A. Sainz

Serologic and Molecular Evaluation of Leishmania infantum in Cats from Central Spain

Infection by different Leishmania spp. in cats has been reported in many countries. In Spain, since the first Leishmania infection described in 1933, sporadic clinical cases in cats have been reported. Various serologic studies performed in other areas of Spain have shown seroprevalences ranging between 1.7 and 60%. The aim of the present study was to determine the prevalence of leishmaniasis in cats from Central Spain (Madrid), and to assess the existence of associations between Leishmania infantum infection and relevant data obtained from each cat. Two‐hundred thirty‐three cats attended at the Veterinary Teaching Hospital in Madrid between September 2005 and June 2006 were tested for L. infantum using the indirect immunofluorescent antibody (IFA) test (cutoff: 1:100) and PCR. PCR testing was performed on the samples to detect Leishmania infection, targeting the kinetoplast DNA (kDNA). Our results showed a seroprevalence of 1.29% (3/233) using IFA test. Another seven cats were also seroreactive to L. infantum one dilution under the cutoff (1:50). Considering all the seroreactive samples, the percentage of positive animals to L. infantum was 4.29%. Only one of the cats (0.43%) included in the study was PCR‐positive. Relative lymphocytosis and an increase in alanine aminotransferase (ALT) value were statistically associated with seroreactivity to L. infantum. Our results demonstrate the presence of cats seroreactive to L. infantum in Central Spain, an endemic area for this disease in dogs.

Guideline for veterinary practitioners on canine ehrlichiosis and anaplasmosis in Europe

Canine ehrlichiosis and anaplasmosis are important tick-borne diseases with a worldwide distribution. Information has been continuously collected on these infections in Europe, and publications have increased in recent years. Prevalence rates are high for Ehrlichia and Anaplasma spp. infections in dogs from different European countries. The goal of this article was to provide a practical guideline for veterinary practitioners on the diagnosis, treatment, and prevention of ehrlichiosis and anaplasmosis in dogs from Europe. This guideline is intended to answer the most common questions on these diseases from a practical point of view.

HEMATOLOGICAL AND PLASMA BIOCHEMICAL REFERENCE INTERVALS IN YOUNG WHITE STORKS

Hematological and plasma chemistry parameters were measured in 129 juvenile white storks (Ciconia ciconia), either wild or captive bred, April to June 1994. Wild storks were members of a colony in the Lozoya River Valley, Madrid, Spain. Red blood cells count, packed cell volume and hemoglobin increased significantly with age. White blood cells count, lymphocytes count and platelets decreased with age. Total solids, total proteins, fibrinogen, albumin, alpha, beta, gamma-globulins and urea increased with age. Differences between captive and wild birds were not notable.

Seropositivity rates for agents of canine vector-borne diseases in Spain : a multicentre study

BackgroundControlling canine vector-borne diseases (CVBD) is a major concern, since some of these diseases are serious zoonoses. This study was designed to determine seropositivity rates in Spain for agents causing the following five CVBD: leishmaniosis (Leishmania infantum: Li), heartworm (Dirofilaria immitis: Di), ehrlichiosis (Ehrlichia canis: Ec), anaplasmosis (Anaplasma phagocytophilum/Anaplasma platys: An) and Lyme disease (Borrelia burgdorferi: Bb).MethodsAnti-An, -Bb, and -Ec antibodies and the Di antigen were determined using the 4DX SNAP® Test (IDEXX Laboratories) and anti-L. infantum (Li) antibodies using the Leishmania SNAP® Test (IDEXX Laboratories) in blood and/or serum samples.ResultsAmong 1100 dogs examined, overall seropositivity rates were: Li (15.7%), Ec (5%), An (3.1%), Di (1.25%) and Bb (0.4%). While seropositivity towards Bb and Di was similar in all geographic regions, rates were significantly higher in the east of Spain (8.3%) for An, significantly higher in the north (20%) for Ec, and significantly higher in the Southeast (46.6%) and South (27.4%), and significantly lower in the north (0%) for Li.No statistical associations were observed between sex and the CVBD analyzed (p ≥ 0.05) while the following associations with other variables were detected: a higher seropositivity to Ec (40%) and Bb (6.7%) in dogs under one year of age compared with adults (p < 0.05); and a higher seropositivity to An and Li in dogs that lived outdoors versus indoors (p = 0.01; p < 0.001, respectively). Seropositivity rates of 2.1%, 0%, 1.7%, 0.5% and 4.2% were recorded respectively for An, Bb, Ec, Di and Li in dogs with no clinical signs (n = 556) versus 3.8%, 0.6%, 7.5%, 1.8% and 25.9% for those with signs (n = 507) suggestive of a CVBD.ConclusionThe data obtained indicate a risk for dogs in Spain of acquiring any of the five CVBD examined. Veterinarians in the different regions should include these diseases in their differential diagnoses and recommend the use of repellents and other prophylactic measures to prevent disease transmission by arthropod vectors. Public health authorities also need to become more involved in the problem, since some of the CVBD examined here also affect humans.

