Topic:Calcium

Diamond burr debridement of calcific band keratopathy in 24 eyes of 22 horses.
Veterinary ophthalmology    March 26, 2024   doi: 10.1111/vop.13212
Jugant S, Regnier A, Douet JY.To evaluate the safety and effectiveness of diamond burr debridement (DBD) for the treatment of calcific band keratopathy (CBK) in horses. Methods: Medical records from horses with CBK were reviewed over a period of 4 years. Diagnosis of CBK was based on slit-lamp examination findings, and DBD was performed on standing sedated horses. Follow-up was obtained by clinical reevaluation of the treated eyes or a telephone survey with the referring veterinarian. Results: Twenty-two horses of different ages (median: 11 years; range: 5-23) representing 24 CBK-affected eyes (14 left eyes and 10 righ...
Nutrient Composition and Feed Hygiene of Alfalfa, Comparison of Feed Intake and Selected Metabolic Parameters in Horses Fed Alfalfa Haylage, Alfalfa Hay or Meadow Hay.
Animals : an open access journal from MDPI    March 13, 2024   Volume 14, Issue 6 889 doi: 10.3390/ani14060889
Ku00f6ninger M, von Velsen-Zerweck A, Eiberger C, Lu00f6ffler C, Tu00f6pper A, Visscher C, Reckels B, Vervuert I.The aim of this study was to examine the nutrient composition and feed hygiene of alfalfa as well as investigate the feed intake, blood, urine and faecal parameters of horses fed alfalfa haylage (AS) compared with alfalfa hay (AH) and meadow hay (MH). A total of 11 geldings were fed ad libitum (2.1% dry matter (DM) of body weight (BW)) with alfalfa haylage, alfalfa hay and meadow hay (MH) in a Latin square design. On days 0 and 21 of the feeding period, blood samples were analysed for kidney and liver parameters. Faecal samples were analysed for pH, DM and short-chain fatty acids (SCFAs). Spon...
Gene co-expression network analysis reveal core responsive genes in Parascaris univalens tissues following ivermectin exposure.
PloS one    February 15, 2024   Volume 19, Issue 2 e0298039 doi: 10.1371/journal.pone.0298039
Dube F, Delhomme N, Martin F, Hinas A, u00c5brink M, Svu00e4rd S, Tydu00e9n E.Anthelmintic resistance in equine parasite Parascaris univalens, compromises ivermectin (IVM) effectiveness and necessitates an in-depth understanding of its resistance mechanisms. Most research, primarily focused on holistic gene expression analyses, may overlook vital tissue-specific responses and often limit the scope of novel genes. This study leveraged gene co-expression network analysis to elucidate tissue-specific transcriptional responses and to identify core genes implicated in the IVM response in P. univalens. Adult worms (n = 28) were exposed to 10-11 M and 10-9 M IVM in vitro for 2...
Differential age-related transcriptomic analysis of ovarian granulosa cells in Kazakh horses.
Frontiers in endocrinology    January 30, 2024   Volume 15 1346260 doi: 10.3389/fendo.2024.1346260
Ren W, Wang J, Zeng Y, Wang T, Meng J, Yao X.The Kazakh horse, renowned for its excellence as a breed, exhibits distinctive reproductive traits characterized by early maturity and seasonal estrus. While normal reproductive function is crucial for ensuring the breeding and expansion of the Kazakh horse population, a noteworthy decline in reproductive capabilities is observed after reaching 14 years of age. Unassigned: In this study, ovarian granulosa cells (GCs) were meticulously collected from Kazakh horses aged 1, 2, 7, and above 15 years old (excluding 15 years old) for whole transcriptome sequencing. Unassigned: The analysis identifie...
Effect of Regular Training on Platelet Function in Untrained Thoroughbreds.
Animals : an open access journal from MDPI    January 27, 2024   Volume 14, Issue 3 414 doi: 10.3390/ani14030414
Miglio A, Falcinelli E, Cappelli K, Mecocci S, Mezzasoma AM, Antognoni MT, Gresele P.Training has a significant effect on the physiology of blood coagulation in humans and in horses. Several hemostatic changes have been reported after exercise in the horse but data available are inconclusive. The aim of this study was to investigate platelet activation and primary platelet-related hemostasis modifications in young never-trained Thoroughbreds in the first incremental training period in order to improve knowledge on this topic. Twenty-nine clinically healthy, untrained, 2-year-old Thoroughbred racehorses were followed during their incremental 4-month sprint exercise training. Bl...
