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1.  Decreased plasma Chromogranin A361-372 (Catestatin) but not Chromogranin A17-38 (Vasostatin) in female dogs with bacterial uterine infection (pyometra) 
Background
Pyometra often induces systemic inflammatory response syndrome (SIRS) and early diagnosis is crucial for survival. Chromogranin A (CgA) is a neuroendocrine secretory protein that is co-released with catecholamines from the adrenal medulla and sympathetic nerve endings. A prognostic value of CgA has been found in humans that are critically ill or that have SIRS associated with infection. CgA has not yet been studied in dogs with bacterial infection. The aim of the study was to investigate CgA, measured by Chromogranin A361-372 (Catestatin; Cst) and Chromogranin A17-38 (Vasostatin; VS) in healthy dogs and in dogs with pyometra.
Results
Fifty dogs with pyometra, sampled prior to surgery and 64 healthy female dogs were included. In 19 pyometra cases, blood samples were also collected postoperatively. Concentrations of Cst and VS were measured in heparinised plasma and Cst also measured in EDTA plasma, by in-house radioimmunoassays. Student’s t-test and Wilcoxon two-sample test was used to test for differences between dog groups. Pre- and postoperative samples in dogs with pyometra were analysed by paired t-test. Pearson correlation was used to investigate associations of laboratory variables and hospitalization. P < 0.05 was considered significant.
Concentrations of Cst were decreased in pyometra dogs (mean ± SE, 1.01 ± 0.05 nmol/L) compared to healthy dogs (mean ± SE, 1.70 ± 0.03 nmol/L) (p ≤ 0.0001). VS concentrations did not differ significantly between dogs with pyometra (0.40 ± 0.04 nmol/L) and healthy dogs (0.42 ± 0.03 nmol/L). Mean ± SE pre- and postoperative concentration of Cst (1.0 ± 0.04 nmol/L and 0.9 ± 0.2 nmol/L) and VS (0.36 ± 0.04 nmol/L and 0.36 ± 0.04 nmol/L) in dogs with pyometra did not differ significantly. Neither Cst nor VS concentrations were associated with duration of hospitalization and were not significantly different in the four dogs with pyometra that had prolonged (≥3 d) postoperative hospitalization.
Conclusion
Concentrations of Cst, but not VS, were decreased in pyometra. Cst and VS concentrations before and after ovariohysterectomy did not differ significantly and were not associated with duration of hospitalization. Further studies are warranted to evaluate a possible diagnostic or prognostic value for Cst and VS.
Electronic supplementary material
The online version of this article (doi:10.1186/s12917-015-0328-6) contains supplementary material, which is available to authorized users.
doi:10.1186/s12917-015-0328-6
PMCID: PMC4318355  PMID: 25636335
Pyometra; Chromogranin A; Catestatin; Vasostatin; C-reactive protein; Biomarker; Dog; Uterine infection
2.  Increased concentrations of Serum amyloid A in dogs with sepsis caused by pyometra 
BMC Veterinary Research  2014;10(1):273.
Background
Sepsis is a serious and potentially life-threatening condition and early diagnosis and appropriate treatment is crucial for survival. Pyometra is one of the most common diseases in intact female dogs. The disease often leads to sepsis (systemic inflammatory response syndrome, SIRS, caused by infection). Diagnostic markers for detecting sepsis are gaining increasing interest in veterinary medicine. Acute phase proteins (APPs) such as C-reactive protein (CRP) are useful for detecting systemic inflammation in dogs. Serum amyloid A (SAA) is another major APP in dogs that is not yet as widely used. Albumin is regarded as a negative APP and has earlier been evaluated for prediction of prognosis in septic dogs. The aim of the present study was to determine SAA, CRP and albumin concentrations in dogs with sepsis and pyometra and to evaluate whether these inflammatory markers are associated with length of postoperative hospitalization.
Results
Thirty-one surgically treated bitches with pyometra were included, whereof 23 septic (SIRS-positive) and eight non-septic (SIRS-negative). Albumin concentrations were analyzed by routine automated methods. SAA and CRP analyses were performed with previously validated commercially available assays (ELISA and immunoturbidimetric).
Mean (±SE) serum concentrations of SAA were significantly higher in septic (130.8 ± 8.0 mg/L) compared to non-septic bitches (88.5 ± 12.5 mg/L). Using a cut-off value for SAA of 109.07 mg/L (n = 31 bitches), the sensitivity and specificity for detecting sepsis was 74% and 50%, respectively. Serum albumin concentrations were not significantly different in septic compared to non-septic bitches (mean ± SE, 25 ± 1 g/L and 26 ± 1 g/L, respectively). CRP concentrations were also not significantly different in septic (mean ± SE 225.6 ± 16.0 mg/L) compared to non-septic bitches (mean ± SE, 176.0 ± 27.1 mg/L). None of these inflammatory markers were associated with the outcome as measured by length of hospitalization.
