Feline diabetes mellitus shares many features with type 2 diabetes in people, regarding clinical presentation, physiology, and pathology. A breed predisposition for type 2 diabetes has been ...identified, with the Burmese breed at a fivefold increased risk of developing the condition compared to other purebred cats. We aimed to characterize the serum metabolome in cats (n = 63) using nuclear magnetic resonance metabolomics, and to compare the metabolite pattern of Burmese cats with that of two cat breeds of medium or low risk of diabetes, the Maine coon (MCO) and Birman cat, respectively. Serum concentrations of adiponectin, insulin and insulin-like growth factor-1 were also measured (n = 94). Burmese cats had higher insulin and lower adiponectin concentrations than MCO cats. Twenty one metabolites were discriminative between breeds using a multivariate statistical approach and 15 remained significant after adjustment for body weight and body condition score. Burmese cats had higher plasma levels of 2-hydroxybutyrate relative to MCO and Birman cats and increased concentrations of 2-oxoisocaproic acid, and tyrosine, and lower concentrations of dimethylglycine relative to MCO cats. The metabolic profile of MCO cats was characterized by high concentrations of arginine, asparagine, methionine, succinic acid and low levels of acetylcarnitine while Birman cats had the highest creatinine and the lowest taurine plasma levels, compared with MCO and Burmese. The pattern of metabolites in Burmese cats is similar to that in people with insulin resistance. In conclusion, the metabolic profile differed between healthy cats of three breeds. Detection of an abnormal metabolome might identify cats at risk of developing diabetes.
Reliable methods for determining whether or not a bitch has ovarian tissue present are needed for cases with unknown neutering status. Vaginal cytology consistent with heat is indicative of ...functional ovarian tissue. Other methods are required when the bitch is not presented in suspected heat. Progesterone can be analyzed during 2 months after suspected heat. During other stages, assays for the analyses of anti-Müllerian hormone (AMH) and luteinizing hormone (LH) have been used. The AMH assay is expected to give detectable concentrations (positive) in bitches with ovarian tissue, and the LH assay should give negative results in intact bitches, except during the pre-ovulatory LH peak. The aim of the present study was to study the diagnostic efficiency for detecting ovarian tissue in bitches using an AMH assay developed for human samples, and a semi-quantitative rapid immune migration (RIM™) LH assay developed for use in dogs. An AMH concentration of ≥0.1 μg/L, and an LH concentration of ≤1 μg/L, was set as the cut-off for presence of ovarian tissue. Client or staff owned bitches were included (N = 125). There were 73 intact bitches that were classified as being in heat (N = 25); in luteal phase (N = 12); or in anestrus (N = 36), and 52 spayed bitches that showed no clinical signs of estrogen influence. In total 64 of the 73 intact bitches (88%) were correctly identified using AMH, and 70/73 (96%) intact bitches were correctly identified using the LH assay. Excluding bitches in heat, the corresponding figures were 42/48 (88%) for AMH and 48/48 (100%) for LH. Of the 52 spayed bitches, 51 (98%) were correctly identified using the AMH assay and 49 (94%) were correctly identified using the LH assay. In this population, the predictive value of a positive AMH for intact bitches was 98%, and of a negative AMH for spayed bitches was 85%. Excluding bitches in heat, the predictive value of a negative LH test for intact bitches was 94%, and the predictive value of a positive LH test for identifying spayed bitches was 100%. It was concluded that analyses of AMH and LH are useful for detecting ovarian tissue in bitches, but that low concentrations of AMH may be obtained in intact bitches, classifying them as spayed. For LH, bitches in suspected estrus should not be tested to avoid the pre-ovulatory LH-surge, that otherwise may cause intact bitches being incorrectly identified as spayed.
•Assays for AMH and LH may be used for determining presence of ovarian tissue in the bitch.•Sensitivity for detecting intact bitches, excluding bitches in heat, was higher with the LH assay than with the AMH assay.•Specificity for detecting spayed bitches was higher with the AMH assay than with the LH assay.
