<?xml version="1.0" encoding="UTF-8"?><!DOCTYPE article  PUBLIC "-//NLM//DTD Journal Publishing DTD v3.0 20080202//EN" "http://dtd.nlm.nih.gov/publishing/3.0/journalpublishing3.dtd"><article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" dtd-version="3.0" xml:lang="en" article-type="research article"><front><journal-meta><journal-id journal-id-type="publisher-id">OJVM</journal-id><journal-title-group><journal-title>Open Journal of Veterinary Medicine</journal-title></journal-title-group><issn pub-type="epub">2165-3356</issn><publisher><publisher-name>Scientific Research Publishing</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="doi">10.4236/ojvm.2023.136007</article-id><article-id pub-id-type="publisher-id">OJVM-125772</article-id><article-categories><subj-group subj-group-type="heading"><subject>Articles</subject></subj-group><subj-group subj-group-type="Discipline-v2"><subject>Medicine&amp;Healthcare</subject></subj-group></article-categories><title-group><article-title>
 
 
  Supplementation Effects of an Alpha-Casozepine and White Fish Muscle Hydrolyzed Complementary Feed on Canine Separation-Related Disorders and Quality of Life of Dogs and Their Pet Caregivers
 
</article-title></title-group><contrib-group><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Gaëlle</surname><given-names>Porcheron</given-names></name><xref ref-type="aff" rid="aff1"><sup>1</sup></xref></contrib><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Marie</surname><given-names>Bodet</given-names></name><xref ref-type="aff" rid="aff2"><sup>2</sup></xref></contrib><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Katell</surname><given-names>Poiron</given-names></name><xref ref-type="aff" rid="aff3"><sup>3</sup></xref></contrib><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Julie</surname><given-names>Brault</given-names></name><xref ref-type="aff" rid="aff4"><sup>4</sup></xref></contrib><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Nicolas</surname><given-names>Massal</given-names></name><xref ref-type="aff" rid="aff5"><sup>5</sup></xref></contrib><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Olivier</surname><given-names>Roy</given-names></name><xref ref-type="aff" rid="aff1"><sup>1</sup></xref></contrib><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Oscar</surname><given-names>Fantini</given-names></name><xref ref-type="aff" rid="aff6"><sup>6</sup></xref><xref ref-type="corresp" rid="cor1"><sup>*</sup></xref></contrib></contrib-group><aff id="aff3"><addr-line>Clinique Vétérinaire LESVET, Lesneven France</addr-line></aff><aff id="aff2"><addr-line>Clinique Vétérinaire de la Victoire, Tours, France</addr-line></aff><aff id="aff5"><addr-line>Clinique Vétérinaire de la Paix, Pau, France</addr-line></aff><aff id="aff1"><addr-line>Cebiphar, Fondettes, France</addr-line></aff><aff id="aff4"><addr-line>Clinique Vétérinaire du Gabereau, Chateuneuf-sur-Loire, France</addr-line></aff><aff id="aff6"><addr-line>Vetoquinol SA, Paris, France</addr-line></aff><pub-date pub-type="epub"><day>13</day><month>06</month><year>2023</year></pub-date><volume>13</volume><issue>06</issue><fpage>68</fpage><lpage>81</lpage><history><date date-type="received"><day>2,</day>	<month>May</month>	<year>2023</year></date><date date-type="rev-recd"><day>22,</day>	<month>June</month>	<year>2023</year>	</date><date date-type="accepted"><day>25,</day>	<month>June</month>	<year>2023</year></date></history><permissions><copyright-statement>&#169; Copyright  2014 by authors and Scientific Research Publishing Inc. </copyright-statement><copyright-year>2014</copyright-year><license><license-p>This work is licensed under the Creative Commons Attribution International License (CC BY). http://creativecommons.org/licenses/by/4.0/</license-p></license></permissions><abstract><p>
 
 
  Background: Separation-related disorders are prevalent in dogs and can result in problem behaviours such as inappropriate elimination, destructive behaviour, and distress vocalizations which occur when a caregiver is absent or perceived as absent. Numerous natural products are marketed for managing these behavioural issues in dogs; however, few, if any, of these products have documented evidence of their positive effects in veterinary behaviour. 
  Purpose: This open, multicentric clinical trial aimed to assess the effect of a new supplement containing alpha-casozepine and fish muscle hydrolysate on the separation-related problem in dogs and the impact on the quality of life of dogs and their caregivers. 
  Material and Methods: Fifty-one dogs with clinical signs compatible with separation-related problems were included. The dogs were supplemented with a complementary feed (Zylkene Plus, Vetoquinol, France) for 30 days. Separation-related behaviour assessment (SRP), emotional disorders assessment (EDED) and the quality of life (QoL) of the pets and their pet caregivers were assessed at baseline and at the end of the study by the veterinarians. In addition, caregivers were asked to rate their dog’s SRP weekly after enrolment using the same questionnaire that the veterinarian used at the beginning of the study. 
  Results: There was a statistically significant improvement in the SRP, EDED, and QoL scores (P &lt; 0.0001). The mean global SRP score decreased by 32% between baseline and D7, 41% between DO and D14, 47% between D0 and D D21, and 49% between D0 and D30. The percentage of dogs in a normal state (EDED &lt; 12) increased during the study from 26% in D0 to 62% at D30, and the percentage of dogs with scores related to anxieties decreased from 40% at D0 to 17% at D30. On day 30, there were significant improvements in QOL scores compared to the baseline. 
