Donato Antonio Raele

We evaluated the presence of antibodies against Orthonairovirus haemorrhagiae , formerly Crimean-Congo haemorrhagic fever virus (CCHFV) in the serum of podolic cattle from southern Italy. A subset of 20 sera (18 positive and 2 negative) from the initial 670 bovine samples screened by commercial ELISA kit were further evaluated by indirect ELISA and virus neutralization tests. Across the three serological assays, antibodies against CCHFV were detected in a proportion of samples, though the results were not fully concordant and varied according to the assay applied. These findings highlight the need for multiple tests to more accurately assess CCHFV circulation in free-range cattle, particularly in non-endemic areas, and emphasize the importance of careful interpretation when relying on serology alone.

The circulation of viruses of medical and veterinary importance is monitored to evaluate risks to both human and animal health. Among the species most commonly used in surveillance programs, the wild boar (Sus scrofa) plays a key role due to its high population density and its contacts with rural swine herds in the areas under investigation. In the present study, molecular (real-time PCR) and serological analyses were conducted on wild boar samples collected in the regions of Apulia, Basilicata, Campania, and Calabria. Specifically, molecular detection was applied to IAVs, FLVs, and Porcine PCV-2, while serological testing was used to assess exposure to IAVs, FLVs, and ADV. Serological results showed an 8.31% positivity for IAVs, whereas viral presence detected by PCR was 1.17%. No active infections were detected for WNV, with only a single seropositive sample (0.30%), while ADV showed a seroprevalence of 2.61%. PCV-2 was detected exclusively by molecular analysis in 66 out of 223 tested animals (29.6%). The overall objective of the study is to deepen the understanding of virus circulation in the wildlife of Southern Italy, using the wild boar as an additional sentinel species for epidemiological surveillance activities. The distinctive feature of the project lies in its multi-pathogen approach applied to a reference population distributed across a particularly wide geographical area, allowing for the simultaneous assessment of the circulation of different viruses over a large territory.

The circulation of viruses of medical and veterinary importance is monitored to evaluate risks to both human and animal health. Among the species most commonly used in surveillance programs, the wild boar (Sus scrofa) plays a key role due to its high popu-lation density and its contacts with rural swine herds in the areas under investigation. In this study, molecular and serological analyses were carried out on wild boar samples collected from the regions of Apulia, Basilicata, Campania, and Calabria. The aim of this study is assessing the circulation of Influenza viruses, porcine Circovirus type 2 (PCV-2), Flaviviruses, and Aujeszky’s disease virus (ADV). Molecular analyses confirmed the presence of Influenza virus and PCV-2 in the wild population, while serological moni-toring detected specific antibodies against Influenza viruses, Flaviviruses, and ADV. The overall objective of the study is to deepen the understanding of virus circulation in the wildlife of Southern Italy, using the wild boar as an additional sentinel species for epidemiological surveillance activities. The distinctive feature of the project lies in its multi-pathogen approach applied to a reference population distributed across a parti-cularly wide geographical area, allowing for the simultaneous assessment of the circu-lation of different viruses over a large territory.

This study investigated the molecular features of Aujeszky's disease in a hunting dog from southern Italy. After consuming meat from infected wild boars, a 5-year-old male dog developed intense itching and died within 48 hours. Pseudorabies was suspected and necropsy was performed at the Experimental Zooprophylactic Institute of Apulia and Basilicata. SuHV-1 DNA was detected in the brain tissue, and the virus was isolated using Vero cells. Histological features and the analysis of genome sequencing revealed a non-suppurative meningoencephalitis caused by Aujeszky's disease. The yielded genome corresponded to a previously reported and unique genome (KU198433) isolated from a hunting dog in Italy. These findings improve understanding of pseudorabies's molecular epidemiology and help assess its epizootic potential.

The Belostomatidae is a family of aquatic Heteropteran insects that normally inhabit swamps of tropical, subtropical and temperate regions where they naturally feed on a large range of prey, mainly fish, amphibians and reptiles. However, these arthropods can occasionally bite humans when provoked, causing pain. The only species of giant water bug known to be found in Europe is Lethocerus (L.) patruelis. We reported the finding of a specimen of female L. patruelis in Southern Italy. The arthropod was collected by a tourist on his body while he lying down on a beach close to a nature reserve. To finalize the giant water bug identification, both morphological and molecular tools were used for the first time in our country. Herein, information on cases of patients bitten by giant water bugs belonging to the Belostomatidae family are reported, and circumstances involving their contact with humans are also discussed.

