Intestinal Parasites and Fecal Cortisol Metabolites in Multi-Unowned-Cat Environments: The Impact of Housing Conditions.

Housing conditions were assessed in different unowned multi-cat management models in order to evaluate their impact on the occurrence of intestinal parasites and fecal cortisol metabolite (FCM) levels. Fresh stool fecal samples were collected from rescue shelters, catteries and feline colonies for coprological analyses in order to detect intestinal parasite patency and fecal cortisol metabolites. A questionnaire provided information about the facilities, management and housing conditions of cats, including information about dog exposure, enclosure size, environment enrichment and changes to group composition. Overall, intestinal parasite infection was detected in 58.2% of fecal samples collected. The occurrence of intestinal parasites detected in free-roaming cats was 82.2%, mainly due to helminth infection. The parasite infection rate was 57.3% in rescue shelters and 34.6% in catteries. In confined cats, protozoa infection was more likely detected in rescue shelters than in catteries (RR = 2.02 (1.30-3.14), p = 0.0012). Although the FCM values were very variable between cats, the enclosure size and parasite infection were correlated with the average FCM. A small enclosure size was correlated with high fecal cortisol metabolites (p = 0.016). Protozoa-positive samples showed higher FCM levels than negative samples (p = 0.0150). High dog exposure was statistically associated with protozoa infection (p = 0.0006). The results indicated that improving housing, especially in terms of floor space and avoiding dog exposure, reduces stress and can thus be applied to make control strategies in multi-unowned-cat environments more efficient, especially when cats are confined.

Detection of Rickettsia massiliae/Bar29 and Rickettsia conorii in red foxes (Vulpes vulpes) and their Rhipicephalus sanguineus complex ticks.

To determine the prevalence of exposure to Rickettsia massiliae/Bar29 and Rickettsia conorii in wild red foxes, we collected blood samples and ticks from 135 foxes shot in different game reserve areas in Catalonia. To detect SFG rickettsia in Rhipicephalus sanguineus complex ticks collected from the foxes, we used real-time polymerase chain reaction (PCR) to screen for ompA gene and a tick-borne bacteria flow chip technique based on multiplex PCR. Serum samples were positive for antibodies against spotted fever group (SFG) rickettsiae in 68 (50.3%). Molecular techniques identified R. massiliae in 107 ticks, R. aeschlimannii in 3 ticks, and R. slovaca in one tick; no R. conorii was identified in any of the ticks analyzed. We conclude that red foxes can carry ticks with SFG rickettsia.

Cardiopulmonary helminths in foxes from the Pyrenees.

The present survey was carried out to investigate the prevalence of cardiopulmonary helminths in red foxes in Pyrenees area and to evaluate the role of foxes in the eco-epidemiology of these nematodes. Hearts and entire respiratory tracts were obtained from 87 foxes from Vall d'Aran region, Pyrenees, Catalonia, north-eastern Spain. The cardiopulmonary tracts were dissected, flushed and examined for nematodes using sedimented flushing water. Of the 87 examined foxes, 53 (61%) were positive for cardiopulmonary helminths. The identified nematodes were Crenosoma vulpis (44.8%), Eucoleus aerophilus (29.9%) and Angiostrongylus vasorum (3.4%). Statistical differences were observed only on comparing age and C.vulpis prevalence, with young foxes being more infected than adults. The high prevalence of cardiopulmonary nematodes suggested that red foxes may play an important role in their transmission and maintenance in the studied area.

The role of cats in the eco-epidemiology of spotted fever group diseases.