  Conclusion: The results of this study support the hypothesis that the combination of alpha-casozepine and fish muscle hydrolysate is a promising and effective option to reduce the frequency and severity of separation-related behaviour in dogs and improve the quality of dogs and their caregivers.
 
</p></abstract><kwd-group><kwd>Alpha-Casozepine</kwd><kwd> Fish Muscle Hydrolysate</kwd><kwd> Canine Separation-Related Disorders</kwd><kwd> Dogs</kwd></kwd-group></article-meta></front><body><sec id="s1"><title>1. Introduction</title><p>Separation-related behaviour problems have been estimated at 20% of the dog population [<xref ref-type="bibr" rid="scirp.125772-ref1">1</xref>] .</p><p>In contrast, dogs with separation-related behaviour problems engage in undesirable behaviours when left alone. The most common are destruction and excessive vocalization, including whining and barking. Less common problem behaviours include inappropriate elimination (urination and defecation), self-injurious behaviour (e.g. over-grooming), increased or repetitive motor activity (e.g. pacing), attempts to escape, trembling, salivation, and depression [<xref ref-type="bibr" rid="scirp.125772-ref2">2</xref>] .</p><p>Dogs that engage in excessive vocalization when left alone are likely to prompt complaints by neighbours and visits from local authorities. Sherman [<xref ref-type="bibr" rid="scirp.125772-ref3">3</xref>] notes that living with a dog with separation-related problems causes emotional and financial distress at home, affecting the quality of life (QoL), which can lead to the dog’s surrender. Indeed, separation-related problem behaviours are commonly cited reasons for relinquishing dogs to animal shelters.</p><p>The management of separation-related problems consists of environmental control, behaviour modification, and medications. However, environmental control and behaviour modification require commitment and regularity in the household routine and often a deliberate weakening of the caregiver-pet bond. In addition, behaviour modifications can sometimes be challenging for the caregiver who has difficulty reducing contact with the dog or in homes with multiple occupants where it is difficult to enforce changes in routine [<xref ref-type="bibr" rid="scirp.125772-ref4">4</xref>] .</p><p>When dogs do not respond to behavioural therapy, pharmacological agents can be prescribed to manage dog behavioural symptoms, such as fluoxetine, benzodiazepine, selegiline amitryptiline, and clomipramine [<xref ref-type="bibr" rid="scirp.125772-ref5">5</xref>] . In addition, due to their cerebral suppressant properties, such treatments can have sedative effects [<xref ref-type="bibr" rid="scirp.125772-ref5">5</xref>] [<xref ref-type="bibr" rid="scirp.125772-ref6">6</xref>] .</p><p>Across veterinary disciplines, there is interest in the use of alternative treatment modalities, including acupuncture [<xref ref-type="bibr" rid="scirp.125772-ref7">7</xref>] [<xref ref-type="bibr" rid="scirp.125772-ref8">8</xref>] , homoeopathy [<xref ref-type="bibr" rid="scirp.125772-ref8">8</xref>] [<xref ref-type="bibr" rid="scirp.125772-ref9">9</xref>] [<xref ref-type="bibr" rid="scirp.125772-ref10">10</xref>] and nutraceuticals [<xref ref-type="bibr" rid="scirp.125772-ref11">11</xref>] for the treatment of common medical and behavioural disorders [<xref ref-type="bibr" rid="scirp.125772-ref11">11</xref>] [<xref ref-type="bibr" rid="scirp.125772-ref12">12</xref>] [<xref ref-type="bibr" rid="scirp.125772-ref13">13</xref>] [<xref ref-type="bibr" rid="scirp.125772-ref14">14</xref>] . Caregivers often perceive these options are safer with fewer side effects.</p><p>Numerous natural products are marketed for managing behavioral issues in dogs; however, few, if any, of these products have documented evidence of efficacy in veterinary behaviour.</p><p>The present study evaluated a new supplement containing alpha-casozepine and fish muscle hydrolysate (Zylkene Plus, Vetoquinol, France) for the management of separation-related problems in dogs. This multicentric clinical trial aimed to assess the effect on signs associated with separation-related problems in dogs and the impact on the quality of life of dogs and their caregivers.</p></sec><sec id="s2"><title>2. Materials and Methods</title><sec id="s2_1"><title>2.1. Design</title><p>This study was designed as an open-label, multicenter, prospective trial.</p><p>A total of 11 veterinary clinics, including general and behaviourist practices, participated.</p></sec><sec id="s2_2"><title>2.2. Study Population</title><p>The dogs were eligible for the study if they met the inclusion and exclusion criteria.</p><p>Inclusion criteria: Occurrence for at least one month of one or more of the following manifestations of separation-related problems: inappropriate urinating and defecating, barking and howling, chewing, digging and destruction, pacing, exaggerated greetings, rearranging the caregiver’s things.</p><p>Exclusion criteria were dogs with severe chronic disease or receiving supplement products, pheromones, or any other psychotropic medication for the behaviour condition in the previous 10 days.</p><p>All dogs had to be in good health to be included in the study, based on history and complete physical examination. Dogs were included in the study regardless of breed, sex, origin (conditions of adoption) or lifestyle.