Antibacterial effects of chitosan-based hydrogels containing Trachyspermum ammi essential oil on pathogens isolated from dogs with otitis externa.

BACKGROUND: Growing antibiotic resistance has made treating otitis externa (OE) increasingly challenging. On the other hand, local antimicrobial treatments, especially those that combine essential oils (EOs) with nanoparticles, tend to be preferred over systemic ones. It was investigated whether Ajwain (Trachyspermum ammi) EO, combined with chitosan nanoparticles modified by cholesterol, could inhibit the growth of bacterial pathogens isolated from OE cases in dogs. In total, 57 dogs with clinical signs of OE were examined and bacteriologically tested. Hydrogels of Chitosan were synthesized by self-assembly and investigated. EO was extracted (Clevenger machine), and its ingredients were checked (GC-MS analysis) and encapsulated in chitosan-cholesterol nanoparticles. Disc-diffusion and broth Micro-dilution (MIC and MBC) examined its antimicrobial and therapeutic properties. RESULTS: Staphylococcus pseudintermedius (49.3%) was the most common bacteria isolated from OE cases, followed by Pseudomonas aeruginosa (14.7%), Escherichia coli (13.3%), Streptococcus canis (9.3%), Corynebacterium auriscanis (6.7%), Klebsiella pneumoniae (2.7%), Proteus mirabilis (2.7%), and Bacillus cereus (1.3%). The investigation into the antimicrobial properties of Ajwain EO encapsulated in chitosan nanoparticles revealed that it exhibited a more pronounced antimicrobial effect against the pathogens responsible for OE. CONCLUSIONS: Using chitosan nanoparticles encapsulated with EO presents an effective treatment approach for dogs with OE that conventional antimicrobial treatments have not cured. This approach not only enhances antibacterial effects but also reduces the required dosage of antimicrobials, potentially preventing the emergence of antimicrobial resistance.

The inhibitory effects of zinc oxide nanoparticles on clinical isolates of Microsporum canis in dogs and cats.

INTRODUCTION: Microsporum canis is the most common dermatophyte infecting pets and their owners, and its long duration of treatment and increasing rate of drug resistance have caused the attention of researchers to be directed towards the use of nanoparticles and new alternatives for treatment. This study investigated the antifungal effects of zinc oxide (ZnO) nanoparticles on clinical isolates of M. canis in dogs and cats and subtilisin 1 (SUB1) gene expression. MATERIALS AND METHODS: Zinc oxide nanoparticles were prepared using the wet chemical method at a concentration of 4000 ppm. Its antifungal potential was evaluated at concentrations of 62.5-4000 ppm by disk diffusion and microdilution methods against 10 isolates of M. canis. The effect of this product on SUB1 gene expression was investigated by quantitative real-time PCR method. RESULTS: The results of the disk diffusion test showed that the highest inhibitory diameter was at the highest concentration of ZnO nanoparticles (34 mm), and the inhibitory zone was observed in dilutions up to 250 ppm. The minimum inhibitory concentration (MIC) of ZnO nanoparticles was between 250 and 500 ppm, and the minimum fungicidal concentration was between 500 and 1000 ppm. There was a significant reduction in SUB1 gene expression in sub-MIC concentration (125-250 ppm) (p < 0.05). CONCLUSION: This study showed that ZnO nanoparticles have a concentration-dependent inhibitory effect on M. canis. Moreover, ZnO nanoparticles could decrease the expression of SUB1, an enzyme involved in fungi adhesion to the epidermis. Nevertheless, more studies must be done in the future to determine the possible side effects and safety of ZnO nanoparticles along with their efficacy in vivo.

Molecular detection of SARS-CoV-2 in domestic cats in close contact with positively-infected owners in Tehran, Iran in 2021.

