Canine Upper Digestive Tract 3D Model: Assessing Its Utility for Anatomy and Upper Endoscopy Learning.

A teaching strategy using 3D-printed models of the canine upper digestive tract (UDT) for anatomy demonstration and upper endoscopy instruction was evaluated. The canine UDT (esophagus-stomach-duodenum) was scanned and 3D-printed molds were manufactured using silicone casting. First-year students were introduced to these 3D models in practical sessions alongside real specimens. Simultaneously, fifth-year students were trained in endoscope handling and anatomical recognition using 3D specimens. Both groups completed an anonymous survey. Results showed that overall, first-year (n = 93) and fifth-year (n = 45) students agreed or strongly agreed that the 3D-printed model was effective for learning purposes. In summary, first-year students highlighted an improved understanding of size, volume, topography, and easier manipulation of the 3D model compared to fresh specimens. Fifth-year students were more enthusiastic, finding the 3D model valuable for spatial vision and clinical training. While both groups were against completely replacing the natural UDT with the 3D model, first-year students were more hesitant. These findings suggest that the 3D model of the canine UDT is an effective tool for hands-on training in clinical endoscopy and a valuable, albeit complementary, resource for teaching anatomy and topography.

Peripheral and intestinal T lymphocyte subsets in dogs with chronic inflammatory enteropathy.

BACKGROUND: Dysregulated T lymphocyte response is thought to play a key role in chronic intestinal inflammation (CIE). OBJECTIVES: To evaluate the presence of changes in peripheral and intestinal T lymphocyte subsets and to describe potential immune and inflammatory biomarkers in dogs with CIE. ANIMALS: Sixteen healthy dogs and 26 dogs were diagnosed with CIE. METHODS: Prospective case-control study evaluating peripheral and intestinal T lymphocytes using flow cytometry and inflammatory markers obtained from complete blood cell counts. RESULTS: Dogs with CIE had higher peripheral activated T helper (Th) lymphocytes (87/µL [18-273] CIE, 44/µL [16-162] healthy control (HC, P = .013) and regulatory T cells (Treg; 108/µL [2-257] CIE, 34/µL [1-114] HC, P = .004). In the intestinal epithelium, CIE dogs presented lower percentages of Th (4.55% [1.75-18.67] CIE, 8.77% [3.79-25.03] HC, P = .002), activated Th cells (0.16% [0.02-0.83] CIE, 0.33% [0.05-0.57] HC, P = .03) and CD4/CD8 ratio (0.08 [0.02-0.39] CIE, 0.21 [0.07-0.85] HC, P = .003). Conversely, higher percentage of activated T cytotoxic cells (20.24% [3.12-77.12] CIE, 12.32% [1.21-39.22] HC, P = .04) and interferon-gamma (IFN-γ) producing T lymphocytes (7.36% [0.63-55.83] CIE, 1.44% [0.00-10.56] HC, P = .01) within the epithelium was observed. In the lamina propria the percentage of Treg lymphocytes was higher (6.02% [1.00-21.48] CIE, 3.52% [0.18-10.52] HC, P = .02). CONCLUSIONS AND CLINICAL IMPORTANCE: Systemic and intestinal immune alterations occur in dogs with CIE suggesting that blood IFN-γ producing T lymphocytes and the systemic immune-inflamation index (SII) could potentially serve as biomarkers for the disease.

Analysis of the culturable gut yeast microbiota of dogs with digestive disorders.

