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18 pages, 722 KiB  
Article
And Child Makes Four: The Impact of a New Child on Dog-Owning Families
by Lori R. Kogan, Jennifer Currin-McCulloch, Cori Bussolari and Wendy Packman
Pets 2024, 1(3), 402-419; https://doi.org/10.3390/pets1030028 - 28 Nov 2024
Viewed by 61
Abstract
Pets are often an integral part of the family structure, with many people viewing their pets as children and thinking of themselves as “pet parents”. Pets have been shown to provide an array of psychological and physical health benefits, but some research suggests [...] Read more.
Pets are often an integral part of the family structure, with many people viewing their pets as children and thinking of themselves as “pet parents”. Pets have been shown to provide an array of psychological and physical health benefits, but some research suggests that the effects of pet ownership are more nuanced than once thought. One common negative aspect of pet ownership is guilt. Drawing from research pertaining to the transition to siblinghood, this study explored pet owner guilt as a result of adding a child to a dog-owning family. It also measured changes in dogs’ behaviors and their impact on owners’ depression and anxiety levels. Results suggest that many parents feel dog-related guilt and struggle with the resultant changes in household dynamics with the arrival of a child. Suggestions to help improve the chances of a successful transition include awareness, preparation, and self-compassion. Planning and preparing can help minimize negative dog behaviors and, as a result, levels of parental depression and anxiety. Parents are encouraged to help their dogs succeed by taking the time to prepare beforehand, thereby minimizing their own guilt and decreasing the potential of a severed human–dog bond. Full article
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<p>Participants’ responses to nine statements pertaining to areas of possible dog-related guilt.</p>
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<p>Participants’ report on 12 dog behavior problems.</p>
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15 pages, 723 KiB  
Systematic Review
A Systematic Review of the Efficacy of Compression Wraps as an Anxiolytic in Domesticated Dogs
by Savannah Mathis, Suzie Schoolfield, Peggy Gross, Margaret Gruen and David C. Dorman
Animals 2024, 14(23), 3445; https://doi.org/10.3390/ani14233445 - 28 Nov 2024
Viewed by 134
Abstract
Many domesticated dogs show signs of anxiety, negatively impacting their own and their owner’s mental and physical health. A systematic review evaluating whether pressure wraps decrease clinical signs of anxiety in dogs was completed to identify relevant studies. The main outcomes of interest [...] Read more.
Many domesticated dogs show signs of anxiety, negatively impacting their own and their owner’s mental and physical health. A systematic review evaluating whether pressure wraps decrease clinical signs of anxiety in dogs was completed to identify relevant studies. The main outcomes of interest included behavioral and physiological measures. A total of 229 studies were identified, of which four met the inclusion criteria. Commercially available pressure wraps and a telemetry vest were used in the reviewed studies. Three experimental studies included brief exposure to recorded firecracker or thunderstorm sounds or separation from their owner as anxiety-invoking stimuli. A non-experimental study evaluated the use of pressure wraps for dogs with thunderstorm phobia living in their home environment exposed to naturally occurring thunderstorms. The risk of bias in domains related to the blinding of investigators or study participants was generally high. Domains with moderate risk of bias often reflected incomplete reporting of research methods and imprecision due to small sample sizes. Most studies reported minimal benefits on behavior or heart rate in the absence of adverse side effects. Our review suggests there is weak and limited evidence to support the beneficial effect of pressure wraps in reducing physiological or behavioral signs of anxiety. Confidence in the available literature is low due to the moderate-to-high risks of bias and inconsistent findings. Further studies are needed regarding the efficacy of these products. Full article
(This article belongs to the Section Human-Animal Interactions, Animal Behaviour and Emotion)
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<p>PRISMA diagram.</p>
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<p>Risk of bias of individual studies. Colors denote low risk of bias (green), unclear risk of bias (yellow), or high risk of bias (red). Cottam [<a href="#B25-animals-14-03445" class="html-bibr">25</a>], Pekkin [<a href="#B26-animals-14-03445" class="html-bibr">26</a>], King [<a href="#B27-animals-14-03445" class="html-bibr">27</a>], Fish [<a href="#B28-animals-14-03445" class="html-bibr">28</a>].</p>
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12 pages, 4187 KiB  
Article
Morbillivirus Canis Infection Induces Activation of Three Branches of Unfolded Protein Response, MAPK and Apoptosis
by Santiago Emanuel Colina, Macarena Marta Williman, Marco Antonio Tizzano, María Soledad Serena, María Gabriela Echeverría and Germán Ernesto Metz
Viruses 2024, 16(12), 1846; https://doi.org/10.3390/v16121846 - 28 Nov 2024
Viewed by 128
Abstract
Morbillivirus canis, commonly named Canine distemper virus (CDV), is a morbillivirus implicated in several signs in the Canidae family. In dogs (Canis lupus familiaris), common signs of infection include conjunctivitis, digital hyperkeratosis and neuropathologies. Even with vaccination, the canine distemper [...] Read more.
