Symptomatic Dogs Research Articles

First molecular detection of Babesia canis in dogs from Bosnia and Herzegovina

Babesia spp. are tick-transmitted protozoan haemoparasites of great economic, veterinary and medical impact worldwide. Herein we reported the very high prevalence of autochthonous babesiosis in symptomatic dogs from Bosnia and Herzegovina in the period from 2014 to 2016. Eighty dogs that did not leave the country were examined using parasitological and molecular analyses and babesiosis was diagnosed in 82.5% and 85.0% of them, respectively (p < 0.001). One species, Babesia canis was identified using molecular methodology (PCR and sequence analysis). Statistical analyses showed that epizootiological characteristics have no influence on the possibility of infection. Agglomerative hierarchical clustering (AHC) analyses used for comparing the symptoms and clinical signs of infection in dogs pointed out that a high degree of anemia, followed by thrombocytopenia (89%), lethargy (100%), loss of appetite (95%), fever (66%) and icterus (61%) was dominant. In addition, results of the statistical analysis performed showed that more dogs with no data of tick prophylaxis (70%) were found Babesia infected. Those results point to further intensified epizootic surveys in the territory of Bosnia and Herzegovina.

Ultrasonographic findings of shoulder teno-muscular structures in symptomatic and asymptomatic dogs.

B-mode sonographic evaluation of shoulder joint in dogs provides qualitative information concerning mainly tendon and muscles structures. Although the sonographic findings of tenomuscular lesions have been described previously, their frequency in symptomatic and asymptomatic patients has not been reported yet. Aim of the study was to describe and compare qualitative ultrasonographic findings of shoulder joint in clinically symptomatic and asymptomatic dogs and to speculate which lesions might be considered major responsible for lameness. Fifty-two dogs with shoulder lameness and 58 asymptomatic dogs (both with unremarkable radiographic findings) underwent ultrasonographic B-mode examination of the scapulohumeral joint. Lesions detected were recorded and statistically compared between groups. Significant differences between groups were observed for the number of ultrasonographic abnormalities detected and for fluid accumulation, biceps brachii tendon (BT) lesions and bone surface irregularities. Sonography was considered useful for the determination of lesions absence and for the detection of BT lesions, fluid accumulation, muscle lacerations and bone surface irregularities. The odds for symptomatic dogs were greater than for asymptomatic patients to ultrasonographically diagnose BT lesions, fluid accumulation and bone surface irregularities. Ultrasound has proven to be a useful tool in the evaluation of tenomusculoskeletal structures of shoulder in dogs with unremarkable radiographic findings. Our results suggest that ultrasonography is useful in the diagnosis of tendon abnormalities, fluid accumulation, muscle lacerations and bone surface irregularities as potential contributors to shoulder lameness in dogs.

Recombinant prohibitin protein of Leishmania infantum acts as a vaccine candidate and diagnostic marker against visceral leishmaniasis

Visceral leishmaniasis (VL) represents a serious public health problem, as Leishmania infantum is one of main disease causative agents in the Americas. In a previous immunoproteomic study, the prohibitin (PHB) protein was identified in L. infantum promastigote and amastigote extracts by antibodies in asymptomatic and symptomatic VL dog sera. This protein was found to be highly conserved between different Leishmania spp., but it presented a low identity with amino acid sequences of other organisms. The aim of the present study was to evaluate the cellular response induced by the recombinant PHB (rPHB) protein in BALB/c mice, as well as in PBMCs purified from untreated and treated VL patients, as well as to evaluate its protective efficacy against an infection by L. infantum promastigotes. Our data showed that there was a Th1 cellular response to rPHB, based on high levels of IFN-γ, IL-12, and GM-CSF in the immunized animals, as well as a proliferative response specific to the protein and higher IFN-γ levels induced in PBMCs from individuals who had recovered from the disease. The protection was represented by significant reductions in the parasite load in the animals’ spleen, liver, bone marrow, and draining lymph nodes, as compared to results found in the control groups. In addition, an anti-rPHB serology, using a canine and human serological panel, showed a high performance of this protein when diagnosing VL based on high sensitivity and specificity values, as compared to results found for the rA2 antigen and the soluble Leishmania antigenic extract. Our data suggest that PHB has a potential application for the diagnosis of canine and human VL through antibody detection, as well as an application as a vaccine candidate to protect against disease.