Despite the increasing interest in studying the gut mycobiota of dogs, the association between fungal colonization and the development of digestive disorders in this species remains largely understudied. On the other hand, the high prevalence of antifungal-resistant yeasts detected in previous studies in samples from animals represents a major threat to public health. We analyzed the presence of culturable yeasts in 112 rectal swab samples obtained from dogs with digestive disorders attended in a veterinary teaching hospital. Our results revealed that Malassezia pachydermatis was frequently isolated from the studied dog population (33.9% of samples), and that the isolation of this yeast was significantly associated to the age of animals, but not to their sex, disease group, or the presence of vomits and/or diarrhea. In contrast, other yeast species were less prevalent (17.9% of samples in total), and their isolation was not significantly associated to any variable included in the analysis. Additionally, we observed that 97.5% of the studied M. pachydermatis isolates (n = 158, 1-6 per positive episode) displayed a minimum inhibitory concentration (MIC) value >4 µg/ml to nystatin, 31.6% had a MIC ≥32 µg/ml to fluconazole, and 27.2% had a MIC >4 µg/ml to amphotericin B. The antifungal susceptibility profiles of non-Malassezia (n = 43, 1-7 per episode) were more variable and included elevated MIC values for some antifungal-species combinations. These results confirm that the intestine of dogs is a reservoir of opportunistic pathogenic yeasts and suggest that the prevalence of M. pachydermatis colonization depends more on the age of animals than on any specific digestive disorder.

Assessing the Quality of Life of Dogs with Inflammatory Bowel Disease and Their Owners.

The aim of the study was to assess the quality of life (QoL) of dogs with inflammatory bowel disease (IBD) and the impact on the life and relationship of the owner. An online questionnaire based on a Likert scale score (1-10) was designed to assess items related to dog QoL, owner QoL, and the owner-dog relationship. Responses from 110 dog owners (30 with IBD and 80 healthy dogs) were included in the study. IBD dogs had significantly lower overall QoL (p < 0.001), health (p < 0.0001), and level of activity (p = 0.049). Owners of dogs with IBD reported lower overall QoL (p < 0.001). The scores for how their dog's QoL might affect their own QoL (p = 0.028), how much their dog limited their social life, leisure time, or daily activities (p = 0.015), and how often they felt burdened by caring for their dog (p = 0.006) were significantly higher in the IBD group when compared to the healthy group. In addition, IBD dog owners were more likely to see their dogs as children (p = 0.0004). IBD has a negative impact on affected dogs and their owners regardless of the severity of the disease.

Characterization of the Fecal and Mucosa-Associated Microbiota in Dogs with Chronic Inflammatory Enteropathy.

Canine chronic inflammatory enteropathy implicates multifactorial pathogenesis where immunological dysregulation and gut microbiota changes have a central role. Most sequencing-based taxonomic studies have been focused on the fecal microbiota. However, the analysis of these samples does not provide complete information regarding the composition of the small intestine affected by this canine disease. Therefore, in this study, we aimed to characterize the intestinal bacterial microbiota in dogs with inflammatory bowel disease (IBD) (n = 34) by means of duodenal biopsies and fecal samples collected at the time of the diagnosis and to compare those to a group of healthy dogs (n = 12) using the 16S ribosomal RNA (16S rRNA) gene-targeted sequencing (Illumina MiSeq platform). Our study showed that IBD dogs presented differences in the fecal bacterial communities when compared with healthy dogs, with a lower relative abundance of Prevotellaceae (p = 0.005), Prevotella (p = 0.002), and Prevotellaceae Ga6A1 group (0.006); Erysipelotrichales (p = 0.019), Candidatus Stoquefichus (p < 0.001), Erysipelotrichaceae (p = 0.011), and Allobaculum (p = 0.003); Lachnospiraceae NK4A136 group (p = 0.015), Sellimonas (p = 0.042), Oscillospirales (p = 0.037), Oscillospiraceae UCG-005 (p < 0.001), Faecalibacterium (p = 0.028), and Fournierella (p = 0.034); Acidaminococcales, Acidaminococcaceae, and Phascolarctobacterium (p = 0.001); Aeromonadales (p = 0.026), Succinivibrionaceae (p = 0.037), and Succinivibrio (p = 0.031). On the other hand, a higher relative abundance of Enterococcaceae (Enterococcus; p = 0.003), Streptococcaceae (Streptococcus, p = 0.021), Enterobacterales (p = 0.027), Enterobacteriaceae (p = 0.008), and Escherichia-Shigella (p = 0.011) was detected. Moreover, when evaluating α-diversity, the dogs with IBD showed lower diversity in terms of richness and abundance of species (observed species [p = 0.031] and Shannon index [p = 0.039]). Furthermore, fecal microbiota in dogs with IBD was significantly different from healthy dogs (p = 0.006). However, only a few taxa relative abundance shifts (lower Rubrobacteria, Rubrobacterales, Rubrobacteriaceae, and Rubrobacter [p = 0.002]; Cyanobacteria [p = 0.010], Vampirivibrionia, Obscuribacterales, and Obscuribacteraceae [p = 0.005]; Neisseriaceae [p = 0.004] and Conchiformibius [p = 0.003]) were observed when assessing duodenal-associated microbiota of dogs with IBD. Thus, even if the bowel inflammation mainly affects the small intestine in the IBD-affected dogs of the study, fecal specimens may constitute a better sample due not only to their easy availability but also in terms of searching for bacterial taxa as biomarkers for canine IBD. The use of different diets in the study can also have a partial influence on the microbiota composition. Future studies encompassing multi-omics approaches should evaluate the functionality in both levels to unravel the pathophysiology of canine IBD.