The Belostomatidae is a family of aquatic Heteropteran insects that normally inhabit swamps of tropical, subtropical and temperate regions where they naturally feed on a large range of prey, mainly fish, amphibians and reptiles. However, these arthropods can occasionally bite humans when provoked, causing pain. The only species of giant water bug known to be found in Europe is Lethocerus (L.) patruelis. We have reported the occasional finding of a specimen of L. patruelis female in Southern Italy. The arthropod was collected by a tourist on his body while he lying down on a beach close to a nature reserve. To finalize the giant water bug identification, both morphological and molecular tools were used for the first time in our country. Herein, information on cases of patients bitten by giant water bugs belonging to Belostomatidae family are reported and circumstances involving their contact with humans are also discussed.

Background: Anopheles sacharovi, a member of the Anopheles maculipennis complex, was a historical malaria vector in Italy, no longer found since the last report at the end of 1960s. In September 2022, within the Surveillance Project for the residual anophelism, a single specimen of An. maculipennis sensu lato collected in Lecce municipality (Apulia region) was molecularly identified as An. sacharovi. This record led to implement a targeted entomological survey in September 2023. Methods: Investigation was conducted in the areas around the first discovery, focusing on animal farms, riding stables and potential breeding sites. Adult and immature mosquitoes were collected, using active search or traps, in several natural and rural sites. Mosquitoes belonging to An. maculipennis complex were identified morphologically and molecularly by a home-made routine quantitative polymerase chain reaction (qPCR) assay, developed specifically for the rapid identification of An. labranchiae, and, when necessary, by amplification and sequencing of the ITS-2 molecular marker. Results: Out of the 11 sites investigated, 6 were positive for Anopheles presence. All 20 An. maculipennis s.l. (7 adults, 10 larvae and 3 pupae) collected in the areas were identified as An. sacharovi by ITS-2 sequencing. Conclusions: The discovery of An. sacharovi, considered to have disappeared from Italy for over 50 years, has a strong health relevance and impact, highlighting an increase in the receptivity of the southern areas. As imported malaria cases in European countries are reported every year, the risk of Plasmodium introduction by gametocyte carriers among travellers from endemic countries should be taken into greater consideration. Our findings allow rethinking and building new models for the prediction and expansion of introduced malaria. Furthermore, to prevent the risk of reintroduction of the disease, the need to strengthen the surveillance of residual anophelism throughout the South should be considered. Graphical

Canine coronavirus (CCoV) is a common agent of gastroenteritis in dogs, although some variants have been found associated with systemic and often fatal diseases. Distinct genotypes (CCoV‐I and CCoV‐II) and subgenotypes (CCoV‐IIa and CCoV‐IIb) are worldwide distributed. In Italy, CCoV infections have been occasionally evaluated, but information about the molecular epidemiology and the genomic features of currently circulating strains is limited. This study reports the detection and molecular characterization of CCoV strains from samples collected from 284 dogs in Italy between 2019 and 2021. CCoV RNA was detected in 39 (13.7%) dogs, as a single viral agent (5 animals, 12.8%) or with other viral pathogens (canine parvovirus types 2a/2b/2c; canine adenovirus type 1; norovirus GIV.2) (34 animals, 87.2%). A total of 48 CCoV strains were detected either alone (CCoV‐I: 51.3%, CCoV‐IIa: 20.5%) or in copresence (CCoV‐I and CCoV‐IIa, 23.1%); surprisingly, CCoV‐IIb was not identified in this study. Five clusters of CCoV‐I were detected, and their spike gene sequences showed the highest nucleotide identities with CCoV‐I strains collected from Greece in 2008/2009 and from China in 2021. CCoV‐IIa spike gene sequences (three variants) had the highest nucleotide identities with CCoV‐IIa strains collected in Greece in 2008/2009 and in Italy in 2009/2011. Given the high CCoV diversity and the variable pathogenicity potential, we underline the need of further surveillance studies to increase our understanding of the epidemiology and evolution of these viruses.

Rickettsia aeschlimannii infection is an emerging human tick-borne disease with only a few recorded cases. We reported a presumable autochthonous case of rickettsiosis in an Italian cattle breeder associated with a Hyalomma marginatum bite. Rickettsia aeschlimannii DNA was detected in both the tick specimen from the patient and the grazing cattle close to his farm.

Linguatula serrata, Frohlich, 1789, is a cosmopolitan zoonotic worm-like parasite of carnivores and other vertebrates including herbivores and omnivores. The adult form of the parasite typically inhabits the upper respiratory system, nares, and frontal sinuses of dogs, wolves, and cats. Infective eggs may be spread by sneezing, nasal secretions, and stool. The immature stages of the parasite are localized in the visceral organs of intermediated hosts, usually ruminants or rodents, and they are orally transmitted to predators during the ingestion of infested viscera. This paper reports the morphological identification and the molecular characterization of L. serrata specimen collected from a gray wolf in the Apulia region (southern Italy) and it also provides epidemiological information on this rarely reported zoonosis.