Objectives: In 2019, COVID-19 emerged in China and has since spread worldwide. Owing to the virus's ability to adhere to specific receptors, cats are susceptible to infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The popularity of pet cats in Iran has sparked fears of human-cat-human transmission of the virus. This study aimed to identify positive cases in cats owned by people infected with SARS-CoV-2, to determine if they remained positive for >3 weeks and to examine the virus genome isolated from a number of cats and one of their owners. Methods: A total of 30 cats were sampled approximately 3 days after their owners tested positive (day 1), and 3 weeks later, in strict accordance with health regulations. Rectal and oropharyngeal samples were collected. All samples were subjected to a qualitative PCR and reverse transcription PCR. The S-gene region was partially sequenced in positive samples and the results were used to create a phylogenetic tree. Results: SARS-CoV-2 was detected in 7/30 (23.3%) cats examined. In the third week, every cat tested negative. The sequence data of positive cats and one of their owners revealed that the retrieved RNAs belonged to the alpha variation. The genetic distance between the samples and the reference sequence (20I/B.1.1.7: OM003849, MZ344997) was minimal, with a 99% similarity. Positive samples of cats had four mutations in gene S. Amino acid substitutions in the spike glycoprotein at positions N501Y, A570D, D614G and P681H were recorded in the isolates compared with 780 other sequences of Iranian strains. Conclusions and relevance: This study confirmed the presence of SARS-CoV-2-infected cats living in close contact with infected owners. Despite cats' susceptibility to COVID-19, the risk of severe infection in these animals is low, as evidenced by the lack of clinical signs in positive cats.

In vitro investigation of canine periodontal ligament-derived mesenchymal stem cells: A possibility of promising tool for periodontal regeneration.

Objectives: Recent investigations indicate that canine periodontal ligament-derived stem cells (cPDLSCs) may reveal a reliable strategy for repair of periodontal tissues via cell-based tissue engineering approaches. Due to limited research, this study aimed to demonstrate the phenotypic characterization of cPDLSc in comparison with canine bone marrow-derived mesenchymal stem cells (cBMSCs) in vitro. Methods: Mesenchymal stem cells (MSCs) were obtained from PDL and BM of five male adult Mongrel dogs. In vitro isolation and expansion as well as biologic characterization including colony unit formation (CFU), osteogenic and adipogenic differentiation, flow cytometric analysis of CD34 and CD44, and RT-PCR of alkaline phosphatase (ALP), osteocalcin (OCN), periostin (POSTN) and S100A4 were performed. Furthermore, electron microscopy analysis was done to complement the comparative research. Results: CFU assay revealed that colonies of cPDLSCs presented 70% confluency with a more finite lifespan than BM-MSCs, showing a significant increase in cPDLSCs. Both types of MSCs showed osteogenic and adipogenic phenotypic characterized with clusters of mineralized depositions and lipid vacuoles, respectively. Both types of MSCs expressed CD44 with limited expression of CD34. RT-PCR of cPDLSCs revealed that expression of ALP, POSTN, OCN and S100A4 genes were significantly higher than those of BMSCs. In addition, comparison of SEM and revealed that cPDLSCs expressed more extracellular collagen fibers. Conclusions: The current study indicated that cPDLSCs show potency as a novel cellular therapy for periodontal regeneration a large animal model.

Effect of site of sample collection on blood glucose concentrations measured with a portable blood glucose meter in healthy and diabetic dogs.

Background: Blood glucose (BG) monitoring with portable blood glucose meters (PBGMs) is a critical aspect of managing canine diabetes mellitus. Some dogs best tolerate sampling from the ear, others from the lip, and others from other body sites. Therefore, it is relevant to know if the choice of the sampling site affects the glucose concentration. Aim: To compare different sampling sites for BG measurement in diabetic and non-diabetic dogs using veterinary PBGM. Moreover, determining the possible impact of body condition score (BCS) on BG concentration. Methods: Thirty-seven healthy and 12 diabetic dogs were included. A veterinary PBGM was used to measure BG concentrations in a total of 196 blood samples collected from the marginal ear vein (MEV), carpal pad, saphenous vein, and cephalic vein. The results obtained from the different sampling sites were compared. Results: The carpal pad, MEV, cephalic vein, and saphenous vein BG values were not significantly different at the different blood collection sites. There was no significant difference between higher and lower BCS in BG measurements in the different sampling sites. Conclusion: Different sampling sites, likewise utilizing either a venous or capillary sample, had no significant effect on BG measurement using veterinary PBGMs. The BCS seems to have no relevant influence on dog BG measurement.

