First detection and molecular identification of Neospora caninum from naturally infected cattle and sheep in North Africa

Neosporosis, caused by the protozoan Neospora caninum , is a major cause of reproductive failure in ruminants causing enormous economic losses. The objective of this study was to estimate the infection rate and molecular identification of N. caninum in Tunisian cattle and sheep. A total number of 348 meat samples were collected from 150 cows and 198 ewes slaughtered in the regional slaughterhouse of Béja (North‐west Tunisia) and tested for the presence of N. caninum ITS 1 gene using PCR followed by sequencing of some PCR products. A phylogenetic tree was then constructed to compare the partial sequences of the ITS 1 gene with GenBank sequences. The overall molecular infection prevalence of N. caninum was significantly higher in cattle than in sheep (22 and 10.6%, respectively, p  = .003). In sheep, the highest prevalence was observed in the northern Béja locality (31.2 ± 16.1), with the Noire de Thibar breed as the most infected sheep breed (31.7 ± 14.2%) (p  < .001). In cattle, there were no differences in the molecular prevalence of N. caninum according to breeds and localities. The association between age and N. caninum molecular prevalence was statistically significant in both species; the highest prevalence was observed in sheep of more than one year of age (19.4 ± 9.1%), and in cattle between two and eight years of age (28.8 ± 10.9%). Comparison of the partial sequences of the ITS 1 gene revealed 96%–100% similarity among our N. caninum amplicon and those deposited in GenBank. To our knowledge, this is the first detection and molecular identification of N. caninum in sheep and cattle in North Africa. This information is pertinent in designing control programmes that would reduce economic losses in the livestock industry.     

First report of peste des petits ruminants virus lineage II in Hydropotes inermis,China

In this study, we investigated an outbreak of peste des petits ruminants (PPR ) at a Hydropotes inermis (water deer) farm in Anhui Province, China. These results demonstrated that PPR viruses (PPRV s) can infect H. inermis and also revealed that virulent lineage II PPRV s exist in China, where they have been responsible for the deaths of wild animals. The government should pay close attention to the threat of PPRV epidemiology in China.     

Molecular Study of Dirofilaria immitis and Dirofilaria repens in Dogs from Tunisia

Dirofilaria immitis and Dirofilaria repens are mosquito‐borne nematodes which infect primarily dogs as their main definitive hosts. They cause cardiopulmonary (D. immitis) or cutaneous (D. repens) dirofilariasis in canids and other carnivores and can accidentally be transmitted to humans where they can induce a variety of clinical outcomes depending on organ localization. Dirofilaria spp. infection in dogs was assessed using molecular methods (PCR and sequencing) to identify the different Dirofilaria species occurring in 200 dogs from Northern and Central Tunisia. The overall molecular prevalence of Dirofilaria spp. was 17.5% (35/200). The prevalence of D. immitis (14.5%) was significantly higher than for D. repens (3%). Molecular prevalence of D. immitis was significantly higher in suburban compared to urban and rural regions. There was no difference in molecular prevalence of D. immitis or D. repens according to the dogs’ (sex or use). Dirofilaria immitis amplicons (accession numbers KR676386 ) fall into the same clade with D. immitis from China, India and Taiwan. Comparison of the partial sequences of D. repensITS2 rDNA gene ( KR676387 ) revealed 99.6% similarity with D. repens reported in dogs from USA. It had also 97.6% similarity with D. repens from mosquitoes in Czech Republic. High dog parasite burdens should motivate both medical doctors and veterinarians to consider these frequent infections.     

