A serological study of canine herpesvirus-1 infection in a population of breeding bitches in Norway

Background

Canine herpesvirus-1 (CHV1) causes a fatal hemorrhagic disease in neonatal puppies and is associated with infertility in female dogs. This study was conducted to assess the status of CHV1 infection in bitches in proestrus or estrus and to investigate possible risk factors by a detailed questionnaire. Blood samples were collected from healthy bitches (n = 193) not vaccinated against CHV1, aged one year or older and admitted for estrus control to the Canine Reproductive Clinical Unit, Norwegian School of Veterinary Science. The serum samples were analysed by immunoperoxidase monolayer assay and serum titers were recorded as the reciprocal value of the highest dilution producing specific cell staining.

Results

Altogether, 85.5% of the dogs had CHV1 titers ≥ 80 and were classified as positive. Mean age for dogs included in the study was 4.2 years (95% CI 4.0-4.5), and there was no difference in age between seronegative dogs vs seropositive dogs. When grouping the seropositive dogs into three categories according to the magnitude of the titer, a total of 38.8% of the bitches displayed a weakly positive titer of 80, 44.8% had moderately positive titers of 160 or 320 and 16.4% of the dogs fell into the strongly positive category with titer of ≥640. No association was demonstrated when comparing CHV1 antibody titers to fertility parameters such as previous matings, pregnancies, whelpings, puppies born or condition of puppies. Further, there was no difference in seroprevalence between bitches that had been abroad for a period of time and dogs only living within a Norwegian environment. Samples from dogs collected in summer and fall displayed moderate to high antibody titers indicating recent infection with CHV1. Season, previous birth, and participation in competitions/shows explained 67-78% of the variation in antibody titer.

Conclusions

This study demonstrates that CHV1 infection is common in breeding bitches in the eastern part of Norway. Associations with putative risk factors were not identified. However, season, previous whelping, and participation in competitions/shows explained 67-78% of the variation in antibody titer.     

Effect of propofol on oxidative stress status in erythrocytes from dogs under general anaesthesia

Background

Alterations of the normal redox balance might be attributed to increase of plasma free-radical concentration and a disruption of the antioxidant defense system. One of the adverse effects of general anaesthetics is the exogen sources of reactive oxygen radicals that are responsible for several diseases. The purposes of the current study were to evaluate the effect of propofol on oxidative stress and to compare the differences between propofol induction only and induction plus continuous infusion on antioxidant status in dogs.

Findings

Beagle dogs were evaluated in the present study. The dogs were assigned randomly to receive three treatments in a crossover model. The three treatments were: group 1 (n = 9), 2% isoflurane; group 2 (n = 9), anaesthesia induced with an intravenous (IV) bolus dose of 6 mg/kg propofol and maintained with 1.5–2% isoflurane; group 3 (n = 9), total IV anaesthesia (induction with 6 mg/kg propofol, infusion with 0.6 mg/kg/min propofol). The results of this study show that dogs exposed to isoflurane had decreased antioxidant enzymes activities, whereas dogs injected with propofol had increased antioxidant enzymes activities.

Conclusions

The results of this study showed that an infusion dose of propofol has antioxidant effects in dogs. These effects may be beneficial to patients in whom free radicals play a role in oxidative stress, such as those with ischemia. Further studies are needed to evaluate whether these antioxidant effects of the anaesthetic are of clinical value.     

Intervertebral disc disease in Dachshunds radiographically screened for intervertebral disc calcifications

Background

Intervertebral disc disease (IDD) is a very common neurological disease, Dachshunds being the breed most often affected. In this breed, IDD has a hereditary background and is associated with intervertebral disc calcification (IDC), an indicator of severe intervertebral disc degeneration. In Finland, spinal radiography is used, when screening for IDC before breeding Dachshunds. We evaluated the association between IDC and IDD in Finnish Dachshunds radiographically screened for IDC.A questionnaire was sent to owners of 193 radiographically screened Dachshunds aged at least ten years. Clinical signs indicative of IDD were compared with IDC grade (grade 0 = no calcifications, grade 1 = 1 – 2 calcifications, grade 2 = 3 – 4 calcifications and grade 3 = 5 or more calcifications) and with age at the time of the radiographic examination. The diagnosis of IDD was confirmed by a veterinarian.

Results

IDD was common in the study population with 31% of dogs being affected. IDD and IDC were clearly connected (P < 0.001); IDD was rare in dogs with no calcifications (grade 0) and common in dogs with severe IDC (grade 3). The IDC grade was strongly positively associated with frequency of back pain periods (P < 0.001), and dogs with IDC grade 3 had frequent periods of pain. Reluctance to jump onto a sofa had a strong positive association with back pain. No association existed between age of the dog at the time of the radiographic examination and clinical signs indicative of IDD.