Morbillivirus canis, commonly named Canine distemper virus (CDV), is a morbillivirus implicated in several signs in the Canidae family. In dogs (Canis lupus familiaris), common signs of infection include conjunctivitis, digital hyperkeratosis and neuropathologies. Even with vaccination, the canine distemper disease persists worldwide so the molecular pathways implicated in the infection processes have been an interesting and promising area in new therapeutic drugs research in recent years. It is known that in the process of virus infection, the endoplasmic reticulum (ER) loses its homeostasis, inducing stress and the subsequent unfolded protein response or UPR in which three ER-trans-membrane proteins are implicated: PERK, IRE1 and ATF6. Moreover, in prolonged ER stress, the apoptosis is induced through the CHOP, as a final step of viral infection. Cell culture and molecular techniques such as RT-qPCR and RT-PCR were used in the present study. We demonstrate the activation in vitro of the three UPR pathways after infection with an attenuated strain of CDV. Also, the implication of a MAPK pathway through the p38 protein and the apoptotic CHOP was demonstrated to contribute to the process of infection. Even more, our study suggested that CDV replication occurs in a PERK-dependent manner. Full article
(This article belongs to the Section Animal Viruses)
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Graphical abstract
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<p>Activation of BIP in VERO CCL-81 cells. An RT-qPCR assay was conducted to evaluate the expression of the GRP78/BIP transcript following the infection with CDVOnd. The induction of ER stress was detected at 8 hpi, reaching its maximum value at 16 hpi. The levels of BIP mRNA were relativised to the mock experiment and represented as the mean ± standard deviation (SD) of three independent experiments. For statistical significance, * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01 and *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Activation of the PERK pathway in CDVOnd infection. The relative levels of the ATF4 transcript indicate the activation of the PERK pathway following 16 hpi with the CDVOnd strain. The data were relativised to the mock well and are presented as means ± SD from three independent experiments. Statistical significance * <span class="html-italic">p</span> &lt; 0.05, *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>The ATF6 pathway is activated by the CDVOnd strain. The ATF6 mRNA levels were detected as early as 8 hpi. All values were relativised to the mock experiment. The values are presented as means ± SD of three independent experiments. Statistical significance values are indicated by * <span class="html-italic">p</span> ˂ 0.05 and ** <span class="html-italic">p</span> ˂ 0.01.</p>
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<p>XBP1 spliced after CDVOnd infection. (<b>a</b>) The images displayed are representative photographs of 2% agarose gels with GAPDH (<b>left</b>) and XBP1 (<b>right</b>) bands for each condition from the same experiment. The triangles indicate the XBP1 transcripts, XBP1u (green) and XBP1s (blue) with a size of approximately 440 bp and 414 bp, respectively. The star symbol denotes a hybrid between the two forms of XBP1 (XBP1H), with an apparent size of 466 base pairs. (<b>b</b>,<b>c</b>) Band densitometry assay. The expression levels of XBP1 species were relativised using a value of 1 for the GAPDH bands. In (<b>b</b>), a comparison was made between the levels of XBP1u and XBP1s. In (<b>c</b>), the ratio XBP1s/XBP1u relativised to the mock condition was evaluated in each condition. Levels are represented as means ± SD of three independent experiments. In all cases, statistical significance is indicated by * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001 and **** <span class="html-italic">p</span> &lt; 0.0001.</p>
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<p>p38⍺ and CHOP induced by CDVOnd infection. The figure illustrates the relative levels of mRNA transcripts for two transcription factors involved in the activation of apoptotic genes. (<b>a</b>) Levels of the p38α mRNA transcript were found to be upregulated following CDV infection. (<b>b</b>) Levels of the CHOP transcript in response to CDV infection, illustrating the upregulation of this gene. The data were relativised to the mock experiment and expressed as means ± SD from three independent experiments. Statistical significance was determined using the following criteria: * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>DNA laddering experiment after CDV infection. The study of apoptosis was conducted through the generation of DNA ladder-like patterns over a period of 16, 24, 48 and 72 h following the infection of CDVOnd. A 100 bp ladder (MW) was employed as a weight molecular marker. (<b>a</b>) Photographs taken at 4X of the mock and 48 hpi (<b>up</b>) and 72 hpi (<b>down</b>) wells. (<b>b</b>) Electrophoretic analysis of genomic DNA in a 1% agarose gel stained with GelGreen.</p>
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<p>Production of viral RNA following CDV infection. The relative RNA level of the CDVOnd strain was investigated using primers to the N gene at different time points. The levels of the viral genome were relativised to the mock experiment and data are presented as means ± SD of three independent experiments. For a statistical significance: * <span class="html-italic">p</span> ˂ 0.05, *** <span class="html-italic">p</span> ˂ 0.001.</p>
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<p>Inhibition of the PERK pathway affects viral progeny. Both experiments were conducted in duplicate in two conditions: without the PERK inhibitor (wo/GSK) and with the PERK inhibitor (w/GSK). (<b>a</b>) The TCID<sub>50</sub> assay was performed to quantify CDVOnd progeny at 48, 72 and 96 hpi. (<b>b</b>) Viral load performed by qPCR targeted the N gene of CDV. The mRNA levels were relativised to mock conditions.</p>
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9 pages, 468 KiB  
Article
First Evidence of Cotinine in Canine Semen Reveals Tobacco Smoke Exposure
by Debora Groppetti, Giulia Pizzi, Elisa Giussani, Alessandro Pecile, Silvia Michela Mazzola, Valerio Bronzo and Eleonora Fusi
Vet. Sci. 2024, 11(12), 598; https://doi.org/10.3390/vetsci11120598 - 26 Nov 2024
Viewed by 284
Abstract
Tobacco smoke has numerous adverse effects on both human and animal health, including impaired reproductive function. Recent research has explored environmental exposure in dogs, investigating various biological matrices. However, no data are currently available on the presence of cotinine, a nicotine metabolite, in [...] Read more.
Tobacco smoke has numerous adverse effects on both human and animal health, including impaired reproductive function. Recent research has explored environmental exposure in dogs, investigating various biological matrices. However, no data are currently available on the presence of cotinine, a nicotine metabolite, in the canine ejaculate. This study aimed to evaluate the detectability of cotinine in the semen of dogs living with smoking owners. Additionally, seminal cotinine concentrations were correlated with those in serum and hair. To further examine the potential impact of smoking on canine fertility, the relationships between seminal cotinine, total sperm concentration, and antioxidant activity in plasma and semen were analyzed in exposed and non-exposed dogs. This study is the first to demonstrate the presence of cotinine in canine ejaculate and its correlation with blood and hair concentrations. While the potential toxic effect of cotinine on seminal parameters and male fertility in dogs requires further investigation, it is crucial to raise awareness among pet owners about the risks associated with domestic smoking for their animals. Full article
(This article belongs to the Section Veterinary Reproduction and Obstetrics)
14 pages, 527 KiB  
Article
Retrospective Safety Evaluation of Combined Chlorambucil and Toceranib for the Treatment of Different Solid Tumours in Dogs
by Yuk-Yin Lai, Rodrigo Dos Santos Horta, Paola Valenti and Antonio Giuliano
Animals 2024, 14(23), 3420; https://doi.org/10.3390/ani14233420 - 26 Nov 2024
Viewed by 278
Abstract
Chlorambucil is used in veterinary medicine for various cancers, while Toceranib, which was licenced for treating canine mast cell tumours, is now used against other solid tumours. Both drugs are generally safe, but their combined use has not been studied. This study aimed [...] Read more.