Vector-Borne Diseases in Client-Owned and Stray Cats from Madrid, Spain

The role of various vector-borne pathogens as a cause of disease in cats has not been clearly determined. The current study evaluated risk factors, clinical and laboratory abnormalities associated with Ehrlichia spp., Anaplasma spp., Neorickettsia spp., Leishmania spp., and Bartonella spp. infection or exposure in 680 client-owned and stray cats from Madrid, Spain. Our results indicate that a large portion (35.1%) of the cat population of Madrid, Spain, is exposed to at least one of the five vector-borne pathogens tested. We found seroreactivity to Bartonella henselae in 23.8%, to Ehrlichia canis in 9.9%, to Anaplasma phagocytophilum in 8.4%, to Leishmania infantum in 3.7%, and to Neorickettsia risticii in 1% of the feline study population. About 9.9% of cats had antibody reactivity to more than one agent. L. infantum DNA was amplified from four cats (0.6%), B. henselae DNA from one cat (0.15%), and B. clarridgeiae DNA from another cat (0.15%).

Distribution and relationships between clinical and biopathological parameters in canine leishmaniasis.

Although the lesions, clinical signs and biochemical alterations observed in the course of canine leishmaniasis have been amply described a thorough definition and characterisation of the affected populations is important in order to detect relationships between parameters which may be involved in the development of this disease and to correctly assess further studies. This study included 61 dogs diagnosed with naturally acquired canine leishmaniasis by means of indirect immunofluorescence assay (IFA). At diagnosis, dogs were classified according to the following qualitative and quantitative variables: gender, breed, age, clinical picture, antibody titre, serum protein electrophoretogram, haemogram (CBC), urea, creatinine and ALT. Our population included dogs of 24 breeds, both sexes and different ages indicating no sex, age or breed predilection. In relation to the clinical picture, eight dogs were asymptomatic, 16 displayed mainly cutaneous signs, 18 presented primarily visceral signs and 19 displayed both cutaneous and visceral signs. Our results indicate that the clinical picture is significantly related to electrophoretogram and to RBC, PCV and haemoglobin. Dogs with mainly cutaneous signs showed the highest eosinophil mean values and those with mainly visceral signs showed the highest α-globulin mean values. This study confirms that the antibody titre is highly correlated with electrophoretogram and with RBC, PCV and haemoglobin. Lymphocytes were not associated or correlated with any other variable considered. PMNC, monocytes and eosinophils, as well as WBC, showed a significant correlation with β-globulins, which is difficult to interpret.