Conclusions

Radiographically detected IDC and IDD are common in Finnish Dachshunds and are strongly associated with one another. Spinal radiography is an appropriate screening tool for breeders attempting to diminish IDC and IDD in Dachshunds. A breeding program that screens dogs and selects against IDC can be expected to reduce the occurrence of IDD in future. Twenty-four to 48 months of age is a suitable age for screening.

Electronic supplementary material

The online version of this article (doi:10.1186/s13028-014-0089-4) contains supplementary material, which is available to authorized users.     

Factors associated with the success of rabies vaccination of dogs in Sweden

Background

United Kingdom, Ireland, Malta and Sweden maintain their national provisions for a transitional period regarding rules concerning rabies vaccination and individual serological test for rabies neutralizing antibodies. The purpose of vaccinating dogs against rabies is to establish pre-exposure immunity and protect individual animals from contracting rabies.The aim of the study was to investigate factors associated with reaching the internationally accepted threshold antibody titre of 0.5 IU/mL after rabies vaccination of dogs.

Methods

The study was a prospective single cohort study including 6,789 samples from Swedish dogs vaccinated with commercially available vaccines in Sweden, and the dog''s antibody responses were determined by the OIE approved FAVN test. Information on potential risk factors; breed, age, gender, date of vaccination, vaccine label and the number of vaccinations, was collected for each dog. Associations between the dependent variable, serological response ≥ 0.5 IU/mL or < 0.5 IU/mL and each of the potential risk factors were investigated using logistic regression analysis.

Results

Of 6,789 vaccinated dogs, 6,241 (91.9%) had an approved test result of ≥ 0.5 IU/mL. The results of the multivariable logistic regression analysis showed that vaccinating with vaccine B reduced the risk of having antibody titres of < 0.5 IU/mL by 0.2 times compared with vaccination using vaccine A. Breed size was found significant as an interaction with number of vaccinations and age at vaccination as an interaction with day of antibody testing after last vaccination. In summary, larger breeds were at higher risk of having antibody titres of < 0.5 IU/mL but if vaccinated twice this risk was reduced. Moreover, there were a increased risk for dogs < 6 months of age and > 5 years of age to have antibody titres of < 0.5 IU/mL, but this was affected by number of days from vaccination till testing.

Conclusions

The probability of success of rabies vaccinations of dogs depends on type of vaccine used, number of rabies vaccinations, the breed size of the dog, age at vaccination, and number of days after vaccination when the antibody titres are tested. The need for a booster vaccination regimen is recommended for larger breeds of dog.     

Evaluation of total oxidant and antioxidant status in dogs under different CO2 pneumoperitoneum conditions

Background

The induction of the pneumoperitoneum increases intraabdominal pressure (IAP), causing splanchnic ischemia, whereas its deflation normalizes IAP and splanchnic blood flow. We investigated the oxidant-antioxidant status of dogs who underwent low pressure (7 mm Hg), standard pressure (12 mm Hg), and high pressure (15 mm Hg) pneumoperitoneum.

Results

Twenty-four beagle dogs (12 males and 12 females), 4–6 years old, weighing 8–11 kg were used. The animals were assigned to one of four groups (n = 6 dogs). Group 1 served as a control; these animals received only anaesthesia for 90 min. In groups 2, 3 and 4, intra-abdominal pressure was increased to 7, 12 and 15 mmHg, respectively, and maintained for 60 min. Total oxidant status (TOS) and total antioxidant status (TAS) were determined in venous blood samples. The percentage ratio of TOS to TAS provided an oxidative stress index (OSI).No significant difference in TOS levels was found among the groups. A significant decrease in TAS levels and an increase in OSI levels were observed at 90 min and 24 h of pneumoperitoneum deflation within group 4. No differences were found among the groups.

Conclusions

A high pressure pneumoperitoneum induced significant changes in TAS and OSI. In addition, TOS and TAS levels are useful markers for evaluating changes in the oxidative status caused by a pneumoperitoneum during laparoscopy. Furthermore, a low-pressure pneumoperitoneum could attenuate oxidative stress induced by CO₂ insufflation in dogs.     