Chlorambucil is used in veterinary medicine for various cancers, while Toceranib, which was licenced for treating canine mast cell tumours, is now used against other solid tumours. Both drugs are generally safe, but their combined use has not been studied. This study aimed to investigate retrospectively the safety profile of the Chlorambucil–Toceranib combination against canine solid tumours. Thirty-eight dogs received this combination. Chlorambucil was administered at a median dose intensity of 15.1 mg/m2 per week, while Toceranib was given at the median dosage of 2.5 mg/kg on a Monday–Wednesday–Friday schedule. Dosages were individually adjusted according to commercially available tablet formulation, co-morbidities, and adverse events (AEs). The resulting clinical benefit rate (CBR) and overall response rate (ORR) were 55.3% and 10.5%, respectively. The median progressive free survival (PFS) and median survival time (MST) were 45.5 (12–537) days and 259 (42–1178) days, respectively. Gastrointestinal AEs occurred in 39.5% of cases (n = 15), 15.8% (n = 6) experienced UPC elevation, while hematological and biochemistry AEs affected 13.2% (n = 5) each. Most of these AEs were grades 1–2 (G1–2). None of the dogs interrupted treatment due to AEs, and the combination appeared safe. Larger prospective clinical trials are required to confirm our findings and investigate its efficacy across various cancers. Full article
(This article belongs to the Section Veterinary Clinical Studies)
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<p>Kaplan–Meyer survival analysis for all dogs. (<span class="html-italic">p</span> &lt; 0.001).</p>
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18 pages, 783 KiB  
Article
The Demography, Longevity and Mortality of Bullmastiffs Attending Veterinary Practices in Australia
by Abigail Carney, Peter Williamson and Rosanne M. Taylor
Animals 2024, 14(23), 3419; https://doi.org/10.3390/ani14233419 - 26 Nov 2024
Viewed by 200
Abstract
The Bullmastiff is a giant breed of dog, and there is evidence the breed is predisposed to cancer and musculoskeletal disorders, though the disease investigation of the breed is limited. This study reports on the demography, longevity and mortality of Bullmastiffs attending veterinary [...] Read more.
The Bullmastiff is a giant breed of dog, and there is evidence the breed is predisposed to cancer and musculoskeletal disorders, though the disease investigation of the breed is limited. This study reports on the demography, longevity and mortality of Bullmastiffs attending veterinary practices in Australia over a ten-year period. VetCompass Australia collects patient data from veterinary practices across Australia for epidemiological analysis. All patient records of Bullmastiffs available in the VetCompass Australia database during this decade period were reviewed, with demographic information on the breed inclusive of coat colour, sex, neuter status, weight and location collated. Standardised veterinary diagnostic (VeNom) codes for the most appropriate cause of death were assigned to deceased dogs. The population comprised 2771 Bullmastiffs with an overall median age of 2.8 years. Within the group, 1259 were female (45.4%), 1491 were male (53.8%), and 21 dogs (0.8%) had no recorded sex or neuter status. Dogs grew rapidly in their first year, with an average gain of approximately 1 kg every 10 days. A slower growth rate continued in their second year, and growth plateaued as adulthood was reached, with the mean body weight of adult male dogs (46.6 kg) being heavier than that of females (40.5 kg). The age at death for the group was 8.5 years. The most common causes of death in the breed were mass lesions (28.2%), old age (9.9%), musculoskeletal-related disease (9.9%) and neurological (5.3%) and behavioural disorders (4.8%). Neutering was protective against mortality from urogenital causes (OR: 0.14; CI: 0.02–0.52; p = 0.003) and had a positive effect on longevity. This study provides demographic and health information on a population of Bullmastiffs attending veterinary practices in Australia, which will benefit evidence-based veterinary decisions for this breed. Additionally, the results may assist owners and breeders in making informed decisions on health risks and breeding programmes in the population. Full article
(This article belongs to the Section Companion Animals)
14 pages, 9528 KiB  
Article
Changes in the Periodontal Gap After Long-Term Tooth Movement into Augmented Critical-Sized Defects in the Jaws of Beagle Dogs
by Kathrin Duske, Mareike Warkentin, Anja Salbach, Jan-Hendrik Lenz and Franka Stahl
Dent. J. 2024, 12(12), 386; https://doi.org/10.3390/dj12120386 - 26 Nov 2024
Viewed by 202
Abstract
Background/Objectives: Extensive and closely coordinated remodeling processes take place in the periodontal ligament (PDL) and the adjacent bone during orthodontic tooth movement. In complex orthodontic cases, it is necessary to move teeth into an augmented bony defect, for example, in patients with cleft [...] Read more.
Background/Objectives: Extensive and closely coordinated remodeling processes take place in the periodontal ligament (PDL) and the adjacent bone during orthodontic tooth movement. In complex orthodontic cases, it is necessary to move teeth into an augmented bony defect, for example, in patients with cleft lip, alveolus, and palate. The important role of the PDL during tooth movement is well accepted but not fully understood. Therefore, the present study investigated the PDL after 23 weeks of tooth movement into an augmented critical-sized defect. Methods: The second molars of four beagle dogs were moved into a critical-sized defect, which was filled with bovine xenograft or nanocrystalline hydroxyapatite. Autogenous bone served as control. After 23 weeks, histological samples were microscopically analyzed, and the dimension of the PDL was measured. For statistical calculations, a Wilcoxon–Mann–Whitney test was used. Results: The PDL was significantly wider on the tension side compared with the compression side for all replacement materials analyzed (p ≤ 0.05). These results apply to both the mesial and distal roots. Conclusions: The remodeling processes reached equilibrium within 23 weeks, resulting in a wider gap on the tension side, which contrasts with the situation a few days after the initial force application. Full article
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<p>Schematic overview of the experimental design with different types of bone replacement materials (BRMs) used to fill critical-sized defects in the mandibles of four beagle dogs (CSD: critical-sized defect; PM: premolar; r: right side of the mandible; l: left side of the mandible; HA: hydroxyapatite; XENO: xenograft; AUTO: autograft).</p>