Serological Reactivity to Ehrlichia canis, Anaplasma phagocytophilum, Neorickettsia risticii, Borrelia burgdorferi and Rickettsia conorii in Dogs from Northwestern Spain

The aim of the present work was to investigate the seroprevalence against Ehrlichia canis (Ec), Anaplasma phagocytophilum (Ap), Neorickettsia risticii (Nr), Rickettsia conorii (Rc), and Borrelia burgdorferi (Bb) in two different clusters of canine samples from Northwestern Spain. Cluster 1 included 479 dogs presented at veterinary clinics located in Ourense and Pontevedra. Cluster II included 170 dogs from the public kennel of Ourense. All 649 canine serum samples were analyzed by immunofluorescent antibody test. Prevalences against the above-mentioned agents in cluster I were: Rc (24.6%), Bb (6.26%), Ec (3.13%), Ap (5.01%), and Nr (1.04%), whereas for cluster II were: Rc (50%), Bb (8.8%), Ec (54.7%), Ap (45.3%), and Nr (4.7%). Rc was significantly associated with age and history of exposure to ticks, and Bb showed a statistical relationship with age and clinical status. Ec and Ap were related to the occupation of the dogs, with stray dogs being the most frequently seropositive. Furthermore, seroreactivity against Ec and Ap was significantly higher in Ourense than in Pontevedra. The univariate analysis demonstrated a significant concomitant seroreactivity between Ec and Ap and between Rc and Ec and Ap antigens. The seroreactivity to Nr must be interpreted very cautiously as this infectious agent has been seldom reported outside North America.

First report of an intraerythrocytic small piroplasm in wild Iberian lynx (Lynx pardinus).

A wild injured Iberian lynx (Lynx pardinus) was taken from the Sierra Morena population. During the health check small intraerythrocytic piroplasms, morphologically indistinguishable from other feline piroplasms, were observed in Wright-Giemsa-stained blood films. Amplification by polymerase chain reaction of a portion of the 18S nuclear small subunit (NSS) rRNA gene and sequencing revealed similarity of the unknown organism with sequences obtained from Pallass cat from Mongolia and from a domestic cat in Spain. In a retrospective (1993–2003) study of 50 Iberian lynx tissue samples, no amplifications of the 18S NSS rRNA gene of the organism were obtained. This is the first report of a naturally occurring erythroparasitemia in the Iberian lynx and the first documented case of naturally occurring piroplasm infection in a free-ranging felid from Europe.

Seroprevalence of Leishmania infantum in Northwestern Spain, an Area Traditionally Considered Free of Leishmaniasis

Abstract: Northwestern Spain has traditionally been considered to be free from leishmaniasis. The aim of this work was to determine the prevalence of canine leishmaniasis in this area and to assess the influence of several risk factors on the incidence of this disease. A total of 479 dogs attended at different veterinary clinics in northwestern Spain were tested for L. infantum with the immunofluorescent antibody (IFA) test. The seroprevalence of L. infantum in this area was 3.7%. Most of the seropositive dogs lived in two locations: Valdcorras (seroprevalence of 29.2%) and Ourense (seroprevalence of 7.5%). The detection of high antibody titers in most of the seropositive dogs (many of which presented clinical signs) coupled with the certainity that some of these dogs had never been outside their home areas indicates the presence of this zoonosis in these two sites. On the other hand, companion dogs were significantly less likely to acquire the disease than sheep dogs, hunting dogs, and those from kennels.

Assessment of Feline Ehrlichiosis in Central Spain Using Serology and a Polymerase Chain Reaction Technique

Abstract: Antibodies to Ehrlichia spp. and inclusion bodies compatible with Ehrlichia spp. in feline blood cells have been previously detected in Spain. The aim of this study was to assess the presence of antibodies to E. canis, N. risticii, and A. phagocytophilum in 122 feline serum samples from Madrid (central Spain). In addition, Ehrlichia genus‐specific, one‐tube, nested polymerase chain reaction (PCR) was performed from blood samples from these cats. Of the cats, 10.6% were seropositive for E. canis, 2.4% were positive for N. risticii, and 4.9% were seropositive for A. phagocytophilum. Two N. risticii‐positive cats and one animal seropositive to A. phagocytophilum were also seropositive for E. canis. Despite these seropositive results, all the blood samples analyzed by PCR were negative. Our results demonstrate reactivity against agents implicated in feline ehrlichiosis in Spain. Further studies should be performed in order to clarify the significance of serology and PCR in the diagnosis of feline ehrlichiosis.