Comparison between tracheal ratio methods used by three observers at three occasions in English Bulldogs

Background

Tracheal hypoplasia is a congenital condition described in mainly brachycephalic breeds and is one component of the brachycephalic obstructive airway syndrome (BOAS). Two radiographic methods have been described to evaluate the dimensions of the tracheal diameter in dogs and to distinguish between hypoplastic and non-hypoplastic tracheas: the tracheal lumen diameter to thoracic inlet distance ratio (TD/TI) and the ratio between the thoracic tracheal luminal diameter and the width of the proximal third of the third rib (TT/3R). The purpose of this study was to compare these two published radiographic methods between observers, different measuring occasions and to investigate the effect on classification of dogs as having hypoplastic or non-hypoplastic tracheas using four previously published mean ratios as cut-offs (<0.11, <0.127 and <0.144 for the TD/TI and <2.0 for the TT/3R method).Three observers evaluated right and left lateral recumbent radiographs from 56 adult English Bulldogs independently on three different occasions. TD/TI and TT/3R ratios were calculated and correlated between measuring occasions. Kappa, observed, positive, and negative agreements were calculated between observers and measuring occasions. Number of hypoplastic and non-hypoplastic dogs for each method and occasion was determined using <0.11, <0.127 and <0.144 as cut-offs for TD/TI and <2.0 for TT/3R.

Results

Intraobserver agreement varied with kappa between 0.45-0.94 for the TD/TI and 0.20-0.86 for the TT/3R method. Interobserver kappa varied between 0.27-0.70 for the TD/TI method and between 0.05-0.57 for the TT/3R method. There was poor agreement in classifying English Bulldogs as tracheal hypoplastic or non-hypoplastic, depending on measuring method, cut-off value and observer.

Conclusions

The diagnostic value of both the TD/TI and TT/3R methods with such poor agreement is questionable, and significantly impacts their reliability for both clinical evaluation of dogs and use in health screening programs.     

Speckle tracking echocardiography in mature Irish Wolfhound dogs: technical feasibility,measurement error and reference intervals

Background

Two-dimensional strain measurements obtained by speckle tracking echocardiography (STE) have been reported in both humans and dogs. Incorporation of this technique into canine clinical practice requires the availability of measurements from clinically normal dogs, ideally of the same breed, taken under normal clinical conditions.The aims of this prospective study were to assess if it is possible to obtain STE data during a routine echocardiographic examination in Irish Wolfhound dogs and that these data will provide reference values and an estimation of measurement error.

Methods

Fifty- four healthy mature Irish Wolfhounds were used. These were scanned under normal clinical conditions to obtain in one session both standard echocardiographic parameters and STE data. Measurement error was determined separately in 5 healthy mature Irish Wolfhounds.

Results

Eight dogs were rejected by the software algorithm for reasons of image quality, resulting in a total of 46 dogs (85.2%) being included in the statistical analysis. In 46 dogs it was possible to obtain STE data from three scanning planes, as well as to measure the rotation of the left ventricle at two levels and thus calculate the torsion of the heart. The mean peak radial strain at the cardiac apex (RS-apex) was 45.1 ± 10.4% (n = 44), and the mean peak radial strain at the base (RS-base) was 36.9 ± 14.7% (n = 46). The mean peak circumferential strain at the apex (CS-apex) was -24.8 ± 6.2% (n = 44), and the mean peak circumferential strain at the heart base (CS-base) was -15.9 ± 3.2% (n = 44). The mean peak longitudinal strain (LS) was -16.2 ± 3.0% (n = 46). The calculated mean peak torsion of the heart was 11.6 ± 5.1 degrees (n = 45).The measurement error was 24.8%, 26.4%, 11.5%, 6.7%, 9.0% and 10 degrees, for RS-apex, RS-base, CS-apex, CS-base, LS and torsion, respectively.

Conclusions

It is concluded that this technique can be included in a normal echocardiographic examination in large breed dogs under clinical conditions. The usefulness of the reference values reported here, given their wide normal range, will ultimately be determined by the values that are obtained from a large numbers of diseased dogs.     

Tumor slices as a model to evaluate doxorubicin in vitro treatment and expression of trios of genes PRSS11, MTSS1, CLPTM1 and PRSS11, MTSS1, SMYD2 in canine mammary gland cancer

Background

In women with breast cancer submitted to neoadjuvant chemotherapy based in doxorubicin, tumor expression of groups of three genes (PRSS11, MTSS1, CLPTM1 and PRSS11, MTSS1, SMYD2) have classified them as responsive or resistant. We have investigated whether expression of these trios of genes could predict mammary carcinoma response in dogs and whether tumor slices, which maintain epithelial-mesenchymal interactions, could be used to evaluate drug response in vitro.