Evaluating the performance of IFAT, RDT, and semi-nested PCR for detection of Leishmania infantum infection on sera samples in symptomatic dogs

Zoonotic visceral leishmaniosis is caused by Leishmania infantum in the Mediterranean region and the Middle East, and dogs are the main reservoirs of the disease. The aim of this study was to estimate the diagnostic results of indirect immunofluorescent antibody test (IFAT), rapid diagnostic test (RDT), and semi-nested PCR to detect L. infantum infection on sera samples of symptomatic dogs in an endemic area of Iran. In this study, 23 clinically infected dogs and 75 clinically suspected dogs were selected for the study and sera samples were examined using IFAT, RDT, and semi-nested PCR. Among clinically suspected dogs, 72% (54/75), 26% (13/50), and 36% (27/75) of sera samples were positive for L. infantum by IFAT, semi-nested PCR, and RDT, respectively. A poor agreement was observed between IFAT and semi-nested PCR. A slight agreement was observed between IFAT with PCR and RT methods, and the worse agreement was observed between semi-nested PCR and RDT. The sensitivity and specificity of RDT and semi-nested PCR were calculated according the IFAT results on clinically suspected dogs. The RDT and semi-nested PCR showed a low sensitivity (35%, 38%), whereas the specificity was the highest (100%) in RDT and high (71%) in semi-nested PCR. The IFAT was shown the best results for the diagnoses of visceral leishmaniosis in symptomatic dogs. The RDT and semi-nested PCR had low sensitivity but high and moderate specificity were observed by RDT and semi-nested PCR, respectively. Based on the high-specificity results, RDT could be used as simple test for diagnoses of visceral leishmaniosis in symptomatic dogs.

Serological tests fail to discriminate dogs with visceral leishmaniasis that transmit Leishmania infantum to the vector Lutzomyia longipalpis

The control of reservoirs for Leishmania infantum -induced zoonotic visceral leishmaniasis requires the identification of dogs posing a population risk. Here, we assessed the performance of several assays to identify Lutzomyia longipalpis infectious dogs. We evaluated 99 dogs that were positive for visceral leishmaniasis based on parasite identification. Serological analyses were performed using an enzyme-linked immunosorbent assay, immunofluorescence antibody tests in 1:40 and 1:80 dilutions, rapid dual path platform tests, immunochromatographic assay with a recombinant rK39 antigen, fast agglutination screening tests, and direct agglutination tests. We also performed PCR to analyze peripheral blood and xenodiagnosis. Forty-six dogs infected at least one L. longipalpis specimen. Although the serological test sensitivities were above 85% for detecting L. longipalpis infectious dogs, none showed a satisfactory performance, as both specificity (0.06 to 13%) and the area under the receiver operating characteristic curve (45 to 53%) were low. The PCR results were also weak, with a sensitivity of 30%, specificity of 72%, and an area under the receiver operating characteristic curve of 51%. The infected L. longipalpis proportion was higher among asymptomatic dogs than symptomatic dogs. Among the symptomatic dogs, those with ulceration-free skin diseases were more infectious, with an odds ratio of 9.3 (confidence interval of 1.10 - 428.5). The larger the number of insects fed, the greater the detected infectiousness. Our study supports the imperative to develop novel technologies for identifying the infectious dogs that transmit L. infantum for the benefit of public health.