Canine microfilaraemia in some regions of Iran.

BACKGROUND: Dirofilaria immitis and Dirofilaria repens are vector-borne zoonotic parasites which affect mainly dogs and humans worldwide. In Iran, information about the distribution of those nematodes is scant in several regions. Therefore, we investigated the prevalence of these filarial parasites in stray dogs from five Iranian provinces where no information about these parasites is available. METHODS: Blood samples were collected from 344 stray dogs in five provinces of Iran (i.e. Mazandaran, Gilan, Esfahan, Qazvin and Loresan). The presence of microfilariae was assessed using direct smear, modified Knott's test, molecular detection of filarial DNA (cox1 gene) and Wolbachia endosymbiont of parasitic nematodes (ftsZ gene) by conventional PCR (cPCR). All of the PCR products were sequenced and phylogenetic analysis was performed. RESULTS: In total, 75 dogs (21.8%) were found to be positive for D. immitis by cPCR. Infection was detected in all provinces, with the highest prevalence in Gilan province (22/28; 78.6%). Acanthocheilonema reconditum was diagnosed in five dogs (1.4%) from three provinces (i.e. Esfahan, Mazandaran, Gilan). Two dogs were infected with both parasites and three were only infected with A. reconditum. Dirofilaria repens infection was not found in the examined population. Representative sequences of the D. immitis cox1 gene from dogs from the northern provinces (Mazandaran, Gilan, Qazvin) were grouped together and distinctly separate from the ones from western and central provinces (Lorestan and Esfahan), suggesting that different nematode populations are present in the country. CONCLUSION: The data reported herein fill existing gaps in knowledge about canine filarial infection in two Iranian provinces and record the highest prevalence of D. immitis ever reported in the country (i.e. 78.6%). A geographical review of the literature about Dirofilaria spp. and A. reconditum infections in dogs and humans has also been summarized, indicating that D. immitis and D. repens are distributed in 22 of 31 provinces in Iran, whereas A. reconditum is present in fewer regions. Effective control strategies are advocated for owned dogs, and a national program for the management of stray dogs is needed to minimize the risk of infection in animals and humans.

Molecular detection of polycystic kidney disease in Persian and Persian-related breeds in Iran.

OBJECTIVES: This study was aimed at detecting feline autosomal dominant polycystic kidney disease in a population of Persian and Persian-related breeds by a molecular method in Iran. METHODS: Buccal swab samples were collected from 47 cats and examined with a touchdown PCR method. Additionally, partial sequencing was performed in two cats with bilateral renal cysts. RESULTS: Twenty-two cats (46.8%) were diagnosed as heterozygous for this mutation. Sequence analysis of two cats showed C to A point mutation in the PKD1 gene, as in previous studies. CONCLUSIONS AND RELEVANCE: Prevalence of this disease is high in Iran, highlighting the need for molecular screening tests before including cats in breeding programmes.

Intestinal microsporidia infection among cat owners and non-pet owners in Iran: a case-control study.

Microsporidia is a group of spore-forming microorganisms with zoonotic potential. This study aimed to compare intestinal microsporidia infections in cat owners and non-pet owners. In total, 210 fecal samples were collected from indoor cats, cat owners, and non-pet owners. DNA extraction was performed and the small subunit ribosomal RNA (SSU rRNA) gene was amplified. To characterize the genotypes, the internal transcribed spacer (ITS) fragment was amplified and sequenced. The phylogenetic trees were drawn to evaluate the relationship among Enterocytozoon bieneusi isolates. Two (2.9%) and one (1.4%) fecal samples from cat owners and one (1.4%) and two (2.9%) fecal samples from non-pet owners were positive for E. bieneusi and Encephalitozoon intestinalis, respectively. E. bieneusi was detected in two cat samples (2.9%). Same infection was not seen between infected cats and their owners. There was no significant difference between the prevalence rate of microsporidia among the cat owners and non-pet owners. Indeed, the genotypes L and type IV were seen in cats, while the genotype D was only detected in human. In this study, E. bieneusi and E. intestinalis were more prevalent among the cat owners and non-pet owners, respectively. Indeed, the higher prevalence of E. bieneusi in cats and their owners might be resulted from the worldwide distribution of this species.