Methods

Tumors from 38 dogs were sliced and cultured with or without doxorubicin 1 μM for 24 h. Tumor cells were counted by two observers to establish a percentage variation in cell number, between slices. Based on these results, a reduction in cell number between treated and control samples ≥ 21.7%, arbitrarily classified samples, as drug responsive. Tumor expression of PRSS11, MTSS1, CLPTM1 and SMYD2, was evaluated by real time PCR. Relative expression results were then transformed to their natural logarithm values, which were spatially disposed according to the expression of trios of genes, comprising PRSS11, MTSS1, CLPTM1 and PRSS11, MTSS1, SMYD2. Fisher linear discrimination test was used to generate a separation plane between responsive and non-responsive tumors.

Results

Culture of tumor slices for 24 h was feasible. Nine samples were considered responsive and 29 non-responsive to doxorubicin, considering the pre-established cut-off value of cell number reduction ≥ 21.7%, between doxorubicin treated and control samples. Relative gene expression was evaluated and tumor samples were then spatially distributed according to the expression of the trios of genes: PRSS11, MTSS1, CLPTM1 and PRSS11, MTSS1, SMYD2. A separation plane was generated. However, no clear separation between responsive and non-responsive samples could be observed.

Conclusion

Three-dimensional distribution of samples according to the expression of the trios of genes PRSS11, MTSS1, CLPTM1 and PRSS11, MTSS1, SMYD2 could not predict doxorubicin in vitro responsiveness. Short term culture of mammary gland cancer slices may be an interesting model to evaluate chemotherapy activity.     

Majority of T. gondii seropositive chickens (Gallus domesticus) in Central Ethiopia carries the infective parasite

Background

The prevalence of Toxoplasma gondii in free range chickens is a good indicator of the prevalence of T. gondii oocysts in the environment. The aim of this study was to isolate T. gondii parasites from heart and brain of seropositive free range (FR) chickens.

Findings

Isolation of T. gondii from pooled heart and brain of 41 direct agglutination test (DAT) positive (≥1:40) free range chickens (Gallus domesticus) was carried out by bioassay in mice. T. gondii specific antibodies in mice were assayed by DAT and microscopy was employed for detection and enumeration of brain tissue cysts. Overall, bioassay was positive in 29 (70.7%) chicken samples. T. gondii tissue cysts were isolated from 59% (24/41) of bioassayed chickens: from 2 of 7 chickens with a titer of 1: ≤ 60, 2 of 5 with titer 1: 180, 6 of 8 with titer 1: 540, 10 of 15 with titer 1: 1620, 1 of 2 with titer 1: 6000, 2 of 3 with titer 1:18000, 1 of 1 with titer 1:54000. None of the isolates was pathogenic for mice. Tissue cysts were detected from 61% of seropositive mice (DAT ≥ 1:40). Generally, tissue cyst counts per brain of mouse were low (mean: 132.7 ± 84.4; range: 47–352).

Conclusions

Majority of T. gondii seropositive chickens (Gallus domesticus) in Central. Ethiopia carries the infective parasite. Tissues from the free range chicken might be a source infection for animals and humans.     

High level of treatment failure with commonly used anthelmintics on Irish sheep farms

Background

In 2013 a Technology Adoption Program for sheep farmers was established to encourage the implementation of best management practices on sheep farms in Ireland. There were 4,500 participants in this programme in 2013. As part of this programme, farmers had the option to carry out a drench test to establish the efficacy of their anthelmintic treatment.

Results

Flock faecal samples were collected before and after treatment administration and gastrointestinal nematode eggs enumerated. In total there were 1,893 participants in the task, however only 1,585 included both a pre- and post-treatment faecal sample. Of those, 1,308 provided information on the anthelmintic product that they used with 46%, 23% and 28% using a benzimidazole (BZ), levamisole (LEV) and macrocyclic lactone (ML) product respectively. The remaining farmers used a product inapplicable for inclusion in the task such as a flukicide or BZ/LEV combination product. Samples were included for analysis of drench efficacy if the pre-treatment flock egg count was ≥200 eggs per gram and the interval post-sampling was 10–14 days for BZ products, 4–7 days for LEV products and 14–18 days for ML products. These criteria reduced the number of valid tests to 369, 19.5% of all tests conducted. If the reduction post-treatment was ≥95% the treatment was considered effective. Only 51% of treatments were considered effective using this criterion. There was a significant difference in efficacy between the anthelmintic drug classes with BZ effective in only 30% of treatments, LEV effective in 52% of cases and ML effective in 76% of cases.

Conclusions

Gastrointestinal nematode anthelmintic treatments, as practiced on Irish farms, have a high failure rate. There was a significant difference between the efficacies of the anthelmintic classes with BZ the least effective and ML the most effective.     

Treatment of canine generalized demodicosis associated with hyperadrenocorticism with spot-on moxidectin and imidacloprid

Background

Canine generalized demodicosis associated with hyperadrenocorticism is often problematic and might be intractable. The aim of this study was to report the efficacy of a weekly application of spot-on moxidectin/imidacloprid in dogs with hyperadrenocorticism and secondary generalized demodicosis.