</p><p>Caregivers were eligible for the study if they were able to provide a stable home environment for the duration of the study (no major changes in household routine, vacation longer than 3 days without the dog, extended guest stays, or change in family members at home (including new pets)). Caregivers also had to commit to complying with the supplement administration.</p></sec><sec id="s2_3"><title>2.3. Supplementation and Administration</title><p>The caregiver administered the supplementation orally daily for 30 &#177; 2 days, 30 minutes to 1 hour before separation. The capsule could be opened, and the powder mixed with food or given as a treat according to <xref ref-type="table" rid="table1">Table 1</xref>.</p><table-wrap id="table1" ><label><xref ref-type="table" rid="table1">Table 1</xref></label><caption><title> Dosage of Zylkene Plus</title></caption><table><tbody><thead><tr><th align="center" valign="middle" >Animal weight</th><th align="center" valign="middle" >Zylkene Plus 75 mg</th><th align="center" valign="middle" >Zylkene Plus 225 mg</th><th align="center" valign="middle" >Zylkene Plus 450 mg</th></tr></thead><tr><td align="center" valign="middle" >[2 - 10] kg</td><td align="center" valign="middle" >2 capsules</td><td align="center" valign="middle" >-</td><td align="center" valign="middle" >-</td></tr><tr><td align="center" valign="middle" >[10 - 15] kg</td><td align="center" valign="middle" >-</td><td align="center" valign="middle" >1 capsule</td><td align="center" valign="middle" >-</td></tr><tr><td align="center" valign="middle" >[15 - 30] kg</td><td align="center" valign="middle" >-</td><td align="center" valign="middle" >-</td><td align="center" valign="middle" >1 capsule</td></tr><tr><td align="center" valign="middle" >[30 - 60] kg</td><td align="center" valign="middle" >-</td><td align="center" valign="middle" >-</td><td align="center" valign="middle" >2 capsules</td></tr></tbody></table></table-wrap><p>Specific behavioural therapy instructions were not given to keep a reproducible environment among the subjects.</p></sec></sec><sec id="s3"><title>3. Outcome Measures</title><p>The dogs were examined as described in the following summary of the protocol (<xref ref-type="fig" rid="fig1">Figure 1</xref>).</p><p>At baseline and D30 &#177; 2, veterinarians performed a clinical examination, an evaluation of dog behaviour related to separation (SRP) (<xref ref-type="table" rid="table2">Table 2</xref>), an evaluation of the dog’s emotional disorders (EDED) (<xref ref-type="table" rid="table3">Table 3</xref>) and an evaluation of the quality of life (QoL) of dogs and their pet caregivers (<xref ref-type="table" rid="table4">Table 4</xref>).</p><p>Weekly after enrolment, caregivers were asked to rate their dog’s SRP using the same questionnaire the veterinarian used at the beginning of the study (<xref ref-type="table" rid="table2">Table 2</xref>). In addition, the caregivers were asked to rate their dog’s behaviour (destructive behaviour, rearranging, excessive vocalization, inappropriate urination, inappropriate defecation, escaping, shadowing, excessive greeting, excessive salivation and pacing) on a 4-point continuous scale from 0 = never to 3 = always, resulting in global SRP ranging from 0 to 42.</p><p>The emotional disorders evaluation (EDED) (Appendix 1) in dogs allows the practitioner to assay basic behaviors of the dog (i.e. eating, drinking, grooming, play, aggression, exploratory behavior) as well organic signs. The scale produces a numeric result of 9 to 45 points, with normal dogs scoring between 9 and 12. Dogs with anxiety-related disorders score between 18 and 30 [<xref ref-type="bibr" rid="scirp.125772-ref15">15</xref>] .</p></sec><sec id="s4"><title>4. Statistics</title><p>Statistical analysis was performed using validated Stata Statistical Software: Release 16; College Station, TX: StataCorp LP.</p><p>For continuous variables, the following descriptive statistics were performed: mean, standard deviation, sample size, minimum, and maximum. For qualitative or binary variables, absolute and relative frequencies were presented.</p><p>Scores at D30 &#177; 2 were compared with the initial score (D0) using the Wilcoxon matched-pairs signed ranks test. Statistical significance was stated if P &lt; 0.05.</p></sec><sec id="s5"><title>5. Results</title><sec id="s5_1"><title>5.1. Participants</title><p>A total of 51 dogs were recruited for the study. Among them, 4 dogs were not included in the analysis (supplement discontinuation, incomplete questionnaire, wrong posology). The dogs ranged in age from 6 months to 13.4 years of age and in weight from 2.5 to 49 kg. Demographic data are shown in <xref ref-type="table" rid="table4">Table 4</xref>.</p><table-wrap id="table2" ><label><xref ref-type="table" rid="table2">Table 2</xref></label><caption><title> Separation-related problem questionnaire. Each question is rated on a 4-point continuous scale from 0 = never to 3 = always, resulting in a global SRP ranging from 0 to 42</title></caption><table><tbody><thead><tr><th align="center" valign="middle" >Separation-related behaviour questionnaires</th></tr></thead><tr><td align="center" valign="middle" >Does your dog insist on following you around when you’re home?