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Pulmonary capillariasis is a parasitic disease caused by the nematode Eucoleus aerophilus which affects wild and domestic carnivores. Currently, there are no anthelmintics approved for use in the treatment of dogs infected with E. aerophilus. The use of several anthelmintics has been reported in a few case reports and field efficacy studies in cats; much less is known on the treatment of dogs infected with E. aerophilus. The paper describes a case of a 4-month-old, mixed breed intact male referred to the Veterinary Teaching Hospital (VTH) of the Department of Veterinary Medical Science of the University of Bologna for a routine vaccination and tested positive for E. aerophilus. The dog has not been responding to three different administered treatments, such as moxidectin, fenbendazole, and milbemycin oxime. Eighteen months after the first fecal examination, owner has brought in the dog for a routine visit; a coprological examination was requested and performed resulting negative for parasites. Veterinary practitioners, parasitologists, diagnostic laboratories, and dog owners need to be aware of the increased danger of possible treatment failure when attempting to control parasitic infections for which there are no approved anthelmintics with established efficacies available for use.

Background In canine leishmaniosis (CanL) endemic areas, pathologists often receive skin biopsies for testing with histopathologic findings suggestive—but not conclusive for a definitive diagnosis—of CanL lesions. I the absence of data on the infective status of animals, the diagnosis can therefore be challenging. The aim of this retrospective study was to evaluate the ability of immunohistochemistry (IHC) and quantitative PCR (qPCR) methods to detect Leishmania infection in skin biopsies with a histopathologic diagnosis of lymphoplasmacytic/histiocytic and/or granulomatous dermatitis and to correlate the pattern, depth and severity of the histopathologic lesions with the parasite load detected by qPCR and IHC. Methods Thirty formalin-fixed, paraffin-embedded skin samples were evaluated by hematoxylin–eosin (H&E) staining, IHC, conventional PCR (cPCR) and qPCR. The severity, pattern and depth of the dermal inflammation and parasite load were graded. Results Leishmania was detected by H&E staining in 8/30 sections (26.66%) and by IHC in 14/30 samples (46.66%). Parasite DNA was detected in 14/30 samples (46.66%) by cPCR and in 21/30 samples (70%) by qPCR, with an extremely variable parasite load (1.32–62.700 copies). The level of agreement was fair between H&E staining and cPCR (κ = 0.32), and moderate between H&E staining and IHC (κ = 0.58). The level of agreement between IHC and cPCR was good (κ = 0.65); between IHC and qPCR, moderate (κ = 0.41); and between cPCR and qPCR, fair (κ = 0.28). A significant association was found between the severity of dermal inflammation and the parasitic skin load by IHC, although with weak linear correlation. Conclusions Our study underlines the difficulty of obtaining a definitive diagnosis of CanL cutaneous lesions, even with the most accurate diagnostic tests currently available. Based on our results, no single test is suitable on its own for the diagnosis of cutaneous lesions caused by Leishmania. However, in the presence of a moderate/severe lymphoplasmacytic/histiocytic and/or granulomatous dermatitis, we suggest performing IHC, as in our study this technique proved to be the method with the highest discriminatory power to estimate the role of the parasite in skin lesions. In mild lesions, IHC loses its discriminatory power and should be effectively combined with techniques such as qPCR. Graphical Abstract

Anaplasma phagocytophilum is a tick-borne pathogen of sheep and other ruminants, humans, horses and dogs. Current data on the prevalence and diffusion of A. phagocytophilum among domestic animals in Italy are fragmentary, especially those related to sheep, considered a natural domestic reservoir. We investigated the prevalence of A. phagocytophilum infection in dairy sheep from Tuscany (Italy), by testing 624 sera with the immunofluorescent antibody (IFA) test. The sera belonged to 33 different flocks distributed throughout the province of Grosseto (Tuscany). Positive results were found in 21 out of the 33 flocks tested (63.63%, 95% IC 47.22–80.05%) with a prevalence rate within flocks ranging from 3.33% to 59.26%. The seroprevalence against A. phagocytophilum within the sheep population was 8.01% (95% IC 6.13–10.41%), and most of the positive cases showed low antibody titres (1/40-1/80). The multivariate analysis identified "small flock" ( Our results reveal a moderate circulation of A. phagocytophilum among sheep flocks in central Italy.