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<p>Intraoral images and X-rays of postoperative situations. Fixed BRMs positioned in the distal direction of the second premolar (PM2) (<b>a</b>). X-rays directly after surgical procedure of the autograft (<b>b</b>), xenograft (<b>c</b>), and hydroxyapatite (<b>d</b>). Bilateral distalization of PM2 started seven weeks after insertion of BRMs with an orthodontic appliance, which corresponded to the Beneslider system (<b>e</b>).</p>
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<p>Resected mandible with marked slicing levels (<b>a</b>). Image (<b>b</b>) shows a slice with the root of the first premolar (PM1) and the mesial and distal root of the second premolar (PM2), which was moved orthodontically into the filled critical-sized defect. Starting from the direction of orthodontic tooth movement, the yellow lines in both roots of PM2 mark the tension side, and the green lines mark the compression side. Light microscopic (<b>c</b>) and scanning acoustic microscopic (<b>d</b>) images through the distal root of PM2, which were moved into the augmented critical-sized defect (Dog 3 left side). Arrows mark periodontal gaps, which were measured at three different points on the compression side (points 1–3) and at three different points on the tension side (points 4–6).</p>
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<p>Light microscopic image of a stained thin section (Toluidine–Giemsa staining) with marked regions of interest (ROIs) (<b>a</b>). After histomorphometric analysis, bone is marked in light blue, cartilage matrix and osteoid in green, and bone marrow in red (<b>b</b>). Scanning acoustic microscopic image (<b>c</b>) of a thin section after histomorphometric analysis with marked ROIs (<b>b</b>). Arrows mark the roots of the PM2, which were moved into the augmented critical-sized defects.</p>
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<p>X-ray images of resected mandibles 30 weeks after implantation. (<b>a</b>,<b>c</b>,<b>e</b>): right side of the Dog 2, Dog 3, and Dog 4, respectively; (<b>b</b>,<b>d</b>,<b>f</b>): left side of the Dog 2, Dog 3, and Dog 4. XENO: xenograft; HA: hydroxyapatite; AUTO: autograft. Note that no residuals of BRMs were detectable. The arrow shows an encapsulated root fragment.</p>
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<p>Exemplary light microscopic images of periodontal gaps of the single root of PM1 and the mesial and distal root of PM2: (<b>a</b>) Dog 3 left side; (<b>b</b>) Dog 2 right side. The periodontal gaps of the single roots of PM1 show smooth and uniform configurations in both animals. Periodontal gaps of mesial and distal roots of PM2 were found to be much wider and irregularly shaped with protuberances around the entire root.</p>
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<p>Periodontal gap dimensions (mean and SD) from both roots of PM2, which was orthodontically moved into the augmented critical-sized defect. Periodontal gaps were measured at three points on the compression and tension sides using light microscopic images (<b>a</b>), as well as scanning acoustic images (<b>b</b>). Mesial and distal roots of PM2 were considered separately. XENO: xenograft; HA: hydroxyapatite; AUTO: autograft; * indicates significant differences between BRMs (<span class="html-italic">p</span> ≤ 0.05).</p>
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<p>Proportion of bone, osteoid, and bone marrow of the mesial and distal ROI shown as boxplots of light microscopy (<b>a</b>) and scanning acoustic microscopy (<b>b</b>). XENO: xenograft, HA: hydroxyapatite, AUTO: autograft. * indicates significant differences (<span class="html-italic">p</span> ≤ 0.05).</p>
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5 pages, 185 KiB  
Brief Report
European EHBP1L1 Genotyping Survey of Dyserythropoietic Anemia and Myopathy Syndrome in English Springer Spaniels
by Sarah Østergård Jensen, Alexandra Kehl and Urs Giger
Vet. Sci. 2024, 11(12), 596; https://doi.org/10.3390/vetsci11120596 - 26 Nov 2024
Viewed by 365
Abstract
Dyserythropoietic anemia and myopathy syndrome (DAMS) with neonatal losses was recently characterized as an autosomal recessive disorder caused by an EHBP1L1 frameshift variant in English Springer Spaniels (ESSPs). The frequency and dissemination of the mutation remained unknown. The EHBP1L1 protein is essential for [...] Read more.
Dyserythropoietic anemia and myopathy syndrome (DAMS) with neonatal losses was recently characterized as an autosomal recessive disorder caused by an EHBP1L1 frameshift variant in English Springer Spaniels (ESSPs). The frequency and dissemination of the mutation remained unknown. The EHBP1L1 protein is essential for muscle function, and the Rab8/10-EHBP1L1-Bin1-dynamin axis participates in nuclear polarization during the enucleation of erythroblasts. Lack of EHBP1L1 function decreases enucleation, leading to increased numbers of nucleated erythrocytes, which are characteristic of DAMS. A genotyping survey for the EHBP1L1 variant was conducted based upon submitted samples of ESSPs from Europe. DNA was extracted, and a real-time PCR assay, with allele-specific TaqMan probes for EHBP1L1 wild-type and frameshift deletion, was applied. Between September 2022 and August 2024, 803 samples were received from 18 European countries. The EHBP1L1 mutant allele frequency was 9.7%, including 4 homozygous dogs and 148 heterozygotes. The mutant EHBP1L1 allele was found in 13 countries. A total of 6 homozygous and 73 heterozygous ESSPs reported on an open database could be tracked to an original common ancestor. Although the survey is biased, it indicates that the mutant EHBP1L1 variant is disseminated in the breed and across Europe. The genotyping of ESSPs is recommended to diagnose DAMS and guide breeders. Full article
(This article belongs to the Section Veterinary Internal Medicine)
15 pages, 1797 KiB  
Article
Rank-Related Differences in Dogs’ Behaviours in Frustrating Situations
by Kata Vékony, Viktória Bakos and Péter Pongrácz
Animals 2024, 14(23), 3411; https://doi.org/10.3390/ani14233411 - 26 Nov 2024
Viewed by 336
Abstract
Dogs are strongly dependent on humans, not only for sustenance, but they also form asymmetrical bonds with us where they rely on assistance from the human partner in the case of difficult situations. At the same time, cohabiting dogs form hierarchies, and their [...] Read more.
