Analyze Diet

Topic:Clinical Study

Clinical studies in equine research involve the systematic investigation of health and disease in horses through structured scientific methods. These studies aim to evaluate the safety and efficacy of treatments, understand disease mechanisms, and improve veterinary care practices. Clinical studies can include randomized controlled trials, observational studies, and case-control studies. They may focus on various aspects such as pharmacokinetics, therapeutic interventions, and diagnostic techniques. This page compiles peer-reviewed research studies and scholarly articles that explore the design, implementation, and findings of clinical studies in equine medicine, providing insights into their impact on horse health and veterinary practices.
Comparison of thoracic ultrasonography and thoracic radiography between healthy adult horses and horses with bacterial pneumonia using a novel, objective ultrasonographic scoring system.
Frontiers in veterinary science    October 13, 2022   Volume 9 991634 doi: 10.3389/fvets.2022.991634
Hepworth-Warren KL, Nelson N, Dembek KA, Young KAS.Thoracic ultrasonography (TUS) is widely used in equine practice but comparison to radiography is limited in horses. Unassigned: To validate a novel, objective scoring system for TUS in adult horses and to compare ultrasonographic and radiographic findings. Unassigned: 13 healthy horses and 9 with confirmed bacterial pneumonia. Unassigned: Prospective study in which TUS and radiography were performed on healthy horses and those with bacterial pneumonia confirmed by clinical signs and results of transtracheal wash analysis. Ultrasonography was scored utilizing a novel scoring system evaluating ...
Therapeutic Exercises for Equine Sacroiliac Joint Pain and Dysfunction.
The Veterinary clinics of North America. Equine practice    October 13, 2022   Volume 38, Issue 3 569-584 doi: 10.1016/j.cveq.2022.07.002
Goff L.This article provides the equine practitioner with a review of sacroiliac joint pain and dysfunction and outlines the importance of providing a specific prescription for a safe and effective therapeutic exercise program. The continuum of clinical dysfunction associated with the sacroiliac region is presented with prescribed interventions. The intent is to encourage the practitioner to perform a thorough assessment of the sacroiliac joint and the adjacent soft tissues and to use sound clinical reasoning to formulate a therapeutic exercise plan.
Screening for Taylorella equigenitalis in Equine Semen: An Exploratory Study.
Journal of equine veterinary science    October 13, 2022   Volume 119 104138 doi: 10.1016/j.jevs.2022.104138
Mawhinney I, Davis N, Carson T, Torrens N, Wales A.The study examined and compared the sensitivity of culture and a quantitative PCR assay for screening equine semen for the presence of Taylorella equigenitalis (CEMO). Chilled semen samples, both raw and treated with extender, from two stallions were spiked with the organism at seven or 23 days postejaculation and prepared in serial dilutions. Culture of the 7-day raw semen readily detected CEMO at all dilutions, but extended semen yielded counts that were two log cycles lower at equivalent dilutions, with the organism being nearly undetectable at the maximal dilutions. By contrast, PCR sensit...
Identification of an ectopic periorbital lymph node in two horses.
Open veterinary journal    October 12, 2022   Volume 12, Issue 5 768-773 doi: 10.5455/OVJ.2022.v12.i5.23
Narinx F, Monclin S, Sauvage A, Vercruysse E, Heimann M, Alloway E, Vandersmissen M, Grauwels M.To describe the clinical presentation and treatment of an ectopic periorbital lymph node in two young horses. Two warm-blood horses were presented at the equine clinic of the University of Liège with a periorbital non-painful mass. Differential diagnosis included neoplasm (lymphoma), (sterile) abscess, cyst, ectopic lacrimal gland tissue, hematoma, adipose tissue, or ectopic lymphoid tissue. Information collected included physical and ophthalmic examination findings, results of the ocular and periocular ultrasound, surgical procedure, histopathology, and follow-up. Masses of 2 × 2 cm and 3 ...
[Immunotherapy with anti-SARS-COV-2 neutralizing F(ab’)2 antibodies from equine serum in the treatment of outpatients with bilateral COVID-19 pneumonia].