</td></tr><tr><td align="center" valign="middle" >Does your dog become anxious at the sound of car keys or when you put on your coat or shoes to go out?</td></tr><tr><td align="center" valign="middle" >Does your dog bark or whine excessively within 30 minutes of your departure?</td></tr><tr><td align="center" valign="middle" >After you leave, does your dog act depressed?</td></tr><tr><td align="center" valign="middle" >After you leave, does your dog have a loss of appetite or ignore treats left out for him?</td></tr><tr><td align="center" valign="middle" >Does your dog destroy property only when you are away (chewing, breaking, tearing, scratching, biting at windows, doors and exits, etc.)?</td></tr><tr><td align="center" valign="middle" >Does your dog urinate at home only when you are away?</td></tr><tr><td align="center" valign="middle" >Does your dog defecate at your home only when you are away?</td></tr><tr><td align="center" valign="middle" >Does your dog attempt to escape from his kennel/pen/room when you are away?</td></tr><tr><td align="center" valign="middle" >Does your dog show rearranging behaviour (i.e. moving household objects around without destruction)?</td></tr><tr><td align="center" valign="middle" >Does your dog have excessive salivation in your absence?</td></tr><tr><td align="center" valign="middle" >Does your dog is pacing (constantly walks in a circle or back and forth) in your absence?</td></tr><tr><td align="center" valign="middle" >Does your dog regularly have diarrhoea, vomit, or lick excessively in your absence?</td></tr><tr><td align="center" valign="middle" >Does your dog exhibit an excessive greeting on your return (jumping, hyperactivity, barking, more than 2 - 3 minutes)?</td></tr></tbody></table></table-wrap><table-wrap id="table3" ><label><xref ref-type="table" rid="table3">Table 3</xref></label><caption><title> Quality of life questionnaire. Each question was scored on a 4-point continuous scale from 0 = not at all to 3 = very much</title></caption><table><tbody><thead><tr><th align="center" valign="middle" >Quality of life questionnaire</th></tr></thead><tr><td align="center" valign="middle" >How severe and disturbing is your dog’s anxiety?</td></tr><tr><td align="center" valign="middle" >How much are your dog’s playing or working activities disturbed by his anxiety?</td></tr><tr><td align="center" valign="middle" >Does your dog’s anxiety have an impact on its relationship with you, other family members or other dogs?</td></tr><tr><td align="center" valign="middle" >How much has your dog’s disease changed its usual habits (change in place where he is allowed to sleep, live, eat, way it which it is walked, etc.)?</td></tr><tr><td align="center" valign="middle" >How much time do you lose for your dog’s anxiety (administration of therapies, home cleaning, veterinary consultations, etc.)?</td></tr><tr><td align="center" valign="middle" >How much effect has your dog’s anxiety on your tiredness?</td></tr><tr><td align="center" valign="middle" >How much are your usual activities and/or those of your family disturbed by your dog’s anxiety (leisure, vacation, walks, work, etc.)?</td></tr><tr><td align="center" valign="middle" >How much impact does your dog’s anxiety have on your expenditure (cost of treatment, veterinarian, etc.)?</td></tr><tr><td align="center" valign="middle" >How much effect does your dog’s anxiety have on causing emotional distress (feeling of guilt, powerlessness, sorrow, regret, anxiety, nuisance, disgust, anger, frustration, etc.)?</td></tr><tr><td align="center" valign="middle" >How much physical uneasiness/discomfort is your experience due to your dog’s anxiety (offending odour, feeling of dirtiness at home, aesthetic nuisance, etc.)?</td></tr><tr><td align="center" valign="middle" >Does your dog’s anxiety have a negative impact on the relationship between family members (between spouses, between parents and sons, with relatives and friends, etc.)?</td></tr></tbody></table></table-wrap><table-wrap id="table4" ><label><xref ref-type="table" rid="table4">Table 4</xref></label><caption><title> Demographic data of included dogs</title></caption><table><tbody><thead><tr><th align="center" valign="middle" >Parameter</th><th align="center" valign="middle" >Statistics</th><th align="center" valign="middle" >Total</th></tr></thead><tr><td align="center" valign="middle" >Number</td><td align="center" valign="middle" >N</td><td align="center" valign="middle" >47</td></tr><tr><td align="center" valign="middle" >Breed</td><td align="center" valign="middle" >N</td><td align="center" valign="middle" >Boxer (2), Shih Tzu (2), Yorkshire Terrier (3), Beagle (1), Beauceron (1), Bichon Frise (1), Bolognese (1), Cane Corso (1), Cavalier King Charles Spaniel (1), Chihuahua (1), Cocker Spaniel (3), Coton du Tulear (1), Dachshund (1), German Shepherd (1), English Bulldog (1), French Bulldog (3), German Pinscher (1), Golden Retriever (1), Griffon (1), Jack Russell Terrier (2), Miniature American Shepherd (1), Miniature Pinscher (1), Parson Russell Terrier (1), Poodle (1), Rottweiler (1), Small Munsterlander (1), Springer Spaniel (1), White Swiss Shepherd (1), Whippet (1), Wire Fox Terrier (1), mixed breed (9)</td></tr><tr><td