BACKGROUND: Mediterranean Spotted Fever (MSF), whose etiological agent is R. conorii, is one of the oldest described vector-borne infectious diseases. Although it is endemic in the Mediterranean area, clinical cases have also been reported in other regions. R. massiliae-Bar29 is related to MSF cases. This strain is distributed worldwide. R. conorii and R. massiliae-Bar29 are transmitted by ticks. Dogs are considered the sentinel of R. conorii infection. Cats could also be involved in their transmission. Rickettsia felis, etiological agent of Flea-borne spotted fever, is mainly transmitted by the cat flea, Ctenocephalides felis. Up to now, the role of cats in its transmission is not entirely elucidated. The aim of the study is to analyze the infection in cats by these microorganisms. METHODS: The study was undertaken in Northeastern Spain. Twenty municipalities of seven regions participated in the study. 212 cats (pets and stray cats) were analyzed. Variables surveyed were: date of collection, age, sex, municipality, source, living place, outdoor activities, health status, type of disease, contact with other animals, and ectoparasite infestation. Sera were evaluated by indirect immunofluorescence antibody assay (IFA). Molecular detection (real-time PCR and sequencing) and cultures were performed on blood samples. RESULTS: There were 59 (27.8%) cats seroreactive to one or more microorganisms. Considering cross-reactions, the seroprevalences were 15.6%-19.5% (R. massiliae-Bar29), 1.9%-6.2% (R. conorii), and 5.2%-7.5% (R. felis). A weak association was observed between SFG seropositivity and tick infestation. Ticks found on seropositive cats were Rhipicephalus pusillus, R. sanguineus and R. turanicus. DNA of Rickettsia was detected in 23 cats. 21 of them could be sequenced. Sequences obtained were identical to those sequences of SFG rickettsiae similar to R. conorii and R. massiliae. No amplification of R. felis was obtained. CONCLUSIONS: Cats can be infected by SFG rickettsiae and produce antibodies against them. Cats may play a role in the transmission cycle of R. conorii and R. massiliae-Bar29, although the role in the R. felis cycle needs further analysis.

Prevalence of intestinal parasites in shelter and hunting dogs in Catalonia, Northeastern Spain.

To compare the prevalence of intestinal parasites in shelter and hunting dogs in Catalonia, Northeastern Spain, fresh faecal samples from 81 shelter dogs and 88 hunting dogs were collected and analysed by faecal flotation. The overall prevalence of intestinal parasites was 71.6% in each population. In the shelter dog group, 67.9% of dogs were positive for intestinal protozoa and 9.8% were positive for helminths. In the hunting dog group, 20.4% of dogs were positive for intestinal protozoa and 63.6% were positive for helminths. A subset of Giardia-positive samples was evaluated by PCR; Giardia assemblages C or D were detected. These results suggest that comprehensive parasite control measures should be implemented in both shelter and hunting dogs in Catalonia.

The role of dogs in the eco-epidemiology of Rickettsia typhi, etiological agent of Murine typhus.

Rickettsia typhi, etiological agent of Murine typhus (MT), is transmitted to humans from an animal reservoir through two cycles: a classic rat-flea-rat cycle, and a peridomestic animal cycle. There are not many studies concerning which animals are involved in the peridomestic cycle, and most of them are focused on cats. The aim of this study was to determine the presence of R. typhi in dogs, not only by serological methods but also by direct methods such as culture and molecular detection. Two hundred and one dog blood samples were collected from Veterinary clinics, kennels, and shelters in Northeastern Spain (2006-2008). Age, sex, municipality, living place, healthy status, contact with animals, and ectoparasite infestations were surveyed. IgG was measured by IFA. Titers ≥ 1/64 were considered positive. Cultures were carried out using samples of dogs with titers ≥ 1/128. The molecular detection was performed by real-time PCR. Nine dogs (4.5%) were positive according to IFA (5: 1/64; 3: 1/128; 1: 1/512). There were no significant differences in the rates of antibodies related to any of the variables. Rickettsial DNA was detected in two cultures. Sequences obtained were identical to those of R. typhi. The results show direct and indirect evidences of the presence of R. typhi infection in dogs.

Evaluation of the presence of Rickettsia slovaca infection in domestic ruminants in Catalonia, Northeastern Spain.

