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Topic:Horses

"Horses" is a broad topic that encompasses various aspects of equine biology, behavior, and management. This category includes studies on the anatomy, physiology, and genetics of horses, as well as their behavior, nutrition, and care. Research in this area may also cover the historical and cultural significance of horses, their roles in agriculture, sport, and therapy, and the challenges associated with their conservation and welfare. The page aggregates peer-reviewed research articles and scholarly studies that explore the multifaceted relationships between humans and horses, examining both scientific and socio-economic perspectives.
Cardiac output affects the response to pulsed inhaled nitric oxide in mechanically ventilated anesthetized ponies determined by CT angiography of the lung.
American journal of veterinary research    April 17, 2023   Volume 84, Issue 6 ajvr.23.01.0005 doi: 10.2460/ajvr.23.01.0005
Auckburally A, Wiklund M, Lord P, Nyman G.To measure changes in regional lung perfusion using CT angiography in mechanically ventilated, anesthetized ponies administered pulsed inhaled nitric oxide (PiNO) during hypotension and normotension. Methods: 6 ponies for anesthetic 1 and 5 ponies for anesthetic 2. Methods: Ponies were anesthetized on 2 separate occasions, mechanically ventilated, and placed in dorsal recumbency within the CT gantry. Pulmonary arterial, right atrial, and facial arterial catheters were placed. During both anesthetics, PiNO was delivered for 60 minutes and then discontinued. Anesthetic 1: hypotension (mean arter...
Sustained-release deslorelin acetate implants disrupt oestrous cyclicity in the mare.
Australian veterinary journal    April 16, 2023   Volume 101, Issue 7 275-278 doi: 10.1111/avj.13244
Joonè CJ, Cavalieri J.There is a need for a safe, effective and practical method of oestrus suppression in the mare. The aim of this study was to monitor ovarian activity in mares exposed to either 9.4 or 28.2 mg deslorelin acetate, a GnRH agonist, in the form of a sustained-release implant. Following oestrus synchronisation, mares were randomly assigned to one of three groups (n = 4 per group) and administered either one (Des1 group; 9.4 mg) or three (Des3 group; 28.2 mg) implants of deslorelin acetate (Suprelorin-12, Virbac Australia) or one blank implant (Control group; Virbac Australia). Mares underwe...
Assessment of Magnetic Resonance Imaging Artefacts Caused by Equine Anaesthesia Equipment: A Cadaver Study.
Journal of equine veterinary science    April 16, 2023   Volume 126 104492 doi: 10.1016/j.jevs.2023.104492
Testa B, Biggi M, Byrne CA, Bell A.Acquisition of magnetic resonance images of the equine limb is still sometimes conducted under general anaesthesia. Despite low-field systems allow the use of standard anaesthetic equipment, possible interferences of the extensive electronic componentry of advanced anaesthetic machines on image quality is unknown. This prospective, blinded, cadaver study investigated the effects of seven standardised conditions (Tafonius positioned as in clinical cases, Tafonius on the boundaries of the controlled area, anaesthetic monitoring only, Mallard anaesthetic machine, Bird ventilator, complete electro...
Equine Brucellosis: Current Understanding and Challenges.
Journal of equine veterinary science    April 16, 2023   Volume 127 104298 doi: 10.1016/j.jevs.2023.104298
Dorneles EMS, Santana JA, Costa ACTRB, Junqueira DG, Heinemann MB, Lage AP.Brucellosis in equines, including horses, donkeys, and mules, is characterized by abscesses in tendons, bursae, and joints. Reproductive disorders, which are common in other animals, are rare in both males and females. Joint breeding of horses, cattle, and pigs was found as the main risk factor for equine brucellosis, with the transmission from equines to cattle or among equines possible, although unlikely. Hence, evaluation of the disease in equines can be considered an indirect indicator of the effectiveness of brucellosis control measures employed for other domestic species. Generally, the ...
Changes in Calprotectin (S100A8-A9) and Aldolase in the Saliva of Horses with Equine Gastric Ulcer Syndrome.