align="center" valign="middle" >Sex</td><td align="center" valign="middle" >N</td><td align="center" valign="middle" >21 females including 15 neutered 26 males including 14 neutered</td></tr><tr><td align="center" valign="middle" >Other pets in house</td><td align="center" valign="middle" >N</td><td align="center" valign="middle" >none (21) 1 pet (15) 2 pets (9) 3 pets (1) 4 pets (1)</td></tr><tr><td align="center" valign="middle" >Lifestyle</td><td align="center" valign="middle" >N</td><td align="center" valign="middle" >indoor and outdoor (38) mostly indoor (9)</td></tr><tr><td align="center" valign="middle"  rowspan="2"  >BW (kg)</td><td align="center" valign="middle" >mean &#177; sd</td><td align="center" valign="middle" >15.1 &#177; 10.9</td></tr><tr><td align="center" valign="middle" >[min. - max.]</td><td align="center" valign="middle" >2.5 - 49.0</td></tr><tr><td align="center" valign="middle"  rowspan="2"  >Age (years)</td><td align="center" valign="middle" >mean &#177; sd</td><td align="center" valign="middle" >4.5 &#177; 3.</td></tr><tr><td align="center" valign="middle" >[min. - max.]</td><td align="center" valign="middle" >0.6 - 13.4</td></tr></tbody></table></table-wrap></sec><sec id="s5_2"><title>5.2. Outcome Measures</title><sec id="s5_2_1"><title>5.2.1. Separation-Related Problem Behaviour Assessment</title><p>To evaluate the improvement, the overall score for each behaviour was evaluated. From Day 7 to the study end, there was a statistically significant improvement in the severity of behavioral symptoms. The mean global SRP score decreased by 32% between D0 and D7, 41% between D0 and D14, 47% between D0 and D21, and 49% between D0 and D30, with a mean total score of 16.7 &#177; 6.0 at D0 and 8.6 &#177; 5.9 at the end of follow-up (<xref ref-type="fig" rid="fig2">Figure 2</xref>).</p></sec><sec id="s5_2_2"><title>5.2.2. Emotional Disorder Evaluation</title><p>At the end of the study, there was a statistically significant improvement on the EDED scale, as shown in <xref ref-type="fig" rid="fig3">Figure 3</xref>. Thirty-four cases (72%) had a lower EDED value at D30 compared to D0, 8 (17%) presented the same value at both visits, and 5 cases (11%) presented a higher EDED value at D30 compared to D0. In addition, the percentage of dogs in a normal state (EDED &lt; 12) increased during the study from 26% at D0 to 62% at D30. The percentage of dogs with scores related to anxieties thus decreased from 40% at D0 to 17% at D30.</p></sec><sec id="s5_2_3"><title>5.2.3. Quality of Life</title><p>The mean QoL score before and after supplementation was 13.34 &#177; 6.4 and 9.5 &#177; 6.7). There was a significant improvement in total QoL scores after the supplement intervention compared to Day 0 (P &lt; 0.0001), as shown in <xref ref-type="fig" rid="fig4">Figure 4</xref>. The QoL was improved by at least 30% for 74% of the dogs.</p><p>When comparing before and after supplement, all questions showed a decrease in scores except questions 8 and 11 (How much impact does your dog’s anxiety have on your expenditure? Does your dog’s anxiety negatively impact the relationship between family members?).</p></sec></sec></sec><sec id="s6"><title>6. Discussion</title><p>This open, non-controlled study aimed to evaluate the effect of a complementary feed containing alpha-casozepine and fish muscle hydrolysate in reducing the frequency and severity of separation-related behaviour signs in dogs and the impact on their quality of life. The effect was evaluated based on three different assessments: 1) separation-related problem behaviour assessment (SRP); 2) emotional disorders scale (EDED); and 3) the quality of life of pets and their pet caregivers (QoL).</p><p>The results of this study support the hypothesis that the combination of alpha-casozepine and whitefish muscle hydrolysate can be an effective method for managing separation-related behavioural signs in dogs. Furthermore, the results show that it can also improve the quality of life of dogs, their caregivers, and their emotional state.</p><p>Analysis of the SRP assessment revealed a significant decrease in the overall score for each behaviour at different times. A statistically significant improvement in the frequency and severity of behavioral symptoms has been observed as soon as D7. The mean global SRP score decreased by 49% at D30 compared to the baseline. Comparing the baseline and final assessments, the percentage improvement for excessive barking, destructiveness, inappropriate urination, escaping and pacing were 48%, 62%, 72% and 60%, respectively.</p><p>Quality of life (QoL) is a term used to evaluate general well-being, and it is defined as “the degree to which an individual enjoys his life”. In medicine, QoL is often assessed in terms of how disease negatively affects it [<xref ref-type="bibr" rid="scirp.125772-ref16">16</xref>] . As in previous studies, the evaluation of QoL can be interesting when studying options for disease management [<xref ref-type="bibr" rid="scirp.125772-ref17">17</xref>] [<xref ref-type="bibr" rid="scirp.125772-ref18">18</xref>] . Although patient care remains the first goal, considering the caregiver feelings can be relevant, as managing many behaviours in pets is time-consuming. When prescribing therapy, the veterinarian must not only take into account the effectiveness of the drugs prescribed but also the compliance [<xref ref-type="bibr" rid="scirp.125772-ref19">19</xref>] .