Association between mast cells, tissue remodelation and parasite burden in the skin of dogs with visceral leishmaniasis

Canine visceral leishmaniosis (CVL) is a zoonosis of major public health impact caused by organisms of the genus Leishmania which is transmitted to human and animals by phlebotomine sand flies. The skin is the first point of contact with Leishmania parasites for sandy fly vectors and it is considered an important reservoir compartment in infected dogs. The aim of this study was to determine the main histophatologic alterations in ear skin of dogs naturally infected by Leishmania infantum with different clinical status and different degrees of parasitism. Therefore, thirty-four dogs naturally infected with L. infantum were grouped according to their clinical status in asymptomatic (AD, n=11), oligosymptomatic (OD, n=11) and symptomatic dogs (SD, n=12) as well as their degrees of parasite load in the skin as low (LP, n=11), median (MP, n=11) and high (HP, n=12) parasitism. Additionally, ten dogs were used as control (CD, n=10). At necropsy, skin samples were collected for further histological and parasitological analysis. The OD and SD groups presented higher parasite burden than AD group. The inflammation was higher in SD group when compared to OD and AD. The LP, MP and HP groups showed an increasing inflammatory process, indicating that a great parasite load is accompanied by a major inflammatory process in the skin. The number of mast cells was higher in the OD and LP groups than CD group, suggesting that these cells may be involved in tissue remodeling, since that an increase of type III collagen fibers and decrease type I collagen fibers were observed in these groups. Taken together, our results enable a better understanding of the alterations in skin of CVL dogs and consequently new insights about the pathogenesis of CVL.

Retrospective Cohort Study to Assess the Risk of Rabies in Biting Dogs, 2013⁻2015, Republic of Haiti.

Background: In canine rabies endemic countries the World Health Organization recommends post-exposure prophylaxis (PEP) be initiated immediately after exposure to an animal suspected to have rabies. Limited capacity in low and middle income countries to assess biting animals for rabies may result in the over prescription of rabies biologics. Few guidelines exist to determine the risk of whether a dog that has bitten someone is rabid. Given PEP cost and access limitations in many countries, accurate and timely assessment of dogs that have bitten people may reduce unwarranted PEP use and improve healthcare seeking behaviors. Methods: Haiti’s animal rabies surveillance program utilizes veterinary professionals to conduct rabies assessments on reported biting dogs and records characteristics of the dog, health outcomes, and laboratory results in a national database. Characteristics of rabid dogs were assessed through a retrospective cohort study of biting dogs investigated during the period from January 2013–December 2015. 1409 biting dogs were analyzed; 1361 dogs that were determined to not have rabies were compared to 48 laboratory-confirmed rabid dogs. Rate ratios, sensitivity, specificity, positive predictive values, negative predictive values, likelihood ratios, quarantine survival of biting dogs, and a risk matrix were developed. Findings: The assessor’s determination that the animal likely had rabies was the most significant predictive factor for a rabid dog (RR = 413.4, 95% CI 57.33–2985, Sn = 79.17, Sp = 91.92). Clinical factors significantly associated with rabid dogs included hypersalivation, paralysis, and lethargy (RR = 31.2, 19.7, 15.4, respectively). Rabid dogs were 23.2 times more likely to be found dead at the time of the investigation compared to case negative dogs (95% CI 14.0–38.6). Rabid dogs were also significantly more likely to lack a history of rabies vaccination or be unowned (RR = 10.3 95% CI 2.5–42.3 and RR = 4.5 95% CI 2.0–10.1, respectively). Rabid dogs were four times more likely to have bitten multiple people (RR = 4.0 95% CI 1.9–8.3). Most rabid dogs died or were killed before quarantine (75%) and all died by day 3 of quarantine, compared to <1% of quarantined case-negatives. The greatest risk of death was predicted to be for persons bitten on the head or neck from symptomatic dogs. Bites from dogs deemed healthy by veterinary assessors and which were available for quarantine presented less than a 0.05% risk of rabies death to the victim. Conclusions: Vaccination of all persons exposed to a suspected rabid dog is a highly effective approach to minimize human rabies deaths. However, this may place undue financial burden on bite victims that have had a low-risk exposure and over-prescription may contribute to regional supply shortages. The results here indicate that in a low-resource country such as Haiti, a well-trained veterinary assessor can provide an accurate risk assessment of biting dogs based on a standard case investigation protocol. In canine rabies endemic countries with limited access to PEP, or where PEP costs may cause undue burden on bite victims, structured risk assessments by trained professionals may be a reliable method of triaging PEP for bite victims. Evaluating rabies risk through a matrix of bite location and risk factor in the dog presents a clear delineation of high and low risk encounters and should be used to develop data-derived PEP recommendations.