Changes in Faecal and Plasma Amino Acid Profile in Dogs with Food-Responsive Enteropathy as Indicators of Gut Homeostasis Disruption: A Pilot Study.

Dogs suffering from food-responsive enteropathy (FRE) respond to an elimination diet based on hydrolysed protein or novel protein; however, studies regarding the amino acid profile in FRE dogs are lacking. The aim of this pilot study was to evaluate whether the plasma and faecal amino acid profiles differed between control and FRE dogs and whether these could serve as indicators of severity of illness. Blood, faecal samples, body condition score, and severity of clinical signs based on the canine inflammatory bowel disease activity index were collected before starting the elimination diet. FRE dogs had lower proportions of plasma Asparagine, Histidine, Glycine, Cystine, Leucine, and branched-chain/aromatic amino acids; however, Phenylalanine increased. In faecal samples, Cystine was greater whereas Phenylalanine was lesser in sick dogs compared to control. Leucine correlated negatively with faecal humidity (r = -0.66), and Leucine and Phenylalanine with faecal fat (r = -0.57 and r = -0.62, respectively). Faecal Phenylalanine (r = 0.80), Isoleucine (r = 0.75), and Leucine (r = 0.92) also correlated positively with total short-chain fatty acids, whereas a negative correlation was found with Glycine (r = -0.85) and Cystine (r = -0.61). This study demonstrates the importance of Leucine and Phenylalanine amino acids as indicators of the disease severity in FRE dogs.

First detection of Anaplasma phagocytophilum and Babesia divergens and high infection rates of Anaplasma marginale and Babesia bigemina in cattle in extensive grazing systems of Central Spain.

Bovine vector-borne diseases have a considerable economic impact worldwide and affect health of humans and animals. However, different aspects of their epidemiology and their pathogenesis remain unclear. Despite the frequent description of clinical cases reported by practitioners attending cattle from Madrid, Central Spain, molecular prevalence of Anaplasma spp. and Babesia spp. has not been described. The aim of this study was to assess the positivity rate of A. phagocytophilum, A. marginale, A. centrale, B. bigemina and B. divergens in livestock of this area and to evaluate the existence of associations between these pathogens and haematological, biochemical and epidemiological data. Babesia divergens and A. phagocytophilum were detected for the first time in cattle from Madrid. Their positivity percentages were low (2.2% ± 1.4% and 1.8% ± 1.2%, respectively), but this description is of special interest, as these agents are potentially zoonotic. Both agents were found in areas of higher altitude and relative humidity and lower temperature. The detection of ticks in livestock during the moment of sampling was confirmed as a risk factor for these infections. Anaplasma marginale showed the highest molecular infection rate (30% ± 4.1%) in this study, followed by B. bigemina (21.9% ± 3.7%). Higher positivity rates of A. marginale and B. bigemina were found in the areas of mountain climate and warm-summer continental Mediterranean climate. The use of ectoparasiticide treatment was found as a risk factor for infection by A. marginale and B. bigemina. This finding could lead to reconsider the ectoparasiticide protocols that are used nowadays. Grazing on pastures with domestic or wild ruminants and the presence of wild carnivores were associated with higher rates of infection by these four agents and coinfections were frequently found.

Isolation and immunophenotyping by flow cytometry of canine peripheral blood and intraepithelial and lamina propria duodenal T lymphocytes.