Acidification is required for calcium and magnesium concentration measurements in equine urine.
BMC veterinary research    January 10, 2024   Volume 20, Issue 1 21 doi: 10.1186/s12917-023-03848-1
Lapsina S, Stirn M, Hofmann-Lehmann R, Schoster A, Riond B.Acidification of equine urine to promote dissociation of ion complexes is a common practice for urine ion concentration measurements. The objective of this study was to evaluate the effect of acidification and storage after acidification on calcium (Ca), magnesium (Mg) and phosphate (P) concentrations and on fractional excretion (FE) of these electrolytes. Thirty-two fresh equine urine samples were analysed between December 2016 and July 2020. Complete urinalysis (stick and sediment) was performed on all samples. Ca, Mg, P and creatinine concentrations were measured in supernatant of centrifug...
Post-mortem feasibility of dual-energy computed tomography in the detection of bone edema-like lesions in the equine foot: a proof of concept.
Frontiers in veterinary science    January 4, 2024   Volume 10 1201017 doi: 10.3389/fvets.2023.1201017
Germonpru00e9 J, Vandekerckhove LMJ, Raes E, Chiers K, Jans L, Vanderperren K.In this proof-of-concept study, the post-mortem feasibility of dual-energy computed tomography (DECT) in the detection of bone edema-like lesions in the equine foot is described in agreement with the gold standard imaging technique, which is magnetic resonance imaging (MRI). Unassigned: A total of five equine cadaver feet were studied, of which two were pathological and three were within normal limits and served as references. A low-field MRI of each foot was performed, followed by a DECT acquisition. Multiplanar reformations of DECT virtual non-calcium images were compared with MRI for the de...
Phospholipase C Zeta 1 (PLCZ1): The Function and Potential for Fertility Assessment and In Vitro Embryo Production in Cattle and Horses.
Veterinary sciences    December 11, 2023   Volume 10, Issue 12 698 doi: 10.3390/vetsci10120698
Gonzalez-Castro RA, Carnevale EM.Phospholipase C Zeta 1 (PLCZ1) is considered a major sperm-borne oocyte activation factor. After gamete fusion, PLCZ1 triggers calcium oscillations in the oocyte, resulting in oocyte activation. In assisted fertilization, oocyte activation failure is a major cause of low fertility. Most cases of oocyte activation failures in humans related to male infertility are associated with gene mutations and/or altered PLCZ1. Consequently, PLCZ1 evaluation could be an effective diagnostic marker and predictor of sperm fertilizing potential for in vivo and in vitro embryo production. The characterization ...
Human neutrophil extracellular traps do not impair in vitro Toxoplasma gondii infection.
Frontiers in immunology    December 5, 2023   Volume 14 1282278 doi: 10.3389/fimmu.2023.1282278
Macedo IS, Lara FA, Barbosa HS, Saraiva EM, Menna-Barreto RFS, Mariante RM., responsible for causing toxoplasmosis, is a prevalent food and waterborne pathogen worldwide. It commonly infects warm-blooded animals and affects more than a third of the global human population. Once ingested, the parasite enters the host's small intestine and rapidly disseminates throughout the body via the bloodstream, infiltrating various tissues. Leukocyte-driven responses are vital against , with neutrophils playing a dual role: swiftly recruited to infection sites, releasing inflammatory mediators, and serving as a replication hub and Trojan horses, aiding parasite spread. Neutrophil...
Erosion behaviour of human, bovine and equine dental hard tissues.
Scientific reports    November 10, 2023   Volume 13, Issue 1 19617 doi: 10.1038/s41598-023-46759-9
Hertel S, Basche S, Schmidt V, Staszyk C, Hannig C, Sterzenbach T, Hannig M.Dental hard tissues from different species are used in dental research, but little is known about their comparability. The aim of this study was to compare the erosive behaviour of dental hard tissues (enamel, dentin) obtained from human, bovine and equine teeth. In addition, the protective effect of the pellicle on each hard tissue under erosive conditions was determined. In situ pellicle formation was performed for 30 min on enamel and dentin samples from all species in four subjects. Calcium and phosphate release was assessed during 120 s of HCl incubation on both native and pellicle-cove...