In golden retriever dogs, a 1 bp deletion in the canine
gene has been shown to cause progressive retinal atrophy (PRA), the canine equivalent of retinitis pigmentosa. In humans,
is also implicated in ...Bardet-Biedl syndrome (BBS). To investigate if the affected dogs only exhibit a non-syndromic PRA or develop a syndromic ciliopathy similar to human BBS, we recruited 10 affected dogs to the study. The progression of PRA for two of the dogs was followed for 2 years, and a rigorous clinical characterization allowed a careful comparison with primary and secondary characteristics of human BBS. In addition to PRA, the dogs showed a spectrum of clinical and morphological signs similar to primary and secondary characteristics of human BBS patients, such as obesity, renal anomalies, sperm defects, and anosmia. We used Oxford Nanopore long-read cDNA sequencing to characterize retinal full-length
transcripts in affected and non-affected dogs, the results of which suggest that three isoforms are transcribed in the retina, and the 1 bp deletion is a loss-of-function mutation, resulting in a canine form of Bardet-Biedl syndrome with heterogeneous clinical signs.
New biomarkers that are directly associated with canine ovulation would be of value to ensure mating on optimal days of heat. In this study, canine plasma and serum were analyzed with liquid ...chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) to quantify a broad range of oxylipins for the purpose of developing a method for biomarker discovery studies in canine reproduction. A majority of the 67 oxylipins probed for were detected at comparable levels in both sample types, but more oxylipins at higher concentrations were detected in serum than in plasma. Nine of the oxylipins were detected in a pilot study of serum at levels that significantly differed (p ≤ 0.1) between time-points before (n = 10), during (n = 10) and after (n = 10) ovulation, and might serve as putative biomarkers for canine ovulation. One oxylipin (20-HETE) was significantly altered after adjusting for multiple comparisons. In conclusion, the results showed that the LC-ESI-MS/MS method was suitable for quantification of canine oxylipins, revealing important similarities and differences between plasma and serum profiles as well as preliminary ovulation-specific changes in a subset of the investigated oxylipins.
Background
Increased plasma concentration of N‐terminal‐prohormone B‐type natriuretic peptide (NT‐proBNP) can be detected in cats with cardiac disease. Potential effects of feline characteristics on ...NT‐proBNP concentration may influence clinical usefulness.
Objectives
To evaluate potential effects of feline characteristics on NT‐proBNP plasma concentration and to compare NT‐proBNP plasma concentrations in healthy cats with results in hypertrophic cardiomyopathy (HCM) cats with or without left atrial enlargement (LAE) using an ELISA and a point‐of‐care test (POCT), and assess if POCT results reflect ELISA results.
Animals
One hundred healthy cats of 3 breeds and 39 HCM cats were included.
Methods
Diseases other than HCM were excluded by physical examination, blood pressure measurement, echocardiography, hematology, and serum biochemistry.
Results
Higher NT‐proBNP concentrations were found in males than in females in healthy (P = .005) and in HCM cats (P = .0021), but breed had no effect on NT‐proBNP concentrations. Using ≥100 pmol/L as a cutoff for abnormal samples, ELISA and POCT had similar sensitivity (SE; 72 and 74%) and specificity (SP; 97 and 98%) for detecting cats with HCM, cats with HCM and LAE (SE, both 100%; SP, 97 versus 98%), and cats with HCM without LAE (SE, both 69%; SP, 97 versus 98%), respectively, when compared to healthy cats.
Conclusions and Clinical Importance
Breed had no effect on plasma NT‐proBNP concentrations, but higher concentrations were found in male than in female cats. The ELISA and POCT had similar SE and SP for detecting HCM. Both tests could identify all HCM cats with LAE but not all HCM cats without LAE.
Background
Measured indirect blood pressure (BP) results in cats in a clinical environment might be affected by stress and characteristics of the cats.
Hypothesis
To investigate the influence of ...clinical setting, cat characteristics, and life situation on BP and pulse rate (PR) in healthy cats.
Animals
Ninety‐four healthy Domestic Shorthair, Birman and Norwegian Forest cats.
