Dog erythrocyte antigens (DEA) 1, 4, 7 and suspected naturally occurring anti-DEA 7 antibodies in Italian Corso dogs.

We sought to determine the prevalence of dog erythrocyte antigen (DEA) 1, 4 and 7 and naturally occurring anti-DEA7 antibodies in Italian Corso dogs. In addition, we correlated DEAs with different epidemiologic variables, compared the prevalence of DEAs against other canine populations and assessed the risk of sensitisation and transfusion reactions (TRs) following unmatched transfusion. Blood samples from 100 Corso dogs were evaluated for DEA 1, 4, 7 and naturally occurring anti-DEA 7 antibodies. Seventy-one percent of samples were DEA 1-negative, 100% tested DEA 4-positive, and 95% tested DEA 7-negative. Suspected anti-DEA7 antibodies were found in 32% dogs. The DEA 1 and 7-negative phenotypes were significantly more common than in most canine populations. When a previously tested Italian canine population was considered as blood donors for Corso dogs, the risk of DEA 1 sensitisation using DEA 1 untyped blood was 29%, and of acute haemolytic TRs after a second untyped DEA 1-incompatible transfusion was 8%. The potential for delayed TRs between DEA 7-negative Corso dogs with suspected naturally occurring anti-DEA 7 antibodies receiving untyped DEA 7-positive blood was 11%. Conversely, when Corso dogs were blood donors for the same population, the risk of DEA 1 sensitisation was 17% and the risk of an acute haemolytic TR after a second DEA 1-incompatible blood transfusion was 3%. Corso dogs can be suitable blood donors. Additional studies are needed to clarify whether the high prevalence of naturally occurring anti-DEA 7 antibodies in this breed could increase their risk of delayed TRs when they are blood recipients.

Detection of 'Candidatus Neoehrlichia mikurensis' and other Anaplasmataceae and Rickettsiaceae in Canidae in Switzerland and Mediterranean countries.

INTRODUCTION: 'Candidatus Neoehrlichia mikurensis' is an emerging tick-borne zoonotic agent that primarily affects immunocompromised human patients. Dogs and foxes are frequently exposed to ticks, and both species are in close proximity to humans. This is the first study to systematically investigate the occurrence of 'Candidatus Neoehrlichia mikurensis' in Canidae in Europa. We analyzed 1'739 blood samples from dogs in Switzerland, Italy, Spain and Portugal and 162 blood samples from free-ranging red foxes (Vulpes vulpes) in Switzerland. All samples were tested using a previously described multiplex real-time PCR for the Anaplasmataceae family, the 'Candidatus Neoehrlichia' genus and the 'Candidatus Neoehrlichia mikurensis' species. All Anaplasmataceae positive samples were subsequently tested using specific real-time PCRs for Anaplasma phagocytophilum, Anaplasma platys, Ehrlichia canis and Rickettsia helvetica. Among the tested animals, one dog from Zurich tested positive for 'Candidatus Neoehrlichia mikurensis'. The 12-year old West Highland white terrier had been splenectomized 3 months prior to the blood collection and presented with polyuria/polydipsia. Fanconi syndrome was diagnosed based on glucosuria with normoglycemia and hyperaminoaciduria. A. platys and E. canis were detected in 14/249 dogs from Sicily and Portugal; two of the dogs were coinfected with both agents. Four Swiss foxes tested positive for A. phagocytophilium. R. helvetica was detected for the first time in a red fox. In conclusion, 'Candidatus Neoehrlichia mikurensis' infection should be considered in sick dogs, particularly when immunocompromised. The pathogen seems not to be widespread in Canidae in the investigated countries. Conversely, other Anaplasmataceae were more readily detected in dogs and foxes.