Symptomatic Raccoon Dogs and Sarcoptic Mange Along an Urban Gradient.

We quantitatively evaluated the effects of landscape factors on the distribution of symptomatic raccoon dogs with sarcoptic mange along an urban gradient. We used 246 camera traps (182 traps from April 2005 to December 2006; 64 traps from September 2009 to October 2010) to record the occurrence of asymptomatic and symptomatic raccoon dogs at 21 survey sites along an urban-rural gradient in the Tama Hills area of Tokyo. Each occurrence was explained in terms of the surrounding forest, agricultural, and grassland areas and additional factors (i.e., seasonal variations and survey methods) at various spatial scales using a generalized additive mixed model (GAMM). In our analysis, a 1000-m radius was identified as the important spatial scale for asymptomatic and symptomatic raccoon dog occurrence. The peak of the predicted occurrence probability of asymptomatic raccoon dogs appeared in the intermediate forest landscape as opposed to non-forest and forest landscapes. However, a high occurrence probability of symptomatic raccoon dogs was detected in non-forest and intermediate forest landscapes (i.e., urban and suburban) as opposed to a forest landscape, presumably because of animals occurring at much higher densities in more urbanized areas. Therefore, our results suggest that human-modified landscapes play an important role in the high occurrence of sarcoptic mange in raccoon dogs.

A Vaccine Therapy for Canine Visceral Leishmaniasis Promoted Significant Improvement of Clinical and Immune Status with Reduction in Parasite Burden.

Herein, we evaluated the treatment strategy employing a therapeutic heterologous vaccine composed of antigens of Leishmania braziliensis associated with MPL adjuvant (LBMPL vaccine) for visceral leishmaniasis (VL) in symptomatic dogs naturally infected by Leishmania infantum. Sixteen dogs received immunotherapy with MPL adjuvant (n = 6) or with a vaccine composed of antigens of L. braziliensis associated with MPL (LBMPL vaccine therapy, n = 10). Dogs were submitted to an immunotherapeutic scheme consisting of 3 series composed of 10 subcutaneous doses with 10-day interval between each series. The animals were evaluated before (T0) and 90 days after treatment (T90) for their biochemical/hematological, immunological, clinical, and parasitological variables. Our major results showed that the vaccine therapy with LBMPL was able to restore and normalize main biochemical (urea, AST, ALP, and bilirubin) and hematological (erythrocytes, hemoglobin, hematocrit, and platelets) parameters. In addition, in an ex vivo analysis using flow cytometry, dogs treated with LBMPL vaccine showed increased CD3+ T lymphocytes and their subpopulations (TCD4+ and TCD8+), reduction of CD21+ B lymphocytes, increased NK cells (CD5−CD16+) and CD14+ monocytes. Under in vitro conditions, the animals developed a strong antigen-specific lymphoproliferation mainly by TCD4+ and TCD8+ cells; increasing in both TCD4+IFN-γ+ and TCD8+IFN-γ+ as well as reduction of TCD4+IL-4+ and TCD8+IL-4+ lymphocytes with an increased production of TNF-α and reduced levels of IL-10. Concerning the clinical signs of canine visceral leishmaniasis, the animals showed an important reduction in the number and intensity of the disease signs; increase body weight as well as reduction of splenomegaly. In addition, the LBMPL immunotherapy also promoted a reduction in parasite burden assessed by real-time PCR. In the bone marrow, we observed seven times less parasites in LBMPL animals compared with MPL group. The skin tissue showed a reduction in parasite burden in LBMPL dogs 127.5 times higher than MPL. As expected, with skin parasite reduction promoted by immunotherapy, we observed a blocking transmission to sand flies in LBMPL dogs with only three positive dogs after xenodiagnosis. The results obtained in this study highlighted the strong potential for the use of this heterologous vaccine therapy as an important strategy for VL treatment.