Animals : an open access journal from MDPI    April 16, 2023   Volume 13, Issue 8 doi: 10.3390/ani13081367
Muñoz-Prieto A, Contreras-Aguilar MD, Cerón JJ, Ayala de la Peña I, Martín-Cuervo M, Eckersall PD, Holm Henriksen IM, Tecles F, Hansen S.Equine gastric ulcer syndrome (EGUS) is a highly prevalent disease that affects horses worldwide. Within EGUS, two different forms have been described: equine squamous gastric disease (ESGD) and equine glandular gastric disease (EGGD). The associated clinical signs cause detrimental activity performance, reducing the quality of life of animals. Saliva can contain biomarkers for EGUS that could be potentially used as a complementary tool for diagnosis. The objective of this work was to evaluate the measurements of calprotectin (CALP) and aldolase in the saliva of horses as potential biomarkers ...
Multilineage Differentiation Potential of Equine Adipose-Derived Stromal/Stem Cells from Different Sources.
Animals : an open access journal from MDPI    April 15, 2023   Volume 13, Issue 8 1352 doi: 10.3390/ani13081352
Stage HJ, Trappe S, Söllig K, Trachsel DS, Kirsch K, Zieger C, Merle R, Aschenbach JR, Gehlen H.The investigation of multipotent stem/stromal cells (MSCs) in vitro represents an important basis for translational studies in large animal models. The study's aim was to examine and compare clinically relevant in vitro properties of equine MSCs, which were isolated from abdominal (abd), retrobulbar (rb) and subcutaneous (sc) adipose tissue by collagenase digestion (ASCs-) and an explant technique (ASCs-). Firstly, we examined proliferation and trilineage differentiation and, secondly, the cardiomyogenic differentiation potential using activin A, bone morphogenetic protein-4 and Dickkopf-1. Fi...
Heat stress in horses: a literature review.
International journal of biometeorology    April 15, 2023   Volume 67, Issue 6 957-973 doi: 10.1007/s00484-023-02467-7
Kang H, Zsoldos RR, Sole-Guitart A, Narayan E, Cawdell-Smith AJ, Gaughan JB.Healthy adult horses can balance accumulation and dissipation of body heat to maintain their body temperature between 37.5 and 38.5 °C, when they are in their thermoneutral zone (5 to 25 °C). However, under some circumstances, such as following strenuous exercise under hot, or hot and humid conditions, the accumulation of body heat exceeds dissipation and horses can suffer from heat stress. Prolonged or severe heat stress can lead to anhidrosis, heat stroke, or brain damage in the horse. To ameliorate the negative effects of high heat load in the body, early detection of heat stress and im...
Risk Factors for Epistaxis in Thoroughbred Flat Races in Japan (2001-2020).
Animals : an open access journal from MDPI    April 14, 2023   Volume 13, Issue 8 doi: 10.3390/ani13081348
Sugiyama F, Takahashi Y, Nomura M, Ebisuda Y, Mukai K, Yoshida T.We investigated the risk factors for epistaxis in Japanese flat races over a 20-year period. The veterinary records of horses identified as having epistaxis by endoscopy on the race day, and the official racing records of all flat races from April to September between 2001 and 2020, were reviewed. The racecourses (n = 10), surface type, surface condition, race class, race distance, race year, sex, age, two training centers, ambient temperature, and body weight on race days were assessed using multivariable logistic regression (p < 0.05). Of 475,709 race starts, 616 (1.30 cases per 1000 starts;...
Identification of telocytes in the oviduct of the mare.
Theriogenology    April 14, 2023   Volume 205 18-26 doi: 10.1016/j.theriogenology.2023.04.012
Etcharren V, Mouguelar H, Aguilar Valenciano JJ.Telocytes (TCs), a recently discovered special type of stromal cells, have been identified in many organs of many species, including the female and male reproductive system, with proposed multiple potential bio-functions such as homeostasis, immunomodulation, tissue remodeling and regeneration, embryogenesis, angiogenesis and even tumorigenesis. The aim of this study was to investigate the existence, and characteristics of telocytes in normal equine oviduct. To identify them, we used routine light microscopy, non-conventional light microscopy (NCLM), transmission electron microscopy (TEM), and...
Lipid metabolites, interleukin-6 and oxidative stress markers in follicular fluid and their association with serum concentrations in mares.
