Evaluation of serum cystatin-C in dogs with visceral leishmaniasis

Serum Cystatin C (sCys-C) is one of the most important serum markers of renal function assessment in dogs. The purpose of this study was to determine the sCys-C concentration in dogs with visceral leishmaniasis (VL). In the study, 16 dogs with VL and 10 clinical healty dogs (control) were used. Mean sCys-C concentration of the infected dogs was significantly higher than that of the control group (p < 0.05). Mean serum creatinine concentration was lower and mean blood urea nitrogen, albumin and globulin concentrations were higher in dogs with VL; however, these changes were not statistically significant. Mean total protein and phosphorus concentrations were found to be higher in dogs with VL than healthy dogs (p < 0.05). No significant correlation had been determined between sCys-C and other variables. Visceral leishmaniasis in dogs has increased sCys-C concentration indicating a possible renal impairment; however, further studies are needed to be performed together with renal biopsies in the investigation sCys-C in dogs with VL.     

Distinct antigen recognition pattern during zoonotic visceral leishmaniasis in humans and dogs

Leishmania infantum is a causative agent of endemic zoonotic visceral leishmaniasis (VL) in regions of South America and the Mediterranean. Dogs are the major reservoirs for L. infantum in these regions, and control of disease in dogs could have a significant impact on human disease. Although dogs share many symptoms of VL with humans as a result of L. infantum infection, they also show some unique clinical manifestations, which are often a combination of visceral and cutaneous leishmaniasis, suggesting different mechanisms of disease development in dogs and humans. Here, we compare antibody responses of dogs and humans with VL to various defined leishmanial antigens. Parasite lysate and K39, the two most commonly used antigens for serodiagnosis of VL, detected the highest levels of antibodies in both humans and dogs with VL, whereas the recognition patterns of these antigens were distinct between the hosts. Among other defined antigens tested, LmSTI1 and CPB detected higher levels of antibodies in dogs and humans, respectively. These results indicate there is a difference between humans and dogs in antigen recognition patterns during VL. We infer that different strategies may need to be used in development of vaccines and diagnostics for humans and for dogs. In addition, we show a correlation between antibody titers to several antigens and severity of clinical symptoms during canine VL.     

Hospital incidence of hypospadias in dogs in North America

A review of medical records from 17 North American university veterinary medical teaching hospitals identified 66 dogs with hypospadias. Males predominated (15:1) and cryptorchidism was the most commonly diagnosed second anomaly, often associated with intersexuality. Fifteen cases, diagnosed at 10 different university hospitals, were Boston terriers, strongly suggesting that this breed has a familial predisposition for hypospadias. Male mongrels, as a group, were seen in approximately the same proportion as their representation in the hospital population, suggesting other components in the aetiology of the condition. If maternal histories are obtained as new cases are identified, a better understanding of the factors associated with the aetiology of the condition may be possible.     

Evaluation of the potential causes of epistaxis in dogs with natural visceral leishmaniasis.

Haemostasis was evaluated in 19 dogs with natural Leishmania infection, six of them with a history of epistaxis, and the results were compared with the results from 24 healthy dogs. In addition, the dogs' blood pressure was measured and biopsies were taken from the nasal mucosa. Buccal mucosa bleeding time was prolonged in the dogs with leishmaniasis (P < 0.002) and most significantly in those with epistaxis (P < 0.005). None of the Leishmania-infected dogs had thrombocytopenia, low levels of plasma von Willebrand factor antigen, a prolonged prothrombin time or activated partial thromboplastin time, a low plasma fibrinogen concentration or high serum fibrin degradation products. These results rule out defects of secondary haemostasis or disseminated intravascular coagulation as significant causes of epistaxis in non-complicated leishmaniasis. Histopathology of the nasal mucosa of 10 of the affected dogs, three of them with epistaxis, revealed ulcerative and inflammatory lesions in all of them.     