Changes in the nutrient profile and the load of mycotoxins, phytoestrogens, and pesticides in horse pastures during spring and summer in Austria.
Journal of equine veterinary science    November 3, 2023   Volume 131 104958 doi: 10.1016/j.jevs.2023.104958
Son V, Penagos-Tabares F, Hollmann M, Khiaosa-Ard R, Sulyok M, Krska R, Zebeli Q.Pastures are used for grazing and the production of conserved roughage in horses. Yet, the nutritional profile of the forage varies from spring to late summer, affecting equine nutrient supply and health. In addition, environmental factors may also favor plant contaminants such as mycotoxins. This study aimed to determine the nutritional profile and contaminant load of selected horse pastures from early spring till late summer. The nutrient composition (main macronutrients, macro elements and trace elements), as well as mycotoxins, metabolites, pesticides, and plant-derived compounds of seven ...
Expression of Cannabinoid Receptors in the Trigeminal Ganglion of the Horse.
International journal of molecular sciences    November 3, 2023   Volume 24, Issue 21 15949 doi: 10.3390/ijms242115949
Zamith Cunha R, Semprini A, Salamanca G, Gobbo F, Morini M, Pickles KJ, Roberts V, Chiocchetti R.Cannabinoid receptors are expressed in human and animal trigeminal sensory neurons; however, the expression in the equine trigeminal ganglion is unknown. Ten trigeminal ganglia from five horses were collected post-mortem from an abattoir. The expression of cannabinoid receptors type 1 (CB1R) and type 2 (CB2R), and the cannabinoid-related receptors like transient receptor potential vanilloid type 1 (TRPV1), peroxisome proliferator-activated receptor gamma (PPARɣ), and G protein-related receptor 55 (GPR55) in the trigeminal ganglia (TG) of the horse were studied, using immunofluorescence on cry...
Arterial Blood Gas, Electrolyte and Acid-Base Values as Diagnostic and Prognostic Indicators in Equine Colic.
Animals : an open access journal from MDPI    October 17, 2023   Volume 13, Issue 20 3241 doi: 10.3390/ani13203241
Viterbo L, Hughes J, Milner PI, Bardell D.The study aimed to investigate if arterial blood analysis in conscious horses presenting with signs of colic and breathing ambient air had diagnostic or prognostic value. Arterial blood samples from 352 horses presenting with colic at a university equine referral hospital were analysed for pH, partial pressure of carbon dioxide (PaCO), partial pressure of oxygen (PaO), concentrations of sodium (Na), potassium (K), ionised calcium (Ca) and chloride (Cl), actual and standardised plasma bicarbonate concentration (HCO (P) and HCO (P, st)), blood and extracellular fluid base excess (Base (B) and Ba...
Review of local extravascular delivery systems for chemotherapeutic agents in small animals and horses.
The Canadian veterinary journal = La revue veterinaire canadienne    October 2, 2023   Volume 64, Issue 10 957-967 doi: 10.1111/vsu.13506
Risselada M, Worth DB.Tumors of various sizes and locations can create a treatment dilemma in achieving adequate surgical margins when sufficient free tissue is not available for closure. Extravascular local adjunctive chemotherapy has been investigated clinically to aid in achieving local tumor control in animals with naturally occurring neoplastic disease. Local chemotherapy can be an alternative primary or a local adjunctive treatment. This is a summary of relevant findings of and studies on local chemotherapeutic delivery through carrier media, together with a summary of outcomes of clinical use of local deli...
Long-term follow-up of equine incisor endodontic treatments using an orthograde technique.
Journal of the American Veterinary Medical Association    September 25, 2023   Volume 261, Issue S2 S96-S101 doi: 10.2460/javma.23.07.0398
du Toit N, Pearce CJ.This retrospective clinical study was performed to determine the suitability and success rate of endodontic treatment of equine incisors presenting with apical and periapical disease. Methods: All horses presented to a referral equine dental clinic between March 2013 and December 2019 specifically selected as candidates suitable for endodontic treatment were included in this study. Methods: Initial clinical and radiographic presentation of incisor disorder cases suitable for endodontic treatment were recorded (88 incisors) and follow-up examination for long-term viability (8 months to 5 years)...
Heterologous Seminal Plasma Reduces the Intracellular Calcium and Sperm Viability of Cryopreserved Stallion Spermatozoa.