A 4-year-old, 20 kg, female mixed breed dog was referred to our Veterinary Teaching Hospital for severe respiratory distress. Thoracic ultrasonography revealed severe pleural effusion with multiple anechoic cystic structures within the fluid. Abdominal cavity presented few small and irregular cystic structures. Diagnosis of infection by larval stages of Mesocestoides spp. on the basis of a PCR approach was performed and therapy with oral fenbendazole was started. Due to an incomplete clinical response, the dog underwent to remove metacestodes cysts by surgical debridement. The dog showed no recurrence of clinical signs for 12 months after the surgery. Pleural larval cestodiasis should be added as a differential diagnosis in the list of diseases causing pleural effusion in dogs.

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.

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.

The presence of viable Toxoplasma gondii was investigated in artisanal cheeses made from milk of naturally infected ewes. Ewe milk was analyzed beforehand for the presence and vitality of T. gondii by loop-mediated isothermal amplification (LAMP) and reverse-transcriptase PCR (RT-PCR), respectively. Cheeses were prepared from raw milk following a traditional cheesemaking process. The cheese obtained from T. gondii-positive milk was analyzed by LAMP to detect Toxoplasma DNA-positive samples. RT-PCR was then carried out to assess the viability of the parasites in T. gondii-positive milk samples and fresh cheese, after 5 and 15 days of ripening. Physical-chemical parameters of cheeses were also investigated. All cheese samples derived from T. gondii-positive milk were positive according to LAMP, at both 5 and 15 days of ripening, while none of the samples were positive according to RT-PCR. Thus, while the presence of the parasite was demonstrated by the detection of specific DNA, the absence of detectable T. gondii RNA supports the hypothesis that changes in the chemical and physical characteristics occurring during the cheesemaking process and ripening period, could be sufficient to inactivate viable T. gondii in milk, minimizing the risk of human infection through consumption of raw sheep milk cheese.

The aim of this study was to evaluate the performance of a commercially available rapid enzyme-linked immonosorbent assay, the Snap® 4Dx test, in the detection of Anaplasma phagocytophilum antibodies in horses. Two hundred apparently healthy horses (asymptomatic) and 244 animals showing clinical symptoms (symptomatic), were tested for A. phagocytophilum immunoglobulin G (IgG) antibodies using both the Snap® 4Dx kit and an indirect fluorescence antibody test (IFAT), with the latter serving as a comparative test. Horses belonging to the symptomatic group were also tested for evidence of active infection with A. phagocytophilum by analysis of IFAT IgM titers and PCR assay amplifying a specific fragment of the 16S rRNA gene. The overall agreement between the results obtained using the two tests, as well as the relative performance exhibited by the Snap® 4Dx test in the two groups, was assessed. Forty of the 45 animals (89%) testing positive for IgG antibodies using IFAT were correctly identified using Snap® 4Dx testing. The agreement between the results of the two tests was very high (k0.9), with almost identical performances in both symptomatic and asymptomatic animals. Conversely, within the symptomatic group, only 44% (no. 11/25) of Snap® 4Dx positives appeared to be associated with a state of active infection, whereas the remaining 56% (no. 14/25) were related both to not infected animals (no. 1) and to horses whose status of infection needed further evaluations to be confirmed (no. 13/25). This study suggests that the Snap® 4Dx test could represent a valid screening method for use during epidemiological surveys of equine populations. Nevertheless, in-clinic application of the test does not appear to be merited.

