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Summary Background Objectives Study design Methods Results Conclusion Regular exercise benefits horses' overall health; yet data on long‐term physiological adaptations in polo horses remains limited.The study aimed to compare the clinical, haematology and serum biochemical profiles of polo and sedentary horses.Cross‐sectional study.Forty adult mares (20 sedentary and 20 polo), aged 10–15 years, were evaluated. The polo horses had competed in three major high‐goal and nine medium/low‐goal tournaments in the previous 12 months and received daily exercise, while the sedentary horses were for leisure and kept in their stalls most of the time with no physical activity. The study evaluated the following clinical parameters at rest: respiratory rate (RR), rectal temperature (RT), blood pressure (BP) and heart rate (HR). Blood samples were collected from the jugular vein via 10 mL syringes and analysed for haematological and biochemical parameters. Independent t test was used for statistical analysis, with a p ≤ 0.05 considered significant.Polo horses showed significantly lower (p < 0.05) resting respiratory rate, heart rate, systolic blood pressure, and mean arterial pressure compared to sedentary horses. Haematologically, polo horses exhibited higher (p < 0.05) packed cell volume (37.2 ± 5.9% vs. 33.1 ± 3.0%), haemoglobin (12.1 ± 2.1 vs. 10.8 ± 1.0 g/dL), red blood cells (5.9 ± 1.1 vs. 5.2 ± 0.8 × 106/μL) and platelets (127.4 ± 17.7 vs. 108.1 ± 20.5 × 109/μL). Biochemically, polo horses demonstrated higher (p < 0.05) total protein, albumin, globulin, creatinine, total bilirubin, creatine kinase, and high‐density lipoprotein, while showing lower (p < 0.05) aspartate transaminase, alkaline phosphatase, blood urea nitrogen, cholesterol, triglycerides and low‐density lipoprotein. No significant differences (p > 0.05) were observed in rectal temperature, diastolic blood pressure, most white blood cell parameters, glucose, electrolytes and several other biochemical markers.Regular polo training induces favourable physiological adaptations. Future research should track these changes longitudinally and inform improved training and care strategies for equine athletes. [ABSTRACT FROM AUTHOR]

Urban wildlife faces unique physiological and behavioral challenges compared to conspecifics which live in less altered natural habitats. Animals in urban habitats are also exposed to urban stressors and commonly make use of inappropriate food sources from trash bins and dumpsters, which may affect their heath status. The goals of this study were to evaluate overall health of an urban population of eastern gray squirrels (Sciurus carolinensis) and to assess seasonal changes in several health parameters. Squirrels (N = 45) were trapped in Durham, North Carolina, USA and briefly anesthetized with inhalant anesthetic for physical examination, body mass collection, and venipuncture via the femoral vein. Collected blood was analyzed for several health parameters via hematology and biochemistry analyses including assessment of lipids and kidney and liver values. Physical examinations were clinically unremarkable and similar across seasons. Females had higher cholesterol values and lower creatinine values compared to males. Seasonal differences occurred in few parameters and were detected only between summer and either fall or spring. Potassium and monocytes were higher in summer compared to spring, and creatinine, BUN and monocytes were higher in summer compared to fall. Overall, health parameters of urban eastern gray squirrels fell within published reference ranges for the species. Together, these results suggest that urban squirrels in this study population can maintain good health in an anthropogenic habitat. [ABSTRACT FROM AUTHOR]

Diabetes mellitus is a major health challenge that has harmful effects on the quality of life globally as a result of its numerous complications. This study aimed to evaluate the positive modulatory effect of Lawsonia inermis (LI) Linn on various haemobiochemical parameters and lipid profiles in streptozotocin-induced diabetic rats. Ten groups of diabetic rats (n = 5) were orally administered 50 and 100 mg/kg of three different fractions of Lawsonia inermis: metformin (500 mg/kg) and glibenclamide (5 mg/kg), whereas untreated hyperglycaemic and normoglycaemic rats received distilled water. The results showed increased red blood cells (RBC) in treated rats compared to untreated diabetic rats. Hb, MCV, MCH, and MCHC decreased nonsignificantly, whereas WBC increased nonsignificantly. Neutrophil increased non-significantly in diabetic, untreated rats, while all the treatment groups decreased non-significantly. Lymphocytes and monocytes increased non-significantly (p > 0.05). Most treatment groups showed a non-significant (p > 0.05) increase in the platelet count. The ALT levels decreased non-significantly (p > 0.05) compared to normoglycemic rats. The AST levels decreased significantly (p < 0.01). ALP decreased non-significantly in both treated and untreated groups, whereas bilirubin did not show any significant changes. Creatinine and urea levels in untreated diabetic rats increased non-significantly, while treatment groups decreased non-significantly. Lawsonia inermis-treated rats showed significant improvement in erythrocytic fragility, while glycated haemoglobin in untreated-diabetic and glibenclamide-treated rats increased significantly (p > 0.001). Triglycerides and high-density lipoprotein decreased non-significantly, while low-density lipoprotein increased non-significantly (p > 0.05) in diabetic untreated rats. All treatment groups showed a non-significant (P > 0.05) decrease in low-density lipoprotein. Lawsonia inermis showed a significant positive modulatory effect on the haemobiochemical changes, glycated Hb, osmotic fragility, and lipid profile in streptozotocin-induced diabetic rats.