Dogs are strongly dependent on humans, not only for sustenance, but they also form asymmetrical bonds with us where they rely on assistance from the human partner in the case of difficult situations. At the same time, cohabiting dogs form hierarchies, and their rank strongly influences their behaviour in various social interactions. In this study, we investigated whether high- and low-ranking dogs would behave differently in non-social and social contexts where a formerly available reward suddenly becomes inaccessible. We hypothesised that dominant and subordinate dogs would show different levels of human dependence; thus, they will show different levels and different signs of frustration depending on the social nature of the context, where the reward was locked either in a closed cage or withheld in the hand of the experimenter. The results showed that while the holistic rank (‘dominant’ vs. ‘subordinate’) of the dogs did not show a significant association with their reactions to frustrating situations, the rank components (‘agonistic’ and ‘leadership’ ranks) were better predictors of the dogs’ behaviour. In the non-social context, the highly resource-oriented ‘agonistic-dominant’ dogs were more persistent with their attempts of getting to the reward. However, in the social context, the dogs with high ‘leadership scores’ behaved more demandingly with the non-complying experimenter. This study provides a first-time indication that the various aspects of dominance in dogs can affect their adaptive reward-oriented behaviours differently, depending on the potentially available human assistance. Full article
(This article belongs to the Section Companion Animals)
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<p>The experimental setup of the non-social frustration test.</p>
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<p>The experimental setup of the social frustration test.</p>
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<p>The results of the between-context (non-social vs. social test) comparisons for separate analyses of dominant and subordinate dogs in the case of (<b>A</b>) time spent gazing at the cage in the non-social test and the experimenter in the social test and (<b>B</b>) time spent gazing at the owner. The box shows the interquartile range between the lower and upper quartiles, divided by the median. The whiskers extend to the minimum and maximum values excluding the outliers; the dots show the outliers (more than 1.5 times the lower/upper quartile beyond the box). NonSoc = non-social test condition; Soc = social test condition; E = experimenter. The asterisks show significance in the following way: ** <span class="html-italic">p</span> &lt;0.01.</p>
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<p>Significant associations in the non-social frustration test. (<b>A</b>) Dogs with higher agonistic scores were more determined to reach the treat; (<b>B</b>) older dogs showed less begging behaviour directed to the experimenter. The yellow dots are the datapoints, the blue band is the confidence interval.</p>
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<p>Significant associations in the social frustration test. (<b>A</b>) Dogs with higher leadership scores showed more demanding behaviour towards the experimenter; (<b>B</b>) older dogs were less focused on the experimenter and the treat. The yellow dots are the datapoints, the blue band is the confidence interval.</p>
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13 pages, 739 KiB  
Article
Cytokinesis-Blocking Micronucleus Assay for Assessing Nuclear Chromatin Integrity Abnormalities in Dog’s Somatic Cells After Exposure to HVAD-Produced Silver Nanoparticles
by Anna Grzesiakowska-Dul, Marek Jan Kasprowicz, Agnieszka Otwinowska-Mindur, Przemysław Baran and Marta Kuchta-Gładysz
Int. J. Mol. Sci. 2024, 25(23), 12691; https://doi.org/10.3390/ijms252312691 - 26 Nov 2024
Viewed by 284
Abstract
The widespread use of silver nanoparticles in many industries is increasing every year. Along with this use, there is growing concern about the potential unintentional exposure of human and animal organisms to these nanomaterials. It has been shown that AgNPs have the ability [...] Read more.
The widespread use of silver nanoparticles in many industries is increasing every year. Along with this use, there is growing concern about the potential unintentional exposure of human and animal organisms to these nanomaterials. It has been shown that AgNPs have the ability to penetrate organisms and can have harmful effects on cells and organs in the body. In order to reduce the effects of AgNPs on living organisms, newer solutions are being investigated, such as particle stabilization or other methods of synthesizing these particles. The physical synthesis of AgNPs using high-voltage arc discharge (HVAD) may be one of these alternatives. To determine the effect of silver nanoparticles obtained by this method, cytogenetic analysis was performed on domestic dog somatic cells using a cytokinesis-blocking micronucleus assay. In the experiments performed, peripheral blood cells of the domestic dog were exposed in vitro for 3 and 24 h to three tested colloidal silver compounds (unstable AgNP-HVAD, sodium citrate-stabilized silver nanoparticles—AgNP+C, and silver nitrate). The toxicity of these compounds was evaluated at concentrations of 5, 10, and 20 µg/L, and the presence of the following cellular abnormalities was analyzed: micronuclei, nuclear buds, nucleoplasmic bridges, or multinucleated cells. The study showed a significant increase in the number of micronuclei compared to the control sample, as well as the presence of nuclear buds and nucleoplasmic bridges in somatic cells of the domestic dog, confirming the genotoxic nature of the particles. However, there was no cytotoxic effect due to the lower number of multinucleated cells and the absence of apoptotic or necrotic cells in the samples analyzed. Further studies are needed to better understand the mechanisms of toxicity of AgNPs produced by the HVAD method and the extent of their effects on mammalian somatic cells. Full article
(This article belongs to the Special Issue Cytotoxicity of the Nanoparticles)
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<p>Cells analyzed by CBMN assay in domestic dog samples: (<b>a</b>) binucleated cell; (<b>b</b>) binucleated cell with one micronucleus; (<b>c</b>) binucleated cell with two micronuclei; (<b>d</b>) binucleated cell with nucleoplasmic bridge; (<b>e</b>) binucleated cell with nuclear bud; (<b>f</b>) multinucleated cell. Giemsa staining, 100× objective magnification, 10 µm scale. Arrows indicate forms of chromosomal instability.</p>
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<p>Cells analyzed by CBMN assay in domestic dog samples: (<b>a</b>) binucleated cell; (<b>b</b>) binucleated cell with one micronucleus; (<b>c</b>) binucleated cell with two micronuclei; (<b>d</b>) binucleated cell with nucleoplasmic bridge; (<b>e</b>) binucleated cell with nuclear bud; (<b>f</b>) multinucleated cell. Giemsa staining, 100× objective magnification, 10 µm scale. Arrows indicate forms of chromosomal instability.</p>
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10 pages, 2705 KiB  
Case Report
Left Ventricular Apical Aneurysm in a Dog—A Case Report
by Ozana Maria Hritcu, Radu Andrei Baisan, Aurelian Sorin Pasca and Florentina Daraban Bocaneti
Animals 2024, 14(23), 3412; https://doi.org/10.3390/ani14233412 - 26 Nov 2024
Viewed by 256
Abstract
The cadaver of a 16-year-old mixed-breed male dog was presented to the Faculty of Veterinary Medicine of IULS, Romania, after having been euthanized in a private practice at the owner’s request following acute cardio-respiratory failure. The necropsy revealed a left ventricular apical aneurysm [...] Read more.