Medicina    October 12, 2022   Volume 82, Issue 5 667-672 
Manganiello CF, Basbus L, Callegari MS, Cayol F, Amaral De Sousa CR, Porta RE.Passive immunotherapy is a therapeutic alternative for patients with COVID-19. Methods: The decision was made to create a prospective database of patients diagnosed with SARS-CoV-2 pneumonia, nonhypoxemic, treated on an outpatient basis at the Hospital de Bolívar, Dr. Miguel Capredoni, province of Buenos Aires, Argentina, with the aim of evaluating the efficacy in reducing severe cases and hospitalizations of treatment with hyperimmune equine serum in this subgroup of patients. We performed a retrospective analysis of the period from 05/26/2021 to 08/28/2021, where a total of 151 patients wer...
Developing advanced therapeutics through the study of naturally occurring immune-mediated ocular disease in domestic animals.
American journal of veterinary research    October 11, 2022   Volume 83, Issue 11 doi: 10.2460/ajvr.22.08.0145
Gilger BC.This review, which is part of the "Currents in One Health" series, describes the importance of the study of immune-mediated ocular disease in the development of innovative therapeutics, such as cell and gene therapy for the eye. Recent examples of cell and gene therapy studies from the author's laboratory are reviewed to emphasize the importance of One Health initiatives in developing innovative therapies for ocular diseases. Spontaneous immune-mediated corneal disease is common in horses, cats, dogs, and humans. Autologous bone marrow-derived mesenchymal stem cells (BM-MSCs) injected subconju...
Radiographic prevalence of juvenile osteochondral conditions of the proximal interphalangeal joint of Australian Thoroughbred racehorse yearlings and associations with sales results and race performance.
Frontiers in veterinary science    October 10, 2022   Volume 9 988826 doi: 10.3389/fvets.2022.988826
Faulkner J, Vanderperren K, Duchateau L, O'Sullivan C.Ascertain the radiographic prevalence and variation in characteristics of juvenile osteochondral conditions (JOC) in the proximal interphalangeal joint (PIPJ) of Australian Thoroughbred racehorse yearlings. Establish whether there are any significant associations with public auction sale results and racing performance. Unassigned: Retrospective evaluation of 1,098 yearling repository radiograph sets. Comparison of sales results and whole career racing performance of the case group with two control groups: maternal siblings ( = 397) and yearlings without PIP JOC ( = 391). Unassigned: 6.3% of ye...
Serum amyloid A and fibrinogen as markers for early detection of surgical site infection associated with internal fixation in the horse.
Frontiers in veterinary science    October 10, 2022   Volume 9 960865 doi: 10.3389/fvets.2022.960865
Thurston CC, Stefanovski D, MacKinnon MC, Chapman HS, Richardson DW, Levine DG.The objective of this study was to determine the diagnostic ability of serum amyloid A (SAA) and fibrinogen for early detection of surgical site infection (SSI) after equine internal fixation. Horses undergoing internal fixation for fracture, arthrodesis, or osteotomy with internal fixation for limb deformity were included in the study. SAA and fibrinogen were measured on blood samples preoperatively and on days 1, 3, 5, 7, 10, and 14 postoperatively. Statistical analysis included use of Spearman's rank correlation, logistic regression, and calculating the area under the receiver operating cha...
Clodronate disodium is neither cytotoxic nor cytoprotective to normal and recombinant equine interleukin-1β-treated joint tissues in vitro.
Veterinary surgery : VS    October 10, 2022   Volume 52, Issue 1 146-156 doi: 10.1111/vsu.13898
Vergara-Hernandez FB, Panek CL, Nielsen BD, Robison CI, Colbath AC.To determine the effects of clodronate disodium (CLO) on control and recombinant equine interleukin-1β (IL-1β)-treated equine joint tissues. Methods: In vitro experimental study. Methods: Cartilage explants, chondrocytes, and synoviocytes (n = 3 horses). Methods: Monolayer cultures of chondrocytes and synoviocytes from three horses were subjected to: control media (CON), 5 ng/ml CLO (C/low), 50 ng/ml CLO (C/med), 100 ng/ml CLO (C/high), with and without IL-1β, and 10 ng/ml IL-1β (IL) alone for 72 hours. Cartilage explants from three horses were subjected to CON, IL, C/low, and C/...
Disease progression, pathologic, and virologic findings of an equine influenza outbreak in rescue donkeys.
