Giulia Morganti

OCCURRENCE OF LUNGWORMS IN EUROPEAN WILDCATS (FELIS SILVESTRIS SILVESTRIS) OF CENTRAL ITALY

Abstract The increasing focus on infections in domestic cats (Felis catus) has raised questions about lungworm distribution in wild hosts. To enhance knowledge of the occurrence of lungworms in enzootic regions of central Italy, we examined the carcasses of 16 European wildcats (Felis silvestris silvestris). Adult nematodes, feces, respiratory flushings, and pulmonary tissues were collected at necropsy and then microscopically and genetically analyzed. Fourteen wildcats had single or mixed lungworm species. Aelurostrongylus abstrusus was the most common parasite retrieved, followed by Troglostrongylus brevior. In addition, three specimens of Angiostrongylus chabaudi were found in the pulmonary arteries of one wildcat. Histologically, the most common lesions were a mild-to-severe chronic catarrhal bronchitis and a chronic interstitial pneumonia with smooth muscle hypertrophy, associated with T. brevior and A. abstrusus, respectively. These results demonstrate that the European wildcats may harbor several species of lungworms that may impair their health and welfare. Also, F. s. silvestris is a potential reservoir for respiratory nematodes in domestic cats.

Occurrence of Giardia duodenalis infection in chinchillas (Chincilla lanigera) from Italian breeding facilities.

The present work investigated the occurrence of Giardia infection in Chinchilla lanigera reared in three Italian breeding facilities and determined their role as potential zoonotic reservoir. One hundred and four fecal samples were tested for the presence of Giardia spp. cysts using a Direct Fluorescent Assay (DFA). A high positivity rate (39.4%) was found despite all animals were asymptomatic at the time of sampling. Thirty-one positive samples were genetically characterized by sequence analysis of the ITS1-5.8S-ITS2 region of the Giardia ribosomal DNA. Assemblages B (29 isolates) and C (two isolates) were identified. These results showed that Giardia infection can be common in chinchillas, thus spurring further molecular epizootiological studies of the infection to assess the zoonotic potential or host specificity of their isolates, to determine the source of infections, to identify the routes of transmission, and to control the infection among animal populations.

Molecular and serological detection of tick-borne pathogens in donkeys (Equus asinus) in Italy

Donkeys, owing to the frequent outdoor activity, are exposed to a high risk of infection with tick-borne pathogens. This work aimed to detect exposure to Theileria equi, Babesia caballi, Anaplasma phagocytophilum and Borrelia burgdorferi s.l. of donkeys reared in Central Italy. For this purpose 122 adult donkeys were selected within 11 herds and submitted to blood collection. IgG antibodies to T. equi, B. caballi, A. phagocytophilum and B. burgdorferi s.l. were detected by IFAT. Conventional PCRs targeting the genes MSP2 and the flagellin were used for the detection of A. phagocytophilum and B. burgdorferi s.l. respectively and a Real Time PCR Sybr Green was used to detect Babesia/Theileria spp…. The species identity was determined by amplicons sequencing. Forty eight (39.3%) and 58 (47.5%) animals tested positive for T. equi and B. caballi antibodies, respectively; nine animals (7.4%) were found positive for antibodies against A. phagocytophilum whereas negative results were obtained for B. burgdorferi s.l. Twenty-six (21.3%) animals showed antibodies for both T. equi and B. caballi. Twenty-three (18.8%) donkeys were positive to Babesia/Theileria spp. PCR assay. Out of 21 sequenced amplicons, 20 were identified as T. equi, belonging to three main groups designated A, B and D and one as B. caballi group A. Neither A. phagocytophilum nor B. burgdorferi PCR results were positive. The study showed a high exposure of donkeys to tick-borne pathogens and provides information on the genetic identity of the T. equi strains circulating in Central Italy.