The gut associated lymphoid tissue (GALT) effector sites play a crucial role on the pathogenesis of many immune-mediated gastrointestinal diseases. The lymphocytes at these effector sites are principally T cells which present important morphological, phenotypical and functional differences. Flow cytometry (FC) is one of the most commonly used techniques to characterize intestinal lymphocytes in human and animal models. Published studies with a focus on dogs for intraepithelial lymphocytes (IEL) immunophenotyping exist in very limited numbers. Moreover, no lamina propria lymphocytes (LPL) isolation protocols in the canine species have been described for FC evaluation. In addition to immune intestinal dysregulation, imbalances in the peripheral blood immune system have been described in both human and animal gastrointestinal disorders. The aim of this study was to provide a protocol for canine IEL and LPL isolation for FC immunophenotyping of T cells subsets. Specifically, T helper, T cytotoxic, activated Th and Tc lymphocytes, regulatory, double negative, double positive, IFN-γ and IL-4 producing T cells, and to compare their respective populations between these effector sites and with the blood stream compartment in healthy dogs. The potential relationship of these cells distributions with age, sex and breed was also evaluated. This study included sixteen healthy dogs of different sexes and breeds with a mean age of 4.55 ± 2.93 years old. The selected protocols for the three immune compartments showed proper cell yield, purity, viability, and the absence of phenotypic and functional disturbances. Histologically, an adequate separation of the duodenal epithelium from the lamina propria was also observed. All the proposed T cells subsets were identified in the three immune compartments studied, showing some statistically significant differences in their distributions at these locations that highlight the importance of their individual evaluation. This study provides an adequate method for canine small intestine IEL and LPL isolation for FC immunophenotyping and is key for future studies on the gastrointestinal immune system associated with different canine diseases.

Effect of chemically modified tetracycline-8 (CMT-8) on hematology, blood chemistry, cytokines and peripheral blood lymphocyte subsets of healthy dogs.

Tetracyclines are antibiotics widely used in human and veterinary medicine. Effects on the immune system and inflammatory response, including effects on blood leukocytes proliferation and function and in cytokines synthesis, have been described. Chemically modified tetracyclines (CMT) have lost their antimicrobial activity, but maintain these other properties. This study analyzes the effect of chemically modified tetracycline-8 (CMT-8) on the evolution of complete blood count, blood chemistry, the mRNA expression of selected cytokines and peripheral blood lymphocyte subpopulations distribution in healthy dogs. CMT-8 at a dose of 10 mg/kg once daily was administered per os to six healthy dogs. A control group of five healthy dogs, living in the same conditions than dogs treated with CMT-8, received placebo with an identical therapeutic regimen. When given at the doses used in this study, no side effects of CMT-8 were detected, suggesting a good tolerance and a limited toxicity of the drug. Dogs treated with CMT-8 showed a gradual increase in mean corpuscular hemoglobin. The administration of CMT-8 in healthy dogs did not affect blood mRNA expression of IFN-γ, TNFα, IL-4, IL-6, IL-10, IL-12 p40 and IL-13. However, the lymphocytes expressing class II MHC on their surface decreased during the first two weeks of CMT-8 treatment and subsequently increased for the next three months. Considering the absence of antimicrobial properties of the drug, the effects of CMT-8 detected in this study seem to be unrelated to the classical antimicrobial activity attributed to tetracyclines.

Pharmacokinetics of doxycycline after oral administration of multiple doses in dogs.