Journal of veterinary internal medicine    October 7, 2022   Volume 36, Issue 6 2230-2237 doi: 10.1111/jvim.16563
Ahearne MM, Pentzke-Lemus LL, Romano AM, Larsen ED, Watson AM, O'Fallon EA, Landolt GA.Equine influenza virus is a common cause of respiratory disease in equids. Few reports describe clinical presentation and disease progression in donkeys. Objective: Describe the clinical and diagnostic findings, outcome, and pathologic lesions associated with influenza pneumonia in donkeys. Methods: Thirteen unvaccinated donkeys ranging from 1 week to 12 years of age and sharing clinical signs and exposure history. Methods: Retrospective case series. Medical records from June to July 2020 at the Colorado State Veterinary Teaching Hospital and collaborating referring veterinary practices wer...
Comparative immune responses after vaccination with the formulated inactivated African horse sickness vaccine serotype 1 between naïve horses and pretreated horses with the live-attenuated African horse sickness vaccine.
Veterinary world    October 7, 2022   Volume 15, Issue 10 2365-2375 doi: 10.14202/vetworld.2022.2365-2375
Chaiyabutr N, Wattanaphansak S, Tantilerdcharoen R, Akesowan S, Ouisuwan S, Naraporn D.African horse sickness (AHS) is a non-contagious, high mortality, and insect-borne disease caused by a double-stranded RNA virus from the genus Orbivirus. The study aimed to develop inactivated vaccines serotype 1 inactivated AHS vaccine (IAV) and to compare the effect of IAV on antibody responses in young naïve horses and adult horses pre-immunized with live-attenuated AHS virus (AHSV) serotypes 1, 3, and 4 live-attenuated vaccine (LAV). Unassigned: A total of 27 horses were vaccinated in two trials. Twelve AHS naïve young horses and 15 adult horses were divided into three groups of 4 and 5...
Study design synopsis: Evidence syntheses-What are they and why do we need them?
Equine veterinary journal    October 6, 2022   Volume 54, Issue 6 1011-1012 doi: 10.1111/evj.13872
Freeman SL.No abstract available
A starch-rich treat affects enteroinsular responses in ponies.
Journal of the American Veterinary Medical Association    October 6, 2022   Volume 260, Issue S3 S94-S101 doi: 10.2460/javma.22.06.0272
Sibthorpe PEM, Fitzgerald DM, Chen L, Sillence MN, de Laat MA.To determine the effect of a starch-rich treat, added to the daily diet of ponies for 10 days, on enteroinsular responses to meal consumption. 10 mixed-breed adult ponies owned by Queensland University of Technology were used in the study. Six ponies were metabolically healthy, and 4 were insulin dysregulated at the start of the study, according to the results of an in-feed oral glucose test. A bread-based treat was offered twice daily for 10 days, adding 0.36 ± 0.04 g/kg body weight (BW) carbohydrates to the daily diet. Before and after treatment, the intestinal capacity for simple carbohydr...
Changes in Head, Withers, and Pelvis Movement Asymmetry in Lame Horses as a Function of Diagnostic Anesthesia Outcome, Surface and Direction.
Journal of equine veterinary science    October 6, 2022   Volume 118 104136 doi: 10.1016/j.jevs.2022.104136
Marunova E, Hoenecke K, Fiske-Jackson A, Smith RKW, Bolt DM, Perrier M, Gerdes C, Hernlund E, Rhodin M, Pfau T.Evaluation of diagnostic anesthesia during equine lameness examination requires comparison of complex movement patterns and can be influenced by expectation bias. There is limited research about how changes in movement asymmetries after successful analgesia are affected by different exercise conditions. Movement asymmetry of head, withers and pelvis was quantified in N = 31 horses undergoing forelimb or hindlimb diagnostic anesthesia. Evaluation on a straight line and a circle was performed with subjective diagnostic anesthesia outcome and quantitative changes recorded. Mixed linear mode...
The post-ovulatory rise in progesterone is lower and the persistence of oestrous behaviour longer during the first compared with the second cycle of the breeding season in mares.
Reproduction in domestic animals = Zuchthygiene    October 6, 2022   Volume 58, Issue 1 141-145 doi: 10.1111/rda.14273
Newcombe JR, Wilsher S, Cuervo-Arango J.Mares are seasonally polyoestrous breeders. Therefore, the first ovulation of the season, following winter anoestrus, is the only cycle in which mares ovulate without the presence of an old CL from the previous cycle. The objective of this study was to compare the length of oestrous behaviour, and plasma progesterone concentrations during the early post-ovulatory period between mares after the first and second ovulation of the breeding season. Overall, 38 mares and 167 oestrous periods were used in the study. From those, 11 mares were used during the first and subsequent oestrous period to mea...