</p><p>Thus, effective treatment of anxiety disorders is not only a necessity for the health and welfare of the pet but also to improve the problem for the caregiver and ultimately restore the bond between caregiver and pet.</p><p>This present study confirmed the hypothesis that supplementation with a complementary feed containing alpha-casozepine and fish muscle hydrolysates significantly improved the QoL of dogs and their caregivers. Furthermore, one month after the beginning of supplementation, there was a significant improvement in total QoL scores compared to baseline.</p><p>At the end of the study, there was a statistically significant improvement in the EDED scale. The percentage of dogs in a normal state (EDED &lt; 12) increased during the study from 26% at D0 to 62% at D30. The percentage of dogs with scores related to anxiety thus decreased from 40% at D0 to 17% at D30.</p><p>Environmental and behavioural modification and pharmacological therapy are often indicated to deal with the problem and address the pet’s well-being [<xref ref-type="bibr" rid="scirp.125772-ref20">20</xref>] . For example, behavioural advice for separation-related problem behaviours can include reducing the dependency of the dog on the caregiver, removing punishment for separation-related behaviours, providing activities for the dog to engage in when left alone, and systematic desensitization [<xref ref-type="bibr" rid="scirp.125772-ref20">20</xref>] . However, pet caregivers received no behavioural advice in this study to avoid confounding factors in interpreting the results.</p><p>The study has some limitations that need to be considered. First, the study was an open, non-controlled study, meaning there was no control group to compare the results with and increase the validity of the results. The lack of a placebo control means that caution is required in interpreting these results, but it does not invalidate them. Second, the sample size was relatively small, which could affect the generalizability of the results. Additionally, the study did not include video recordings of the dog’s behaviour after the caregiver left. Video recordings can provide valuable insights into the dog’s behaviour when left alone and help monitor and assess the frequency and severity of separation-related problems.</p></sec><sec id="s7"><title>7. Conclusion</title><p>Despite the limitations mentioned above, the study’s findings suggest that Zylkene Plus may be a promising supplementation option for dogs with separation-related problems. The decrease in the frequency and severity of symptoms indicates that the product may effectively manage this behavioural problem. Furthermore, the potential improvement in dog and pet caregivers’ quality of life is an essential consideration when evaluating the efficacy of any complementary feed.</p></sec><sec id="s8"><title>Conflicts of Interest</title><p>The authors declare no conflicts of interest regarding the publication of this paper.</p></sec><sec id="s9"><title>Cite this paper</title><p>Porcheron, G., Bodet, M., Poiron, K., Brault, J., Massal, N., Roy, O. and Fantini, O. (2023) Supplementation Effects of an Alpha-Casozepine and White Fish Muscle Hydrolyzed Complementary Feed on Canine Separation-Related Disorders and Quality of Life of Dogs and Their Pet Caregivers. Open Journal of Veterinary Medicine, 13, 68-81. https://doi.org/10.4236/ojvm.2023.136007</p></sec><sec id="s10"><title>Appendix 1</title><p>EDED scale: Centripetal, internal factors; centrifugal, external stimuli; PU, polyuria; PD, polydipsia.</p><p><sup>1</sup>Hyperphagia with regurgitation and reingestion. The dog eats rapidly, display spasms, followed by vomiting. It then reingests what it has just expelled and resumes its meal. This behaviour appears regularly (1 meal in 2).</p><p><sup>2</sup>Carries empty water bowl around. The dog moves or carries its bowl toward one or several family members. This behaviour stops as soon as the bowl is filled.</p><p><sup>3</sup>Chews at water without swallowing it (dipsomania). The dog nibbles the water and spreads it around its bowl while swallowing very little.</p><p><sup>4</sup>Excessive licking, nibbling. A dos that is licking or nibbling itself, then spontaneously stops.</p><p><sup>5</sup>Stereotypic nibbling, dizziness. When there is licking or nibbling that does not stop spontaneously (the caregivers must stop the dog or divert its attention) or else dizziness or any other stereotypy (e.g. licking of the face, jumping, wandering).</p><p><sup>6</sup>Increase in sleep, hypersomnia. When the duration of sleep is longer than the age norm (+25%).</p><p><sup>7</sup>Insomnia, during sleep. Awakenings appear more than 90 minutes after going to sleep.</p><p><sup>8</sup>Wakes up shortly after going to sleep, anxiety at time of going to sleep. The dog awakes in the 30 - 45 minutes following going to sleep. Prior to going to sleep the dog may display moans, excitement, and a search for contacts, as if it is afraid of going to sleep.</p><p><sup>9</sup>In order to take the physical examination into account, all the manifestations observed must be scored and counted.