Methods

Dogs with hyperadrenocorticism and secondary generalized demodicosis were included. The condition of hyperadrenocorticism was treated and stabilized with trilostane before and throughout the study period in all dogs.

Results

Average total live adult mite counts before treatment and after four, eight and 12 weeks of spot-on moxidectin/imidacloprid (2.5/10 mg/kg) applications were 20.1 ± 6.3 (range, 13–33), 0.5 ± 0.7 (range, 0–2; 6/11 were negative), 0.2 ± 0.4 (range, 0–1; 9/11 were negative), 0.2 ± 0.4 (range, 0–1; 9/11 were negative) and 0.1 ± 0.3 (range, 0–1; 10/11 were negative) respectively; this difference was significant (P < 0.001). Ten of 11 dogs (90.1%) achieved clinical remission, as demonstrated by the absence of demodectic mites at any life stage at monthly scrapings for eight consecutive weeks, and maintained remission throughout the 12-month follow-up period.

Conclusion

The weekly application of spot-on moxidectin/imidacloprid appeared to be effective and safe against generalized adult onset canine demodicosis associated with hyperadrenocorticism.     

Basal Serum Cortisol Concentration as a Screening Test for Hypoadrenocorticism in Dogs

Background

Measurement of basal serum or plasma cortisol concentration is used as a screening test for hypoadrenocorticism in dogs, but is not well characterized.

Objectives

To evaluate the sensitivity and specificity of basal serum cortisol to detect hypoadrenocorticism in a population of dogs with a clinical suspicion of hypoadrenocorticism.

Animals

Four hundred and fifty dogs with nonadrenal gland illness and 14 dogs with naturally occurring hypoadrenocorticism were included.

Methods

Retrospective case‐control study. The records of all dogs having had an ACTH stimulation test performed between January 2005 and September 2011 at the University of Bristol were reviewed. Dogs were included if the test was performed as a screening for hypoadrenocorticism. The sensitivity and specificity of basal serum cortisol concentration to detect dogs with hypoadrenocorticism were calculated using 2 cut‐offs and compared to the gold standard ACTH stimulation test.

Results

Using a cut‐off of ≤2 μg/dL (≤55 nmol/L), the sensitivity and specificity of basal cortisol to detect hypoadrenocorticism were 100% and 63.3%, respectively, whereas for a cut‐off of ≤1 μg/dL (≤28 nmol/L), the sensitivity and specificity were 85.7% and 91.8%, respectively.

Conclusions and Clinical Importance

Measurement of basal serum cortisol is useful as a screening test for hypoadrenocorticism in dogs using a cut‐off of ≤2 μg/dL (≤55 nmol/L), and the disease is unlikely with a basal serum cortisol >2 μg/dL (>55 nmol/L). A basal serum cortisol ≤2 μg/dL (≤55 nmol/L) cannot be used to diagnose hypoadrenocorticism, and an ACTH stimulation test should be performed in these cases.     

Detection of circulating microfilariae in canine EDTA blood using lens-free technology: preliminary results

Dirofilaria immitis causes life-threatening heart disease in dogs, thus screening of dog populations is important. Lens-free technology (LFT) is a low-cost imaging technique based on light diffraction that allows computerized recognition of small objects in holographic images. We evaluated an algorithm capable of recognizing microfilariae in canine whole blood using the LFT. We examined 3 groups of 10 EDTA blood specimens, from dogs with microfilaremia (group A), healthy dogs (B), and dogs with hematologic modifications other than microfilaremia (C). The LFT analyzer photographed repeated series of 5 images of all samples. The algorithm declared a sample positive if a microfilaria was detected on ≥1, ≥2, or ≥3 of the 5 images of a series. Microfilariae were detected visually in the images in 9 of 10 cases in group A; no microfilariae were seen in the images from groups B and C. Of the 30 cases, there were 14, 4, and only 3 false-positives with the 1 of 5, 2 of 5, and 3 of 5 image cutoffs, respectively. There were no false-negatives, regardless of cutoff. LFT seems useful for detecting microfilaria and could have application in clinical pathology.     

Epidemiology of methicillin resistant Staphylococcus pseudintermedius in guide dogs in Finland

Background

Methicillin resistant Staphylococcus pseudintermedius (MRSP) and Staphylococcus aureus (MRSA) are common multi-drug resistant (MDR) bacteria in dogs. In 2012–2013 three dogs of the Guide Dog School of the Finnish Federation of the Visually Impaired were found to be MRSP positive. Guide dogs have regular contact with each other during their first year of life and prolonged contact when in training. Since dogs are placed in different parts of Finland after training, there is a risk for national spread of MDR bacteria. In this study the prevalence of MRSP and MRSA, as well as the risk factors for MRSP were determined in the Finnish guide dog population. MRSP isolates were investigated using molecular methods and compared to the earlier isolates.