High Prevalence of Anaplasma spp. in Small Ruminants in Morocco

The prevalence of infection by Anaplasma spp. (including Anaplasma phagocytophilum) was determined using blood smear microscopy and PCR through screening of small ruminant blood samples collected from seven regions of Morocco. Co‐infections of Anaplasma spp., Babesia spp, Theileria spp. and Mycoplasma spp. were investigated and risk factors for Anaplasma spp. infection assessed. A total of 422 small ruminant blood samples were randomly collected from 70 flocks. Individual animal (breed, age, tick burden and previous treatment) and flock data (GPS coordinate of farm, size of flock and livestock production system) were collected. Upon examination of blood smears, 375 blood samples (88.9%) were found to contain Anaplasma‐like erythrocytic inclusion bodies. Upon screening with a large spectrum PCR targeting the Anaplasma 16S rRNA region, 303 (71%) samples were found to be positive. All 303 samples screened with the A. phagocytophilum‐specific PCR, which targets the msp2 region, were found to be negative. Differences in prevalence were found to be statistically significant with regard to region, altitude, flock size, livestock production system, grazing system, presence of clinical cases and application of tick and tick‐borne diseases prophylactic measures. Kappa analysis revealed a poor concordance between microscopy and PCR (k = 0.14). Agreement with PCR is improved by considering microscopy and packed cell volume (PCV) in parallel. The prevalence of double infections was found to be 1.7, 2.5 and 24% for Anaplasma‐Babesia, Anaplasma‐Mycoplasma and Anaplasma‐Theileria, respectively. Co‐infection with three or more haemoparasites was found in 1.6% of animals examined. In conclusion, we demonstrate the high burden of anaplasmosis in small ruminants in Morocco and the high prevalence of co‐infections of tick‐borne diseases. There is an urgent need to improve the control of this neglected group of diseases.     

A serological Survey of Brucella spp., Salmonella spp., Toxoplasma gondii and Trichinella spp. in Iberian Fattening Pigs Reared in Free‐Range Systems

Zoonotic agents such as Brucella spp., Salmonella spp., Toxoplasma gondii and Trichinella spp., all considered high‐risk zoonotic pathogens by the European Food Safety Agency (EFSA), may cause no symptoms of infection in free‐range pigs yet still have a significant public health impact. A serological survey was therefore performed to determine the history of occurrence of these pathogens in such pigs in southern Spain. A total of 709 serum samples were collected at abattoir from pigs from 79 farms and analysed for specific antibodies against the above pathogens using commercially available ELISA kits. Encysted Trichinella spp. larvae were also sought following the artificial digestion method of diaphragm pillar muscle. The results showed Salmonella spp. to be widely distributed among the sampled herds [73.42%, 95% confidence interval (CI₉₅) 65.6–81.78] and Toxoplasma gondii to be present in over half (58.23%, CI₉₅ 47.33–69.07). The seroprevalence of Brucella spp. was very low (3.8%, CI₉₅ 0.18–7.42), and antibodies against Trichinella spp. were not detected. No encysted Trichinella spp. larvae were microscopically detected.     

Sero‐prevalence and epidemiology of peste des petits ruminants in Libya

We conducted a cross‐sectional study during 2013 to quantify the serological prevalence of peste des petits ruminants ( PPR) infection and to investigate host factors associated with PPR infection in small ruminants in Libya. A two‐stage sampling design was carried out. A total number of 148 flocks owning at least 100 heads each were randomly selected. Sixteen to forty‐eight samples were collected from each selected flock. A total number of 3,508 serum samples from unvaccinated animals were collected and analysed at IZSLER Brescia, Italy, by using competitive ELISA, IDvet innovative diagnostics (IDvet 310, France). The overall serological prevalence among SR was 33% (95% CI: 31.4–34.5). Significant differences between the prevalence in the geographical branches were observed. The lowest prevalence level was observed in Zawiyah branch (16.1%), whereas the highest value was obtained for the Sabha branch (56.8%). Considering the age, a serological prevalence of 24.7%, 31.5% and 42.1% was observed in SR <1 year, between 1 and 2 years and more than 2 years, respectively. Statistically significant differences (p  <  .001) in the sero‐prevalence levels were also observed between the age groups. Our findings suggest that the southern part of Libya could be more exposed to the infections coming from the neighbouring countries and this should be better investigated to correctly identify wherever specific entry points can be considered at higher risk than others. The results also confirmed the endemic status of PPR in Libya, with a constant exposure to the infection of the animals during their life. In the framework of the global strategy for control and eradication of PPR, our results, even if obtained by a preliminary study, can contribute to the assessment of the epidemiological situation of PPR in Libya as required by the Stage 1 of the plan.     

First Molecular Identification and Genetic Characterization of Theileria lestoquardi in Sheep of the Maghreb Region

Theileria lestoquardi is the most prominent Theileria species in small ruminants that causes malignant theileriosis of sheep in Africa and Asia. In the present survey, blood samples and ticks were collected in Kebili (southern Tunisia) from 166 Queue Fine de l'Ouest sheep. Giemsa‐stained blood smears, immunofluorescent antibody test (IFAT) and PCR were performed. The DNA was extracted from blood and analysed by PCR targeting 18S rRNA gene of Theileria spp. and then sequenced. A total number of 140 ticks were collected from a total number of 166 sheep during the four seasons. The ticks belonged to two genera and 4 species; the most frequent tick was Hyalomma excavatum 84.3% (118/140) and then Rhipicephalus spp. 15.7% (22/140). Only two animals had positive Giemsa‐stained blood smears, and they were also positive by IFAT. The amplicons had 99.3 and 99.6% homology with the BLAST published T. lestoquardi amplicons. To our knowledge, this is the first report of T. lestoquardi in small ruminants within the Maghreb region.     