The cadaver of a 16-year-old mixed-breed male dog was presented to the Faculty of Veterinary Medicine of IULS, Romania, after having been euthanized in a private practice at the owner’s request following acute cardio-respiratory failure. The necropsy revealed a left ventricular apical aneurysm with a bulging fibrous sac measuring about 0.7 cm/0.3 cm. The lesion involved all three structures of the cardiac wall, with mineralization areas and neovascularization. Immunohistochemistry showed strong positivity for desmin and weak positivity for CD-31 in the cardiac muscle fibers, positivity for α-sma in the smooth muscle cells of the vascular walls and positivity for vimentin in the cardiac stroma. MMP-2 and MMP-9 showed strong expression in the cardiomyocytes, whereas TIMP-1 showed positive immunoexpression only in the Eberth’s lines. To the authors’ knowledge, left ventricular apical aneurysm has only been reported once in dogs. Full article
(This article belongs to the Special Issue New Advances in Canine and Feline Diagnostics—2nd Edition)
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<p>Gross pathology of the cardiac apical aneurysm. (<b>A</b>) Saccular region of the apical aneurysm without any pressure exerted on it (white arrow). (<b>B</b>) Saccular region of the apical aneurysm with pressure exerted on it, causing it to bulge out (white arrow). (<b>C</b>,<b>D</b>) Apical aneurysm transversally sectioned. The saccular region is so thinned out that it is barely visible (white arrows).</p>
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<p>Cardiac apical aneurysm in a dog. (<b>A</b>) Thinned apical wall with fibrosis and no cardiomyocytes (black arrow), Masson’s trichrome stain, 40X; (<b>B</b>) Inset—epicardium, endocardium and thinned myocardium in the saccular region of the aneurysm, with visible fibrosis and a low number of cardiomyocytes (blue arrow), Masson’s trichrome stain, 100X. (<b>C</b>) Detail of the saccular region of the aneurysm. Fibrosis and dystrophic calcification (yellow arrow); Masson’s trichrome stain, 400X. (<b>D</b>) Multiple capillaries formed in the saccular region of the aneurysm (red arrow) and fibrosis, Masson’s trichrome stain, 100X.</p>
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<p>Desmin, α-sma, vimentin and CD-31 immunostaining in a dog aneurysm. (<b>a</b>) Strong immunosignal for desmin in cardiomyocytes; Hematoxylin background, 400X. (<b>b</b>) Strong immunosignal for α-sma in the smooth muscle fibers in the myocardial capillaries; Hematoxylin background, 400X. (<b>c</b>) Strong vimentin immunosignal in the endothelial cells of myocardial capillaries; Hematoxylin background, 400X. (<b>d</b>) Weak CD-31 immunosignal in cardiomyocytes; Hematoxylin background, 400X.</p>
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<p>MMP-2, MMP-9 and TIMP-1 immunostaining in a dog aneurysm. (<b>a</b>) Strong immunosignal for MMP-2 in cardiomyocytes; Hematoxylin background, 400X. (<b>b</b>) Strong immunosignal for MMP-9 in cardiomyocytes; Hematoxylin background, 400X. (<b>c</b>) Weak immunosignal for TIMP-1 in cardiomyocytes. Strong immunosignal in the Eberth’s lines; Hematoxylin background, 400X. (<b>d</b>) Negative control; Hematoxylin background, 400X.</p>
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22 pages, 1781 KiB  
Article
Preliminary Analysis of Collar Sensors for Guide Dog Training Using Convolutional Long Short-Term Memory, Kernel Principal Component Analysis and Multi-Sensor Data Fusion
by Devon Martin, David L. Roberts and Alper Bozkurt
Animals 2024, 14(23), 3403; https://doi.org/10.3390/ani14233403 - 26 Nov 2024
Viewed by 349
Abstract
Guide dogs play a crucial role in enhancing independence and mobility for people with visual impairment, offering invaluable assistance in navigating daily tasks and environments. However, the extensive training required for these dogs is costly, resulting in a limited availability that does not [...] Read more.
Guide dogs play a crucial role in enhancing independence and mobility for people with visual impairment, offering invaluable assistance in navigating daily tasks and environments. However, the extensive training required for these dogs is costly, resulting in a limited availability that does not meet the high demand for such skilled working animals. Towards optimizing the training process and to better understand the challenges these guide dogs may be experiencing in the field, we have created a multi-sensor smart collar system. In this study, we developed and compared two supervised machine learning methods to analyze the data acquired from these sensors. We found that the Convolutional Long Short-Term Memory (Conv-LSTM) network worked much more efficiently on subsampled data and Kernel Principal Component Analysis (KPCA) on interpolated data. Each attained approximately 40% accuracy on a 10-state system. Not needing training, KPCA is a much faster method, but not as efficient with larger datasets. Among various sensors on the collar system, we observed that the inertial measurement units account for the vast majority of predictability, and that the addition of environmental acoustic sensing data slightly improved performance in most datasets. We also created a lexicon of data patterns using an unsupervised autoencoder. We present several regions of relatively higher density in the latent variable space that correspond to more common patterns and our attempt to visualize these patterns. In this preliminary effort, we found that several test states could be combined into larger superstates to simplify the testing procedures. Additionally, environmental sensor data did not carry much weight, as air conditioning units maintained the testing room at standard conditions. Full article
(This article belongs to the Special Issue The Science of Working and Sporting Dog Performance)
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<p>(<b>a</b>) Smart collar device, (<b>b</b>) smart collar implemented, and (<b>c</b>) dog undergoing IFT with smart collar on. Examples of objects corresponding to labels are shown in (<b>c</b>).</p>
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<p>ConvLSTM architecture strongly based on [<a href="#B29-animals-14-03403" class="html-bibr">29</a>].</p>
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<p>KPCA architecture.</p>
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<p>Manifold against action. Top figures are walking cycles and bottom figures are jogging cycles. Left figures show <span class="html-italic">Y</span>-axis against <span class="html-italic">X</span>-axis and the right is an alternative view showing <span class="html-italic">Z</span>-axis against <span class="html-italic">X</span>-axis [<a href="#B31-animals-14-03403" class="html-bibr">31</a>].</p>
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<p>Effects of orientation on manifolds.</p>
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<p>Autoencoder architecture.</p>
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<p>Relation between LSTM train time and dataset length.</p>
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<p>LSTM training curves for interpolated 10-state IMU-only dataset.</p>
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<p>Examples of (<b>left</b>) the 10-state confusion matrix and (<b>right</b>) a 50-state confusion matrix for Conv-LSTM.</p>
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<p>Relation between KPCA training (<b>a</b>) space vs. dataset length and (<b>b</b>) time vs. dataset length.</p>
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<p>Sankey diagram showing common IFT state confusion. The null state indicates confusion with many other states.</p>
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<p>Autoencoder training of the interpolated 10-state IMU-only dataset.</p>
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<p>Six examples of autoencoder input sequence (black) and corresponding output sequence (red) from the interpolate-50-IMU dataset.</p>
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<p>Generated sequences’ latent space (<b>a</b>) and corresponding sequences (<b>b</b>) indicated by color. Ran on subsample set.</p>
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<p>Generated sequences’ latent space (<b>a</b>) and corresponding sequences (<b>b</b>) indicated by color. Ran on interpolation set.</p>
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<p>(<b>a</b>) Acceleration and (<b>b</b>) position sequences derived from <math display="inline"><semantics> <msup> <mi>ϕ</mi> <mo>*</mo> </msup> </semantics></math> using the decoder from the autoencoder. Color indicates which sequence corresponds to the same sequence in <a href="#animals-14-03403-f015" class="html-fig">Figure 15</a>.</p>
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17 pages, 9364 KiB  
Article
Computed Tomographic Findings in Dogs with Hepatic Bacterial Parenchymal Infection and Abscessation
by Luis Maté de Haro, Andrea Vila, Andrea Di Bella, Claudia Mallol, Carlo Anselmi, Jose-Daniel Barreiro-Vazquez, Danica Pollard, Raquel Salgüero, Ella Fitzgerald and Beatriz Moreno-Aguado
Animals 2024, 14(23), 3399; https://doi.org/10.3390/ani14233399 - 25 Nov 2024
Viewed by 310
Abstract
Bacterial liver parenchymal infections in dogs are rarely documented, and their imaging characteristics are scarce in the veterinary literature, especially in Computed Tomography (CT). This retrospective multicentric study aimed to describe the CT characteristics of parenchymal bacterial liver infection and abscessation in dogs [...] Read more.