The aim of this study was to determine the pharmacokinetic parameters of doxycycline in dogs and assess the efficacy of an oral drug dosage regimen of 10 mg/kg daily for 28 days through Pharmacokinetic/Pharmacodynamic (PK/PD) target analysis based on Monte Carlo simulation, using previously published data for the zoonotic pathogen Staphylococcus pseudintermedius. After a multiple-dosage regimen, the accumulation index was 1.88 ± 0.82. The Cmaxss and Cminss values were 5.18 ± 1.81 µg/ml and 1.91 ± 1.35 µg/ml, respectively. There were statistically significant differences for Cmax, Cmin at 24 hr, MRTt, AUCt and AUC∞ between days 1 and 28. The Cminss value was over the MIC of the principal pathogens, and Cmaxss was higher than the resistance values (>2 µg/ml). For AUC/MIC indices of 12, 25 and 40, the cumulative fraction responses (CFR) were 94.01%, 69.55% and 60.86%, respectively; for an MIC value of 2 µg/ml, the corresponding probability of target attainment (PTA) was 99.94%, 84.78% and 45.16%, respectively. Doxycycline was used against numerous localized infections in different organs and tissues. For the strains with MIC < 1 µg/mL, PTA was close to 100%, even for the most demanding ones, specifically 94.98% for an index of 40% and 99.9% for an index of 25.

Short-Chain and Total Fatty Acid Profile of Faeces or Plasma as Predictors of Food-Responsive Enteropathy in Dogs: A Preliminary Study.

The aim of this study was to evaluate differences in short-chain fatty acids (SCFAs) and the total fatty acid profile of faeces or plasma as possible indicators of FRE in comparison with healthy dogs. FRE dogs had a lower concentration (p = 0.026) of plasma α-tocopherol as an indicator of the oxidative status of the animal, and lower C20:5n-3 (p = 0.033), C22:5n-3 (p = 0.005), polyunsaturated fatty acids (PUFA) (p = 0.021) and n-6 (p = 0.041) when compared with the control dogs; furthermore, sick dogs had higher proportions of plasma C20:3n-6 (p = 0.0056). The dogs with FRE showed a decrease in the production of faecal levels of SCFAs, mainly propionic acid (C3) (p = 0.0001) and isovaleric acid (iC5) (p = 0.014). FRE dogs also had a lower proportion of C15:0 (p = 0.0003), C16:1n-9 (p = 0.0095), C16:1n-7 (p = 0.0001), C20:5n-3 (p = 0.0034) and monounsaturated fatty acids (p = 0.0315), and tended to have lower n-3 (p = 0.058) and a reduced desaturase activity index in the stool when compared with the control group. However, the dogs with chronic enteropathy tended to have greater C20:4n-6 (p = 0.065) in their faeces as signs of damage at the intestinal level. The faecal parameters were better predictors than plasma. The highest correlations between faecal odd-chain, medium- or long-chain fatty acids and SCFAs were observed for C15:0 that correlated positively with faecal acetic acid (C2) (r = 0.72, p = 0.004), propionic acid (r = 0.95, p = 0.0001), isobutyric acid (iC4) (r = 0.59, p = 0.027) and isovaleric acid (r = 0.64, p = 0.0136), as well as with total SCFAs (r = 0.61, p = 0.02). Conversely, faecal C20:4n-6 showed a high inverse correlation (r = -0.83, p = 0.0002) with C2 and C3 (r = -0.59, p = 0.027). Canine inflammatory bowel disease (IBD) activity (CIBDAI) index correlated negatively mainly with faecal measurements, such as C3 (r = -0.869, p = 0.0005) and C15:0 (r = -0.825, p = 0.0018), followed by C16:1/C16:0 (r = -0.66, p= 0.0374) and iC5 (r = -0.648, p = 0.0310), which would indicate that these fatty acids could be good non-invasive indicators of the chronic inflammatory status, specifically FRE.

Stray dogs in Nepal have high prevalence of vector-borne pathogens: a molecular survey.