Biomarkers of Kidney Disease in Horses: A Review of the Current Literature.
Animals : an open access journal from MDPI    October 5, 2022   Volume 12, Issue 19 2678 doi: 10.3390/ani12192678
Galen GV, Olsen E, Siwinska N.Creatinine only allows detection of kidney disease when 60 to 75% of the glomerular function is lost and is therefore not an ideal marker of disease. Additional biomarkers could be beneficial to assess kidney function and disease. The objectives are to describe new equine kidney biomarkers. This systematic review assesses the available literature, including the validation process and reference values, following which the authors suggest recommendations for clinical use. SDMA may have some potential as equine kidney biomarker, but there is currently a lack of evidence that SDMA offers any advan...
Equine Odontoclastic Tooth Resorption and Hypercementosis.
Journal of veterinary dentistry    October 5, 2022   Volume 39, Issue 4 346-357 doi: 10.1177/08987564221120715
James O.Equine odontoclastic tooth resorption and hypercementosis (EOTRH) is a recently described painful and progressive condition of unknown etiology that occurs in middle-aged and geriatric equines. It predominantly affects the permanent incisor and canine teeth and, less commonly, the premolar and molar dentition. EOTRH was first reported in peer-reviewed literature in 2008, with subsequent publications of case reports, histological studies and retrospective case series. There have been few significant research developments related to this disease. The existing studies have primarily involved sing...
Short-term administration of flunixin meglumine or firocoxib does not alter viscoelastic coagulation profiles in healthy horses.
Journal of the American Veterinary Medical Association    October 4, 2022   Volume 260, Issue 15 1963-1966 doi: 10.2460/javma.22.08.0367
Bishop RC, McCoy AM, Kemper AM, Stewart RM, Wilkins PA.To evaluate the effect of the cyclooxygenase-2-selective NSAID firocoxib, compared to the nonselective NSAID flunixin meglumine on viscoelastic coagulation parameters in healthy horses. 12 healthy adult mixed-breed horses. Following a crossover protocol, horses were administered flunixin meglumine (1.1 mg/kg, IV, q 12 h for 5 days), allowed a 6-month washout period, and then administered firocoxib (0.3 mg/kg, PO, once, then 0.1 mg/kg, PO, q 24 h for 4 days). Omeprazole (1 mg/kg, PO, q 24 h) was administered concurrently with each NSAID. Viscoelastic coagulation profiles and traditional coagula...
Acute diarrhea in horses: A multicenter Canadian retrospective study (2015 to 2019).
The Canadian veterinary journal = La revue veterinaire canadienne    October 4, 2022   Volume 63, Issue 10 1033-1042 
Gomez DE, Leclere M, Arroyo LG, Li L, John E, Afonso T, Payette F, Darby S.This study describes the clinicopathological findings, diagnostic approach, treatment, and factors associated with non-survival of diarrheic horses admitted to 4 Canadian university teaching hospitals between 2015 and 2019. A total of 300 horses, ≥1-year-old, with acute diarrhea were included and represented 1.6% (300/18 481; range: 0.7 to 3%) of admissions during that period, 70% of the horses survived to discharge. Testing for enteropathogens was limited to a single fecal culture for Salmonella spp. in most cases. An enteropathogen was identified in 14% (42/300) of the horses, but in the h...
Retrospective review of atrial fibrillation in Standardbred racehorses at a tertiary care facility in Atlantic Canada.
The Canadian veterinary journal = La revue veterinaire canadienne    October 4, 2022   Volume 63, Issue 10 1051-1056 doi: 10.1111/evj.13551
Burns JJ, MacMillan KM, John EE.Atrial fibrillation (AF) is the most clinically relevant cardiac arrhythmia identified in the Standardbred racehorse. However, there is limited literature regarding athletic ability in Standardbred racehorses following AF conversion. To investigate this issue, the objectives of this review were to: i) determine the success rate in using quinidine sulphate to treat AF in a population of hospitalized equine patients in Atlantic Canada between January 2008 and December 2019; and ii) measure return to athletic ability (using racetrack earnings and top-3 finishes) following cardioversion with quini...