Evaluation of pulmonary hypertension and clinical status in dogs with heartworm by Right Pulmonary Artery Distensibility Index and other echocardiographic parameters

BackgroundPulmonary hypertension (PH) is a frequent and severe phenomenon in heartworm disease caused by Dirofilaria immitis, mainly caused by intimal proliferation of the arteries and pulmonary thromboembolisms. Transthoracic echocardiography is the method of choice for diagnosing PH in dogs although the diagnosis is often based on indirect and subjective parameters. The Right Pulmonary Artery Distensibility Index (RPAD Index) is a method that has been recently validated to estimate the presence and severity of PH in heartworm-infected dogs. This study compared some echocardiographic parameters commonly used to estimate PH in 93 dogs infected by D. immitis and evaluated the impact of the parasite burden, microfilaremia, sex or origin of the dog (client-owned/shelter).ResultsNone of the studied echocardiographic variables seemed useful in the estimation of the evaluated clinical aspects, except for the PA/Ao ratio for parasite burden. The RPAD Index was determined in 88 of the dogs; of these, 70.4% had PH (mild: 37.5%, moderate: 19.3%, severe: 13.6%). This Index showed non-significant differences according to microfilaremia, sex, origin or parasite burden. Symptomatic dogs showed PH more often and displayed more severe PH, in addition the presence of symptoms was greater among dogs with high burden; on the other hand 64.4% of asymptomatic dogs had some degree of PH according to the RPAD Index. Apart from the PA/Ao ratio, the other evaluated echocardiographic variables were not useful in evaluating of the hypertensive status of the heartworm-infected dog compared to the RPAD Index.ConclusionsThe estimation of most common indirect parameters is not useful in predicting PH in heartworm-infected dogs. The results confirm the RPAD Index as an objective and supportive test in the monitoring and evaluation of PH in the heartworm-infected dog, and show a potential diagnostic value for the detection of PH in asymptomatic animals.

Parasitism and inflammation in ear skin and in genital tissues of symptomatic and asymptomatic male dogs with visceral leishmaniasis

Canine visceral leishmaniasis (CVL) is transmitted through vector, although venereal transmission has been suggested. This study aimed to compare the parasitic loads and inflammatory processes in genital tissues with ear skin from seropositive male dogs. Forty-five seropositive dogs were separated into groups containing symptomatic (n=23) and asymptomatic (n=22) animals. The control group (n=2) healthy animals with seronegative and negative results in direct parasitological test. Samples of ear tip skin, prepuce, glans penis, testis, epididymis, and prostate were collected for evaluation of parasitic load and inflammatory infiltrate. Although ear tip skin was the most intensely parasitized, prepuce and epididymis revealed no difference in parasitism when compared with ear tip skin (P>0.05). Parasitic loads in testis and prostate were lower than other tissues (P<0.05). Parasitism in glans penis was high, similar to prepuce and epididymis, but lower than ear tip skin. High parasitism was more frequent in symptomatic dogs than asymptomatic animals. Severe inflammatory processes were more frequent within the symptomatic animals compared with asymptomatic and more predominant in prepuce and epididymis. Ear tip skin and genital tissues presented signs of chronic inflammation. There were weak and moderate positive correlations between parasitic loads and inflammatory processes. Our results demonstrate that, likewise with the ear tip skin, the genital of seropositive dogs can carry a large number of Leishmania infantum amastigotes and this process are more intense in symptomatic animals. These data have important implications for understanding the possibility of venereal transmission of CVL.

Barrett’s Disease and Helicobacter Spp. Detection in Dogs with Gastritis

Barrett’s disease (BE) is a metaplastic disorder of the esophageal mucosa in which specialized columnar epithelium replaces healthy squamous epithelium; it is the most common cause of esophageal carcinoma in humans. The dog has been used as a model of Barrett’s esophagus, although the pathology is very rare in the species. In man, infection with Helicobacter pylori resulted inversely associated with BE. The role of Helicobacter spp. in dogs gastritis has not been fully elucidated, as in esophageal disease. Aim of this work was to evaluate esophageal and gastric biopsies of symptomatic dogs in association with the isolation of Helicobacter spp. and to describe and discuss a case of BE.