Experimental visceral leishmaniasis in the opossum

Adult wild-trapped opossums were infected with Leishmania donovani (Khartoum strain, WR 378) and evaluated as an animal model of visceral leishmaniasis. All infected opossums died within 32 days. Loss of body fat, hepatomegaly, and petechiae of skin and abdominal musculature were seen at necropsy. Microscopically, numerous amastigote-laden macrophages were seen in histologic sections of liver, spleen, and lymph nodes; fewer parasite-laden macrophages were in the bronchial-associated lymphoid tissues and renal glomeruli. Hematological findings included thrombocytopenia (terminal), neutropenia, and lymphopenia. Blood lymphocyte blastogenesis in response to concanavalin A and phytohemagglutinin was decreased markedly at day 24 post-infection (PI). Serum antibodies (1:40 dilution) to promastigotes of L. donovani were detected in five of eight infected opossums tested on days 10 and 24 PI. Total bilirubin concentrations and alanine aminotransferase and aspartate aminotransferase activities were increased after day 25 PI. Activated partial thromboplastin times and one-stage prothrombin times were prolonged before death. Concurrently, factors V, VIII, and XII activities were decreased.     

Diagnosis of anatoxin-a poisoning in dogs from North America.

Anatoxin-a, a toxin produced by several genera of blue-green algae, is considered a potent neurotoxin. Ingestion of water contaminated with the toxin results in acute neurological signs and often death. This report describes fatal cases of anatoxin-a ingestion in 6 dogs, with confirmation of anatoxin-a exposure by liquid chromatography/tandem mass spectrometry (LC-MS/MS/MS). In 1 outbreak, 3 dogs developed seizures and died within an hour after swimming in a river in California, while the other outbreak involved 3 dogs that died within 1 hour after swimming in a pond in Ontario. Anatoxin-a poisoning is rarely reported in dogs as a cause of acute neurological signs and death. However, increased occurrences of blue-green algae blooms in North America make this neurotoxin an important consideration in the diagnosis of sudden death associated with environmental water exposure. This brief communication reports on the isolation and detection of anatoxin-a from environmental water sources and the stomach contents of North American dogs dying of acute neurotoxicosis. This demonstrates the first documented cases of anatoxin-a poisoning in dogs in North America and the importance of LC-MS/MS/MS in identifying neurotoxins responsible for sudden death in cases of suspected blue-green algae toxicosis; especially those cases showing no gross or histological lesions.     

Prevalence of the breed-related glaucomas in pure-bred dogs in North America

Objective To determine the prevalence of the breed‐related glaucomas in pure‐bred dogs presented to the veterinary medical teaching hospitals in North America that participate in the Veterinary Medical Data Base (VMDB). Materials and methods In this retrospective study, age of first diagnosis, breed, and gender data for all breeds of dogs were collected from the VMDB with the clinical diagnosis of primary glaucoma (glaucoma‐NOS) at 5–10 year intervals from 1964 to 2002. The prevalence for each breed (affected dogs compared to all dogs of each breed), any changes over the 38 years, and any gender differences for these glaucomas were determined. Results The prevalence of the primary breed‐related glaucomas has gradually increased from 0.29% (1964–1973); 0.46% (1974–1983); 0.76% (1984–1993); to 0.89% (1994–2002). Breeds that consistently featured among the highest 10 for glaucoma prevalence from four different periods (1964 to 2002) included American Cocker Spaniel, Basset Hound, Wire Fox Terrier, and Boston Terrier. During the last observation period (1994–2002), 22 different breeds had 1% or higher prevalence of the glaucomas. The highest prevalence of glaucomas in 1994–2002 by breed included: American Cocker Spaniel (5.52%); Basset Hound (5.44%); Chow Chow (4.70%); Shar‐Pei (4.40%); Boston Terrier (2.88%); Wire Fox Terrier (2.28%); Norwegian ElkHound (1.98%); Siberian Husky (1.88%); Cairn Terrier (1.82%); and Miniature Poodle (1.68%). A predominance of females with glaucoma occurred in the American Cocker Spaniel, Basset Hound, Cairn Terrier, Chow Chow, English Cocker Spaniel, Samoyed, and perhaps the Siberian Husky, and a predominance of males in the Australian Cattle dog and St Bernard. Age affected the time for first presentation of the glaucomas in the pure‐bred dog. In the majority of breeds the glaucomas were presented for initial diagnosis in dogs between 4 and 10 years of age. Conclusion Breed‐related glaucomas in pure‐bred dogs are frequently presented to the veterinary medical teaching hospitals in North America. The prevalence of the breed‐related glaucomas in the dog appears similar to humans, and in some breeds exceeds that in humans. In many breeds the high prevalence of the glaucomas suggests a genetic basis.     