INTRODUCTION: 'Candidatus Neoehrlichia mikurensis' est un agent de zoonose transmis par les tiques qui gagne en importance et concerne principalement les patients immunosupprimés. Les chiens comme les renards sont souvent concernés par des morsures de tiques et vivent en contact étroit avec les êtres humains. Dans le présent travail, nous étudions pour la première fois systématiquement la présence de 'Candidatus Neoehrlichia mikurensis' chez les canidés en Europe. Les échantillons sanguins analysés provenaient de 1'739 chiens de Suisse, d'Italie, d'Espagne et du Portugal ainsi que de 162 renards (Vulpes vulpes) de Suisse. Tous les échantillons ont été examinés avec un test de PCR multiplex en temps réel déjà publié quant à la présence d'agents de la famille des Anaplasmataceae, du genre 'Candidatus Neoehrlichia' et de l'espèce 'Candidatus Neoehrlichia mikurensis'. Les échantillons positifs aux Anaplasmataceae ont ensuite été testés avec un test PCR en temps réel spécifique quant à Anaplasma phagocytophilum, Anaplasma platys, Ehrlichia canis und Rickettsia helvetica. Parmi les échantillons examinés se trouvait celui d'un chien de Zürich qui était infecté par 'Candidatus Neoehrlichia mikurensis'. Ce West Highland White Terrier de 12 ans avait été présenté pour polyurie/polydipsie; il avait été splénectomisé trois mois avant la prise de l'échantillon. Au vu d'une glycosurie et d'une hyperaminoacidurie accompagnées d'une glycémie normale, on a posé le diagnostic de syndrome de Fanconi. A. platys et E. canis ont été mis en évidence chez 14/249 chiens provenant de Sicile et du Portugal; deux chiens étaient infectés par les deux agents pathogènes. Quatre renards suisses étaient positifs à A. phagocytophilium et R. helvetica a été trouvé pour la première fois chez un renard. En résumé, on peut dire qu'une infection à 'Candidatus Neoehrlichia mikurensis' chez un chien malade doit être prise en considération comme diagnostic différentiel, particulièrement chez les anomaux immunosupprimés. Toutefois cet agent n'est pas très répandu chez les canidés des pays examinés, contrairement aux autres Anaplasmataceae spp. qui ont été trouvées plus souvent chez les chiens et les renards.

Cross-sectional survey on Tritrichomonas foetus infection in Italian cats.

The feline genotype of Tritrichomonas foetus is a widespread cause of large-bowel diarrhoea in cats. The aim of this study was to determine the prevalence of the T. foetus infection in cat populations across Italy. Fresh, individual faecal samples were collected from 267 cats, kept in different environments (i.e., private households, breeding structures, municipal catteries and colonies) in three different sites across Italy. The faecal samples were tested by PCR to detect T. foetus. Moreover, the same samples were subjected to a concentration-flotation technique and a commercial direct fluorescent-antibody (DFA) test to detect additional enteric parasites, including Giardia duodenalis. The overall prevalence of T. foetus infection was 5.2%. All the infected cats showed diarrhoea at the time of sampling: 9 out of 14 positive cats were co-infected with G. duodenalis, 1 with Toxocara cati and 3 with Dipylidium caninum. The risk factor analysis showed that not only the breed, but also co-infections with G. duodenalis and Dipylidium caninum were significantly associated with the presence of T. foetus. This study confirms the presence of T. foetus in cats living in Italy, suggesting that this protozoan parasite should always be included in the differential diagnosis of patients referred with large-bowel disease symptoms, especially if they were purebred animals, or affected by other enteric protozoa, such as G. duodenalis.

Molecular detection of Bartonella henselae and Bartonella clarridgeiae in clinical samples of pet cats from Southern Italy.