</p></sec></body><back><ref-list><title>References</title><ref id="scirp.125772-ref1"><label>1</label><mixed-citation publication-type="other" xlink:type="simple">Bradshaw, J. (2011) In Defence of Dogs: Why Dogs Need Our Understanding. Penguin Press, London.</mixed-citation></ref><ref id="scirp.125772-ref2"><label>2</label><mixed-citation publication-type="other" xlink:type="simple">Sargisson, R.J. (2014) Canine Separation Anxiety: Strategies for Treatment and Management. Veterinary Medicine: Research and Reports, 5, 143-151. https://doi.org/10.2147/VMRR.S60424</mixed-citation></ref><ref id="scirp.125772-ref3"><label>3</label><mixed-citation publication-type="other" xlink:type="simple">Sherman, B.L. and Mills, D.S. (2008) Canine Anxieties and Phobias: An Update on Separation Anxiety and Noise Aversions. Veterinary Clinics of North America: Small Animal Practice, 38, 1081-1106. https://doi.org/10.1016/j.cvsm.2008.04.012</mixed-citation></ref><ref id="scirp.125772-ref4"><label>4</label><mixed-citation publication-type="other" xlink:type="simple">Tod, E., Brander, D. and Waran, N. (2005) Efficacy of Dog Appeasing Pheromone in Reducing Stress and Fear Related Behaviour in Shelter Dogs. Applied Animal Behaviour Science, 93, 295-308. https://doi.org/10.1016/j.applanim.2005.01.007</mixed-citation></ref><ref id="scirp.125772-ref5"><label>5</label><mixed-citation publication-type="other" xlink:type="simple">King, J.N., Simpson, B.S., Pageat, P., Overall, K.L., Appleby, D., Pageat, P., Ross, C., Chaurand, J.P., Heath, S., Beata, C., Weiss, A.B., Muller, G., Paris, T., Bataille, B.G., Parker, J., Petit, S., Wren, J. and The CLOCSA Group (2000) Treatment of Separation Anxiety in Dogs with Clomipramine: Results from a Prospective, Randomized, Double-Blind, Placebo-Controlled, Parallel-Group, Multicenter Clinical Trial. Applied Animal Behaviour Science, 67, 255-275. https://doi.org/10.1016/S0168-1591(99)00127-6</mixed-citation></ref><ref id="scirp.125772-ref6"><label>6</label><mixed-citation publication-type="other" xlink:type="simple">Sari, A., Fukuda, Y., Sakada, T., Maekawa, T. and Ishikawa, T. (1975) Effects of Psychotropic Drugs on Canine Cerebral Metabolism and Circulation Related to EEG—Diazepam, Clomipramine and Chlorpromazine. Journal of Neurology, Neurosurgery, and Psychiatry, 38, 838-844. https://doi.org/10.1136/jnnp.38.9.838</mixed-citation></ref><ref id="scirp.125772-ref7"><label>7</label><mixed-citation publication-type="other" xlink:type="simple">Sanchez-Araujo, M. and Puchi, A. (2011) Acupuncture Prevents Relapses of Recurrent Otitis in Dogs: A 1-Year Follow-Up of a Randomized Controlled Trial. Acupuncture in Medicine, 29, 21-26. https://doi.org/10.1136/aim.2010.002576</mixed-citation></ref><ref id="scirp.125772-ref8"><label>8</label><mixed-citation publication-type="other" xlink:type="simple">Kidd, J.R. (2012) Alternative Medicines for the Geriatric Veterinary Patient. Veterinary Clinics of North America: Small Animal Practice, 42, 809-822. https://doi.org/10.1016/j.cvsm.2012.04.009</mixed-citation></ref><ref id="scirp.125772-ref9"><label>9</label><mixed-citation publication-type="other" xlink:type="simple">Chapman, S.F. (2011) Homeopathic and Integrative Treatment for Feline Hyperthyroidism—Four Cases 424 (2006-2010). Homeopathy, 100, 270-274. https://doi.org/10.1016/j.homp.2011.05.004</mixed-citation></ref><ref id="scirp.125772-ref10"><label>10</label><mixed-citation publication-type="other" xlink:type="simple">Neumann, S., Stolt, P., Braun, G., Hellmann, K. and Reinhart, E. (2011) Effectiveness of the Homeopathic Preparation Zeel Compared with Carprofen in Dogs with Osteoarthritis. Journal of the American Animal Hospital Association, 47, 12-20. https://doi.org/10.5326/JAAHA-MS-5483</mixed-citation></ref><ref id="scirp.125772-ref11"><label>11</label><mixed-citation publication-type="other" xlink:type="simple">Gingerich, D.A. and Strobel, J.D. (2003) Use of Client-Specific Outcome Measures to Assess Treatment Effects in Geriatric, Arthritic Dogs: Controlled Clinical Evaluation of a Nutraceutical. Veterinary Therapeutics, 4, 376-386.</mixed-citation></ref><ref id="scirp.125772-ref12"><label>12</label><mixed-citation publication-type="other" xlink:type="simple">Araujo, J.A., Landsberg, G.M., Milgram, N.W. and Miolo, A. (2008) Improvement of Short-Term Memory Performance in Aged Beagles by a Nutraceutical Supplement containing Phosphatidylserine, Ginkgo biloba, Vitamin E and Pyridoxine. Canadian Veterinary Journal, 49, 379-385.</mixed-citation></ref><ref id="scirp.125772-ref13"><label>13</label><mixed-citation publication-type="other" xlink:type="simple">Kato, M., Miyaji, K., Ohtani, N. and Ohta, M. (2012) Effects of Prescription Diet on Dealing with Stressful Situations and Performance of Anxiety-Related Behaviors in Privately Owned Anxious Dogs. Journal of Veterinary Behavior, 7, 21-26. https://doi.org/10.1016/j.jveb.2011.05.025</mixed-citation></ref><ref id="scirp.125772-ref14"><label>14</label><mixed-citation publication-type="other" xlink:type="simple">Wells, D.L. (2006) Aromatherapy for Travel-Induced Excitement in Dogs. Journal of the American Veterinary Medical Association, 229, 964-967. https://doi.org/10.2460/javma.229.6.964</mixed-citation></ref><ref id="scirp.125772-ref15"><label>15</label><mixed-citation