Results

Out of 132 tested dogs 4 were MRSP positive thus giving the prevalence estimate of 3% (95% CI: 1–8%) for MRSP in the target population. MRSA was not detected (prevalence estimate 0%, 95% CI: 0–3%). Risk factors associated with MRSP were being a breeding bitch (OR = 8.4; 95% CI: 1.1–64.1, P = 0.012), the number of veterinary visits (OR = 1.23; 95% CI: 1.0–1.5, P = 0.025) and number of antimicrobial courses (OR = 1.63; 95% CI: 1.0–2.55; P = 0.035). Identified MRSP isolates belonged to five different sequence types (ST45, 71, 402, 403 and 404). All ST71 isolates carried SCCmec II-III, while the SCCmec type of the ST45 and ST402 (a single locus variant of ST45) isolates were non-typeable with the method used.

Conclusions

MRSP and MRSA had low prevalence in the studied dog population despite the close contact between dogs, and the MRSP population was heterogenic. Antimicrobial therapy and veterinary visits are risk factors for MRSP even among a small case group.

Electronic supplementary material

The online version of this article (doi:10.1186/s13028-015-0129-8) contains supplementary material, which is available to authorized users.     

The use of the rapid osmotic fragility test as an additional test to diagnose canine immune-mediated haemolytic anaemia

Background

Diagnosing canine immune-mediated haemolytic anaemia (IMHA) is often challenging because all currently available tests have their limitations. Dogs with IMHA often have an increased erythrocyte osmotic fragility (OF), a characteristic that is sometimes used in the diagnosis of IMHA. Since the classic osmotic fragility test (COFT) is time-consuming and requires specialized equipment, an easy and less labour-intensive rapid osmotic fragility test (ROFT) has been used in some countries, but its diagnostic value has not yet been investigated.This study aimed to evaluate erythrocyte osmotic fragility in dogs with and without IMHA, to compare results of the classic (COFT) and rapid (ROFT) test and to assess the value of the ROFT as diagnostic test for canine IMHA.Nineteen dogs with IMHA (group 1a), 21 anaemic dogs without IMHA (group 1b), 8 dogs with microcytosis (group 2), 13 hyperlipemic dogs (group 3), 10 dogs with lymphoma (group 4), 8 dogs with an infection (group 5) and 13 healthy dogs (group 6) were included.In all dogs, blood smear examination, in-saline auto-agglutination test, Coombs’ test, COFT and ROFT were performed. In the COFT, OF5, OF50 and OF90 were defined as the NaCl concentrations at which respectively 5, 50 and 90% of erythrocytes were haemolysed.

Results

Compared with healthy dogs, OF5 and OF50 were significantly higher in group 1a (P < 0.001) and OF5 was significantly higher in group 3 (P = 0.0266). The ROFT was positive in 17 dogs with IMHA, 10 hyperlipemic dogs, one anaemic dog without IMHA and one healthy dog.

Conclusions

Osmotic fragility was increased in the majority of dogs with IMHA and in dogs with hyperlipidemia, but not in dogs with microcytosis, lymphoma or an infection. Although more detailed information was obtained about the osmotic fragility by using the COFT, the COFT and ROFT gave similar results. The ROFT does not require specialized equipment, is rapid and easy to perform and can be used easily in daily practice. Although, the ROFT cannot replace other diagnostic tests, it may be a valuable additional tool to diagnose canine IMHA.     

Alternative classification and screening protocol for transitional lumbosacral vertebra in German shepherd dogs

Background

Lumbosacral transitional vertebra (LTV) is a common congenital and hereditary anomaly in many dog breeds. It predisposes to premature degeneration of the lumbosacral junction, and is a frequent cause of cauda equina syndrome, especially in German shepherd dogs. Ventrodorsal hip radiographs are most often used in diagnosis of LTV in screening programs. In this study, value of laterolateral lumbar spine radiographs as additions to ventrodorsal radiographs in diagnosis of LTV, and characteristics of LTV and the eighth lumbar vertebra (L8) in laterolateral radiographs were studied. Additionally, computed tomography (CT) features of different types of LTV were elucidated.