Molecular diagnosis of Hepatozoon canis in symptomatic dogs in the city of Goiania, Goiás, Brazil

ABSTRACT More than 300 species have been described in the genus Hepatozoon, occurring in different vertebrates. Among these, only Hepatozoon canis and Hepatozoon americanum are seen in dogs. Different methods may be used for laboratory diagnosis. The most common of these is direct parasitological examination of parasite stages in blood smears. The aim of this investigation was to conduct a phylogenetic study on Hepatozoon isolates from symptomatic dogs in the city of Goiânia, Goiás, Brazil. Blood samples were obtained from 40 symptomatic dogs that had been referred to the Veterinary Hospital of the Federal University of Goiás. Among these, only two samples were positive for Hepatozoon spp. using the direct parasitological method. These samples were then subjected to a DNA extraction process and amplification of a fragment of the 18S rRNA by means of PCR. Subsequently, the PCR products from each sample were purified and sequenced. The sequences obtained were then analyzed using the BLASTn algorithm, which identified both sequences of this study as Hepatozoon canis. By applying the Mega4 software, it was confirmed that these isolates of H. canis from dogs in Goiânia are similar to other reference isolates of the same species from other regions of Brazil and worldwide.

Multiplex PCR-based identification of Streptococcus canis, Streptococcus zooepidemicus and Streptococcus dysgalactiae subspecies from dogs.

Streptococcus canis (S. canis), Streptococcus equi subspecies zooepidemicus (S. zooepidemicus) and Streptococcus dysgalactiae subspecies (S. dysgalactiae subspecies) are β-haemolytic Gram positive bacteria infecting animals and humans. S. canis and S. zooepidemicus are considered as two of the major zoonotic species of Streptococcus, while more research is needed on S. dysgalactiae subspecies bacteria. In this work, a multiplex-PCR protocol was tested on strains and clinical samples to detect S. canis, S. dysgalactiae subspecies and S. equi subspecies bacteria in dogs. All strains were correctly identified as S. canis, S. equi subspecies or S. dysgalactiae subspecies by the multiplex-PCR. The main Streptococcus species isolated from symptomatic dogs were confirmed S. canis. The multiplex-PCR protocol described is a rapid, accurate and efficient method for identifying S. canis, S. equi subspecies and S. dysgalactiae subspecies in dogs and could be used for diagnostic purposes and for epidemiological studies.

Retraction Note: Detection of Helicobacter spp. DNA in the colonic biopsies of stray dogs: molecular and histopathological investigations.

In dogs, the gastric Helicobacter spp. have been well studied, but there is little information regarding the other parts of the alimentary system. The incidence of Helicobacter spp. infection in dogs is largely unknown and to our knowledge there are no data about their potential pathogenic role. In light of these considerations, the aims of this study were (i) to assess the prevalence of Helicobacter spp. in colonic biopsies of healthy and symptomatic stray dogs also (ii) we isolate and characterize helicobacters in canine colonic biopsies to compare the commonly used tests for the identification of Helicobacter spp. and to determine the occurrence of these species in dogs. Tissues from fifteen stray dogs (8 males and 7 females, age 6 months –10 years) were used in this study. From each stray dog, multiple colonic biopsies were taken for PCR. Biopsies for PCR of cecum and colon were immediately frozen and stored at -20°C until DNA extraction. Samples for histological examination were fixed in 10% neutral buffered formalin and embedded in paraffin wax. In the cecum and colon, Helicobacter spp. DNA was detected in all dogs. H.canis, H.bizzozeronii, H. bilis, H.felis, H.salomonis and H.pylori Identified by specific polymerase chain reaction. Histopathology demonstrated that Helicobacter organisms were localized within the surface mucus and the intestinal crypts. Dogs with heavy Helicobacter spp. colonization were significantly in younger as well as had a higher level of mucosal fibrosis/atrophy than dogs with uncolonized or poorly colonized biopsies (p < 0.05). We have indicated that the crypts of the cecum and colon of healthy and symptomatic dogs are heavily colonized by Helicobacter spp.. Combined molecular and histological approaches demonstrated that enterohepatic Helicobacter spp. infection is rather common in colonic biopsies of healthy and symptomatic stray dogs, with Helicobacter spp. specialy H. canis, H.bizzozeroni, H.billis, H.felis and H. salomonis identified as the most common species. The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1957989294118782 .