Rickettsia slovaca is the etiological agent of the human disease tick-borne lymphadenopathy (TIBOLA) transmitted by Dermacentor spp. ticks. In our area, Dermacentor marginatus is the most important tick vector; adult ticks feed on mammals, especially ungulates such as wild boars and domestic ruminants. The epidemiology of tick-transmitted diseases describes a wild cycle and a domestic cycle and both are connected by ticks. To identify the role of domestic ruminants in the transmission and maintenance of R. slovaca infection, blood samples from sheep (n=95), goats (n=91), and bullfighting cattle (n=100) were collected during a herd health program, and livestock grazing was selected to ensure tick contact. Samples were analyzed by serology using an indirect immunofluorescent assay (IFA) and molecular techniques (real-time PCR). Seroprevalence was 15.7% in sheep, 20.8% in goats, and 65.0% in bullfighting cattle. On the basis of molecular methods, R. slovaca infection was demonstrated in a goat blood sample with an antibody titer of 1:160. This is the first time that R. slovaca has been identified in a goat blood sample. These results suggest that domestic ruminants are exposed to R. slovaca infection and, because the domestic cycle is close to the human environment, this could increase the risk of transmitting the pathogen to human beings.

Rickettsia felis in fleas from Catalonia (Northeast Spain).

INTRODUCTION: Rickettsia felis produces a syndrome indistinguishable from murine typhus, which has been described in Spain. R. felis is transmitted to humans by fleas. Although no clinical case has been described so far, serologic evidence of infections in humans, cats, and dogs has been obtained in our area. However, no study has been conducted regarding its presence in vectors. Recognition of routes of transmission is of great importance to prevent infection in humans. Taking into account these results, R. felis seems to be present in animals that are in contact with humans. The aim of this study was to determine the presence of R. felis in the fleas of cats and dogs from Northeast Spain, to show the presence of peridomestic cycle in our area. MATERIALS AND METHODS: Between May 2006 and July 2008, 78 fleas were collected. Sixty-three fleas were recovered from kennels. Most of them were collected from cages and a few of them on dogs and cats living in kennels. Fifteen fleas were collected from dogs and cats attended at a veterinary clinic. Fleas were rinsed with ethanol, dried, identified, and stored at 4°C. DNA was extracted from each flea individually. Rickettsial DNA was determined by quantitative real-time polymerase chain reaction. OmpB-specific primers and molecular beacon probes targeting specifically R. felis were used. RESULTS: All 78 fleas were identified as Ctenocephalides felis. R. felis was detected in 34 (43.6%) fleas. No nucleic acids were amplified from negative controls and expected results were obtained from positive controls. Eight positive samples were also confirmed by sequencing. CONCLUSIONS: R. felis was found in a high percentage of Ct. felis from cats and dogs. It seems that there is a peridomestic cycle in Northeast Spain, which would allow contact of R. felis with humans.

Anti-HEV antibodies in domestic animal species and rodents from Spain using a genotype 3-based ELISA.

A truncated ORF2 capsid HEV antigen derived from a genotype 3 strain was developed in insect cells and insect larvae, and compared with the Sar55 antigen and a commercial ELISA. The antigen expressed in insect cells showed a better correlation with Sar55 (kappa value (k)=0.84) than the insect larvae antigen (k=0.69), and a better reproducibility as indicated by the intra and interplate variation coefficients. Commercial ELISA designed for human diagnosis but adapted to animal use using specific secondary antibodies demonstrated to have a very low sensitivity. The insect cell expressed antigen was used to develop an ELISA to detect anti-HEV-IgG in serum samples of different domestic animal and rodents. Seropositivity in the studied animal populations was 71.4% for pigs, 0.60% for goats, 1.92% for sheep, and 11.11% for cats. None of the 1170 cattle samples or 166 rodent samples analyzed was positive.

Seroprevalence of antibodies to Neospora caninum in dogs from Spain.

The prevalence of Neospora caninum antibodies was determined in sera of 139 dogs from Catalonia (northeastern Spain) using the indirect immunofluorescence antibody test (IFAT). Antibodies in the IFAT were found in 17 of 139 dogs (12.2%) with titers ranging from 1:50 to 1: 1,600. Seroprevalence was higher in dogs over 1 yr old compared with dogs younger than 1 yr (P < 0.05). No statistical difference was observed when sex, breed, purpose, or modus vivendi was compared with seropositivity. Most dogs had low antibody titers, which indicated subclinical infection in the area studied. No neosporosis-related disease was reported from any dog, although a German shepherd with an antibody titer of 1:800 showed pododermatitis. All sera were also screened using a commercial direct agglutination test (DAT). The DAT showed a similar specificity but a lower sensitivity when compared with IFAT as a reference technique.