First Detection of Co‐circulation of West Nile and Usutu Viruses in Equids in the South‐west of Tunisia

In the last fifteen years, West Nile Virus (WNV) has dramatically expanded its geographic range and is now considered the most widespread arbovirus in the world. In Tunisia, West Nile Fever (WNF) outbreaks were reported in humans in 1997, 2003 and 2012. Usutu Virus (USUV), which is a ‘new’ emerging Flavivirus antigenically close to WNV, has never been reported in Tunisia. A serological investigation in 284 equids was conducted in 2012 in the southern west region of the country to assess the presence and prevalence of the WNV and USUV infection. Of the 284 samples tested by competitive enzyme‐linked immunoassay, 129 were positive. Of these, 120 (42.3%) had WNV‐specific neutralizing antibodies. The prevalence was significantly higher in areas closer to the oasis compared with that of the surrounding arid areas. Antibody titres against USUV were also reported in 10 equids. This was the first evidence of USUV circulation in Tunisia. Data recorded by this study indicate that WNV and USUV have circulated/are circulating in the region and that there is an urgent need to adapt the current surveillance programmes to this new scenario.     

Concurrent infection of Bluetongue and Peste‐des‐petits‐ruminants virus in small ruminants in Haryana State of India

Bluetongue (BT ) and peste‐des‐petits‐ruminants (PPR ) are major transboundary diseases of small ruminant, which are endemic in India. Testing of bluetongue virus (BTV ) and peste‐des‐petits‐ruminants virus (PPRV ) from recent outbreaks (2015–2016) in different regions of Haryana State of India revealed that 27.5% of the samples showed the presence of dual infection of BTV and PPRV . Analysis of Seg‐2 of BTV (the serotype‐determining protein) showed the presence of BTV ‐12w in several isolates. However, analysis of N gene fragment amplicons showed that viruses belong to lineage IV were most closely related to a pathogenic strain of PPRV from Delhi. This is the first report of co‐circulation of PPRV lineage IV and bluetongue virus serotype 12 in the state.     

Evaluation of an IGM‐specific ELISA for early detection of bluetongue virus infections in domestic ruminants sera

Competitive‐ELISA (c‐ELISA) is the most widely used serological test for the detection of Bluetongue virus (BTV) viral protein 7 (VP7) antibodies (Ab). However, these BTV c‐ELISAs cannot to distinguish between IgG and IgM. IgM Ab are generated shortly after the primary immune response against an infectious agent, indicating a recent infection or exposure to antigens, such as after vaccination. Because the BTV genome or anti‐VP7 Ab can be detected in ruminant blood months after infection, BTV diagnostic tools cannot discriminate between recent and old infections. In this study, we evaluated an IgM‐capture ELISA prototype to detect ruminant anti‐BTV VP7 IgM on 1,650 serum samples from cattle, sheep, or goats. Animals were BTV‐naive, infected, or/and vaccinated with BTV‐1, ‐2, ‐4, ‐8, ‐9, ‐16, or ‐27, and we also included 30 sera from cattle infected with the Epizootic haemorrhagic disease virus (EHDV) serotype 6. Results demonstrated that this ELISA kit is specific and can detect the presence of IgM with satisfactory diagnostic specificity and sensitivity from 1 to 5 weeks after BTV infection in domestic ruminants (for goats and cattle; for sheep, at least up to 24 days). The peak of anti‐VP7 IgM was reached when the level of infectious viruses and BTV RNA in blood were the highest. The possibility of detecting BTV‐RNA in IgM‐positive sera allows the amplification and sequencing of the partial RNA segment 2 (encoding the serotype specific to VP2) to determine the causative BTV serotype/strain. Therefore, BTV IgM ELISA can detect the introduction of BTV (or EHDV) in an area with BTV‐seropositive domestic animals regardless of their serological BTV status. This approach may also be of particular interest for retrospective epidemiological studies on frozen serum samples.     