Prevalence of Angiostrongylus vasorum in southern Belgium, a coprological and serological survey.

BackgroundCanine angiostrongylosis, a gastropod-borne helminthic infection, is increasingly being described in North America and is now reported in many European countries. In dogs, Angiostrongylus vasorum may cause a wide spectrum of clinical signs. Respiratory distress such as coughing and dyspnoea are the most frequently described manifestations. The aim of the present study was to gain additional information on the distribution, prevalence and risk factors associated with A. vasorum infection in dog from southern Belgium through the combined used of a commercially available in-clinic assay for detection of circulating antigen (Angio Detect™, IDEXX, Westbrook, USA) and coprology in two different canine populations: dogs with clinical signs compatible with angiostrongylosis and asymptomatic dogs or dogs presented for unrelated conditions (control).ResultsA total of 979 dogs were enrolled in the study from November 2014 until February 2016. Seven hundred fifty-seven dogs were included in the control group, whereas 222 dogs had clinical signs compatible with angiostrongylosis. Forty-six dogs out of 979 (4.7 %) had A. vasorum circulating antigen. There was a highly significant difference between the two populations (3.6 % (27/747) and 8.6 % (19/222) in control and symptomatic dogs, respectively) (P = 0.00379). First stage larvae (L1) of A. vasorum were found in seven out of 24 serologically positive control dogs and in six out of 17 serologically positive symptomatic dogs. Interestingly, L1 of Crenosoma vulpis were detected by Baermann technique in one control and nine symptomatic dogs, respectively. Out of 17 Angio Detect™ (IDEXX, Westbrook, USA) positive dogs with negative (14) or not performed Baermann test (three), one dog was positive in both in-house ELISAs (Ag and Ab) and one dog was positive for Ag. Statistical analysis was unable to detect any risk factors associated with the direct and/or indirect detection of A. vasorum.ConclusionsThis seroepidemiological study demonstrated for the first time a high seroprevalence in Southern Belgium for A. vasorum. The Angio Detect™ was found to be suitable in this context as the collection, preservation and examination of stools were difficult. Nevertheless, discrepancies were observed between the different available tests. Additional research is clearly needed. Also, coproscopy remains a very useful tool in dogs infected for less than nine weeks and for the identification of other canine lung nematodes such as C. vulpis. This study also demonstrates that asymptomatic dogs may shed A. vasorum L1 in their faeces and therefore contribute to the maintenance of A. vasorum life-cycle.

Recovery of antigen-specific T cell responses from dogs infected with Leishmania (L.) infantum by use of vaccine associated TLR-agonist adjuvant

Visceral leishmaniasis (VL), caused by infection with the obligate intracellular protozoan parasite Leishmania infantum, is a fatal disease of dogs and humans. Protection against VL requires a T helper 1 (Th1) skewed CD4+ T response, but despite this knowledge, there are currently no approved-to-market vaccines for humans and only three veterinary-use vaccines globally. As VL progresses from asymptomatic to symptomatic, L. infantum–specific interferon gamma (IFNγ) driven-Th1 responses become dampened and a state of immune exhaustion established. T cell exhaustion and other immunoregulatory processes, starting during asymptomatic disease, are likely to hinder vaccine-induced responses if vaccine is administered to infected, but asymptomatic and seronegative, individuals. In this study we evaluated how immune exhaustion, shown previously by our group to worsen in concert with VL progression, effected the capacity of vaccine candidate antigen/toll-like receptor (TLR) agonist combinations to promote protective CD4+ T cell responses during progressive VL. In conjunction with Th1 responses, we also evaluated concomitant stimulation of immune-balanced IL-10 regulatory cytokine production by these vaccine products in progressive VL canine T cells. Vaccine antigen L111f in combination with TLR agonists significantly recovered CD4+ T cell IFNγ intracellular production in T cells from asymptomatic VL dogs. Vaccine antigen NS with TLR agonists significantly recovered CD4+ T cell production in both endemic control and VL dogs. Combinations of TLR agonists and vaccine antigens overcame L. infantum induced cellular exhaustion, allowing robust Th1 CD4+ T cell responses from symptomatic dogs that previously had dampened responses to antigen alone. Antigen-agonist adjuvants can be utilized to promote more robust vaccine responses from infected hosts in endemic areas where vaccination of asymptomatic, L. infantum-infected animals is likely.