Changes in Perioperative Antimicrobial and Anti-Inflammatory Drugs Regimens for Colic Surgery in Horses: A Single Center Report.
Veterinary sciences    October 4, 2022   Volume 9, Issue 10 546 doi: 10.3390/vetsci9100546
Gandini M, Cerullo A, Franci P, Giusto G.Reducing postoperative incisional infection is the main reason to administer postoperative antimicrobials (AMD) after emergency laparotomy in horses, while reducing inflammation and providing analgesia are the reasons to administer anti-inflammatory drugs (AID). The basis for postoperative AMD and AID administration is empirical and only recently has been questioned. Empirical approaches can be changed, and these changes, along with the description of their outcomes, can help produce appropriate stewardship. The aim of this study is to report the changes in AMD and AID regimens in horses under...
Retrospective Study on Risk Factors and Short-Term Outcome of Horses Referred for Colic from 2016 to 2022.
Veterinary sciences    October 3, 2022   Volume 9, Issue 10 545 doi: 10.3390/vetsci9100545
Straticò P, Varasano V, Palozzo A, Guerri G, Celani G, Revelant O, Petrizzi L.(1) Background: colic syndrome is a multifactorial life-threatening condition in equids. Awareness of predisposing and prognostic indicators is useful to adequately inform the owner of the outcome and plan the best treatment. The aim of the study was to identify the variables associated with short-term survival in a cohort of horses referred for colic to a veterinary teaching hospital; (2) Methods: medical records of horses referred for colic from 2016 to 2022 were retrospectively reviewed. Univariate and multivariate regression models were built to assess the odds ratios of discharge from the...
Ultrasonography of the Metacarpal/Tarsal-Phalangeal Joints in Healthy Racehorses: Normal Appearance, Breed-Related and Age-Related Features.
Animals : an open access journal from MDPI    October 3, 2022   Volume 12, Issue 19 2657 doi: 10.3390/ani12192657
Nocera I, Puccinelli C, Sgorbini M, Bagnoli E, Citi S.In adult horses, specific ultrasound (US) features and reference values have been reported for the appearance of the joint cartilage and thickness according to the type of joint, such as femoropatellar and tarsocrural. The US appearance of the fetlock has been described in several diseases. The present research evaluates the US features of the metacarpal/tarsal-phalangeal joints in healthy racehorses according to age and breed, since no information is available in the literature. Seventy-one fetlocks in 28 healthy horses (15/28 thoroughbreds and 13/28 standardbreds) were assessed. The horses w...
Generic approach for the discovery of drug metabolites in horses based on data-dependent acquisition by liquid chromatography high-resolution mass spectrometry and its applications to pharmacokinetic study of daprodustat.
Analytical and bioanalytical chemistry    October 1, 2022   Volume 414, Issue 28 8125-8142 doi: 10.1007/s00216-022-04347-2
Ishii H, Shibuya M, Kusano K, Sone Y, Kamiya T, Wakuno A, Ito H, Miyata K, Sato F, Kuroda T, Yamada M, Leung GN.In drug metabolism studies in horses, non-targeted analysis by means of liquid chromatography coupled with high-resolution mass spectrometry with data-dependent acquisition (DDA) has recently become increasingly popular for rapid identification of potential biomarkers in post-administration biological samples. However, the most commonly encountered problem is the presence of highly abundant interfering components that co-elute with the target substances, especially if the concentrations of these substances are relatively low. In this study, we evaluated the possibility of expanding DDA coverag...
Body Condition Score Is Not Correlated to Gastric Ulcers in Non-Athlete Horses.
Animals : an open access journal from MDPI    September 30, 2022   Volume 12, Issue 19 doi: 10.3390/ani12192637
Busechian S, Turini L, Sgorbini M, Bonelli F, Pisello L, Pieramati C, Orvieto S, Rueca F.Equine Gastric Ulcer Syndrome (EGUS) is a worldwide disease of the stomach that can be found in different categories of horses. Different clinical signs may be present, but a large number of horses are asymptomatic. The aim of this study was to identify a possible correlation between body condition score (BCS) and EGUS in a population of horses. A total of 203 non-athlete horses were submitted for gastroscopies, and the presence and severity of lesions of the glandular and squamous mucosa were recorded. A board-certified veterinarian blinded to the gastroscopy results assessed the BCS of the h...