Outbreaks of abortions by Coxiella burnetii in small ruminant flocks and a longitudinal serological approach on archived bulk tank milk suggest Q fever emergence in Central Portugal

Q fever is a worldwide zoonotic infectious disease caused by Coxiella burnetii and sheep and goats are known to be the main reservoir for human infection. This study describes the epidemiological and laboratory findings of C. burnetii outbreaks affecting sheep and goat flocks and also provides the results of a prospective serosurvey in bulk tank milk samples to assess C. burnetii circulation in a population of sheep living in close contact to the human population in Central Portugal. In the epizooties, C. burnetii was identified in tissues of the resulting abortions by qPCR . As for the serological survey, 10.2% (95%CI : 4.5‐19.2) of the 78 bulk tank milk samples collected in 2015 presented IgG antibodies against C. burnetii . The same farms were visited and sampled in 2016 and 25.6% (95%CI : 16.4‐36.8) were positive. This steep increase in the number of anti‐C. burnetii farms between the 2015 and 2016 collections showed to be statistically significant (p  = 0.020) and is strongly suggestive of Q fever emergence in Central Portugal. Measures on animal health and on disease spread control to the human population should be considered.     

First Experience Using Contrast‐enhanced Ultrasound to Evaluate Vascularisation of Acellular Dermal Matrices after Implant‐Based Breast Reconstruction

Acellular dermal matrices (ADM) have been used frequently in therapeutic and prophylactic breast procedures. To date there have been no reports on vascularisation of ADMs and formation of tissue around them as seen with modern non‐invasive imaging techniques such as contrast‐enhanced ultrasound (CEUS). In this case series, we used CEUS to investigate the features of ADM in relation to vascular ingrowth and scaffold for “new” tissue formation. This is a retrospective evaluation of patients who underwent successful skin‐ and nipple‐sparing mastectomy (SSM, NSM) with immediate IBBR using ADM from May 31, 2010, through December 28, 2012. Over a 24‐month period, 16 patients, with an average age of 44 years (range 27–70 years), were evaluated with CEUS. No contrast agent allergies or side effects were reported for the ultrasound examination. After contrast agent injection (1–18 months postoperatively), homogeneous normal enhancement in the ADM and peripheral region with physiological tissue formation was seen in all patients. In this small study, the most obvious contribution of CEUS is the in vivo evaluation of vascular ingrowth and tissue formation after IBBR with ADM after follow‐up of 1–18 months postoperatively. Level of Evidence III: Retrospective cohort or comparative study; case–control study; or systematic review of these studies.     

Epidemiological Study of Lumpy Skin Disease Outbreaks in North‐western Iran

Lumpy skin disease (LSD) is a highly contagious transboundary disease of cattle with major economic losses. This study was undertaken to address the emergence and epidemiological features of LSD in four north‐western provinces of Iran. These provinces have extensive borders with others country including Iraq, Turkey, Azerbaijan and Armenia. A population of 683 cattle from 91 farms were examined during LSD outbreak in Iran during 2014–2016. The information of the farms including the population size, gender, age, vaccination status, clinical signs and the number of death because of LSD were recorded in the designed questionnaires. A number of 234 blood samples were collected randomly from animals with and without clinical signs of LSD. DNA was extracted from blood samples, and they were used for amplifying a fragment of 434 bp in size coupled with restriction fragment length polymorphism (RFLP) for molecular detection of lumpy skin disease virus (LSDV). The estimated prevalence, cumulative mortality and case fatality were 17.9%, 3.5% and 19.7%, respectively. There was no significant difference in occurrence of the disease between male and female cattle. LSD occurrence in age groups above 5 years old and below 6 months old showed highest and lowest relative frequencies, respectively. Vaccination was significantly decreased the occurrence of clinical disease. The developed PCR–RFLP technique was able to differentiate between LSDV, sheep pox virus (ShPV) and goat pox virus (GPV). It was concluded that LSD was entered into Iran probably from Iraq via uncontrolled animal movements along common land borders between two countries. Developed PCR–RFLP could be used as a rapid and inexpensive method for differentiating Capripoxviruses (CaPVs).     