Methods

The ventrodorsal pelvic and laterolateral lumbar spine radiographs of 228 German shepherd dogs were evaluated for existence and type of LTV. Morphology of transverse processes was used in classification of LTV in ventrodorsal radiographs. The relative length of sixth (L6) and seventh (L7) vertebrae (L6/L7) was used in characterization of these vertebrae in laterolateral radiographs. CT studies were available for 16 dogs, and they were used for more detailed characterization of different types of LTV. Non-parametric χ² statistics, generalized logit model for multinomial data, and one-way analysis of variance was used for statistical analyses.

Results

In all, 92 (40%) dogs had a LTV, the most common type being separation of first spinous process from the median crest of the sacrum in 62 dogs (67% of LTV). Eight dogs had eight lumbar vertebrae. Those dogs with LTV had longer L7 in relation to L6 than dogs with normal lumbosacral junctions. When L6/L7 decreased by 0.1 units, the proportion of dogs belonging to the group with L8 was 14-fold higher than in the group with normal lumbosacral junctions. L8 resembled first sacral vertebra (S1) in length and position and was therefore classified as one type of LTV. With CT it was shown that categorizing LTV, based on shape and visibility of transverse processes seen in ventrodorsal radiographs, could be misleading.

Conclusions

We suggest that L8 be included as a part of the LTV complex, and the laterolateral radiographs of the lumbar spine be considered as an addition to ventrodorsal projections in the screening protocols for LTV.     

Concentrations of cardiac Troponin I before and after ovariohysterectomy in 46 female dogs with pyometra

Background

Canine pyometra is a common disease in countries where routine spaying of young dogs is not common practice. This disease is known to lead to systemic inflammation potentially affecting multiple organs in the body, including the heart. Cardiac-specific Troponin I (cTnI) is a sensitive marker of myocardial cell damage, which can result from ischemia, trauma, toxins or inflammation. Dogs with pyometra are also exposed to anaesthesia which can potentially result in myocardial cell damage. The aims of the study were 1) to evaluate the occurrence of myocardial cell damage as indicated by increased serum concentrations of cTnI in dogs with pyometra and relate these to presence of systemic inflammation and 2) to evaluate the change in cTnI-concentrations after anaesthesia and surgery.

Methods

Serum cTnI concentration was measured preoperatively and one day after surgery in 46 female dogs with pyometra and 15 female dogs that underwent surgery for other reasons (ovariohysterectomy and mammary tumours).

Results

Forty-six female dogs of different breeds diagnosed with pyometra were included. The dogs had a median age of 8.5 years (IQR 7.5–10) and a median weight of 29 kg (IQR 9–32). Of the 46 dogs, 37 (80%) fulfilled the chosen criteria for systemic inflammatory response syndrome (SIRS) at inclusion. Thirteen (28%) of the dogs had increased cTnI concentrations (> 0.2 μg/l) before surgery and 18 (39%) had increased cTnI-concentrations the day after surgery. The cTnI concentrations in the 13 dogs with increased preoperative cTnI concentrations decreased in 8 dogs, increased in 4 dogs, and was unchanged in one dog. Seven dogs with nondetectable preoperative cTnI concentrations had increased postoperative concentrations. The only significant association between the studied laboratory or clinical variables (including SIRS) and cTnI concentration was preoperative percentage band neutrophils (PBN) and postoperative cTnI concentration (P = 0.016). In total, 20 dogs (43%) had increased pre- or postoperative cTnI concentrations. Seven dogs (15%) had pre-or postoperative concentrations of cTnI of 1.0 μg/l or higher.

Conclusion

Mild to moderate increases in cTnI appears to be common in dogs with pyometra before and after surgery, but the clinical importance of this finding is uncertain. None of the studied clinical variables were found to reliably predict increased preoperative cTnI concentrations. Because of the pre- and postoperative variation in cTnI concentrations, it was not possible to identify a negative effect of anaesthesia and surgery on myocardial cell integrity.     

Diurnal variation in fecal concentrations of acid-detergent insoluble ash and alkaline-peroxide lignin from cattle fed bermudagrass hays of varying nutrient content

Background

The effect of time of fecal sampling on the accuracy of acid-detergent insoluble ash (ADIA) and alkaline-peroxide lignin (APL) for the prediction of fecal output (FO) in cattle was evaluated. Eight ruminally cannulated cows (594 ± 35.5 kg) were allocated randomly to 4 bermudagrass [Cynodon dactylon (L.) Pers.] hay diets markedly different in crude protein concentration (79–164 g/kg) with 2 replicates per diet for 3 periods. Cows were offered hay individually at 20 g DM/kg of body weight daily in equal feedings at 08:00 and 16:00 h for a 10-d adaptation period followed by 5-d of total fecal collection. Fecal grab samples also were taken each day during the fecal collection period at 06:00, 12:00, 18:00, and 24:00 h either directly from the rectum or from freshly voided feces. Samples were composited within cow and time across the 5 d total fecal collection period. Additionally, forage, ort, and fecal samples were analyzed for concentrations of APL and ADIA.