Biopreservation and biobanking    July 19, 2023   doi: 10.1089/bio.2022.0197
Talluri TR, Kumaresan A, Paul N, Elango K, Raval K, Nag P, Legha RA, Pal Y.Despite the vital role of seminal plasma (SP) in maintaining sperm function and aiding gamete interaction in many species, SP is usually removed before cryopreservation of stallion sperm to improve cryosurvival of sperm. The present study assessed if the vital sperm functional parameters of genetically superior stallions producing poor quality semen can be enhanced by the supplementation of heterologous SP from the stallion producing high quality semen. Spermatozoa from poor quality semen producing stallions were divided into three aliquots: two aliquots were supplemented with SP obtained from...
Assessment of calcium, phosphorus, magnesium, vitamin D and PTH levels in sera of lame horses.
Veterinary medicine and science    July 19, 2023   doi: 10.1002/vms3.1198
Etemadi F, Tabatabaei Naeini A, Aminlari M.Minerals play vital roles in physiologic processes. Calcium, phosphorus and magnesium are common minerals. Secretion, absorption and homeostasis of these minerals are affected by associations between the active vitamin D and parathyroid hormone (PTH). Objective: The aim of this study was to assess concentrations of vitamin D, PTH and minerals, such as calcium, phosphorus and magnesium in horse sera, as well as associations of these values with lameness in horses. In the references, the reasons for lameness can be deficiency and imbalance of minerals, and clinical observations also confirm this...
Study of Pansalt® or Laminaria Ochroleuca seaweed powder as potential NaCl replacers in dry-cured foal “cecina”.
Meat science    June 15, 2023   Volume 204 109253 doi: 10.1016/j.meatsci.2023.109253
Cittadini A, Domu00ednguez R, Sarriu00e9s MV, Pateiro M, Lorenzo JM.This study aimed to evaluate the effect of the total or partial replacement of NaCl by a commercial low-salt mixture Pansalt® (T1) or Kombu (L. ochroleuca) seaweed powder (T2), respectively, on the quality properties of dry-cured foal "cecina". Salt reformulation unaffected fat percentages, decreased (P < 0.001) moisture and ash contents, while protein amounts increased (P < 0.001) in the T2 batch. Considering the physicochemical variables, any significant differences were observed among batches, except for a*, water activity, and springiness values. T1 and T2 samples presented a si...
Effect of Physical Exercise in Hemogasometric and Electrolytic Profiles of Young Mangalarga Marchador Horses Beginning Training for Gait Competitions.
Journal of equine veterinary science    May 23, 2023   Volume 127 104823 doi: 10.1016/j.jevs.2023.104823
Nogueira WA, Ferreira Padilha FG, Angelo Luiz M, da Silva Salomu00e3o N, Monteiro Fonseca AB, Reis Ferreira AM.This study aimed to analyze effect of physical exercise in hemogasometric and electrolytic profiles of young Mangalarga Marchador horses beginning training for gait competitions. Six Mangalarga Marchador gaited horses with 6 months of training were evaluated. The ages ranged from three and a half to five years and included four stallions and two mares, with mean (±S.D.) body weight of 435±30Kg. Venous blood samples were collected from the horses, rectal temperature and HR were measured before and immediately after the gait test, and blood samples were used for hemogasometric and laboratory a...
Effects of calcium supplementation to resuscitation fluids in endurance horses: A randomized, blinded, clinical trial.
Journal of veterinary internal medicine    May 2, 2023   Volume 37, Issue 3 1216-1222 doi: 10.1111/jvim.16715
Fielding CL, Deane EL, Major DS, Mayer JR, Love JC, Peralez MS, Magdesian KG.The addition of calcium to resuscitation fluids is a common practice in horses, but studies evaluating the effects of calcium supplementation are limited. In healthy horses, decreases in heart rate and changes in serum electrolyte concentrations have been reported. Objective: Calcium gluconate administration at a rate of 0.4 mg/kg/min to eliminated endurance horses with metabolic problems will affect heart rate, gastrointestinal sounds, and serum electrolyte concentrations. Methods: Endurance horses eliminated from the Tevis Cup 100-mile (160 km) endurance ride for metabolic problems and req...
Concentration of mineral and heavy metals in raw mare (horse) milk consumed in Yazd, Iran: A risk assessment study.