BACKGROUND: Population of stray dogs is significant in large cities of Nepal, such as Kathmandu. Most of stray dogs suffer a lack of basic health care. Considering the clinical relevance, the broad distribution and the lack of information of canine vector borne diseases (CVBD) in Nepal, the aim of this study was to evaluate the prevalence of different vector-borne pathogens (VBP) in stray dogs living in the metropolitan area of Kathmandu, and to assess different traits as possible risk factors. METHODS: A total of 70 canine blood samples from stray dogs attended at the Kathmandu Animal Treatment Centre during August 2017 were collected on filter paper (Flinders Technology Associates (FTA) cards). Data regarding signalment, clinical signs and epidemiological characteristics were recorded for each animal. Real-time polymerase chain reaction assays were performed for Leishmania spp., Ehrlichia spp./Anaplasma spp., Babesia spp./Theileria spp. and Hepatozoon canis. RESULTS: The overall prevalence detected was 31.43% for Hepatozoon canis, 31.43% for Anaplasma platys, 27.14% for Ehrlichia canis, 18.57% for Leishmania donovani species complex, 12.86% for isolates corresponding to Theileria spp., 12.86% for Babesia vogeli and 2.86% for B. gibsoni. A total of 81.43% of the dogs were positive to at least one of the VBP tested. Co-infections were detected in 41.43% of the dogs. Dogs positive to any of the VBP tested, and particularly to E. canis, were older than those that were negative. CONCLUSIONS: To our knowledge, this is the first molecular detection of VBP in stray dogs from Kathmandu, Nepal. The high prevalence of VBP detected highlights the need to implement a surveillance programme and control strategies for these CVBD in the population of stray dogs in this area.

Serologic and fecal markers to predict response to induction therapy in dogs with idiopathic inflammatory bowel disease.

BACKGROUND: Little information is available of markers that assess the disease course in dogs with idiopathic inflammatory bowel disease (IBD). OBJECTIVES: Evaluate relationship between disease severity and serum and fecal biomarkers in dogs with idiopathic IBD before and after treatment. ANIMALS: Sixteen dogs with idioptahic IBD and 13 healthy dogs. METHODS: Prospective case control study. Canine IBD activity index (CIBDAI) clinical score, serum concentrations of C-reactive protein (CRP), perinuclear antineutrophil cytoplasmic antibodies (pANCA), and serum and fecal canine calprotectin (cCP) were measured before and after 21 days of treatment. RESULTS: Serum CRP (median 3.5 mg/L; range: 0.1-52.4 mg/L), fecal cCP (median 92.3 µg/g; range: 0.03-637.5 µg/g), and CIBDAI scores significantly increased in dogs with IBD before treatment compared with serum CRP (median 0.2 mg/L; range: 0.1-11.8 mg/L; P < .001), fecal cCP (median 0.67 µg/g; range: 0.03-27.9 µg/g; P < .001) and CIBDAI (P < .001) after treatment. No significant associations between CIBDAI scores and before or after treatment serum biomarkers. There was a significant association between fecal cCP and CIBDAI scores before treatment (rho = 0.60, P = .01). CRP and fecal cCP significantly decreased after treatment (median 3.5 mg/L v. 0.2 mg/L; P < .001 and 92.3 µg/g v. 0.67 µg/g; P = .001, respectively). CONCLUSIONS AND CLINICAL IMPORTANCE: Our data indicate that measurement of fecal cCP concentration is a useful biomarker for noninvasive evaluation of intestinal inflammation. Dogs with severe signs of GI disease more often have abnormal markers than dogs having less severe disease.

Epidemiological study of hemotropic mycoplasmas (hemoplasmas) in cats from central Spain.

BACKGROUND: Hemotropic mycoplasmas (hemoplasmas) have been found infecting cats worldwide. However, studies about feline hemoplasma infections in Spain are scarce. Therefore, the purpose of the research was to evaluate the prevalence of feline hemotropic mycoplasmas and to characterize risk factors and clinical findings associated with these infections in a cat population from the Madrid area, Spain. METHODS: Polymerase chain reaction (PCR) was employed to detect Mycoplasma haemofelis (Mhf), "Candidatus Mycoplasma haemominutum" (CMhm) and "Candidatus Mycoplasma turicensis" (CMt) in blood samples from 456 client-owned and 138 stray cats from Madrid. In order to assess associations between these hemoplasma infections and epidemiological parameters, data regarding signalment, environment, prophylaxis measures, retrovirus status, clinical signs and laboratory findings were compiled, whenever possible. RESULTS: DNA of feline hemoplasmas was detected from the blood of 63 out of 594 cats (10.6%), with a prevalence of 3.7% (22/594) for Mhf, 8.1% (48/594) for CMhm and 0.5% (3/594) for CMt. Stray cats had statistically higher prevalences of feline hemoplasmas (15.9%) and, specifically, of Mhf (8.7%) than client-owned cats (9 and 2.2%, respectively). A total of seven cats (1.17%) were co-infected with "Candidatus M. haemominutum" and M. haemofelis, two (0.33%) with "Candidatus M. haemominutum" and "Candidatus M. turicensis" and another one (0.17%) with M. haemofelis and Candidatus "M. turicensis". Male gender, collection of blood during warm months and FeLV/FIV positivity status were associated with hemotropic mycoplasma infection in cats from Madrid. Additionally, within the group of client-owned cats, hemoplasma infection was associated with adult age, outdoor access, and the existence of low haematocrit, erythrocyte count and haemoglobin concentration values. CONCLUSIONS: To our knowledge, this is the first epidemiological survey of feline hemoplasmas performed in central Spain (Madrid). Our study confirms that "Ca. Mycoplasma haemominutum", Mycoplasma haemofelis and "Ca. Mycoplasma turicensis" are infecting client-owned and stray cats in this region of Spain, "Ca. Mycoplasma haemominutum" being the most prevalent species. More studies are necessary to help understand the role of the natural infection by these species of hemoplasma in cats.