Diabetes mellitus is a major health challenge that has harmful effects on the quality of life globally as a result of its numerous complications. This study aimed to evaluate the positive modulatory effect of Lawsonia inermis (LI) Linn on various haemobiochemical parameters and lipid profiles in streptozotocin-induced diabetic rats. Ten groups of diabetic rats (n = 5) were orally administered 50 and 100 mg/kg of three different fractions of Lawsonia inermis: metformin (500 mg/kg) and glibenclamide (5 mg/kg), whereas untreated hyperglycaemic and normoglycaemic rats received distilled water. The results showed increased red blood cells (RBC) in treated rats compared to untreated diabetic rats. Hb, MCV, MCH, and MCHC decreased nonsignificantly, whereas WBC increased nonsignificantly. Neutrophil increased non-significantly in diabetic, untreated rats, while all the treatment groups decreased non-significantly. Lymphocytes and monocytes increased nonsignificantly (p > 0.05). Most treatment groups showed a non-significant (p > 0.05) increase in the platelet count. The ALT levels decreased non-significantly (p > 0.05) compared to normoglycemic rats. The AST levels decreased significantly (p<0.01). ALP decreased non-significantly in both treated and untreated groups, whereas bilirubin did not show any significant changes. Creatinine and urea levels in untreated diabetic rats increased non-significantly, while treatment groups decreased non-significantly. Lawsonia inermis-treated rats showed significant improvement in erythrocytic fragility, while glycated haemoglobin in untreateddiabetic and glibenclamide-treated rats increased significantly (p > 0.001). Triglycerides and high-density lipoprotein decreased non-significantly, while lowdensity lipoprotein increased non-significantly (p > 0.05) in diabetic untreated rats. All treatment groups showed a non-significant (P > 0.05) decrease in low-density lipoprotein. Lawsonia inermis showed a significant positive modulatory effect on the haemobiochemical changes, glycated Hb, osmotic fragility, and lipid profile in streptozotocin-induced diabetic rats. [ABSTRACT FROM AUTHOR]

Simple Summary: Canine hemangiosarcoma and hemangioma are neoplasms of blood vessel-lining cells commonly found in dogs. Most tumors are often asymptomatic; however, ruptures of the affected organs, especially the spleen, frequently lead to hemorrhage or even hemorrhagic shock before diagnosis. This retrospective study used blood as a practical specimen and routine assessments, including hematology, serum biochemistry, and coagulation profiles, to develop predictive models for the early detection of canine vascular neoplasms. Analyses revealed associations between anemia and lymphopenia with hemangioma diagnosis, while anemia, lymphopenia, and hyperfibrinogenemia were associated with hemangiosarcoma diagnosis. These findings emphasize the importance of integrating comprehensive laboratory data with clinical information to facilitate early diagnosis and management of these critical conditions. Vascular neoplasms, including hemangiosarcoma (HSA) and hemangioma (HMA), are more common in dogs than other domestic animal species; however, comprehensive laboratory screening tests for early diagnosis are currently limited. The aims of this study were to investigate general signalments, anatomic locations, and clinicopathological abnormalities of dogs diagnosed with vascular neoplasms and to determine the diagnostic significance of these abnormalities. Retrospective data of dogs with HMA, HSA, and healthy dogs were analyzed. Dogs with HMA and HSA were seniors, with mixed breeds being most affected. HMA affected predominantly non-visceral sites, while HSA was more common in visceral sites, particularly the spleen. In multivariate model analyses, the odds of HMA diagnosis were 5.5 times higher in anemic dogs and 33.0 times higher in lymphopenic dogs compared to dogs without the abnormalities. The odds of HSA diagnosis were 42.5 times higher in anemic dogs, 343 times higher in lymphopenic dogs and 92.7 times higher in dogs with hyperfibrinogenemia compared to dogs without the abnormalities. The study suggested that these identified abnormalities were nonspecific and commonly observed in various chronic diseases, and hence their combination with clinical information, such as diagnostic imaging and histopathology, is important to facilitate a more precise diagnosis of canine vascular neoplasms. [ABSTRACT FROM AUTHOR]