Methods
Blood pressure measured by high‐definition oscillometry in 3 settings: cat placed in its own carrier with veterinarian present; cat placed in carrier with owner alone present; and cat placed on table with veterinarian present. Statistical analyses were performed using mixed linear models.
Results
Systolic BP (SBP) did not differ among settings. Higher mean arterial pressure (MAP), diastolic BP (DBP), and PR were found when measurements were performed with cat placed on table, rather than in carrier. Coefficients of variation (CVs) higher for SBP, MAP, DBP, and PR when measured with cat placed on table than in carrier. Birman cats had lower BP than other breeds. Systolic BP, MAP, DBP, and PR increased with age. Cats allowed outdoors had lower PR than cats living strictly indoors.
Conclusion and Clinical Importance
No difference in SBP was found among settings, but measuring BP with the cat placed on the examination table gave higher MAP, DBP, PR, and CV than measuring BP with the cat in its carrier. Breed affected BP, with lower BP in Birman cats than other breeds. Blood pressure increased with age. Pulse rate was lower in cats allowed outdoors than cats living strictly indoors.
Today immunoassays are widely used in veterinary medicine, but lack of species specific assays often necessitates the use of assays developed for human applications. Mass spectrometry (MS) is an ...attractive alternative due to high specificity and versatility, allowing for species-independent analysis. Targeted MS-based quantification methods are valuable complements to large scale shotgun analysis. A method referred to as parallel reaction monitoring (PRM), implemented on Orbitrap MS, has lately been presented as an excellent alternative to more traditional selected reaction monitoring/multiple reaction monitoring (SRM/MRM) methods. The insulin-like growth factor (IGF)-system is not well described in the cat but there are indications of important differences between cats and humans. In feline medicine IGF-I is mainly analyzed for diagnosis of growth hormone disorders but also for research, while the other proteins in the IGF-system are not routinely analyzed within clinical practice. Here, a PRM method for quantification of IGF-I, IGF-II, IGF binding protein (BP) -3 and IGFBP-5 in feline serum is presented. Selective quantification was supported by the use of a newly launched internal standard named QPrEST™. Homology searches demonstrated the possibility to use this standard of human origin for quantification of the targeted feline proteins. Excellent quantitative sensitivity at the attomol/μL (pM) level and selectivity were obtained. As the presented approach is very generic we show that high resolution mass spectrometry in combination with PRM and QPrEST™ internal standards is a versatile tool for protein quantitation across multispecies.
Bernese mountain dogs are a large dog breed formed in the early 1900s in Switzerland. While originally farm dogs that were used for pulling carts, guarding, and driving cattle, today they are ...considered multi-purpose companion and family dogs. The breed is predisposed to several complex diseases, such as histiocytic sarcoma, degenerative myelopathy, or hip dysplasia. Using whole-genome sequencing (WGS) data, we assessed the genomic architecture of 33 unrelated dogs from four countries: France, Sweden, Switzerland, and the United States. Analysis of runs of homozygosity (ROH) identified 12,643 ROH with an average length of 2.29 Mb and an average inbreeding coefficient of 0.395. Multidimensional scaling analysis of the genetic relatedness revealed limited clustering of European versus USA dogs, suggesting exchanges of breeding stock between continents. Furthermore, only two mtDNA haplotypes were detected in the 33 studied dogs, both of which are widespread throughout multiple dog breeds. WGS-based ROH analyses revealed several fixed or nearly fixed regions harboring discreet morphological trait-associated as well as disease-associated genetic variants. Several genes involved in the regulation of immune cells were found in the ROH shared by all dogs, which is notable in the context of the breed's strong predisposition to hematopoietic cancers. High levels of inbreeding and relatedness, strongly exaggerated in the last 30 years, have likely led to the high prevalence of specific genetic disorders in this breed.