Results

Fecal concentrations of ADIA and APL were not affected by sampling time (P ≥ 0.22), even though diet affected (P < 0.01) fecal ADIA and APL concentrations. There were no diet × sampling time interactions (P ≥ 0.60). Estimates of FO and dry matter digestibility (DMD) from ADIA and APL were not affected (P ≥ 0.16) by sampling time or the diet × sampling time interaction (P ≥ 0.74). Estimates of FO and DMD from markers from different sampling times or all different combinations of sampling time were not different (P ≥ 0.72) from those of total collection among internal markers.

Conclusion

Little variation in concentrations of ADIA and APL in daily fecal excretion across time increases flexibility in fecal grab sampling schedules for predicting FO and DMD.     

Cross-reactivity of commercially available anti-human monoclonal antibodies with canine cytokines: establishment of a reliable panel to detect the functional profile of peripheral blood lymphocytes by intracytoplasmic staining

Background

The process for obtaining monoclonal antibodies against a specific antigen is very laborious, involves sophisticated technologies and it is not available in most research laboratories. Considering that most cytokines remain partially conserved among species during evolution, the search for antibody cross-reactivity is an important strategy for immunological studies in veterinary medicine. In this context, the amino acid sequence from human and canine cytokines have demonstrated 49–96 % homology, suggesting high probability of cross-reactivity amongst monoclonal antibodies. For this, 17 commercially available anti-human monoclonal antibodies [IL-1α, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8 (#1, #2), IL-10, IL-12, IL-13, IL-17A, IFN-γ (#1, #2), TNF-α (#1, #2) and TGF-β], were evaluated in vitro for intracellular cytokine detection in a stimulated canine blood culture by flow cytometry and confocal microscopy. Lymphocytes from peripheral blood of healthy and two unhealthy dogs were analyzed.

Results

Eleven anti-human mAbs [IL-1α, IL-4, IL-5, IL-6, IL-8 (#1, #2), IL-12, IL-17A, TNF-α (#1, #2) and TGF-β] cross-reacted against canine intracellular cytokines. The specificity of the assays was not affected after Fc-blocking. Three anti-human cytokine mAbs [IL-4, IL-8 (#2) and TGF-β] when evaluated by confocal microscopy also cross-reacted with intracellular canine cytokines. The identification of human mAbs that cross-reacted with canine cytokines may support their use as immunological biomarkers in veterinary medicine studies.

Conclusion

The identification of these 11 anti-human cytokine mAbs that cross-reacted with canine cytokines will be useful immunological biomarkers for pathological conditions by flow cytometry and fluorescence microscopy in dogs.     

Prevalence of thermophilic Campylobacter species in Swedish dogs and characterization of C. jejuni isolates

Background

The aims of this study were to investigate the prevalence of Campylobacter species in Swedish dogs, to identify the species of the Campylobacter isolates and to genotype the C. jejuni isolates. Young and healthy dogs were targeted and the sampling was performed at 11 veterinary clinics throughout Sweden from October 2011 to October 2012. Faecal swab samples were collected and sent to the laboratory at the National Veterinary Institute (SVA) for isolation of Campylobacter, speciation and genotyping.

Results

Campylobacter spp. were isolated from 67 of the 180 sampled dogs which yields an overall prevalence of 37%. The most prevalent species of Campylobacter among the participating dogs was C. upsaliensis with 52 of the 67 identified isolates. A lower prevalence was observed for C. jejuni with seven identified isolates and one isolate was identified as C. helveticus. Multi-locus sequence typing (MLST) was carried out on the seven C. jejuni isolates and all sequence types that were found are also commonly found in humans. The dogs were divided into three age groups; 1) under 12 months, 2) 12 to 23 months and 3) 24 months and older. The highest prevalence was found in the two younger age groups. Dogs shedding C. jejuni were between 3–12 months of age while dogs shedding C. upsaliensis were found in all ages.

Conclusions

The present investigation finds that Campylobacter spp. known to cause campylobacteriosis in humans are present in Swedish dogs. The results suggest an age predisposition where dogs under 2 years of age are more likely to shed Campylobacter spp. than older dogs. The most commonly isolated species was C. upsaliensis followed by C. jejuni, which was only detected in dogs up to 12 months of age. All C. jejuni isolates identified in the present study were of the same MLST types that have previously been described both in humans and in animals. The awareness of the Campylobacter risk of healthy young dogs may be an important way to reduce the transmission from dogs to infants, young children and immunocompromised adults.