Comparative measurement of FeLV load in hemolymphatic tissues of cats with hematologic cytopenias.

BACKGROUND: Feline leukemia virus (FeLV) is a serious viral infection in cats. FeLV is found in some tissues, such as spleen, lymph nodes and epithelial tissues. However, there is controversy about the organ in which the virus can be reliably detected in infected cats. The purpose of this study was to determine the level of viral infection in hemolymphatic tissues, including blood, bone marrow and spleen by reverse-transcriptase quantitative polymerase chain reaction (RT-qPCR). RESULTS: A total of 31 cats with clinical signs of FeLV infection associated with at least a single lineage hematologic cytopenia were included in this study. Peripheral blood, bone marrow and spleen samples were obtained from each cat. Complete blood counts, biochemical tests, and a rapid test to detect FeLV p27 antigen in blood samples of cats were performed. Of 31 cats, 9 had anemia alone, 4 had thrombocytopenia alone, 2 had neutropenia alone, 9 had bicytopenia of anemia and thrombocytopenia, 3 had bicytopenia of anemia and neutropenia, and 4 had pancytopenia. FeLV RNA was then detected by RT-qPCR in the whole blood, bone marrow and spleen. Viral RNA copy numbers were detected in all cats by RT-qPCR whereas 24 out of 31 cats were positive for the serum FeLV antigen. We detected a significantly greater number of viral RNA in the spleen compared with the whole blood and bone marrow. CONCLUSION: Spleen is a site where FeLV is most frequently detected in cats with hematologic cytopenias.

Association between DLA-DRB1.2 allelic diversity and development of mammary gland tumors in dogs.

BACKGROUND: The major histocompatibility complex (MHC) is the best-characterized genetic region related to resistance/susceptibility to a wide range of infectious and immune-mediated diseases. Evidences suggest that MHC class II genes may play an important role in developing different types of tumors including breast cancer. Canine mammary gland tumors (CMTs) are the most common neoplasms in female dogs. In the current study, the association of canine MHC class II DLA-DRB1.2 genotypes with development of mammary gland tumor profiles in dogs was investigated. DLA-DRB1.2 allelic diversity was determined in 40 dogs (18 CMT cases and 22 controls) using HRM technique and DNA sequencing. Association of the DLA-DRB1.2 genotypes with CMT profiles was expressed as odds ratio (OR). RESULTS: Based on the histopathological typing of tumors, CMT cases were categorized into 4 groups: simple carcinoma, complex carcinoma, carcinoma arising in a benign tumor and special types of carcinoma. A total of eight HRM profiles (A to H) were identified in dogs sampled. The association study revealed a significant correlation between DLA-DRB1.2 genotypes with different CMT profiles. The E genotype was significantly associated with increased risk of carcinoma arising in a benign tumor, and the B genotype represented a positive correlation with complex carcinoma. Significant association was also observed between the heterozygosity of DLA-DRB1.2 genotypes and decreased risk of developing tumor in dogs. CONCLUSIONS: These results provide additional support for the association between DLA-DRB1 genes and development of mammary gland tumors in dogs and could potentially be used for early diagnosis of neoplasia and identifying susceptible dogs.

Molecular Detection and Genotyping of Intestinal Microsporidia from Stray Dogs in Iran.