Conclusions
SAA concentrations were increased in dogs with sepsis induced by pyometra and may be useful as an adjunctive diagnostic marker for sepsis. To evaluate the full potential of SAA as a marker for sepsis also in other diseases, further studies are warranted.
doi:10.1186/s12917-014-0273-9
PMCID: PMC4247870  PMID: 25430894
SAA; CRP; SIRS; Bitch; Acute phase protein
3.  Characterisation of bacterial growth and antimicrobial susceptibility patterns in canine urinary tract infections 
BMC Veterinary Research  2014;10(1):217.
Background
Bacterial urinary tract infection (UTI) is a common reason for antimicrobial therapy in dogs.
A reported increase in multi-drug resistance in canine bacterial pathogens, including resistance to extended-spectrum cephalosporins (ESC) is of concern as antimicrobial resistance complicates therapy in dogs. In addition, it is a possible public health concern.
The objectives of this study were to investigate the relative prevalence of pathogens in urine samples from dogs with urinary tract infection sampled at referral hospitals, clinics and mixed veterinary practices and to investigate if this was influenced by sample material or by contamination of the culture. The second objective was to assess the susceptibility patterns to clinically relevant antimicrobials and to investigate if this was influenced by whether the samples originated from smaller clinics or from referral hospitals and to perform active screening for the presence of Enterobacteriaceae resistant to ESC.
Results
Escherichia coli was the most frequently isolated pathogen (68%) followed by staphylococci (11%).
E. coli isolates were found significantly more often in pure culture than in contaminated samples. Staphylococcus pseudintermedius and Staphylococcus aureus isolates were significantly more prevalent in pre-incubated samples compared to samples submitted as non-incubated media.
Susceptibility to the majority of the tested first-line antimicrobials was common. Multiresistance was rare, and these isolates were all susceptible to at least one relevant antimicrobial. Isolates in samples from small animal clinics or mixed veterinary practices were less likely to be susceptible compared to isolates originating from referral animal hospitals. ESC-resistant Enterobacteriacae isolates were found in one per cent of the positive cultures. Bacteria with transferable ESC resistance were confirmed in one dog. The gene demonstrated was blaCMY2.
Conclusions
Choice of sample material might influence the possibility of detecting Staphylococcus pseudintermedius and Staphylococcus aureus isolates in clinical cases of UTI in dogs. Based on the study results, use of first-line antimicrobials is a rational empirical antimicrobial therapy for the studied dog population.
E. coli was the most prevalent pathogen, but prevalence of infection with ESC resistant Enterobacteriaceae including E. coli was low, as such isolates were found in only one per cent of the positive cultures.
doi:10.1186/s12917-014-0217-4
PMCID: PMC4180317  PMID: 25249356
Urinary tract; Bacterial infection; Dog; Antimicrobial resistance; Extended spectrum cephalosporins
5.  The first case of Brucella canis in Sweden: background, case report and recommendations from a northern European perspective 
Infection with Brucella canis has been diagnosed in Sweden for the first time. It was diagnosed in a three-year-old breeding bitch with reproductive disturbances. Fifteen in-contact dogs were tested repeatedly and all of them were negative for B. canis. The source of infection could not be defined. The present article describes the case and the measures undertaken and gives a short review over B. canis. Recommendations on how to avoid the infection in non-endemic countries are given.
doi:10.1186/1751-0147-54-18
PMCID: PMC3349463  PMID: 22452858
Brucella canis; Canine abortion; Reproduction; Canine brucellosis
6.  Carriage of methicillin-resistant Staphylococcus pseudintermedius in dogs--a longitudinal study 
Background
Methicillin-resistant S. pseudintermedius strains (MRSP) are reported with increasing frequency in bacterial cultures from dogs. The objectives of this study were to determine whether MRSP could be found in dogs several months after a clinically apparent infection and whether the length of carriage varied depending on systemic antimicrobial treatment, diagnosis at time of the first positive MRSP culture and the presence of skin disease or wounds. Thirty-one dogs previously diagnosed with a clinical infection were sampled repeatedly for a minimum of eight months or, with the exception of two dogs, until two consecutive negative results were obtained. Five specified locations were sampled, and the results were evaluated to determine future recommendations concerning sample strategies when screening for MRSP carriage. Information was collected from medical records and questionnaires to evaluate factors that may influence length of carriage.
Results
The overall median length of MRSP carriage was 11 months (48 weeks). The presence of wounds and signs of dermatitis did not influence length of carriage. Systemic treatment for three weeks or longer with antimicrobial agents to which the bacterium was resistant was associated with prolonged carriage compared to dogs treated for a shorter period of time. Three of five dogs treated with an antimicrobial to which their MRSP-isolates were susceptible (tetracycline) were found to still be MRSP-positive when sampled after the end of treatment. Wound samples had the highest positive MRSP yield (81%) for the positive sample sites, compared to less than 70% for each of the other four sample sites. Cultures from the nostrils were less likely to detect MRSP carriage relative to the pharynx, perineum, wounds and the corner of the mouth.