Genetic diversity of Bartonella spp. in vampire bats from Brazil

Recently, an increasing number of Bartonella species have been emerged to cause human diseases. Among animal reservoirs for Bartonella spp., bats stand out due to their high mobility, wide distribution, social behaviour and long‐life span. Although studies on the role of vampire bats in the epidemiology of rabies have been extensively investigated in Latin America, information on the circulation and genetic diversity of Bartonella species in these bat species is scarce. In the present work, 208 vampire bats, namely Desmodus rotundus (the common vampire bat; n = 167), Diphylla ecaudata (the hairy‐legged vampire bat; n = 32) and Diaemus youngii (the white‐winged vampire bat; n = 9) from 15 different states in Brazil were sampled. DNA was extracted from liver tissue samples and submitted to real‐time PCR (qPCR) and conventional PCR (cPCR) assays for Bartonella spp. targeting five genetic loci, followed by phylogenetic and genotype network analyses. Fifty‐one out of 208 liver samples (24.51%) were positive for Bartonella DNA in the ITS real‐time PCR assay [40 (78.43%) of them were from D. rotundus from 11 states, and 11 (21.57%) samples from D. ecaudata from three states. Eleven genotypes were found for each gltA and rpoB genes. Several ITS sequences detected in the present study clustered within the lineage that includes B. bacilliformis and B. ancachensis. The Bayesian phylogenetic inference based on the gltA gene positioned the obtained sequences in six different clades, closely related to Bartonella genotypes previously detected in D. rotundus and associated ectoparasites sampled in Latin America. On the other hand, the Bartonella rpoB genotypes clustered together with the ruminant species, B. schoenbuchensis and B. chomelii. The present study describes for the first time the molecular detection of Bartonella spp. in D. ecaudata bats. It also indicates that Bartonella spp. of vampire bats are genetically diverse and geographically widespread in Brazil.     

Distinction Between Persistent and Transient Infection in a Bovine Viral Diarrhoea (BVD) Control Programme: Appropriate Interpretation of Real‐Time RT‐PCR and Antigen‐ELISA Test Results

Control of bovine viral diarrhoea (BVD) in Belgium is currently implemented on a voluntary basis at herd level and mainly relies on detection and culling of persistently infected (PI) animals. The present field study was conducted during the winter of 2010/2011 to assess the performances of diagnostic assays used in the testing scheme for BVD as proposed by the two Belgian regional laboratories. Individual blood samples were collected from 4972 animals, and individual samples from the same herd were pooled (maximum of 30 individual samples per pool) and screened for the presence of Bovine Viral Diarrhoea Virus (BVDV)‐specific RNA using a commercial real‐time RT‐PCR test (ADIAGENE). Individual samples from positive pools were then tested in parallel with the same RT‐PCR test and with an antigen‐capture ELISA test (IDEXX) to detect viremic animals. This study demonstrated that individual results differed according to the type of assay used (P < 0.001): 140 animals (2.8%) were positive by RT‐PCR and 72 (1.4%) by antigen‐ELISA. A second blood sample was taken 40 days later from 74 PCR positive animals to detect persistent viremia: 17 (23%) of these were still PCR positive and considered to be PI and the 57 that no longer tested positive were assumed to be transiently infected (TI) animals. All PI animals were positive also by antigen‐ELISA at both time points. Among TI animals, 10 (16%) were positive by antigen‐ELISA at the first but none at the second sampling. A highly significant difference in cycle threshold (C_t) values obtained by RT‐PCR was observed between PI and TI animals. ROC analysis was performed to establish thresholds to confirm with high probability that an animal is PI, based on the result of RT‐PCR test performed on a single individual blood sample.     

Post‐epidemic investigation of Schmallenberg virus in wild ruminants in Slovenia

Schmallenberg virus (SBV) is a vector‐borne virus belonging to the genus Orthobunyavirus within the Bunyaviridae family. SBV emerged in Europe in 2011 and was characterized by epidemics of abortions, stillbirths and congenital malformations in domestic ruminants. The first evidence of SBV infection in Slovenia was from an ELISA‐positive sample from a cow collected in August 2012; clinical manifestations of SBV disease in sheep and cattle were observed in 2013, with SBV RNA detected in samples collected from a total of 28 herds. A potential re‐emergence of SBV in Europe is predicted to occur when population‐level immunity declines. SBV is also capable of infecting several wild ruminant species, although clinical disease has not yet been described in these species. Data on SBV‐positive wild ruminants suggest that these species might be possible sources for the re‐emergence of SBV. The aim of this study was to investigate whether SBV was circulating among wild ruminants in Slovenia and whether these species can act as a virus reservoir. A total of 281 blood and spleen samples from wild ruminants, including roe deer, red deer, chamois and European mouflon, were collected during the 2017–2018 hunting season. Serum samples were tested for antibodies against SBV by ELISA; the overall seroprevalence was 18.1%. Seropositive samples were reported from all over the country in examined animal species from 1 to 15 years of age. Spleen samples from the seropositive animals and serum samples from the seronegative animals were tested for the presence of SBV RNA using real‐time RT‐PCR; all the samples tested negative. Based on the results of the seropositive animals, it was demonstrated that SBV was circulating in wild ruminant populations in Slovenia even after the epidemic, as almost half (23/51) of the seropositive animals were 1 or 2 years old.     