Bartonella henselae is considered an emerging pathogen of veterinary and medical interest that can be occasionally transmitted to humans. Cats are considered to be the only reservoir host for B. henselae. In this study, we used a nested-PCR assay to investigate the prevalence of B.henselae and Bartonella clarridgeiae DNA in peripheral blood samples, fine needle lymph node aspirate specimens and oral swabs from 85 cats in order to develop an easy diagnostic strategy for the selection of infection-free cats that are being considered as pets, especially for immunocompromised patients. Overall, molecular analysis showed that 71 cats (83.5%) tested PCR positive for the presence of B. henselae DNA. PCR amplification of DNA B. henselae produced positive products from lymph node aspirate specimens (62/85; 72.9%) similar to those obtained from blood samples (60/85; 70.6%) and higher than those from oral swabs (51/85; 60%) of cats. No PCR product was obtained for B. clarridgeiae. The simultaneous analysis of three different clinical samples in our study increased the diagnostic possibilities for B. henselae infection in the examined cats from 60-72.9% to 83.5%. Lymph node aspirates were found to be the most effective clinical samples for the detection of B. henselae and blood samples were the next best. Oral swab samples were used in this study with good results when considered in combination with blood and/or lymph node aspiration. The use of nested-PCR assay on these three clinical samples may enhance the diagnostic sensitivity for bartonellosis in cats irrespective of the clinical status of animals.

Directions for the diagnosis, clinical staging, treatment and prevention of canine leishmaniosis.

Canine leishmaniosis (CanL) due to Leishmania infantum is a life threatening zoonotic disease with a wide distribution in four continents and importance also in non-endemic regions. The purpose of this report is to present a consensus of opinions on the diagnosis, treatment, prognosis and prevention of CanL in order to standardize the management of this infection. CanL is a disease in which infection does not equal clinical illness due to the high prevalence of subclinical infection among endemic canine populations. The most useful diagnostic approaches include serology by quantitative techniques and PCR. High antibody levels are associated with severe parasitism and disease and are diagnostic of clinical leishmaniosis. However, the presence of lower antibody levels is not necessarily indicative of disease and further work-up is necessary to confirm CanL by other diagnostic methods such as cytology, histopathology and PCR. We propose a system of four clinical stages, based on clinical signs, clinicopathological abnormalities and serological status. Suitable therapy and expected prognosis are presented for each of the stages. The combination of meglumine antimoniate and allopurinol constitutes the first line pharmaceutical protocol. However, although most dogs recover clinically after therapy, complete elimination of the parasite is usually not achieved and infected dogs may eventually relapse. Follow-up of treated dogs with blood counts, serum biochemistry, urinalysis, serology and PCR is essential for prevention of relapses. Protection against sand fly bites by topical insecticides is effective in reducing infection, and recent development of vaccines has indicated that prevention by vaccination is feasible.

Pet cats as carriers of Arcobacter spp. in Southern Italy.

AIMS: To evaluate the presence of Arcobacter spp. in different biological samples from domestic cats in Southern Italy by using a species-specific PCR assay and thus to elucidate their potential significance as sources of human infection. METHODS AND RESULTS: We investigated the prevalence of Arcobacter DNA in oral swabs, in peripheral blood samples and fine needle lymph node aspirate specimens from 85 cats of which 17 were clinically healthy and 68 had clinical signs of oral disease or lymphadenomegaly. Overall, molecular analysis has shown that Arcobacter-specific DNA was found in 78.8% (67 of 85) of all the cats. In the 67 Arcobacter-positive cats, 66 (77.6%) and 29 (34.1%) were found positive for Arcobacter butzleri and Arcobacter cryaerophilus, respectively. None of the examined samples gave a PCR product for Arcobacter skirrowii. CONCLUSIONS: This study demonstrates that pet cats commonly carry Arcobacter in the oral cavity. According to the clinical data, the Arcobacter detection results showed no significant difference between cats with oral pathology and those suffering from other different pathologies. SIGNIFICANCE AND IMPACT OF THE STUDY: Pet cats harbour Arcobacter spp. and may play a role in their dissemination in the domestic habitat. The high prevalence in a limited number of cat samples in this study may be of significance.

Efficacy of oxfendazole and fenbendazole against tortoise (Testudo hermanni) oxyurids.