Parasitic load and histological aspects in different regions of the spleen of dogs with visceral leishmaniasis

Leishmania infantum causes from subclinical infection to severe disease in humans and dogs. The spleen is one of the organs most affected by the infection. Although evidence exists that the parasitic load distribution and histological alterations may not be homogeneous in the affected organs of naturally infected individuals, it has not been formally demonstrated using the current techniques used for studying the disease.In six dogs naturally infected with Leishmania, parasitic load and histological changes were compared in samples collected from the lower, middle and upper third of the spleen.Parasitic load in the spleen of the group of dogs was variable, revealing a difference of 61 times between animals with the lowest and the highest parasitism. The set of parasitic load values of each dog showed a cluster trend, when compared to the other animals. Nevertheless, the parasitic load values of each dog showed a variation ranging from 3.2 to 34.7 times between lowest and highest value. Histological changes showed recognizable variation in frequency (granulomas) or intensity (perisplenitis) in the spleen of 2 out of the 6 dogs. The agreement of histological findings between samples collected from the different thirds of the spleen was good (kappa coeficient, 0.61–0.80) very good (0.81–0.99) or perfect (1.00), for most of the parameters analyzed.Variability of parasitic load and, to a lesser extent, histological changes in spleen of dogs with visceral leishmaniasis is observed. Such variability may be taken in account in the design of studies on pathogenesis, vaccine and therapeutic drug development.     

Clinical and serological aspects of visceral leishmaniasis in northeast Brazilian dogs naturally infected with Leishmania chagasi

Human visceral leishmaniasis is endemic in the northeast of Brazil, where the domestic dog is an important parasite reservoir in the infectious cycle of Leishmania chagasi. In this study, we evaluated the clinical signs of canine visceral leishmaniasis (CVL), serum protein profile and the antileishmanial IgG antibody production in 86 dogs living in northeast endemic areas of leishmaniasis. Thirty dogs from a leishmaniasis-free area were used as a control group. The major clinical signs of CVL seen were emaciation and skin ulcers (80%), followed by onychogryphosis and conjunctivitis (73%). Depilation was observed in 60% of animals while lymphadenomegaly, splenomegaly, liver enlargement or kidney involvement was less frequent (< or =20%). VL seropositive dogs presented with serum hyperproteinemia, hypoalbuminemia, hypergammaglobulinemia and decreased albumin/globulin ratio. A lower sensitivity and higher specificity was observed for promastigote indirect fluorescent antibody test (IFAT) (83 and 100%, respectively) compared with enzyme-linked immunosorbent assay (ELISA) (94 and 90%), which uses a crude extract of Leishmania. There was a positive correlation between IFAT and ELISA titers of antileishmanial IgG antibodies (Spearman test, P < 0.05), which was augmented in CVL dogs. This study found that the determination of serum protein, A/G ratio and the use of two different leishmanial serological tests like IFAT and ELISA are essential in CVL screening.     