Evaluation of Immunofluorescence Antibody Test Used for the Diagnosis of Canine Leishmaniasis in the Mediterranean Basin: A Systematic Review and Meta-Analysis.

With an expected sensitivity (Se) of 96% and specificity (Sp) of 98%, the immunofluorescence antibody test (IFAT) is frequently used as a reference test to validate new diagnostic methods and estimate the canine leihmaniasis (CanL) true prevalence in the Mediterranean basin. To review the diagnostic accuracy of IFAT to diagnose CanL in this area with reference to its Se and Sp and elucidate the potential causes of their variations, a systematic review was conducted (31 studies for the 26-year period). Three IFAT validation methods stood out: the classical contingency table method, methods based on statistical models and those based on experimental studies. A variation in the IFAT Se and Sp values and cut-off values was observed. For the classical validation method based on a meta-analysis, the Se of IFAT was estimated in this area as 89.86% and 31.25% in symptomatic and asymptomatic dogs, respectively. The Sp of IFAT was estimated in non-endemic and endemic areas as 98.12% and 96.57%, respectively. IFAT can be considered as a good standard test in non-endemic areas for CanL, but its accuracy declines in endemic areas due to the complexity of the disease. Indeed, the accuracy of IFAT is due to the negative results obtained in non-infected dogs from non-endemic areas and to the positive results obtained in sera of symptomatic dogs living in endemic areas. But IFAT results are not unequivocal when it comes to determining CanL infection on asymptomatic dogs living in endemic areas. Statistical methods might be a solution to overcome the lack of gold standard, to better categorize groups of animals investigated, to assess optimal cut-off values and to allow a better estimate of the true prevalence aiming information on preventive/control measures for CanL.

Dogs attending primary-care practice in England with clinical signs suggestive of Chiari-like malformation/syringomyelia

Chiari-like malformation/syringomyelia (CM/SM) in dogs describes a developmental disorder that can cause pain and reduced quality of life. This retrospective study aimed to report the period prevalence, clinical signs and...

Longitudinal Study of the Relationship among Craniocervical Morphology, Clinical Progression, and Syringomyelia in a Cohort of Cavalier King Charles Spaniels.

BackgroundCraniocervical junction (CCJ) anomalies and secondary syringomyelia are commonly diagnosed in Cavalier King Charles spaniel (CKCS). Familiarity with the natural history of these abnormalities is vital to understanding the disease syndrome.ObjectiveTo evaluate magnetic resonance imaging (MRI) predictors of worsening clinical signs, syringomyelia, and morphology in CKCS longitudinally.AnimalsFifty‐four client‐owned CKCS, 5–13 years old; 50% currently symptomatic.MethodsLongitudinal observational study. We enrolled CKCS with an MRI of the CCJ performed ≥3 years earlier. We used questionnaires and neurologic examinations to grade initial and current clinical status. Dogs that could be anesthetized were reimaged. Morphologic assessments included the presence and severity of: Chiari‐like malformations, medullary position, atlantooccipital overlapping (AOO), dorsal atlantoaxial bands, and syringomyelia. Cranial cavity volumes and foramen magnum height were measured.ResultsClinical status was evaluated in 54 dogs; 36/54 were reimaged. Mean follow‐up was 71 months. Of initially asymptomatic dogs, 32% were symptomatic at re‐evaluation. Of initially symptomatic dogs, 56% had worsened; 13% had improved with medical management. The morphology of the CCJ at initial imaging did not predict development of either new or worsened signs or syringomyelia by the time of re‐evaluation.ConclusionCraniocervical junction anomalies assessed in this study did not appear predictive of future clinical status or syringomyelia in our cohort. The impacts of syringomyelia, AOO, and atlantoaxial bands on future clinical status merit further study in larger groups of CKCS. Clinical progression in our cohort of medically managed CKCS did not differ substantially from published reports of those treated surgically.