Veterinary research communications    April 14, 2023   Volume 47, Issue 4 2221-2228 doi: 10.1007/s11259-023-10122-0
Hedia M, Leroy JLMR, Govaere J, Van Soom A, Smits K.The application of trans-vaginal ovum pick up (OPU) and intracytoplasmic sperm injection (ICSI) is well established for commercial in vitro embryo production in horses. These assisted reproductive techniques are especially applied during the non-breeding season of the mare. However, little is known about how the health of the oocyte donor may affect the biochemical composition of the follicular fluid (FF) in small and medium-sized follicles routinely aspirated during OPU. This study aimed to investigate associations between systemic and FF concentrations of interleukin-6 (IL-6), total choleste...
Detecting fatigue of sport horses with biomechanical gait features using inertial sensors.
PloS one    April 14, 2023   Volume 18, Issue 4 e0284554 doi: 10.1371/journal.pone.0284554
Darbandi H, Munsters C, Parmentier J, Havinga P.Detection of fatigue helps prevent injuries and optimize the performance of horses. Previous studies tried to determine fatigue using physiological parameters. However, measuring the physiological parameters, e.g., plasma lactate, is invasive and can be affected by different factors. In addition, the measurement cannot be done automatically and requires a veterinarian for sample collection. This study investigated the possibility of detecting fatigue non-invasively using a minimum number of body-mounted inertial sensors. Using the inertial sensors, sixty sport horses were measured during walk ...
Development of a promising antigenic cocktail for the global detection of Babesia caballi in horse by ELISA.
PloS one    April 14, 2023   Volume 18, Issue 4 e0284535 doi: 10.1371/journal.pone.0284535
El-Sayed SAE, Rizk MA, Baghdadi HB, Ringo AE, Sambuu G, Nugraha AB, Igarashi I.In this study, we designed novel truncated Babesia caballi (B. caballi) recombinant proteins from the previously used B. caballi proteins; 134-Kilodalton Protein (rBC134) and Merozoite Rhoptry 48 Protein (rBC48). Then, we evaluated the diagnostic performance of the newly designed proteins when used as a single antigen or when used as cocktail antigen consists of rBC134 full length (rBC134f) + newly designed rBC48 (rBC48t) or newly designed rBC134 (rBC134t) + rBC48t for the detection of B. caballi infection in horse using indirect enzyme-linked immunosorbent assay (iELISA). We used one dose and...
Flow-controlled expiration reduces positive end-expiratory pressure requirement in dorsally recumbent, anesthetized horses.
Frontiers in veterinary science    April 14, 2023   Volume 10 1135452 doi: 10.3389/fvets.2023.1135452
Brandly JE, Midon M, Douglas HF, Hopster K.Equine peri-anesthetic mortality is higher than that for other commonly anesthetized veterinary species. Unique equine pulmonary pathophysiologic aspects are believed to contribute to this mortality due to impairment of gas exchange and subsequent hypoxemia. No consistently reliable solution for the treatment of peri-anesthetic gas exchange impairment is available. Flow-controlled expiration (FLEX) is a ventilatory mode that linearizes gas flow throughout the expiratory phase, reducing the rate of lung emptying and alveolar collapse. FLEX has been shown to improve gas exchange and pulmonary me...
Direct and culture-enriched 16S rRNA sequencing of cecal content of healthy horses and horses with typhlocolitis.
PloS one    April 13, 2023   Volume 18, Issue 4 e0284193 doi: 10.1371/journal.pone.0284193
Zakia LS, Gomez DE, Caddey BB, Boerlin P, Surette MG, Arroyo LG.Next generation sequencing has demonstrated that alpha diversity of the fecal microbiota is significantly altered in horses with typhlocolitis. The objective of this study was to evaluate the bacterial composition of the cecum content of horses with and without typhlocolitis through direct and culture-enriched 16S gene sequencing of six healthy horses and six horses with acute typhlocolitis; a case-control study design. Cecal content was collected after euthanasia. An aliquot was used for direct 16S gene sequencing. Another was serially diluted with brain heart infusion (BHI) and plated onto f...
Water stabilizes an alternate turn conformation in horse heart myoglobin.
Scientific reports    April 13, 2023   Volume 13, Issue 1 6094 doi: 10.1038/s41598-023-32821-z
Bronstein A, Marx A.Comparison of myoglobin structures reveals that protein isolated from horse heart consistently adopts an alternate turn conformation in comparison to its homologues. Analysis of hundreds of high-resolution structures discounts crystallization conditions or the surrounding amino acid protein environment as explaining this difference, that is also not captured by the AlphaFold prediction. Rather, a water molecule is identified as stabilizing the conformation in the horse heart structure, which immediately reverts to the whale conformation in molecular dynamics simulations excluding that structur...