Experimental visceral leishmaniasis in the owl monkey

Visceral leishmaniasis developed in eight owl monkeys (Aotus trivirgatus) after intravenous inoculation with a Khartoum strain (WR378) of Leishmania donovani. Six monkeys died within 93 days, and two monkeys recovered from the disease. Clinically, signs were weight loss, anemia, and hepatosplenomegaly. Hematologic findings included anemia, granulocytopenia, thrombocytopenia, and lymphocytosis. Analysis of serum or plasma revealed hyperbilirubinemia, azotemia, hyperglobulinemia, hypoalbuminemia, and altered hemostasis. All monkeys developed positive antibody titers to promastigotes of L. donovani and had increases in immunoglobulins M and subsequently G. Liver, spleen, bone marrow, and lymph nodes were the principal organs containing numerous parasitized macrophages. The owl monkey was highly susceptible to L. donovani infection and should be a useful animal model for the study of visceral leishmaniasis.     

Pancytopenia in a cat with visceral leishmaniasis

Abstract: A 4‐year‐old, domestic shorthair, female spayed cat was presented for decreased appetite and depression. Severe pancytopenia with erythrocyte autoagglutination was found. The cat was seronegative for feline immunodeficiency and leukemia viruses. Immune‐mediated hemolytic anemia was suspected but no response to treatment with a blood transfusion, enrofloxacin, and prednisone was observed. Blood and bone marrow smears obtained 11 days later contained Leishmania amastigotes in the cytoplasm of neutrophils and macrophages, respectively. Serologic and PCR testing of peripheral blood confirmed infection with Leishmania infantum. Despite treatment, the cat worsened clinically and was euthanized. At necropsy, visceral dissemination of the parasite was confirmed. The findings in this case indicate that visceral leishmaniasis should be considered as a differential diagnoses in cats with pancytopenia in areas endemic for Leishmania. In addition, amastigotes may be observed in peripheral blood neutrophils.     

Transmission of visceral leishmaniasis through blood transfusions from infected English foxhounds to anemic dogs.

OBJECTIVE: To conduct serologic surveillance for Leishmania spp in English foxhounds from a kennel, as well as recipients of blood from these dogs, and determine whether L infantum organisms could be transmitted via blood transfusion. DESIGN: Serologic prevalence survey. ANIMALS: 120 English foxhounds and 51 dogs of various breeds receiving blood from these donors. PROCEDURE: Foxhound blood donors, foxhound nondonors, and nonfoxhound blood recipient dogs were evaluated serologically for Leishmania spp by indirect fluorescent antibody testing. Dogs that received packed RBC (PRBC) transfusions from foxhound donors from mid-1996 through mid-2000 were identified. Furthermore, dogs were serologically evaluated if they had received fresh frozen plasma (FFP) transfusions in 1999 and 2000 from seropositive foxhound blood donors. RESULTS: Thirty percent of the English Foxhounds were seropositive for Leishmania spp (titer > or = 1:16), although the degree of seropositivity varied considerably during the period. Furthermore, 57 foxhounds had been used as donors from 1996 to 2000, and 342 units of PRBC had been transfused to at least 227 patients. All 25 dogs screened that received PRBC from seronegative foxhound donors tested negative, whereas 3 of 7 dogs that received PRBC from seropositive donors tested positive. All 9 dogs that received FFP from seropositive foxhound donors remained seronegative. CONCLUSIONS AND CLINICAL RELEVANCE: To our knowledge, this report documents the first transmission of Leishmania spp by blood transfusion. The use of foxhounds as blood donors may not be advisable in North America.     

Biology and control of ticks infesting dogs and cats in North America

At least 15 different infectious agents and diseases are transmitted or produced by ticks parasitizing dogs and cats in North America. Those infectious agents and diseases are distributed among eight different tick species that commonly infest dogs and cats. All but one of these species are hard ticks with a three-host development cycle in which each motile stage (larva, nymph, and adult) feeds on a different host after molting. Tick species, disease occurrence, and peak activity of each tick life stage can vary dramatically depending on geographic and climatic conditions. Gaining an understanding of tick distribution, tick ecology, and seasonal occurrence of different tick life stages can help with the management of tick infestations and reduce the incidence of tick-transmitted diseases in dogs and cats. Control should be based on an understanding and management of ecologic factors responsible for tick infestations and selection of appropriate acaricides. Occasionally, topical acaricides will appear not to have prevented tick infestations. This lack of control may be real or perceived based on reinfestation rates and/or pet owner expectations of product performance.     