Bacterial liver parenchymal infections in dogs are rarely documented, and their imaging characteristics are scarce in the veterinary literature, especially in Computed Tomography (CT). This retrospective multicentric study aimed to describe the CT characteristics of parenchymal bacterial liver infection and abscessation in dogs and compare them with the human literature. Twenty dogs met the inclusion criteria. All dogs, except one, showed discrete hepatic lesions consistent with pyogenic liver abscess (19/20). A single case showed diffuse liver changes, which was diagnosed with granulomatous bacterial hepatitis (1/20). Multifocal lesions were associated with the presence of abdominal pain (p = 0.023). CT characteristics of pyogenic liver abscesses in our study resemble those described in the human literature, with multifocal (14/19) or single (5/19), round or ovoid (19/19), hypoattenuating hepatic lesions, which are better visualised in post-contrast images. Pyogenic liver abscesses can also show features such as the “cluster sign” (8/19), transient arterial segmental enhancement (6/10), rim enhancement (6/19), and intralesional gas (4/19). Additional CT findings, such as local lymphadenomegaly (18/20), peritoneal fat stranding (14/20), and peritoneal fluid (13/20), are also commonly observed. Full article
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<p>Classification of the liver changes.</p>
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<p>Transverse unenhanced (<b>A</b>) and venous phase (<b>B</b>) images at the cranial abdomen (window level 50 HU, window width 350 HU). Diffuse liver changes were present, characterised by irregular margination of the liver lobes and homogeneous contrast enhancement. Peritoneal effusion was present (*).</p>
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<p>Transverse unenhanced (<b>A</b>) and venous phase (<b>B</b>) images of a hypoattenuating lesion that deforms the liver contour (white arrows) in the left lateral liver lobe (window level 50 HU, widow width 350 HU). Note the presence of adjacent focal peritoneal fat stranding (*).</p>
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<p>Transverse unenhanced (<b>A</b>), arterial phase (<b>B</b>), and venous phase (<b>C</b>) images of a large lesion showing the “cluster sign” (white arrows) and septal enhancement in the left lateral liver lobe (window level 50 HU, widow width 350 HU). Note the presence of adjacent focal peritoneal fat stranding (*).</p>
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<p>Transverse unenhanced (<b>A</b>), arterial phase (<b>B</b>), and venous phase (<b>C</b>) close-up images at the level of the right medial liver lobe (window level 50 HU, widow width 350 HU). A cluster sign, rim enhancement in the arterial phase, and intralesional gas were noted (black arrows).</p>
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<p>Transverse unenhanced (<b>A</b>) and portal phase (<b>B</b>) images at the left lateral liver lobe (window level 50 HU, widow width 350 HU). A hypoattenuating lesion with intralesional gas was present (thick white arrows). Note also the presence of pneumoperitoneum (black arrows).</p>
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<p>Transverse unenhanced (<b>A</b>), arterial phase (<b>B</b>), and venous phase (<b>C</b>) images at the right medial liver lobe (window level 50 HU, widow width 350 HU). Transient segmental arterial enhancement is present (black arrows).</p>
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11 pages, 1801 KiB  
Article
Correlation of Red Blood Cell Morphology with Serum Cobalamin and Folate Concentration in Dogs with Chronic Diarrhea: A Retrospective Study
by Argyrios Ginoudis, Anna Maria Ioannidou, Dimitra Pardali, Asteria Tsikna and Zoe Polizopoulou
Metabolites 2024, 14(12), 657; https://doi.org/10.3390/metabo14120657 - 25 Nov 2024
Viewed by 321
Abstract
Background/Objectives: Chronic diarrhea in dogs is a prevalent condition that significantly impacts canine health, often leading to weight loss, dehydration, and malnutrition. Diagnosing and treating chronic diarrhea is challenging due to its multifactorial nature, necessitating collaboration among veterinarians across various specialties. Measuring [...] Read more.