BACKGROUND: Microsporidia as one of the most important pathogens in veterinary and agricultural settings, have emerged in immunocompromised patients in Iran. To date, different Enterocytozoon bieneusi genotypes have been identified in humans and animals, supporting the possibility of zoonotic zoonosis transmission potential. The aim of this study was to evaluate the distribution of E. bieneusi genotypes among overpopulated stray dogs in vicinity of Tehran, the capital city of Iran. METHODS: Totally, 75 stool and 75 urine samples were obtained from 75 stray dogs during the time period from Mar 2015 to Oct 2015. DNA extraction was performed on all the samples and specific fragment of small subunit ribosomal RNA gene of E. bieneusi and Encephalitozoon spp. was amplified. Furthermore, specific primers targeting the internal transcribed spacer region of E. bieneusi were applied to determine the genotype of the microorganism. RESULTS: Microsporidia was detected in 5.3% of stool samples, while none of the urine samples was positive for microsporidia species. Overall, 440 bp fragment of E. bieneusi was amplified in all the samples and there was no amplification for Encephalitozoon spp. The results of sequencing of 410 bp fragment of internal transcribed spacer region showed that all the E. bieneusi were genotype D. CONCLUSION: E. bieneusi was the most prevalent microsporidian species in the stray dogs and all the positive isolates were characterized as genotype D.

Transplantation of Bone Marrow-Derived Mesenchymal Stem Cells, Platelet-Rich Plasma, and Fibrin Glue for Periodontal Regeneration.

Appropriate regeneration of periodontal tissues is the primary purpose of periodontal disease treatment. The present study assessed the impact of three key regenerative elements-platelet-rich plasma (PRP), canine bone marrow mesenchymal stem cells (cBM-MSCs), and fibrin glue-on periodontal regeneration. In each of the study's five dogs, Class II furcation defects were established on the buccal surface of five teeth. One tooth (five total) from each dog was placed into one of five groups: (1) PRP + fibrin glue, (2) PRP + fibrin glue + cBM-MSCs, (3) fibrin glue, (4) cBM-MSCs + fibrin glue, and (5) control (no treatment). Histologic and histometric evaluations were performed to assess the formation of new bone, cementum, and the periodontal ligament. Different types of new bone and cementum, the maximum thickness of new cementum and the periodontal ligament (PDL), the vitality of bone, and the presence of inflammation or foreign-body reactions were also elucidated. The histologic and histometric evaluations revealed substantial differences in all groups between the observed maximum thickness of newly formed cementum and PDL. The percentage of bone and cement formation drastically increased with the combined presence of stem cells, fibrin glue, and PRP. The results showed that the inherent regenerative capacity of periodontal tissues can be sufficient if their latent self-repair mechanisms are stimulated.

Bone marrow or adipose tissue mesenchymal stem cells: Comparison of the therapeutic potentials in mice model of acute liver failure.

Acute liver failure (ALF) is a lethal disease with limited life-saving therapy. Because lack of whole organ donors for liver transplantation, a substitute treatment strategy is needed for these patients. Preclinical and clinical findings have proved that treatment with mesenchymal stem cells (MSCs) is beneficial for recovery from ALF. In this approach, however, the appropriate sources of these cells are unclear. In the present study, we investigated and compared the therapeutic potentials of bone marrow-mesenchymal stem cells (BM-MSC) with those of adipose tissue (AT-MSC) in carbon tetrachloride (CCL4)-induced acute liver failure in mice. Murine BM- and AT-MSCs obtained from normal mice were cultured and labelled. The cells were transplanted to CCL4-induced ALF mice models intravenously. After cell transplantation, blood samples and liver tissues were collected daily for 72 h to analyze liver enzymes and liver histopathology, respectively. We found that survival rate of AT-MSC transplanted (AT-TR) mice was significantly higher than that of control (ALF) group. Liver histopathology was superior in the AT-TR mice, but not significantly, compared to that in BM-MSC transplanted (BM-TR) ones. Furthermore, in the AT-TR mice the level of aspartate aminotransferase (AST) and alanine aminotransferase (ALT), in some time points were significantly less than those of BM-TR. Taken together, these data suggest that in comparison to BM-MSC, AT-MSCs is an appropriate choice for cell therapy in the case of acute liver failure.