Tracking the Source of Human Q Fever from a Southern French Village: Sentinel Animals and Environmental Reservoir.
Microorganisms    April 13, 2023   Volume 11, Issue 4 1016 doi: 10.3390/microorganisms11041016
Laidoudi Y, Rousset E, Dessimoulie AS, Prigent M, Raptopoulo A, Huteau Q, Chabbert E, Navarro C, Fournier PE, Davoust B., also known as the causal agent of Q fever, is a zoonotic pathogen infecting humans and several animal species. Here, we investigated the epidemiological context of from an area in the Hérault department in southern France, using the One Health paradigm. In total, 13 human cases of Q fever were diagnosed over the last three years in an area comprising four villages. Serological and molecular investigations conducted on the representative animal population, as well as wind data, indicated that some of the recent cases are likely to have originated from a sheepfold, which revealed bacterial c...
A Systematic Review of Current Applications of Fecal Microbiota Transplantation in Horses.
Veterinary sciences    April 13, 2023   Volume 10, Issue 4 doi: 10.3390/vetsci10040290
Tuniyazi M, Wang W, Zhang N.Fecal microbiota transplantation (FMT) is a technique involving transferring fecal matter from a healthy donor to a recipient, with the goal of reinstating a healthy microbiome in the recipient's gut. FMT has been used in horses to manage various gastrointestinal disorders, such as colitis and diarrhea. To evaluate the current literature on the use of FMT in horses, including its efficacy, safety, and potential applications, the authors conducted an extensive search of several databases, including PubMed, MEDLINE, Web of Science, and Google Scholar, published up to 11 January 2023. The authors...
Conservative management of equine tarsal collateral ligament injuries may allow return to normal performance.
Journal of the American Veterinary Medical Association    April 11, 2023   Volume 261, Issue 7 995-1003 doi: 10.2460/javma.22.12.0597
Fraschetto C, Dancot M, Vandersmissen M, Denoix JM, Coudry V.To describe type and distribution of tarsal collateral ligament (CL) injuries and to assess the long-term outcome in horses treated conservatively. Methods: 78 horses (median age, 7 years [IQR, 4 to 9.75 years]) of different breeds and disciplines. Methods: Retrospective analysis (2000 through 2020) of horses with tarsal CL lesions diagnosed on ultrasound. The resting time, ability to return to work, and performance level after the injury were compared between horses having a single ligament (group S) or multiple ligaments (group M) affected and according to the case severity. Results: Most of...
Homocysteine-Potential Novel Diagnostic Indicator of Health and Disease in Horses.
Animals : an open access journal from MDPI    April 11, 2023   Volume 13, Issue 8 1311 doi: 10.3390/ani13081311
Gołyński M, Metyk M, Ciszewska J, Szczepanik MP, Fitch G, Bęczkowski PM.Homocysteine is an endogenous, non-protein sulfuric amino acid, an intermediate metabolite formed by the methionine transmethylation reaction. Its elevated serum concentration in humans, hyperhomocysteinemia, is a sensitive indicator and a risk factor for coagulation disorders, cardiovascular diseases and dementia. However, the role of homocysteine in veterinary species has not been unequivocally established. Although some research has been conducted in dogs, cats, cattle and pigs, relatively few studies on homocysteine have been conducted in horses. So far, it has been established in this spe...
Comparison of Trap and Equine Attraction to Mosquitoes.
Insects    April 11, 2023   Volume 14, Issue 4 374 doi: 10.3390/insects14040374
Dilling SC, TenBroeck SH, Hogsette JA, Kline DL.Mosquitoes are pests of horses, but mosquito trap efficacy data, especially the ability of traps to protect horses, are lacking. Studies were conducted to investigate the comparative attraction between traps and horses, increase trap attraction by adding horse odors to the airstream of a trap, determine the spatial distribution of adult mosquitoes, estimate the numbers of mosquitoes feeding on horses, determine the relative attraction of horses to mosquitoes, and estimate the range of mosquitoes' attraction between two horses. When a horse and a mosquito trap were placed 3.5 m apart, there was...
Development and evaluation of a blocking ELISA for serological diagnosis of equine infectious anemia.