Analysis of immune responses in dogs with canine visceral leishmaniasis before, and after, drug treatment.

The incidence of canine visceral leishmaniasis (CVL) is increasing in the Mediterranean region. Many drugs have been tested for treatment of CVL, but little is known regarding their effect on test immune responses. In our study, three dogs naturally infected with Leishmania infantum and five dogs experimentally infected with the same strain, were treated with dimethasulfonate pentamidine (Lomidine) and the immune response evaluated before, and after, treatment. After the last injection, animals began to gain weight and the major clinical signs disappeared. Antibody titers gradually decreased to low levels, six months after treatment. At the same time, antigen specific lymphoproliferation reappeared in the sampled animals. This study shows that, after treatment, immune cellular responses to leishmanial antigens, involved in protection against Leishmania infection, were established.     

Autochthonous Dirofilaria (Nochtiella) repens infection in dogs in Kuwait

Human dirofilariasis caused by Dirofilaria repens is a rare but increasingly reported zoonosis in the Middle East. However, eligible animal reservoirs seldom have been investigated in the area until now. Seven (1.8 percent) out of 381 dogs examined in Kuwait carried D. repens microfilariae showing compatible cutaneous clinical signs and positive response to the adulticide and microfilaricide therapies. This study indicates that dogs from Kuwait are natural hosts of D. repens and therefore act as an animal reservoir for the zoonotic condition.     

Autochthonous infections with Angiostrongylus vasorum in dogs in Switzerland and Germany

Angiostrongylus vasorum is endemic in foxes an other carnivores in the South and south East of France, Denmark and Great Britain. The reddish nematode is present in the Arteria pulmonalis and the right side of the heart and causes respiratory and cardiovascular symptoms. From 1999 to 2004, A. vasorum was diagnosed in 5 dogs from northern Switzerland, in 1 dog from southern Germany and in 3 dogs from south Ticino. Clinical signs in the affected dogs varied and ranged from cough, tachypnoea and dyspnoea to neurological symptoms in 2 of the dogs. Four dogs died and in 3 of the 4 dogs adults and larvae were found after digestion of formalin-fixed tissue. Diagnosis in the other 5 dogs was achieved by detecting the larvae coproscopically. Based on anamnestic data, these 9 dogs are probably autochthonous cases.     

Evaluation of renal and hepatic functions in dogs naturally infected by visceral leishmaniasis submitted to treatment with meglumine antimoniate

The present study aimed to evaluate the renal and hepatic responses in eight dogs with visceral leishmaniasis submitted to treatment with meglumine antimoniate and to verify the occurrence of possible side effects. Urinalysis, hepatic and renal function tests were carried out in all animals at up to seven moments. After the end of a six-month observation period, all dogs were euthanized. Before the beginning of the experiment urinary and biochemical alterations were observed in four dogs due to the changes caused by the parasite itself. These alterations included the presence of renal cells, cylindruria, proteinuria, azotemia, hyperproteinemia and hypoalbuminemia. One dog died on the third day after treatment because an aggravation of the clinical picture, probably due to the medication. During the course of the study, an increase in hepatic enzymes was verified in two animals. Sixty days after the beginning of the treatment four dogs showed remission of clinical signs. The other three were asymptomatic with persistent biochemical alterations. From these, two presented recurrence of clinical signs about 150 days after the beginning of the treatment while in the other, hyperproteinemia persisted. Meglumine antimoniate was not efficient to treat dogs with severe renal dysfunction and the side effects observed were pain at the site of injection and the probable transient hepatotoxicity, evidenced by biochemical examinations, but without the presence of clinical signs.