Identification of Helicobacter and Wolinella spp. in Oral Cavity of Toy Breed Dogs With Periodontal Disease.

Periodontal diseases are the most common oral cavity infectious diseases in adult dogs. We aimed in this study to identify Helicobacter and Wolinella spp. in saliva and dental plaque of dogs with periodontitis. Sixty-two small-breed pet dogs, aged more than 6 years from both sexes, were categorized into healthy and periodontitis groups. Samples from saliva and dental plaques were collected, and Helicobacter and Wolinella were identified on genus and species levels using polymerase chain reaction. Our results showed significant increase in infection rate of Wolinella spp. in periodontitis compared with healthy dogs (P = .002). Furthermore, infection rate of Helicobacter genus was significantly higher in periodontitis compared with healthy dogs (P = .007). Infection with Wolinella spp. showed higher rate than Helicobacter spp. in dogs with periodontitis. According to species-specific polymerase chain reaction results, Helicobacter felis (9.76%) was the main Helicobacter spp. in dogs with periodontitis compared with healthy dogs (P < .001). Oral cavity of pet dogs with periodontitis could be considered as an important source of Wolinella and Helicobacter spp. infections.

EGFP reporter protein: its immunogenicity in Leishmania-infected BALB/c mice.

Optical reporter genes such as green fluorescent protein (GFP) and luciferase are efficiently and widely used in monitoring and studying the protective/therapeutic potential of candidate agents in leishmaniasis. But several observations and controversial reports have generated a main concern, whether enhanced GFP (EGFP) affects immune response. To address this issue, we studied the immunogenicity of EGFP in vivo by two lines of stably transfected parasites (Leishmania major (EGFP) or L. major (EGFP-LUC)) in BALB/c model and/or as a recombinant protein (rEGFP) produced in vitro by bacteria in parallel. Disease progression was followed by footpad swelling measurements and parasite burden in draining lymph nodes using microtitration assay and real-time PCR, and immune responses were also evaluated in spleen. EGFP-expressing parasites generated larger swellings in comparison with wild-type (L. major) while mice immunized with rEGFP and challenged with wild-type parasite were quite comparable in footpad swelling with control group without significant difference. However, both conventional and molecular approaches revealed no significant difference in parasite load between different groups. More importantly, no significant inflammatory responses were detected in groups with higher swelling size measured by interferon-γ (IFN-γ), interleukin (IL)-10, IL-5, and nitric oxide against frozen and thawed lysate of parasite as stimulator. Altogether, these results clearly revealed that EGFP protein expressed in prokaryotic and eukaryotic hosts is not an immunological reactive molecule and acts as a neutral protein without any side effects in mice. So, EGFP expressing Leishmania could be a safe and reliable substitution for wild-types that simplifies in situ follow-up and eliminates the animal scarification wherever needed during the study.

Occurrence of Chlamydophila felis, feline herpesvirus 1 and calcivirus in domestic cats of Iran.

BACKGROUND AND OBJECTIVES: Feline herpesvirus-1, feline calicivirus and Chlamydophila felis are the main causes of feline upper respiratory tract disease. This study was conducted to identify of FeHV-1, FCV and C. felis infections in domestic cat population and also to estimate the prevalence of each specific infection in Iran. MATERIALS AND METHODS: The ocular conjunctiva and oropharyngeal specimens obtained from 80 cats were examined using PCR and reverse transcription PCR. RESULTS: FeHV-1 was detected in 23 (28.8%), FCV in 2 (2.5%) and C. felis in 16 (20%) cats. Twelve cats(15%) had co-infection with 2 or 3 of the mentioned pathogens. Ocular lesions were the most common clinical signs in the FeHV-1 and C. felis infections whereas respiratory lesions were more observed with the FCV infections. It seems that there is an age-related tendency in the infected cats, meaning that the age of the C. felis positive cats was less than those with FeHV-1 and FCV infections. CONCLUSION: These results confirm the presence and show the prevalence of three major pathogens associated with upper respiratory tract disease for the first time in Iran.

Immunological comparison of DNA vaccination using two delivery systems against canine leishmaniasis.