Applied microbiology and biotechnology    April 11, 2023   Volume 107, Issue 10 3305-3317 doi: 10.1007/s00253-023-12504-5
Hu Z, Guo K, Du C, Sun J, Naletoski I, Chu X, Lin Y, Wang X, Barrandeguy M, Samuel M, Wang W, Lau PI, Wernery U, Raghavan R, Wang X.Equine infectious anemia (EIA) is an important viral disease characterized by persistent infection in equids worldwide. Most EIA cases are life-long virus carriers with low antibody reactions and without the appearance of clinical symptoms. A serological test with high sensitivity and specificity is required to detect inapparent infection. In this study, a B-cell common epitope-based blocking ELISA (bELISA) was developed using a monoclonal antibody together with the EIAV p26 protein labelled with HRP. The test has been evaluated against the standard and with field serum samples globally. This ...
Molecular Identification of Culicoides Species and Host Preference Blood Meal in the African Horse Sickness Outbreak-Affected Area in Hua Hin District, Prachuap Khiri Khan Province, Thailand.
Insects    April 8, 2023   Volume 14, Issue 4 doi: 10.3390/insects14040369
Kamyingkird K, Choocherd S, Chimnoi W, Klinkaew N, Kengradomkij C, Phoosangwalthong P, Thammasonthijarern N, Pattanatanang K, Inpankaew T, Phasuk J....African horse sickness (AHS) was reported as an outbreak in Thailand in 2020. Hematophagous insects from the genus Culicoides are the suspected vector responsible for AHS transmission. Horses in Hua Hin district, Prachuab Khiri Khan province, Thailand, were affected and died from AHS in 2020. However, the potential Culicoides species and its host preference blood meal in the affected areas are unknown. To investigate the potential vectors of AHS, Culicoides were collected using ultraviolet light traps placed near horse stables. Six horse farms, including five farms with AHS history and one far...
A case of chronic schistosomiasis in a Dongola stallion (Equus ferus caballus Linnaeus, 1758) from Nigeria.
Journal of parasitic diseases : official organ of the Indian Society for Parasitology    April 8, 2023   Volume 47, Issue 2 442-450 doi: 10.1007/s12639-023-01581-y
Abalaka SE, Audu Z, Kolawole VO, Adeyemo BT, Idoko IS, Okafor ROS, Oyelowo-Abdulraheem FO, Tags SZ, Ogbe AO, Sanni S, Jegede OC, Sani NA, Tenuche OZ....Schistosomiasis is a worldwide snail-borne parasitic infestation of man and animals with acute or chronic phases having devastating sequelae. The present case report focused on post-mortem examination of a cachexic Dongola stallion ( Linnaeus, 1758) in Abuja, Nigeria, that failed to respond to treatment. Typical dense collagenous granulomatous lesions with marked inflammatory responses and fibrosis occurred in the liver and several visceral organs of the horse along with other lesions indicative of systemic collapse. Special Ziehl-Neelsen and Periodic Acid-Schiff staining as well as microbial ...
The Effect of Insect Bite Hypersensitivity on Movement Activity and Behaviour of the Horse.
Animals : an open access journal from MDPI    April 8, 2023   Volume 13, Issue 8 1283 doi: 10.3390/ani13081283
Söderroos D, Ignell R, Haubro Andersen P, Bergvall K, Riihimäki M.Insect bite hypersensitivity (IBH) associated with biting midges is a common allergic skin disease in horses, reducing the welfare of affected horses. This study investigated the effect of IBH on animal welfare and behaviour and assessed a new prophylactic insect repellent. In total, 30 horses were recruited for a prospective cross-over and case-control study. Clinical signs of IBH, inflammatory markers in skin biopsies and behavioural data (direct observations, motion index) were scored longitudinally during two consecutive summers. No differences were observed in the total number of itching...
Endoscopic-guided diagnosis of and interventional stent placement for nasolacrimal duct obstruction in a horse.