Endocrine and ovarian responses to combined estradiol benzoate-sulpiride in seasonally anovulatory mares treated with kisspeptin.
Animal reproduction science    September 30, 2022   Volume 247 107087 doi: 10.1016/j.anireprosci.2022.107087
Bailey VN, Sones JL, Camp CM, Gomes VCL, Oberhaus EL.The objective of this experiment was to determine if incorporation of kisspeptin-10 (Kp10) into treatment with estradiol benzoate (EB) and sulpiride to induce early cyclicity would result in greater endocrine responses and a greater number of mares responding with either follicle(s) > 30 mm or ovulation within 25 days of treatment. Eighteen anestrous mares were blocked by breed, body condition, and age before random assignment to treatment or control. All mares received 50 mg EB before receiving osmotic minipumps containing either saline (n = 9) or Kp10 (50 μg/hour; n = 9) one day ...
Animal-assisted therapy in eating disorder treatment: A systematic review.
Eating behaviors    September 30, 2022   Volume 47 101673 doi: 10.1016/j.eatbeh.2022.101673
Fennig MW, Weber E, Santos B, Fitzsimmons-Craft EE, Wilfley DE.Eating disorders (EDs) are deadly illnesses with high relapse rates, highlighting need for better interventions. Animal-assisted therapy (AAT) has been implemented supplementally for EDs, with horses utilized at many residential facilities. AAT shows promise with meta-analyses of randomized control trials (RCTs) showing significant decreases in depression, anxiety, and negative affect; however, no review to date has evaluated efficacy for EDs. Therefore, this study conducted a systematic review of primary literature to investigate the efficacy of AAT for EDs. A systematic review was conducted ...
Enhancement of chronic wound healing with maltodextrin/ascorbic acid gel: a clinical evaluation of distal limb wounds in horses.
Journal of animal science and technology    September 30, 2022   Volume 64, Issue 5 997-1007 doi: 10.5187/jast.2022.e52
Helal IE, Al-Abbadi HA, El-Daharawy MH, Ahmed MF.Delayed healing associated with distal limb wounds is highly challenging in equine clinical practice. This study aimed to evaluate healing rates between chronic non-granulating wounds of horse distal limbs that were treated with maltodextrin/ascorbic acid gel alone or in combination with povidone-iodine 1% solution and those treated with povidone-iodine 1% only throughout the study period (35 days) in clinical settings. The study was conducted on 18 adult horses (3-15 years old). Based on the treatment regimen utilized, the horses were divided into three groups (n = 6), with each group having ...
The effect of manually facilitated flexion of the thoracic spine on the interspinous space among horses with impinging dorsal spinous processes of the thoracic vertebrae.
Veterinary journal (London, England : 1997)    September 29, 2022   Volume 289 105909 doi: 10.1016/j.tvjl.2022.105909
O'Sullivan S, McGowan CM, Junnila J, Hyytiäinen HK.Impinging dorsal spinous processes (IDSP) are typically diagnosed and graded using radiography, during which the effect of the horses' thoracolumbar posture on the interspinous spaces is not commonly considered. Posture can be altered from a spontaneous, relatively extended, or lordotic, position by manual facilitation of thoracic flexion ('thoracic lift'). This study aimed to ascertain if the thoracic vertebral interspinous space distances were increased by using facilitated thoracic flexion to alter the posture in horses diagnosed with IDSP. Seven horses of mixed breed and sex, median age 9....
Slow Cooling is Beneficial for Storage of Frozen-Thawed Equine Spermatozoa.
Journal of equine veterinary science    September 29, 2022   Volume 118 104132 doi: 10.1016/j.jevs.2022.104132
van Heule M, Verstraete M, Blockx Z, De Blende P, Dini P, Daels P.Cooled storage of semen after thawing can expand the use of frozen semen, providing the possibility of thawing and evaluating the semen at the storage site and subsequently shipping the semen. Our objectives were (1) to examine the motility and viability of frozen-thawed semen after cooled storage and (2) to compare two cooled-storage protocols for frozen-thawed semen. The samples (n = 31) were either placed immediately in a passive cooling box for 8 or 24 hours (CB) or placed in a refrigerator at 4°C for 30 minutes and then transferred to a passive cooling box (REF). Total and progressive ...
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