PCR-based assay for the mitochondrial cox1 specific amplification of Eucoleus böhmi

Eucoleus böhmi (syn. Capillaria boehmi) is a trichuroid nematode affecting the epithelium of the nasal turbinates, frontal and paranasal sinuses of wild and domestic canids. Knowledge of the geographic distribution of nasal eucoleosis is fragmentary, despite the infection has been described from Europe and North America. Moreover, gaps exist in information available on the importance of the disease in canine clinical practice. The lack of knowledge on E. böhmi is likely due to limitations inherent to diagnostic methodologies. The aim of the present work was to assess a PCR-based assay instrumental to the amplification of a species-specific region of the mitochondrial DNA (mtDNA) gene encoding for the subunit 1 (cox1) gene of E. böhmi. Adult worms of E. böhmi from red foxes and dogs from Norway, Serbia and Italy and individual fecal samples from naturally infected dogs from Italy were included in the study. Stool samples from dogs negative for E. böhmi, but positive for other common parasites in both single and mixed infections, and adult stages of common dog parasites, were used to assess the specificity of this genetic assay. Using the panel of faecal samples, the assay showed a sensitivity of 85.14% and a specificity of 100%.

A pilot trial evaluating the efficacy of a 10% imidacloprid/2.5% moxidectin spot-on formulation in the treatment of natural nasal capillariosis in dogs.

The efficacy and safety of a spot-on formulation containing 10% imidacloprid and 2.5% moxidectin (Advocate(®), Bayer Animal Health GmbH, Leverkusen, Germany) were evaluated in a pilot trial for the treatment of canine nasal capillariosis caused by Capillaria boehmi (syn. Eucoleus boehmi). Sixteen dogs copromicroscopically positive for C. boehmi eggs were confirmed, either by rhinoscopy or species-specific PCR-coupled sequencing assays, as being affected by nasal capillariosis. The animals were randomly allocated to two different study groups, i.e. one (Group T) treated with Advocate(®) and one (Group C) left untreated, in a ratio of 1:1. The animals underwent clinical examination and quantitative copromicroscopy for C. boehmi eggs on Days -6 and -2 (baseline) and Day 28 ± 2 (post-baseline). Animals in Group T received Advocate(®) on Day 0. On Day 28 ± 2 the efficacy of the treatment (Group T) or the persistence of the infection (Group C) was confirmed by rhinoscopy or, alternatively, by molecular procedures. Seven of the eight dogs in Group T were negative on Day 28 ± 2 (reduction of baseline faecal egg counts by 99.14%), while for one dog a second treatment on Day 28 ± 2 was necessary to clear the infection, as demonstrated on Day 56 ± 2 (reduction of baseline faecal egg counts by 100% in Group T). Seven animals in Group C received a rescue dose of Advocate(®) on Day 28 ± 2 and scored microscopically and molecularly negative for the parasite on Day 56 ± 2, thus increasing the reduction of post-baseline egg counts to 99.57% after a single administration. These promising results show that Advocate(®) spot-on is an effective formulation for the treatment of canine nasal capillariosis under field conditions.

Natural infection of Anaplasma platys in dogs from Umbria region (Central Italy).

Anaplasma platys is a tick-borne pathogen causing the Infectious Canine Cyclic Thrombocytopenia. The pathogenesis of this disease is not yet well understood, due to the wide variety of clinico-pathological patterns described worldwide and to the high prevalence of co-infections with other vector-borne pathogens occurring in endemic areas. The present paper reports 3 cases of infection by A. platys occurring in dogs native to Central Italy, considered a non-endemic area to date. Infections were initially diagnosed based on clinical data and observation of morulae within platelets and then confirmed by biomolecular techniques. Moreover, two dogs showed an immune-mediated hemolytic anemia, as yet not described in literature in association with A. platys infection. The symptoms and the pathological findings observed will be discussed, as well as the importance to include this pathogen in the differential diagnosis of tick-borne diseases even in Central Italy.

Evaluation of the Clinical Efficacy and Safety of a Spot-on Combination of Imidacloprid 10 % / Moxidectin 2.5 % (Advocate®, Advantage® Multi) in Comparison to an Untreated Control Group in the Treatment of Capillaria boehmi in Naturally Infected Dogs