Background/Objectives: Chronic diarrhea in dogs is a prevalent condition that significantly impacts canine health, often leading to weight loss, dehydration, and malnutrition. Diagnosing and treating chronic diarrhea is challenging due to its multifactorial nature, necessitating collaboration among veterinarians across various specialties. Measuring cobalamin and folate levels is a crucial diagnostic step for all dogs with chronic diarrhea. The role of these vitamins in erythropoiesis is well-documented in human medicine, where deficiencies are linked to erythropoietic disorders and megaloblastic anemia. This study explores the relationship between cobalamin and folate concentrations with hematologic parameters in dogs with chronic diarrhea to develop novel diagnostic methods that facilitate timely decision making. Methods: Forty-seven adult dogs with a history of chronic diarrhea (2019–2023) were included in the study. Upon presentation, complete blood count and measurement of cobalamin and folate concentrations were performed. The correlation of cobalamin and folate levels with erythrocytic parameters, including hematocrit (HCT), hemoglobin concentration (HGB), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), red cell distribution width (RDW), and reticulocyte count, as well as morphological changes in the blood smear were examined. Results: Serum cobalamin was significantly correlated with RBC (p = 0.032), HGB (p = 0.006), HCT (p = 0.005), and MCV (p = 0.022). Anisocytosis was significantly correlated with hypocobalaminemia (p = 0.002), while acanthocytosis correlated with normal cobalamin levels (p = 0.046). No correlation was found between serum folate and erythrocytic parameters or morphological changes. Conclusions: These findings emphasize cobalamin’s potential role in canine erythropoiesis, highlighting the need for routine evaluation and supplementation when necessary. Conversely, the lack of association with folate suggests it plays a less significant role in this species. These results underscore the importance of complete blood count in the diagnostic investigation of dogs with chronic diarrhea. Full article
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<p>Scattergram illustrating the relationship between serum cobalamin concentration (B12, pmol/L) and red blood cell count (RBC, ×10<sup>3</sup>/µL). Each point represents an individual measurement, where the x-axis corresponds to the B12 concentration and the y-axis corresponds to the RBC concentration. The dotted line represents the trend line (linear regression) indicating the correlation between B12 and RBC.</p>
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<p>Scattergram illustrating the relationship between serum cobalamin concentration (B12, pmol/L) and hemoglobin concentration (HGB, g/dL). Each point represents an individual measurement, where the x-axis corresponds to the B12 concentration and the y-axis corresponds to the HGB concentration. The dotted line represents the trend line (linear regression) indicating the correlation between B12 and HGB.</p>
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<p>Scattergram illustrating the relationship between serum cobalamin concentration (B12, pmol/L) and hematocrit (HCT, %). Each point represents an individual measurement, where the x-axis corresponds to the B12 concentration and the y-axis corresponds to the HCT. The dotted line represents the trend line (linear regression) indicating the correlation between B12 and HCT.</p>
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<p>Scattergram illustrating the relationship between serum cobalamin concentration (B12, pmol/L) and mean corpuscular volume (MCV, fL). Each point represents an individual measurement, where the x-axis corresponds to the B12 concentration and the y-axis corresponds to the MCV. The dotted line represents the trend line (linear regression) indicating the correlation between B12 and MCV.</p>
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16 pages, 1188 KiB  
Article
Evaluation of Serum YKL-40 in Canine Multicentric Lymphoma: Clinical and Diagnostic Implications
by Chien-Chun Kuo, Jih-Jong Lee, Shang-Lin Wang, Yuan-Yuan Xia and Albert Taiching Liao
Animals 2024, 14(23), 3391; https://doi.org/10.3390/ani14233391 - 25 Nov 2024
Viewed by 243
Abstract
YKL-40, a secretory glycoprotein, is known as a prognostic biomarker in human cancers, but its role in canine multicentric lymphoma is not well understood. This study aimed to investigate serum YKL-40 levels in thirty dogs with multicentric lymphoma to determine their prognostic value, [...] Read more.
YKL-40, a secretory glycoprotein, is known as a prognostic biomarker in human cancers, but its role in canine multicentric lymphoma is not well understood. This study aimed to investigate serum YKL-40 levels in thirty dogs with multicentric lymphoma to determine their prognostic value, association with patient characteristics, and potential to predict chemotherapy response. Serum samples were collected before, during, and after chemotherapy, and YKL-40 level was measured using ELISA. The results showed that the pretreatment serum YKL-40 levels were significantly higher in dogs with multicentric lymphoma (394.0 pg/mL, n = 30) than in healthy controls (218.6 pg/mL, n = 11) (p = 0.012). While a cutoff value of 445.1 pg/mL was observed, further studies are needed to clarify its diagnostic utility. Dogs with clinical stage V had the highest YKL-40 levels (p = 0.027), potentially reflecting disease severity. Furthermore, YKL-40 levels decreased after chemotherapy (p = 0.030). However, YKL-40 levels showed no significant association with progression-free survival (PFS) (HR = 0.93, p = 0.830) or overall survival (OS) (HR = 0.99, p = 0.267). In conclusion, serum YKL-40 levels may potentially detect the disease severity, but its prognostic role remains uncertain. Further studies are required to evaluate serum YKL-40 levels as a potential indicator of treatment response or disease recurrence. Full article
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<p>Comparison of serum YKL-40 levels between dogs with multicentric lymphoma and healthy controls. The scatter plot presents individual data points for each group, with the median represented by a central horizontal line and IQR shown around the median. The serum YKL-40 levels significantly differed between the dogs with multicentric lymphoma (<span class="html-italic">n =</span> 30) and the healthy control (<span class="html-italic">n =</span> 11). * <span class="html-italic">p =</span> 0.012.</p>
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<p>ROC curve of serum YKL-40 levels for differentiating dogs with multicentric lymphoma from healthy dogs. The ROC curve demonstrates the diagnostic accuracy of serum YKL-40 levels in distinguishing between dogs with multicentric lymphoma (<span class="html-italic">n =</span> 30) and healthy dogs (<span class="html-italic">n =</span> 11). The area under the curve (AUC) is 0.766. A cut-off value for YKL-40 was determined to be 445.1 pg/mL, yielding a sensitivity of 43.3% and a specificity of 100%.</p>
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<p>Scatter plot with a bar chart showing pretreatment serum YKL-40 levels in dogs with different WHO clinical stages. Each data point represents an individual dog, with dots, squares, triangles, and inverted triangles representing dogs at clinical stages II, III, IV, and V, respectively. The median values for each stage are shown at the top of the bars, and IQR is represented as horizontal lines around the median. Significant differences are found between the dogs with stage IV and stage V. ** <span class="html-italic">p =</span> 0.035.</p>
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<p>Box and whiskers plot showing comparisons of serum YKL-40 levels across different treatment time points. The plot presents the distribution of serum YKL-40 levels at each time point, with boxes indicating the IQR around the median, and whiskers extending to minimum and maximum values. Significant differences were observed between pretreatment and post-treatment follow-up. * <span class="html-italic">p =</span> 0.0312.</p>
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