Increased knowledge on serum protein profiles during early pregnancy in dogs would be valuable for several reasons, including animal welfare. Inflammatory changes during this period have been ...described. Today, mass spectrometry (MS) is a well-established technique to perform unbiased qualitative and quantitative studies of proteins in body fluids regardless of species. In the present study, a shotgun proteomic analysis based on nano-liquid chromatography-MS was performed to identify proteins of altered abundance during canine pregnancy, and, thereafter, a targeted parallel reaction monitoring (PRM)-method was developed and applied to absolutely quantify the concentrations of a selection of these proteins. Among the 32 proteins found altered between pregnant and non-pregnant dogs in the initial analysis, 12 were selected based on their changes in concentration and known biological importance, and these were analyzed using the PRM method. The PRM method showed good linearity, repeatability and sensitivity, and confirmed the higher concentration of Fibrinogen A, protein S alpha and C-reactive protein at early time points in pregnant bitches. In conclusion, the combination of both methods allowed the identification of several altered proteins, and the quantification and description of the concentration patterns for a selection of them during the early stage of dog pregnancy.
MS is a powerful technique that allows the investigation of protein variations in samples from different origin, such as serum from dogs. The application of a shotgun proteomic analysis as a screening method has revealed the alteration of several proteins after fifteen days of pregnancy in dogs. The complementary development of a PRM MS-based method for several of these proteins has enabled the absolute quantification of their concentrations at five different time points during early pregnancy. With the MS technique, a combination of proteins can be studied with lower limits of detection than with immunoassays. Care should be taken not to interpret the observed changes in pregnant dogs as signs of disease.
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•Investigation of proteins with altered concentrations during early pregnancy in dogs.•Shotgun proteomics screening of altered proteins during canine pregnancy.•PRM method development for absolute quantification using heavy isotope-labeled peptides.•Protein concentration patterns may help in early diagnosis of dog pregnancy.
Pregnancy is considered a pro-inflammatory state that requires physiologic adaptation of the immune system of the mother. The aim of the present study was to study inflammatory and hormonal changes ...during canine pregnancy. Studies included analyses of peripheral concentrations of the acute phase proteins fibrinogen and C-reactive protein (CRP), the hormones progesterone and insulin-like growth factor I (IGF-I), hemoglobin, and analyses of the total leukocyte numbers and expression of cell surface antigens. Twenty bitches were included in the present study; 12 pregnant bitches and eight non-pregnant control bitches that were followed during the corresponding phase of the oestrous cycle. Blood samples were collected at the day of optimal mating (day 0) and then on days 7, 14, 21, 28 and 42. Progesterone, IGF-I and CRP were analysed in serum and fibrinogen in EDTA plasma. Haematology and leukocyte expression of a panel of inflammation-associated adhesion molecules (CD 11a, CD 18 and CD 49d) were evaluated from EDTA blood. The data were analyzed as repeated-measures data, using a mixed model approach. Progesterone varied with time in both pregnant and control bitches, and IGF-I varied with time in pregnant bitches. Both fibrinogen and CRP increased significantly with time for the pregnant bitches, but no significant change was detected for the control bitches. Increases were seen from day 21. The hemoglobin concentration decreased significantly with time in both pregnant and non-pregnant bitches. The neutrophil and monocyte numbers increased significantly in pregnant but not in control bitches. Pregnancy induced increased granulocyte expression of cell surface marker CD 18, increased monocyte expression of CD 18 and CD 49d, and increased lymphocyte expression of CD 49d. In conclusion, we describe inflammatory changes during canine pregnancy that are manifested as increases in concentrations of CRP and fibrinogen, an increase in neutrophils and monocytes, and in activation of granulocytes, monocytes and lymphocytes. The changes should be taken into account when evaluating concentrations of APPs and WBC in bitches during pregnancy. A variation in IGF-I concentrations was detected during pregnancy.
•Canine pregnancy is associated with inflammatory changes.•Fibrinogen and CRP increased from day 21 in pregnant bitches.•Neutrophil and monocyte numbers increased in pregnant bitches.•Granulocyte expression of cell surface marker CD 18 increased during pregnancy.•Monocyte expression of CD 18 and CD 49d, and lymphocyte expression of CD 49d increased in pregnancy.
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