Molecular, epidemiological, haematological and biochemical evaluation in asymptomatic Theileria annulata infected cattle from an endemic region in Spain.

Mediterranean theileriosis is one of the most fatal theilerioses, with considerable economic impact on livestock production. The potential consequences of asymptomatic infection on the animal's health and on the epidemiology in endemic regions are still unclear. The objectives of this study were to determine the molecular prevalence of T. annulata in a representative population of asymptomatic cattle in extensive management in Madrid, Central Spain, an area where practitioners frequently report cases of clinical theileriosis, and to evaluate the existence of associations between infection by this pathogen and haematological, biochemical and epidemiological data. T. annulata DNA was detected in 22.4% of the study population. The age was statistically associated with T. annulata prevalence rates, with a higher prevalence in cows older than 8 years (26.1%). Introduction of new cattle to the farm, grazing on pastures with other herds of cattle and previous history of clinical cases were statistically related to a higher prevalence of T. annulata infection. Herds with more than one ectoparasiticide treatment per year and that used more than one drug had significantly lower prevalences of infection with T. annulata. The location of farms in areas with a mean temperature higher than 17.1°C and mean altitude lower than 962m was statistically associated with the presence of T. annulata. In our study, the mean values of haematological parameters were within the normal adult range, but it is noteworthy that some T. annulata-infected animals presented low values for red blood cell parameters.

Molecular detection of Hepatozoon spp. and Cytauxzoon sp. in domestic and stray cats from Madrid, Spain.

BACKGROUND: Different species of apicomplexan protozoans of the genera Hepatozoon and Cytauxzoon can infect domestic cats, but their epidemiology and clinical relevance are not fully understood. The aim of this study was to assess the molecular prevalence of Hepatozoon spp. and Cytauxzoon spp. and to identify associated risk factors and clinical and laboratory abnormalities in a population of cats from Madrid, Spain. METHODS: Six hundred and forty-four client-owned and stray cats from Madrid, Spain, were included in this study. DNA samples were analyzed by two polymerase chain reaction (PCR) tests to detect a partial sequence of the 18S rRNA gene of Hepatozoon spp. and Cytauxzoon spp. In order to evaluate possible associations between infection by these protozoans and epidemiological or clinical parameters, data were collected related to: the season of sample collection, age, gender, spayed/neutered status, breed, living area, lifestyle, outdoor access, contact with other animals, prey on wild animals, history of tick or flea infestation, travel history, ectoparasiticide treatment, previous blood transfusion, previous tetracycline administration in the last 60 days, Feline Leukemia virus (FeLV) and Feline Immunodeficiency virus (FIV) status, positivity to other vector-borne diseases, the presence or absence of clinical signs and hematological or biochemical alterations. RESULTS: DNA of Hepatozoon spp. and Cytauxzoon sp. was amplified from the blood of 10 (1.6%) and 8 (1.2%) cats, respectively. Previous treatment with tetracyclines in the last 60 days, previous administration of blood transfusion, a decrease in haematocrit and an increase in creatinine were associated with Hepatozoon spp. infection. Cytauxzoon sp. infection was more frequent in samples collected during the winter months and in cats living in rural areas. This infection was associated with a FIV-positive status. Some of the cats that were positive for Hepatozoon spp. or Cytauxzoon sp. had been exposed to other vector-borne pathogens, such as Ehrlichia canis and Bartonella henselae. CONCLUSIONS: Our results indicate that cats from Madrid, central Spain, are infected with Hepatozoon spp. and Cytauxzoon sp., although with a low prevalence. Further studies are needed to determine the virulence of these agents in Spanish cats.

