program. The mosquito collections showed the noteworthy occurrence of Anopheles hyrcanus s.l. (n = 156; 35.8% of the whole sample), a relatively rare mosquito species in Italy, with molecular analyses confirming that these mosquitoes belong to An. hyrcanus s.s. The same catching site was characterized by a relevant richness in mosquito species, in particular the following were found: Anopheles maculipennis s.l. (n = 146; 33.5% composed by 80% Anopheles melanoon, 13.3% Anopheles maculipennis s.s. and 6.7% Anopheles atroparvus), Culex pipiens s.l. (n = 116; 26.6% composed by 90% Cx. p. pipiens, 5% Cx. p. molestus and 5% Cx. p. pipiens/molestus), Culiseta annulata (n = 10; 2.3%), Aedes albopictus (n = 3; 0.7%), and Anopheles claviger (n = 5; 100%).

Ochlerotatus caspius is a mosquito of medical and veterinary relevance both for its synanthropy a... more Ochlerotatus caspius is a mosquito of medical and veterinary relevance both for its synanthropy and for its potential role in transmission of viruses and nematodes in the areas that it inhabits. Due to its wide range and the marked variability in the adult colour pattern, some authors have recognized Ochlerotatus caspius as a complex of species. In this study, we purposed to evaluate the possible taxonomic heterogeneity between 2 chromatic forms by using both morphological and molecular approaches. The preliminary results based on the identity of the rRNA internal transcribed spacer 2 (ITS‑2) lead us to believe the 2 forms as a single species with a chromatic polymorphism.

Molecular characterization and antimicrobial susceptibility of Pasteurella multocida strains isol... more Molecular characterization and antimicrobial susceptibility of Pasteurella multocida strains isolated from hosts affected by various diseases in Italy. Vet Ital. Summary Pasteurella multocida is a widespread pathogen associated with major animal diseases of economic significance. Despite this, little is known about the capsular types, virulence gene pattern, and antimicrobial susceptibility of isolates from hosts affected by different diseases, and no data are available in Italy. One hundred eighty six isolates of P. multocida, were taken from different species in different states of health in several Italian regions, and were tested for genes encoding for capsular types (cap) and major virulence factors (tbpA, toxA, hgbB and pfhA). Antimicrobial susceptibility was investigated with the agar diffusion test. The majority of isolates was capA+. However, the distribution differed according to species and disease of origin, with a greater heterogeneity in isolates from rabbits; capE was never found, while capB was detected once. Only capA+ and capF+ strains tested positive for pfhA. Conversely, almost all capD+ isolates were hgbB+. In bovine respiratory disease, pfhA+/tbpA+/capA+ isolates predominated, while tbpA+/toxA+/capD+ isolates predominated in sheep. Overall, low levels of resistance were found, with full susceptibility to ceftiofur and florfenicol. Lower susceptibility to older antimicrobials was recorded, since only approximately 1/3 of the isolates showed susceptibility to tylosin and erythromycin, and resistance to tetracycline (7.5%), and trimethoprim-sulphametoxazole (4.8%) was also observed.

The skin is the most requested product of ratite slaughter and ‘first-rate’ ratite’s ... more The skin is the most requested product of ratite slaughter and ‘first-rate’ ratite’s skin has
a high economic market. Ostrich hide is tanned in South Africa and sold all over the
world. Besides, the ostrich meat is considered a niche production in the marketplace, thus
allowing for another profit for the ostrich breeding. Theorical analysis of skinning process
has been carried out, and an automatic measurement system has been designed to record
the drawing-force on the animal’s body during skinning tests. The aim of this study was to
indentify the best animal position and provide values of drawing-force in order to design
a specific machine for ostrich skinning. Experimental results partially confirm the proposed
analysis. They point out that the angle of inclination of drawing-force with back-line mostly
influences the value of drawing-force and that this influence tends to increase as the angle
is near to 90° or exceeds this limit. In any case, the applied drawing-force must not exceed
350.0 daN. Therefore, with respect to their anatomy structure and the tensile strenght of
the hide, ostrich skinning should be done with vertical-drawing machines and the animal
should be suspended by its wings at the rail. In case of low working capacity (8-10 heads
per hour), given the difficoulties highlighted during the experiments run in this study, the
animal should be skinned suspended by the legs.

Campylobacter spp. are ubiquitous bacteria and campylobacteriosis is the most frequently reported... more Campylobacter spp. are ubiquitous bacteria and campylobacteriosis is the most frequently
reported zoonotic disease in humans in Europe, since 2005. Handling or consuming
contaminated/undercooked meat (especially poultry) are the most important sources of
human campylobacteriosis. However, in recent years, the role of dogs as a source of infection
for humans has been amply demonstrated. Approximately 6% of human campylobacteriosis
cases are due to contact with pets. This review analyses the current literature related to risk
factors at the dog‑human interface.

In this study, the prevalence of Campylobacter spp. in poultry fresh meat and ready‑to‑cook produ... more In this study, the prevalence of Campylobacter spp. in poultry fresh meat and ready‑to‑cook
products was evaluated. Seventy‑three samples were collected at retail level from
supermarkets and discount stores, obtaining 61.6% positivity. Of 133 Campylobacter isolates,
86 strains (Campylobacter coli, 58.1% and Campylobacter jejuni, 41.9%) were selected for
characterisation on the basis of their SmaI and kpnI pulsed field gel‑electrophoresis (PFGE)
profiles, to exclude clonal replicates. Campylobacters resulted highly resistant to tetracycline,
ciprofloxacin, and nalidixic acid (79.1%, 72.1% and 65.1%, respectively); 50% of C. coli and
13.9% of C. jejuni were resistant to ciprofloxacin and erythromycin, the most important
antimicrobials for human campylobacteriosis therapy. Five C. coli were resistant to 5/7 of
the tested antimicrobials. HS4c was the prevailing C. jejuni serotype group (22.3%), whereas
8 other serotypes were identified in low percentages. SmaI and kpnI profiles showed a
wide variability. The survey showed a high Campylobacter contamination of poultry meat
and poultry products at retail level in Tuscany, Italy. A wide strains’ heterogeneity and a
remarkable level of strains’ antimicrobial resistance have been reported, confirming the need
for an improvement of specific preventive measures along the production chain.

Bovine tuberculosis (bTB) is a chronic infectious disease that affects both domestic animals and ... more Bovine tuberculosis (bTB) is a chronic infectious disease that affects both domestic animals
and wildlife. Veterinary epidemiology studies evaluate bTB using geographic information
systems (GIS), which can characterise the spatial and temporal distribution of diseases and
identify the geographic areas and animal populations at risk of contracting a disease. The
present study used space‑time permutation scan statistic to identify the spatial and temporal
distribution of bTB from 2005 to 2010 in the State of Mexico with the goal of creating a
similarity model using Mahalanobis Distance to identify areas suitable for bTB occurrence.
Three significant clusters were identified using space‑time permutation scan statistic and
the similarity model identified several areas with suitable environmental and demographic
characteristics. The results demonstrate that the occurrence of bTB in the State of Mexico is
not randomly distributed.