Capillaria boehmi affects the upper respiratory tract of domestic and wild canids. The aim of the present study was to investigate the efficacy of imidacloprid 10 % / moxidectin 2.5 % spot-on (Advocate®, Advantage® Multi, Bayer) in dogs naturally infected by C. boehmi. Twenty dogs infected with C. boehmi were randomly allocated to two groups: T1 (10 dogs) received a single treatment of Advocate® using the recommended dose on day 0 and T2 (10 dogs) served as an untreated control group. The reduction of the faecal egg counts (EPG) from baseline (days -6 ± 2 and -2 ± 2) to study completion was set as the primary efficacy criterion; clinical assessments of the upper respiratory tract and a rhinoscopy to visualize the parasites were used as secondary efficacy criteria. Eight dogs in T1 were not shedding eggs on days 28 ± 2 (reduction of EPG 99.66 %). A second treatment was administered to two dogs still positive on days 30 ± 3. A second efficacy evaluation was performed on days 42 ± 2 (study completion), when the two dogs tested negative. The mean number of EPG at study completion was 0 in T1 and 368.49 in T2. The difference between the groups was statistically significant (P < 0.01). Treatment efficacy at study completion was 100 %. None of the T1 dogs showing clinical signs on day 0 were symptomatic on days 28 ± 2. No adverse events occurred. The results show that Advocate® is safe and effective in the treatment of canine nasal capillariosis. Original Article WAAVP_2017.indb 65 29.06.17 15:33 S66 EndoparasitEs EndoparasitEs

Frankfurters made with pork meat, emmer wheat ( Triticum dicoccum Schübler) and almonds nut ( Prunus dulcis Mill.): evaluation during storage of a novel food from an ancient recipe

The physical, chemical and microbiological characteristics of reformulated meat-based frankfurters, derived from an ancient Roman cookbook and produced from pork meat, emmer wheat, almond, fish sauce and spices, were investigated during storage. Two different formulations were considered, with pork thigh and pork shoulder muscles, respectively. The products contain 13-16% protein, 14-19% fat, 14% carbohydrates, >7% fibre and a polyunsaturated/saturated fatty acids ratio above 0.4. During the 24 days of storage, both products evidenced a decrease in the pH and increases in the total volatile nitrogen content and TBARs value, whereas, the aw remained stable. From a microbiological perspective, increases in the total viable count and lactic acid bacteria up to 4.8 log cfu/g occurred during storage, but no pathogens were found. Sensory analyses revealed a change in odour and flavour at 18 days, with the detection of a fermented and rancid taste. Survival sensory analysis defined a shelf life of the products of between 18.6 and 22.7 days.

A first record of the tortoise tick, Hyalomma aegyptium (Linnaeus 1758) on Malta

ABSTRACT Tortoises of the genus Testudo are commonly kept pets on the Maltese Islands. This is a first record of the presence of Hyalomma aegyptium (Linnaeus 1758), also known as the “tortoise tick,” on Malta. Two male H. aegyptium were collected from an adult Mediterranean spur-thighed tortoise (Testudo graeca Linnaeus 1758) and a further two male H. aegyptium were collected from an adult female Egyptian tortoise (Testudo kleinmanni Lortet 1883). These tortoises were believed to be illegally imported, wild-caught specimens from North Africa. The presence of H. aegyptium on Malta is of both biological and medical relevance since these ticks are known potential vectors for numerous zoonotic diseases. This is also believed to be the first record of T. kleinmanni as a host for H. aegyptium.

Microbiological and parasitological investigation on chelonians reared in Italian facilities

The rapid rise in the number of pet chelonians and their illegal trade can modify the ecology, involving exotic pets, humans, and microbiological agents. Therefore, different epidemiological situations and the related risk to introduce and spread infectious diseases, especially zoonotic agents, have to be considered. The aim of this study was to investigate the microbiological and parasitological situation in 2 chelonian facilities (a private breeding of tortoises and a shelter for turtles) collecting oral/cloacal swabs and cloacal flushes to research viruses, bacteria, and parasites. No Chelonian Herperviruses, Cryptosporidium spp., and Giardia spp. infections were found. Salmonella spp. were detected in 8% of tortoises and in 37.5% of turtles and oxyurid eggs in 23.7% of tortoises and 15% of turtles; ascarid eggs were present only in tortoises. Moreover, 6 turtles showed cutaneous lesions, where Aeromonas sobria was isolated as main pathogen. Further studies should be performed to understand the zoonotic and infectious risk in each chelonian facility and to characterize the variables that could influence the microbiological patterns.