Conclusions
Dogs can carry MRSP for more than a year after a clinically apparent infection. Systemic antimicrobial treatment of infections with antimicrobial agents to which the MRSP-bacteria are resistant should be avoided when possible in dogs with possible or confirmed MRSP carriage or infection, since it may prolong time of MRSP carriage. Simultaneous sampling of pharynx, perineum, and the corner of the mouth as well as wounds when present is recommended when screening for MRSP. Cultures from nostrils were shown to be less likely to detect MRSP carriage.
doi:10.1186/1746-6148-8-34
PMCID: PMC3325892  PMID: 22444911
MRSP; Methicillin resistant Staphylococcus pseudintermedius; Dog; Carriage; Bacterial infection
7.  An investigation on the presence of Chlamydiaceae in Swedish dogs 
Background
Bacteria belonging to the family Chlamydiaceae cause a broad spectrum of diseases in a wide range of hosts, including man, other mammals, and birds. Upper respiratory and genital diseases are common clinical problems caused by Chlamydiaceae. Very little is known about chlamydial infections in dogs. Few clinical reports on natural disease in dogs describe mainly conjunctival and upper respiratory signs, and the role of Chlamydiaceae in genital disease is unclear. The present study aimed at studying the prevalence of Chlamydiaceae in healthy dogs and in dogs with genital or upper respiratory disease, including conjunctivitis.
Methods
A real-time polymerase chain reaction (PCR) for Chlamydiaceae was used to detect any chlamydial species within this family. Swab samples from the conjunctiva and the mucosal membranes of the oropharynx, rectum and genital tract were taken from 79 dogs: 27 clinically healthy dogs, 25 dogs with clinical signs from the genital tract and 28 dogs with conjunctivitis. There were 52 female and 27 male dogs. From 7 of the male dogs, additional semen samples were analysed.
Results
No Chlamydiaceae were detected from any dog.
Conclusions
Although the number of dogs that was included is limited, the results suggest that cases of Chlamydiaceae in dogs probably are related to infection from other species, and that dogs in general do not harbour Chlamydiaceae. Bacteria belonging to the family Chlamydiaceae do not seem to be of major importance for genital or ocular disease in Swedish dogs.
doi:10.1186/1751-0147-52-63
PMCID: PMC2993712  PMID: 21078208
8.  Application of a zona pellucida binding assay (ZBA) in the domestic cat benefits from the use of in vitro matured oocytes 
Background
Zona pellucida binding assays (ZBAs) have proven useful in determining the fertilising ability of spermatozoa in several species. Most ZBAs use fresh or salt-stored oocytes collected from fresh ovaries but because ovaries are not easy to obtain on a regular basis, chilled and frozen-thawed ovaries have been tested, with varying results. The present study tested the hypothesis that cat spermatozoa, either fresh or frozen-thawed, can bind to homologous zona pellucida of oocytes retrieved from frozen-thawed queen ovaries to a similar extent as they can bind to the zona pellucida of fresh, in vitro matured oocytes.
Methods
Ovaries were collected from queens after routine ovario-hysterectomy and either stored in NaCl at -20°C until use (treatment animals), or used fresh (controls). Cumulus-oocyte complexes (COCs) were retrieved by ovarian slicing from either source and used directly (immature oocytes from frozen-thawed ovaries; treatment animals) or after in vitro maturation (IVM) (fresh ovaries; controls) for 24 hours in TCM 199, supplemented with 1 IU hCG/mL and 0.5 IU eCG/mL and 0.5% bovine serum albumin (BSA). The oocytes were incubated for 4 hours in 5% CO2 in air at 38°C and 100% humidity in the presence of 5 × 106 fresh or frozen-thawed spermatozoa/mL. Representative samples of oocytes were processed for scanning electron microscopy (SEM).
Results
Both fresh and frozen-thawed spermatozoa bound to the in vitro matured zona pellucida but significantly fewer, or no, spermatozoa bound to frozen-thawed, immature zona pellucida (P < 0.001). Also, more fresh spermatozoa than frozen-thawed spermatozoa bound to the zona pellucida (P < 0.001). The zona pellucida surface differed in morphology (SEM), with in vitro matured oocytes showing a dense surface with few fenestrations in contrast to their frozen-thawed, immature counterparts, where fenestrations were conspicuously larger.
Conclusion
In conclusion, under the conditions of the present study, immature oocytes recovered from ovaries frozen immersed in NaCl at -20°C are less suitable for use in feline ZBA.
doi:10.1186/1751-0147-49-28
PMCID: PMC2092430  PMID: 17908298

Results 1-8 (8)