Thirty-six tortoises (Testudo hermanni) with naturally acquired oxyurids infections were used to assess the anthelmintic efficacy of oxfendazole (Dolthene; Merial) and fenbendazole (Panacur; Hoechst Roussel Vet). Animals were randomly assigned to three groups (A, B, and C) based on sex and weight. Animals in group A (seven males and six females) were orally treated with oxfendazole at dose rate of 66 mg/kg, group B animals (nine males and eight females) were orally treated with fenbendazole at dose rate of 100 mg/kg, and group C animals (three males and three females) were not treated and served as controls. All animals were individually stabled in plexiglas boxes under controlled conditions of temperature, humidity, and light beginning 7 days pretreatment and continuing for the duration of the trial. Individual tortoises feces were examined daily by the McMaster technique and drugs efficacy was assessed by the fecal eggs count reduction (FECR) test. Both drugs showed 100% of FECR. However, oxfendazole reached this level 12 days after treatment, whereas 31 days after treatment were necessary to obtain the same stable result with fenbendazole. The two drugs were well tolerated by all the animals and no adverse reactions were observed after treatment.

Factors associated with upper respiratory tract disease caused by feline herpesvirus, feline calicivirus, Chlamydophila felis and Bordetella bronchiseptica in cats: experience from 218 European catteries.

A full history of the management practices and the prevalence of upper respiratory tract disease (URTD) at 218 rescue shelters, breeding establishments and private households with five or more cats was recorded. Oropharyngeal and conjunctival swabs and blood samples were taken from 1748 cats. The prevalences of feline herpesvirus (FHV), feline calicivirus (FCV), Chlamydophila felis and Bordetella bronchiseptica were determined by PCR on swab samples. An ELISA was applied to determine the prevalence of antibodies to B. bronchiseptica. The rates of detection by PCR of each pathogen in the cats in catteries with and without ongoing URTD were, respectively, FHV 16 per cent and 8 per cent; FCV 47 per cent and 29 per cent; C. felis 10 per cent and 3 per cent; and B. bronchiseptica 5 per cent and 1.3 per cent; the seroprevalences of B. bronchiseptica were 61 per cent and 41 per cent, respectively. There was evidence that FHV, FCV and B. bronchiseptica played a role in URTD. The risk factors associated with the disease were less than excellent hygiene, contact with dogs with URTD, and larger numbers of cats in the cattery or household.

Efficacy of the treatment of dogs with leishmaniosis with a combination of metronidazole and spiramycin.

Twenty-seven dogs infected naturally with Leishmania infantum were used in a randomised controlled trial to compare the clinical and parasitological efficacy of an oral treatment with a combination of metronidazole and spiramycin (13 dogs) with the efficacy of conventional treatment with meglumine antimonate and allopurinol (14 dogs) as controls. In the test group one dog had to be withdrawn from the treatment because it developed pemphigus foliaceus; 10 of the dogs were clinically responsive but none was cured parasitologically. In the control group four dogs were withdrawn from the treatment because of side effects; eight of the dogs were clinically responsive but none was cured parasitologically. The control group showed signs of improvement after an average of 30 days, whereas the test group did not show signs of improvement until after an average of 45 days.

Serological analysis and identification of feline calicivirus strains isolated in Sicily.

The isolation and characterization of calicivirus strains from symptomatic cats are reported. The correlations between the feline calicivirus strains isolated and the vaccinal strain FCV F9 were investigated by a virus-neutralization test, suggesting a strong antigenic variability.

Activity and postantibiotic effect of marbofloxacin, enrofloxacin, difloxacin and ciprofloxacin against feline Bordetella bronchiseptica isolates.