Isolation of Clostridium difficile from dogs with digestive disorders, including stable metronidazole-resistant strains.

The prevalence of Clostridium difficile in 107 dogs with diverse digestive disorders attended in a Spanish veterinary teaching hospital was assessed. The microorganism was isolated from 13 dogs (12.1%) of different disease groups. Isolates belonged to PCR ribotypes 078, 106, 154 and 430 (all of them toxigenic) and 110 (non-toxigenic), and were resistant to several antimicrobial drugs. Notably, seven isolates obtained from different dogs displayed stable resistance to metronidazole. The results of this study provide further evidence that dogs can act as a reservoir of C. difficile strains of epidemic ribotypes with resistance to multiple antibiotics.

Comparative study of peripheral blood leukocytes in healthy dogs and in dogs with cancer and inflammatory diseases.

In the present study, the peripheral blood lymphocyte subset distribution was compared between healthy dogs and dogs with chronic gastrointestinal disease, dental and skin conditions, and cancer. The immunophenotype of the group with chronic gastrointestinal disease and the group with dental and skin conditions showed no statistically significant differences with other groups of healthy or diseased dogs. When compared with healthy dogs, animals with cancer showed significantly lower absolute values of T cytotoxic cells (CD3+, CD8+) and lymphocytes that express major histocompatibility complex (MHC) class II (MHC-II+) in peripheral blood. The results suggest that peripheral blood immunophenotype is mainly altered in dogs with cancer but not in other diseases. Further studies are required to evaluate the clinical relevance of these findings.

Evaluation of lymphocyte populations in dogs naturally infected by Ehrlichia canis with and without clinical signs.

Immune response elicited by the host during ehrlichial infections could influence the clinical signs and laboratory and pathological findings. Twenty-eight dogs naturally infected by Ehrlichia canis were included in this study. Twenty of them presented only laboratory findings traditionally associated with canine monocytic ehrlichiosis (CME), whilst 8 dogs also showed clinical signs classically associated with CME (pale mucous membranes, fever, lymphadenopathy, weight loss, anorexia, lethargy or signs attributable to bleeding tendencies). A multiparametric flow cytometric study was performed to analyze the distribution of the main lymphocyte subsets (T, Th, Tc, B, and those that express MHC class II) in the peripheral blood. Statistically significant differences between dogs naturally infected by E. canis in a clinical or subclinical stage were not detected when evaluating lymphocyte subsets in peripheral blood samples. Dogs with clinical signs showed lower relative and absolute values of B lymphocytes than dogs without clinical signs, although the differences were not statistically significant.

Early-life longitudinal survey of peripheral blood lymphocyte subsets in Beagle dogs.

A longitudinal study of peripheral blood lymphocyte subsets from 7 to 15 months of age was performed in Beagle dogs employing a multiparametric flow cytometry. The data were compared with data obtained from adult Beagle dogs that were housed in the same animal facilities and that were subjected to the same controls during the 34 weeks of the study. Absolute counts of total lymphocytes and CD3+ T, CD3+CD4+ Th and CD21+ B lymphocytes decreased during the entire 34 weeks period of the study in the young dogs group. The same was observed with regard to the percentage of CD3+CD4+ Th lymphocytes and the CD4/CD8 ratio, while the percentage of CD3+CD8+ Tc lymphocytes increased from 7 to 15 months of age. These age-related changes found in lymphocyte subsets distribution of young dogs led to level the absolute and relative values of adult dog lymphocytes. The observations of this longitudinal study illustrate the changes related to maturation of lymphocyte subsets that occur during early life in Beagle dogs.