Visceral leishmaniasis (VL) is a fatal disease caused by the intracellular protozoan parasite Leishmania infantum. Dogs are the primary reservoirs of this parasite, and vaccination of dogs could be an effective method to reduce its transfer to humans. In order to develop a vaccine against VL (apart from the choice of immunogenic candidate antigens), it is necessary to use an appropriate delivery system to promote a proper antigen-specific immune response. In this study, we compared two vaccine delivery systems, namely electroporation and cationic solid-lipid nanoparticle (cSLN) formulation, to administer a DNA vaccine containing the Leishmania donovani A2 antigen, and L. infantum cysteine proteinases of type I (CPA) and II (CPB) without its unusual C-terminal extension. The protective potencies of these two vaccine delivery systems were evaluated against L. infantum challenge in outbred dogs. Our results show that the administration of pcDNA-A2-CPA-CPB(-CTE)GFP vaccine as a prime-boost by either electroporation or cSLN formulation protects the dogs against L. infantum infection. Partial protection in vaccinated dogs is associated with significantly (p<0.05) higher levels of IgG2, IFN-γ, and TNF-α and with low levels of IgG1 and IL-10 as compared to the control group. Protection was also correlated with a low parasite burden and a strong delayed-type hypersensitivity (DTH) response. This study demonstrates that both electroporation and cSLN formulation can be used as efficient vaccine delivery systems against visceral leishmaniasis.

Evaluation of Live Recombinant Nonpathogenic Leishmania tarentolae Expressing Cysteine Proteinase and A2 Genes as a Candidate Vaccine against Experimental Canine Visceral Leishmaniasis.

Canine Visceral Leishmaniasis (CVL) is a major veterinary and public health problem caused by Leishmania infantum (L. infantum) in many endemic countries. It is a severe chronic disease with generalized parasite spread to the reticuloendothelial system, such as spleen, liver and bone marrow and is often fatal when left untreated. Control of VL in dogs would dramatically decrease infection pressure of L. infantum for humans, since dogs are the main domestic reservoir. In the past decade, various subunits and DNA antigens have been identified as potential vaccine candidates in experimental animal models, but none has been approved for human use so far. In this study, we vaccinated outbreed dogs with a prime-boost regimen based on recombinant L. tarentolae expressing the L. donovani A2 antigen along with cysteine proteinase genes (CPA and CPB without its unusual C-terminal extension (CPB-CTE) and evaluated its immunogenicity and protective immunity against L. infantum infectious challenge. We showed that vaccinated animals produced significantly higher levels of IgG2, but not IgG1, and also IFN-γ and TNF-α, but low IL-10 levels, before and after challenge as compared to control animals. Protection in dogs was also correlated with a strong DTH response and low parasite burden in the vaccinated group. Altogether, immunization with recombinant L. tarentolae A2-CPA-CPB-CTE was proven to be immunogenic and induced partial protection in dogs, hence representing a promising live vaccine candidate against CVL.

Evaluation of gallbladder volume and contraction index with three-dimensional ultrasonography in healthy dogs.

Three-dimensional (3D) ultrasonography has been shown to be an accurate and appropriate tool for measurement of gallbladder volume in humans. Therefore, we applied this novel technique for the first time to study fasting and postprandial gallbladder volume in 10 healthy dogs and compared the results with those of 2-dimensional (2D) ultrasonography. Fasting gallbladder volumes determined by 3D ultrasonography were significantly higher than corresponding volumes determined by 2D ultrasonography (P<0.01). Additionally, gallbladder volumes were significantly decreased in the postprandial state compared with the fasting state using 3D ultrasonography (P<0.001), but 2D ultrasonography showed no significant difference (P=0.189). The Gallbladder contraction index was higher in 3D ultrasonography than 2D ultrasonography; however, it did not reach statistical significance (P=0.25). In conclusion, 3D ultrasonography was able to measure gallbladder volume in healthy dogs in this study. It is suggested that 3D ultrasonography can be used to accurately estimate gallbladder volume and contractility.