Koenhemsi, Lora/0000-0002-4979-170X; or, erman/0000-0002-8764-1956 WOS:000475487800011 Equine piroplasmosis (EP) is a tick-borne disease caused by Babesia caballi and Theileria equi, which causes a significant economic loss. The aim of this study was to determine the disease-related changes of serum trace elements that play an important role in the immune system. Our study was conducted on 20 horses with piroplasmosis and 20 healthy horses. It was performed to evaluate some of the serum trace element levels (chromium (Cr), copper (Cu), iron (Fe), magnesium (Mg) manganese (Mn), selenium (Se), zinc (Zn), arsenic (As), boron (B), and nickel (Ni)) in equine piroplasmosis. The elements were analyzed in serum using 1CP-OES. Infected horses had significantly higher serum Mg and Mn levels. In conclusion, changes of serum trace elements in infected horses can provide valuable information about pathophysiological aspects and they are also good indicators for diagnosis, prognosis and treatment evaluation. Much more scientific research is required to evaluate the trace elements related to equine piroplasmosis.

Sheep coccidiosis is a pathology caused by protozoan parasites of the genus Eimeria spp. with clinical and economic effect especially in young animals (Ambrosi 1995; Pellérdy 1974). A field study was conducted to evaluate the efficacy of a metaphylactic treatment with toltrazuril 5 % suspension (Baycox®, Bayer) in comparison with diclazuril (Vecoxan®, Janssen-Cilag) and untreated controls against naturally acquired Eimeria infections in housed lambs. A total of 170 animals, aged 24 to 34 days and randomly divided in three homogeneous groups, were included in the study. The assessment of treatment efficacy was based on total faecal oocyst excretion (opg) and count reduction (FOCR) in the two groups of animals treated with toltrazuril (TOLT) and diclazuril (DICL) compared with untreated control group (C). The animals treated with toltrazuril showed a considerably lower mean opg to that of group C (5.78 opg versus 144.62 opg) (p0.05) and a FOCR of 97.7 %. The higher efficacy (99.23 %) was observed at 15 days post treatment; however, the average efficacy of the drug remained extremely high (90 %) for all the study. The lambs treated with diclazuril showed an intense, persistent oocyst excretion with average levels of 97.54 opg, (p0.05). This study demonstrates the good efficacy of toltrazuril administered orally to lambs in the prepatent period in subclinical natural Eimeria infections in housed lambs.

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.

Eucoleus bohmi (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. bohmi 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 ( cox 1) gene of E. bohmi . Adult worms of E. bohmi 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. bohmi , 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%.

In the last decade an increase of the number of red foxes in anthropized habitats across European countries, including Italy, has been observed. This pones implications in terms of disease transmission between wildlife and domestic animals; in fact, there are evidences of the role of foxes as reservoirs and amplifiers of a broad spectrum of parasites infecting pets. The present study evaluated the prevalence of Eucoleus boehmi, an emerging extra-intestinal nematodes of the Capillariinae subfamily, in red foxes. The nasal passages and sinuses of 179 red foxes culled from several areas of northern and central Italy were inspected and the mucosal surfaces were scrapped and examined for adult nematodes and eggs, microscopically and genetically identified. Overall 55 foxes (30.7%) were found to be infected with E. boehmi, i.e. 27 on inspection of the nasal passages and sinuses and 28 on mucosal flush and scraping. The occurrence of E. boehmi was significantly (p < 0.05) correlated to the sampling location, the age and gender of the animals examined; the higher rates of prevalence were observed in animals culled in Piedmont (43.3%) and in female (60.6%) and adult (38.1%) subjects. A total of 184 adult parasites were recovered, with a mean intensity of infection of 3.34, and a more frequent localization of E. boehmi in the nasal passages rather than in the sinuses. A significant (p < 0.05) relationship was found between the parasite burden and body condition and age of the animals; the intensity of infection was significantly higher in juveniles (mean: 6.3 specimens) and in animals showing poor fox body condition (mean: 7.8 specimens). These results show that E. boehmi is highly prevalent in the red fox populations of certain areas of Italy. Epidemiological implications are discussed, with a special focus on the role that this wild canid may have in the increasing transmission of nasal eucoleosis to domestic dogs.