Veterinary medicine and science    April 17, 2023   Volume 9, Issue 4 1592-1598 doi: 10.1002/vms3.1138
Alipour A, Akrami Mohajeri F, Javdan G, Pourramezani F, Fallahzadeh H, Khalili Sadrabad E.In recent years, the mare's milk has been introduced as a rich source of nutrients with hypoallergic characteristics which is widely used for Iranian infants. The present study aimed to investigate the heavy metal concentration of mare's milk and its consumption risk assessment. About 88 mare's milk was collected from Yazd, the centre of Iran, during the summer of 2020. The raw mare's milk was digested and analysed for mineral and heavy metal content (As, Ca, Cd, Co, Cu, Fe, Mg, Mn, Ni, P, Pb and Zn) by ICP-OES. To estimate the health hazard for consumers the Estimated Daily Intake (EDI), Haza...
Milk from Halari Donkey Breed: Nutritional Analysis, Vitamins, Minerals, and Amino Acids Profiling.
Foods (Basel, Switzerland)    February 16, 2023   Volume 12, Issue 4 853 doi: 10.3390/foods12040853
Garhwal R, Bhardwaj A, Sangwan K, Mehra R, Pal Y, Nayan V, Iquebal MA, Jaiswal S, Kumar H.This current research set out to characterize Halari donkey milk by investigating its nutritional constituents, including its proximate analysis, water activity, titratable acidity, energy, and microbiological analysis. A comprehensive profiling of vitamins, minerals, and amino acids was also carried out. It was found that the composition of Halari donkey milk was consistent with previously published donkey milk literature and was comparable to that of human milk. Halari donkey milk has low 0.86 ± 0.04% fat content, 2.03 ± 0.03% protein content, 0.51 ± 0.05% ash content, and high 5.75 ± 0....
Hydroelectrolytic and Acid-Base Parameters after 80 to 115 km Endurance Races (Raid Uruguayo) and Their Association with the Comfort Index.
Animals : an open access journal from MDPI    February 14, 2023   Volume 13, Issue 4 670 doi: 10.3390/ani13040670
Marichal G, Trigo P, Soto C, Meikle A, Suu00e1rez G.The Raid Uruguayo (RAID) is an equestrian endurance competition. This study characterized the hydroelectrolytic parameters (Na, K, Cl, tCa, and iCa), hematocrit (Ht), total plasma protein (TPP), and blood pH from 900 equine athletes (finishers and non-finishers) competing over distances of 80 to 115 km under different climate conditions. Paired blood samples were taken prior to the start of the competition (sample 1) and at the end of the race or at the time of leaving the competition (sample 2). The association of the comfort index (CI: low, moderate, and high) with blood parameters was evalu...
Sample stability and heparin interference in ionized calcium and ionized magnesium measurements in horses using the Stat Profile Prime Plus co-oximetry electrolyte analyzer.
Veterinary clinical pathology    February 6, 2023   Volume 52, Issue 2 252-260 doi: 10.1111/vcp.13200
Sanmartu00ed J, Robles-Guirado JA, Jose-Cunilleras E, Bassols A.The determination of iCa and iMg is important in veterinary medicine, but their immediate determination in whole blood is not always possible. Their stability in other sample types and the existence of interferences must be evaluated before its use. Objective: We aimed to analyze the effects of storage time on the stability of iCa, iMg, and other analytes in whole blood, plasma, and serum samples in horses and assess the interference of heparin in these measurements. Methods: Whole blood, heparin-plasma, and serum samples from 10 horses were stored at 4°C and analyzed 1, 2, 3, 4, 5, 6, 7, 8, ...
Donkey Milk: An Overview of its Chemical Composition and Main Nutritional Properties or Human Health Benefit Properties.
Journal of equine veterinary science    January 14, 2023   Volume 121 104225 doi: 10.1016/j.jevs.2023.104225
The donkey milk has a remarkable similarity to human milk, in addition to its valuable nutritional composition and content of numerous immune factors. The donkey milk is the subject of research worldwide, and data from the literature suggest significant differences with respect to the contents of individual components. However, some basic characteristics of donkey milk have been established: low contents of fat and cholesterol, total proteins and casein and high contents of lactose, whey proteins, calcium, selenium, and Vitamin D3. The donkey milk is rich in various protective proteins (α-lac...