publication-type="other" xlink:type="simple">Makawey, A., Iben, C. and Palme, R. (2020) Cats at the Vet: The Effect of Alpha-s1 Casozepin. Animals, 10, Article 2047. https://doi.org/10.3390/ani10112047</mixed-citation></ref><ref id="scirp.125772-ref16"><label>16</label><mixed-citation publication-type="other" xlink:type="simple">Miclo, L., Perrin, E., Driou, A., Papadopoulos, V., Boujrad, N., Vanderesse, R., Boudier, J.F., Desor, D., Linden, G. and Gaillard, J.L. (2001) Characterization of α-Casozepine, a Tryptic Peptide from Bovine α&lt;sub&gt;s1&lt;/sub&gt;-Casein with Benzodiazepine-Like Activity. The FASEB Journal, 15, 1780-1782. https://doi.org/10.1096/fj.00-0685fje</mixed-citation></ref><ref id="scirp.125772-ref17"><label>17</label><mixed-citation publication-type="other" xlink:type="simple">Messaoudi, M., Lefranc-Millot, C., Desor, D., Demagny, B. and Bourdon, L. (2004) Effects of a Tryptic Hydrolysate from Bovine Milk α&lt;sub&gt;s1&lt;/sub&gt;-Casein on Hemodynamic Responses in Healthy Human Volunteers Facing Successive Mental and Physical Stress Situations. European Journal of Nutrition, 44, 128-132. https://doi.org/10.1007/s00394-004-0534-7</mixed-citation></ref><ref id="scirp.125772-ref18"><label>18</label><mixed-citation publication-type="other" xlink:type="simple">V&amp;#248;ls, K.K., Heden, M.A., Kristensen, A.T. and Sand&amp;#248;e, P. (2017) Quality of Life Assessment in Dogs and Cats Receiving Chemotherapy—A Review of Current Methods. Veterinary Comparative Oncology, 15, 684-691. https://doi.org/10.1111/vco.12242</mixed-citation></ref><ref id="scirp.125772-ref19"><label>19</label><mixed-citation publication-type="other" xlink:type="simple">Dinel, A.L., Lucas, C., Le Faouder, J., Bouvret, E., Pallet, V., Layé, S. and Joffre, C. (2021) Supplementation with Low Molecular Weight Peptides from Fish Protein Hydrolysate Reduces Acute Mild Stress-Induced Corticosterone Secretion and Modulates Stress Responsive Gene Expression in Mice. Journal of Functional Foods, 76, Article ID: 104292. https://doi.org/10.1016/j.jff.2020.104292</mixed-citation></ref><ref id="scirp.125772-ref20"><label>20</label><mixed-citation publication-type="other" xlink:type="simple">Bernet, F., Montel, V., No&amp;#235;l, B. and Dupouy, J.P. (2000) Diazepam-Like Effects of a Fish Protein Hydrolysate (Gabolysat PC60) on Stress Responsiveness of the Rat Pituitary-Adrenal System and Sympathoadrenal Activity. Psychopharmacology, 149, 34-40. https://doi.org/10.1007/s002139900338</mixed-citation></ref><ref id="scirp.125772-ref21"><label>21</label><mixed-citation publication-type="other" xlink:type="simple">Braat, K.M. and Brinker, C.J. (2004) GABA (B) Receptors: Pharmacology and Therapeutic Potential. Current Opinion in Pharmacology, 4, 12-18.</mixed-citation></ref><ref id="scirp.125772-ref22"><label>22</label><mixed-citation publication-type="other" xlink:type="simple">Pageat, P. (1995) Pathologie du comportement du chien. Le Point Veterinaire, Paris.</mixed-citation></ref><ref id="scirp.125772-ref23"><label>23</label><mixed-citation publication-type="other" xlink:type="simple">Noli, C. (2019) Assessing Quality of Life for Pets with Dermatologic Disease and Their Owners. Veterinary Clinics of North America: Small Animal Practice, 49, 83-93. https://doi.org/10.1016/j.cvsm.2018.08.008</mixed-citation></ref><ref id="scirp.125772-ref24"><label>24</label><mixed-citation publication-type="other" xlink:type="simple">Wojciechowska, J.I. and Hewson, C.J. (2005) Quality-of-Life Assessment in Pet Dogs. Journal of the American Veterinary Medical Association, 226, 722-728. https://doi.org/10.2460/javma.2005.226.722</mixed-citation></ref><ref id="scirp.125772-ref25"><label>25</label><mixed-citation publication-type="other" xlink:type="simple">Noli, C., Colombo, S., Cornegliani, L., Ghibaudo, G., Persico, P. and Vercelli, A. (2011) Quality of Life of Dogs with Skin Disease and of Their Owners. Part 2: Administration of a Questionnaire in Various Skin Diseases and Correlation to Efficacy of Therapy. Veterinary Dermatology, 22, 344-351. https://doi.org/10.1111/j.1365-3164.2011.00956.x</mixed-citation></ref><ref id="scirp.125772-ref26"><label>26</label><mixed-citation publication-type="other" xlink:type="simple">Maddison, J., Cannon, M., Davies, R., Farquhar, R., Faulkner, B., Furtado, T., Harvey, G., Leathers, F., Snowden, A. and Wright, I. (2021) Owner Compliance in Veterinary Practice: Recommendations from a Roundtable Discussion. Companion Animal, 26, S1-S11. https://doi.org/10.12968/coan.2021.0029</mixed-citation></ref><ref id="scirp.125772-ref27"><label>27</label><mixed-citation publication-type="other" xlink:type="simple">Craven, A.J., Pegram, C., Packer, R.M.A., Jarvis, S., McGreevy, P.D., Warnes, C., Church, D.B., Brodbelt, D.C. and O’Neill, D.G. (2022) Veterinary Drug Therapies Used for Undesirable Behaviours in UK Dogs under Primary Veterinary Care. PLOS ONE, 17, e0261139. https://doi.org/10.1371/journal.pone.0261139</mixed-citation></ref><ref id="scirp.125772-ref28"><label>28</label><mixed-citation publication-type="other" xlink:type="simple">Sargisson, R. (2014) Canine Separation Anxiety: Strategies for Treatment and Management. Veterinary Medicine: Research and Reports, 5, 143-151. https://doi.org/10.2147/VMRR.S60424</mixed-citation></ref></ref-list></back></article>