The minimum inhibitory concentration (MIC) and postantibiotic effect (PAE) of marbofloxacin, enrofloxacin, difloxacin and ciprofloxacin were evaluated in vitro against 43 feline-source Bordetella bronchiseptica strains. All strains tested were susceptible to marbofloxacin and enrofloxacin (MIC90 0.5mg/l), while 93 and 84% of the strains were susceptible, respectively, to ciprofloxacin and difloxacin with MIC(90) values of, respectively, 1 and 8mg/l. The PAE was studied in 10 strains by exposure of bacteria to marbofloxacin, enrofloxacin, difloxacin and ciprofloxacin at 5 and 10 times minimum inhibitory concentration (MIC) for 1 and 2h. Regrowth was determined by measuring the viable counts after drug removal by a 10(3) dilution procedure. PAEs increased as a function of concentration and exposure time. The mean duration of PAEs varied between 1.1 and 8.2h, showing the following order: marbofloxacin>enrofloxacin>ciprofloxacin>difloxacin. These data are encouraging since fluoroquinolones have a possible role in the clinical treatment of B. bronchiseptica infections, and the strong PAE caused by quinolones may contribute to the in vivo efficacy of these drugs.

Activity of nine fluoroquinolones against strains of Bordetella bronchiseptica.

Thirty-two strains of Bordetella bronchiseptica were tested for their antimicrobial susceptibilities to nine fluoroquinolones. The most active agents were fleroxacin, temafloxacin, ciprofloxacin (MIC90 1 microg/ml), ofloxacin, lomefloxacin and enoxacin (MIC90 2 microg/ml). Pefloxacin and norfloxacin were active only against 59.3 and 83.1%, respectively, of the strains tested, whereas rufloxacin lacked activity against all the strains of B. bronchiseptica tested.

Protein degradation and apoptotic death in lymphocytes during Fiv infection: activation of the ubiquitin-proteasome proteolytic system.

The movement of a cell through the sequential phases of apoptosis is accompanied by a progressive decrease in cell size with loss in protein mass. In lymphocytes from Hiv-infected persons, protein loss during apoptosis is due to increased protein degradation rather than decreased synthesis. To identify and characterize the proteolytic enzymes or enzyme systems involved in this process, we studied several features of protein turnover in lymphocytes from peripheral blood and lymph nodes during the natural and experimental infection by feline immunodeficiency virus (Fiv). This animal model allowed us to integrate in vivo results with in vitro observations of protein damage. Here we report that protein breakdown in apoptotic cells is concomitant with the activation of the ATP and ubiquitin-dependent multicatalytic system (proteasome). We suggest that proteasome activation is part of the proteolytic cascade in the execution phases of apoptosis in AIDS.

Infection of small ruminants with Ehrlichia spp. in Sicily.

In 1991, the experimental infection of a goat with pooled blood from goats that were positive for anti-Ehrlichia canis, E. risticii, E. equi and E. phagocytophila antibodies was monitored (physical examination, cell blood count, microscopical examination of blood smears, serology) for 180 days. The infection produced a clinical condition characterized by intermittent fever, anaemia and leukopenia with neutropenia during the first 40 days. Recurrent leukocytosis with lymphocytosis was noticed afterwards. A permanent high-level thrombocytosis appeared after the 18th day. During the first week, cytoplasmic basophilic inclusion bodies were seen in smears of peripheral venous blood stained with May-Grunwald-Giemsa, first in mononuclear cells and then in neutrophils (in max 3% of circulating leukocytes). Seroconversion occurred during the 2nd week and the highest antibody titre (IFAT) was registered vs E. equi (10,240) at the 19th day, vs E. canis (320) at the 24th and vs E. risticii (80) at the 30th day. At the end of the observation period the infected goat was still positive for E. equi (titre 160) and E. canis (titre 10) only. The preinoculation serum of the infected goat was reactive with E. phagocytophila antigen (serum was tested for IF antibodies to E. phagocytophila at 1:200 dilution only, because of the limited quantities of antigen available), but the qualitative evaluation of fluorescence showed an increase from the 7th day, maximum intensity between the 14th and the 40th day and passed to negative from the 74th day. Although it was based on microscopy and serology only and not carried out in a SPF goat, the above experiment gave evidence of the existence of species of the E. phagocytophila genogroup in Italy for the first time.