Tracing Acid-Base Variables in Exercising Horses: Effects of Pre-Loading Oral Electrolytes.
Animals : an open access journal from MDPI    December 24, 2022   Volume 13, Issue 1 73 doi: 10.3390/ani13010073
Waller AP, Lindinger MI.Oral electrolyte supplementation may influence acid-base state during exercise due to the intestinal absorption of administered water and electrolytes used to mitigating sweat losses. This study examined the effect of pre-exercise electrolyte supplementation (3 and 8 L) on plasma acid-base variables at rest, during moderate intensity exercise and during recovery. It was hypothesized that electrolyte supplementation will result in improved acid-base state compared to the alkalosis typical of prolonged exercise. In randomized crossover fashion, four horses were administered 3 L or 8 L of a hypot...
In vitro aging of stallion spermatozoa during prolonged storage at 5°C.
Cytometry. Part A : the journal of the International Society for Analytical Cytology    December 22, 2022   Volume 103, Issue 6 479-491 doi: 10.1002/cyto.a.24712
Umair M, Claes A, Buijtendorp M, Cuervo-Arango J, Stout TAE, Henning H.Artificial insemination with chilled stallion semen is hampered by a limited period of maximum fertility maintenance (24-48 h). This study used multiparametric flow cytometry to simultaneously measure reactive oxygen species (ROS) production, mitochondrial function or [Ca ] and plasma membrane fluidity in viable, acrosome-intact spermatozoa, with the aim of providing insight into changes in sperm function during storage at 5°C. High proportions of viable and acrosome-intact spermatozoa (71 ± 8%) remained after 96 h of storage demonstrating that the basic integrity of the cells was we...
Changes in haematological and biochemical parameters in blood serum of horses during exposition to workload stress.
Heliyon    December 13, 2022   Volume 8, Issue 12 e12241 doi: 10.1016/j.heliyon.2022.e12241
Massu00e1nyi M, Halo M, Massu00e1nyi P, Mlynekovu00e1 E, Greu0144 A, Formicki G, Halo M.Health state of animals undergoing experimental procedures is an important topic nowadays, as even the small changes can influence the outcome of entire outcomes. Main aim of the study was to evaluate the influence of horse training on variety of blood parameters including mineral profile, energy profile, hepatic profile and haematology. In the experiment, the studied group of horses underwent training programme which consisted of transportation, lounging, riding, jumping, racing, treadmill training and shoeing. Blood samples were collected and later evaluated at the beginning, in the middle a...
Measuring Biochemical Variables and Serum Amyloid A (SAA) in Working Mules in Central Italy.
Animals : an open access journal from MDPI    October 16, 2022   Volume 12, Issue 20 2793 doi: 10.3390/ani12202793
Bazzano M, Arfuso F, Bonfili L, Eleuteri AM, McLean A, Serri E, Spaterna A, Laus F.According to FAO reports, the global mule population counts about 9 million mules. This hybrid cross of a male donkey and a female horse is mainly used for draft purposes because they are thought to be strong and hardy animals. Most consider mules to be less susceptible to disease and fatigue compared to horses. Therefore, the aim of this study was to investigate the effects of fieldwork on biochemical variables and serum amyloid A in working mules. Blood samples were collected from 10 healthy, female, working mules before and after 8 h of fieldwork. According to statistical analysis, a signif...
Effects of total parenteral nutrition associated with glutamine, enteral fluid therapy with or without glutamine, and fluid therapy on the acid-base and electrolyte balance of horses starved after exploratory laparotomy.
Brazilian journal of veterinary medicine    October 14, 2022   Volume 44 e003222 doi: 10.29374/2527-2179.bjvm003222
Melo UP, Palhares MS, Ferreira C, Leme FOP, Gheller VA.This study aimed to evaluate the effects of the total parenteral nutrition associated with glutamine, enteral fluid therapy with or without glutamine, and fluid therapy on the acid-base and electrolyte balance of horses starved after exploratory laparotomy. Sixteen healthy male and female adult horses of mixed breed, aged between 4 and 14 years, and having a mean body weight of 248.40 ± 2.28 kg and a body score index of 3-4 (scale of 1-5) were divided into four groups with four animals per group. After an adaptation period of 30 days, they were randomly divided into four experimental groups: ...
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