Molecular detection and genetic diversity of Bartonella species in large ruminants and associated ectoparasites from the Brazilian Cerrado

Currently, five Bartonella species and an expanding number of Candidatus Bartonella species have globally been reported in ruminants. Likewise, different Bartonella genotypes were identified. However, studies relating to ruminant‐associated Bartonella in Brazil are scarce. The current study aimed to assess the prevalence and genetic diversity of Bartonella in cattle, buffaloes and associated ectoparasites in Brazil. For this purpose, EDTA‐blood samples from 75 cattle and 101 buffaloes were sampled. Additionally, 128 Rhipicephalus microplus and one Amblyomma sculptum ticks collected from cattle, and 197 R. microplus, one A. sculptum and 170 lice (Haematopinus tuberculatus) collected from buffaloes were included. Bartonella DNA was initially screened through an HRM real‐time PCR assay targeting the 16S–23S internal transcribed spacer (ITS), and the positive samples were submitted to an additional HRM assay targeting the ssrA gene. The HRM‐positive amplicons were sequenced, and the nucleotide identity was assessed by BLASTn. Bartonella spp.‐positive DNA samples were analysed by conventional PCR assays targeting the gltA and rpoB genes, and then, the samples were cloned. Finally, the phylogenetic positioning and the genetic diversity of clones were assessed. Overall, 21 of 75 (28%) cattle blood samples and 13 of 126 (10.3%) associated ticks were positive for Bartonella bovis. Out of 101 buffaloes, 95 lice and 188 tick DNA samples, one (1%) buffalo and four (4.2%) lice were positive for Bartonella spp. Conversely, none of the ticks obtained from buffaloes were positive for Bartonella. The Bartonella sequences from buffaloes showed identity ranging from 100% (ITS and gltA) to 94% (ssrA) with B. bovis. In contrast, the Bartonella DNA sequences from lice were identical (100%) to uncultured Bartonella sp. detected in cattle tail louse (Haematopinus quadripertusus) from Israel in all amplified genes. The present study demonstrates the prevalence of new B. bovis genotypes and a cattle lice‐associated Bartonella species in large ruminants and their ectoparasites from Brazil. These findings shed light on the distribution and genetic diversity of ruminant‐ and ectoparasite‐related Bartonella in Brazil.     

Molecular detection and characterization of carnivore parvoviruses in free‐ranging Eurasian otters (Lutra lutra) in southern Italy

The most important Italian population of the Eurasian otter (Lutra lutra) occurs in the southern part of the peninsula with two isolated sub‐populations of about 250 adult individuals. The Eurasian otter is considered to be near threatened and it is a fully protected species. The aims of this study were to investigate for the first time the occurrence and characterize the parvoviruses included in the species Carnivore protoparvovirus 1 in seven carcasses of road‐killed Eurasian otters from the southern Italy. Carnivore protoparvovirus 1 are responsible for acute gastroenteritis and leukopenia in pets and free‐ranging carnivores. Initial screening of tissue samples by real‐time PCR revealed CPV/FPV DNA in tissue samples of five Eurasian otters; three of them, showed co‐infections by both CPV and FPV. Among the five positive Eurasian otters, we successfully obtained six DNA sequences from four individuals including two CPV‐2a, one CPV‐2b, one CPV‐2c, and two FPV sequences. Comparison of these sequences with 250 VP2 gene sequences deposited in the GenBank database, showed 10 nt differences resulting in two synonymous and eight non‐synonymous substitutions. On the basis of these results, two sequences here found were characterized as new CPV‐2a, one was characterized as new CPV‐2b variant, and one was characterized as FPV‐like mutant. The last two sequences belong to a FPV and CPV‐2c strain respectively. Carnivore protoparvovirus 1 is reported for the first time in the Eurasian otter showing high infection value in southern Italy. Occurrence of this infection should be studied further to understand its possible pathogenicity and virulence to the fragile and isolate Eurasian otter population which live in southern Italy.