Veterinary ophthalmology    April 8, 2023   Volume 26, Issue 3 256-261 doi: 10.1111/vop.13094
Smith LH, Adams L, Derré M, Townsend WM.To describe the successful use of endoscopy to visualize and place a soft canine ureteral stent to relieve a chronic nasolacrimal duct (NLD) obstruction in a horse. Methods: A 7-year-old, Quarter horse gelding. Methods: Under general anesthesia, retrograde nasolacrimal endoscopy was performed using an 8.5 Fr Storz Flex XC ureteroscope through the nasal punctum (NP). An obstructive web of fibrous tissue was visualized approximately 20 cm proximal to the NP. A 0.035″/150 cm hydrophilic guidewire was passed normograde from the ventral lacrimal punctum and used to puncture the stenotic tissu...
Performance assessment of an equine metabolomics model for screening a range of anabolic agents.
Metabolomics : Official journal of the Metabolomic Society    April 7, 2023   Volume 19, Issue 4 38 doi: 10.1007/s11306-023-01985-0
Cloteau C, Dervilly G, Loup B, Delcourt V, Kaabia Z, Bagilet F, Groseille G, Dauriac K, Fisher S, Popot MA, Garcia P, Le Bizec B, Bailly-Chouriberry L.Despite their ban, Anabolic Androgenic Steroids (AAS) are considered as the most important threat for equine doping purposes. In the context of controlling such practices in horse racing, metabolomics has emerged as a promising alternative strategy to study the effect of a substance on metabolism and to discover new relevant biomarkers of effect. Based on the monitoring of 4 metabolomics derived candidate biomarkers in urine, a prediction model to screen for testosterone esters abuse was previously developed. The present work focuses on assessing the robustness of the associated method and def...
Administration safety of medical-grade honey (MGH) in septic synovial structures in horses: 3 cases. Terschuur JA, Coomer RPC, McKane SA.The objective of this study was to investigate the postoperative use of intrasynovial honey as an antimicrobial after treatment for synovial sepsis in horses. One colt and 2 mares were presented with acute lameness, with or without an associated wound. All 3 cases were initially managed with surgical endoscopic or tenoscopic debridement and lavage for treatment of different synovial structures. Collection of synovial fluid was consistent with synovial sepsis and this was diagnosed in each case. All horses subsequently underwent arthroscopic lavage under general anesthetic and intraarticular or...
Effects of concentrated fecal microbiota transplant on the equine fecal microbiota after antibiotic-induced dysbiosis. Di Pietro R, Arroyo LG, Leclere M, Costa M.Bacterial imbalances are observed in intestinal diseases and fecal microbiota transplantation (FMT) has been used to restore the intestinal microbiota of horses. However, there is evidence that the current methods proposed for FMT in horses have limited efficacy. The objective of this study was to concentrate the bacteria present in the donor stool by centrifugation, and to test the effect in horses with antibiotic-induced dysbiosis. One healthy 11-year-old horse was selected as a fecal donor and 9 horses were given trimethoprim sulfadiazine (TMS) for 5 days to induce dysbiosis. Horses receive...
Plasma UCHL-1 as a Biomarker of Brain Injury in Hospitalized Foals With Neonatal Encephalopathy.
Journal of equine veterinary science    April 7, 2023   Volume 126 104490 doi: 10.1016/j.jevs.2023.104490
Ryan CA, Giguère S, Morresey PR.A plasma biomarker such as ubiquitin carboxyl-terminal hydrolase L1 (UCHL-1) to distinguish neonatal encephalopathy (NE) from other disorders and provide prognostic information would be useful for equine practitioners. In this prospective study, plasma UCHL-1 was measured in 331 hospitalized foals ≤4 days of age. Clinical diagnoses of neonatal encephalopathy only (NE group, n = 77), sepsis only (Sepsis group, n = 34), concurrent sepsis and NE (NE+Sepsis group, n = 85), or neither sepsis nor NE (Other group, n = 101) were made by the attending veterinarian. Plasma UCHL-1 concentration...
Effect of dietary iron supplementation on the equine fecal microbiome. Arantes JA, Borges AS, Zakia LS, Surette MG, Weese JS, Costa MC, Arroyo LG.Iron is an essential element for all living organisms, including bacteria, as several virulence factors and replication components are influenced by iron concentration. The objective of this study was to determine whether the composition and diversity of the fecal microbiota of adult horses are affected by supplemental dietary iron. Ten clinically healthy horses were randomly divided into a control and an iron-supplemented group ( = 5). The treated group was supplemented with oral ferrous sulphate monohydrate (720 ppm of iron), whereas the control group received 320 ppm of iron daily for 15 d....