Researchers and veterinarians often use hematology and clinical chemistry to evaluate animal health. These biomarkers are relatively easy to obtain, and understanding how they change across healthy aging is critical to clinical care and diagnostics for these animals. We aimed to evaluate how clinical biomarkers from a chemistry profile and complete blood count (CBC) change with age in common marmosets (Callithrix jacchus). We assessed blood samples collected during routine physical exams at the Southwest National Primate Research Center and the University of Texas Health San Antonio marmoset colonies from November 2020–November 2021. We found that chemistry and CBC profiles varied based on facility, sex, and age. Significant changes in albumin, phosphorus/creatinine ratio, albumin/globulin ratio, amylase, creatinine, lymphocyte percent, hematocrit, granulocytes percent, lymphocytes, hemoglobin, red cell distribution width, and platelet distribution width were all reported with advancing age. Aged individuals also demonstrated evidence for changes in liver, kidney, and immune system function compared with younger individuals. Our results suggest there may be regular changes associated with healthy aging in marmosets that are outside of the range typically considered as normal values for healthy young individuals, indicating the potential need for redefined healthy ranges for clinical biomarkers in aged animals. Identifying animals that exhibit values outside of this defined healthy aging reference will allow more accurate diagnostics and treatments for aging colonies. Research Highlights: The parameters that were reduced in peri‐geriatric or geriatric marmosets compared with younger marmosets were albumin, phosphorus/creatinine ratio, albumin/globulin ratio, calcium, glucose, lymphocytes, red blood cells, red cell distribution width, mean platelet volume, red cell distribution standard deviation, plateletcrit, platelet distribution width, and platelet distribution standard deviation.The parameters that were increased in peri‐geriatric and geriatric compared with young marmosets were amylase, creatinine,anion gap, granulocytes percent, and platelets.Our results suggest normal changes associated with healthy aging marmosets indicating the potential need for refinement of normal ranges for clinical biomarkers in aged marmosets. [ABSTRACT FROM AUTHOR]

ABSTRACT The selection of an appropriate production system plays a crucial role in ensuring the health, welfare, and productivity of American Pekin ducks. The aim of this study was to investigate the influence of various rearing production systems on the performance of American Pekin Ducks. A total of 180 straight run ducklings (10 days old) were randomly allocated into three experimental groups: intensive production system (IPS), free-range production system (FRPS), and pool with yard production system (PYPS). The distribution was carried out following a completely randomized design. We investigated the morphological parameters, growth performance, serum chemistry, meat cholesterol and triacylglycerol, cellular immunity, intestinal morphology, and carcass traits of Pekin ducks. The results indicate that PYPS birds exhibited significantly better morphometric parameters compared to IPS and FRPS ones. Ducks raised in PYPS had the highest growth rate till the 75th day as compared to those reared in IPS and FRPS. Triacylglycerol, total lipids, blood and meat fat and triacylglycerol were higher in IPS; whereas albumin, AG ratio, total proteins, glucose, and globulin were higher in PYPS. Similarly, cellular immunity (phagocytic activity, eosinophils, basophils, and monocytes) was increased in PYPS. Intestinal morphology and overall carcass traits were better in birds reared under PYPS than in those of IPS and FRPS. American Pekin ducks raised in PYPS showed improved growth, serum chemistry, cellular immunity, and carcass traits. These results suggest that the PYPS could be a promising production system for raising ducks in sub-tropical regions.

The current study aimed to compare the utilization efficiency of iron (Fe) feed additives from either bulk or nano sources in Nile tilapia, Oreochromis niloticus diets on growth, haematological, immunity, anti-oxidative, and intestinal topography capacities. Five isonitrogenous and isoenergetic diets were performed; the basal diet served as a control with no Fe added, whereas the experimental diets were shaped by adding bulk-Fe2O3 and nano-Fe2O3 to the basal diet to preserve Fe levels at 0.2 and 0.4 mg kg−1, respectively. Results indicated that superior growth performance was recorded in fish-fed diets supplemented with 0.4 nano-Fe2O3 mg kg−1 diet. In addition, the highest (P ≤ 0.05) survival rate, absorption area of villous (AAV), mucosal to serosal amplification ratio (MSR), and villi parameters (height and width) were noticed in fish fed diet enrichment with either bulk or nano-Fe2O3 source. However, the superiority observed in nano-Fe2O3 fish groups. Also, the highest values of plasma albumin, total protein, high-density lipoprotein cholesterol (HDL-C), white blood cells (WBCs), and lymphocyte absolute count (LYM) (P ≤ 0.05) recorded in fish fed a diet supplemented with nano-Fe2O3 versus the basal diet. Moreover, the highest values of catalase (CAT), glutathione peroxidase (GPx), superoxide dismutase (SOD), and plasma lysozyme activity (P ≤ 0.05) were observed in fish fed 0.4 mg/kg−1 nano-Fe2O3, while the lowest value was recorded in fish fed the control diet. The best value of malondialdehyde activity (P ≤ 0.05) recorded in a fish-fed diet supplemented with 0.4 mg/kg−1 nano-Fe2O3. The current findings emphasize the importance of including Fe to improve fish growth, immunity, antioxidant capabilities, and intestinal structure, primarily with a nano-Fe source, which demonstrated a more effective function in satisfying Nile tilapia dietary Fe requirements and improving the aforementioned parameters. [ABSTRACT FROM AUTHOR]

The present study is the first record of deducing age-wise and sex-wise normal reference intervals (RIs) for various serum biochemical attributes in apparently healthy Sipli sheep being reared under intensive farming system at Livestock Farm of the Faculty of Veterinary and Animal Sciences (FV&AS), The Islamia University of Bahawalpur (IUB). Blood samples were aseptically collected, serum was harvested and analyzed for serum chemistry analytes through semiautomatic chemistry analyzer using commercial kits. For the sake of analysis, groups were assigned as per age i.e. G1= up till 1year (n= 41), G2= from 1 to 2 years (n= 46), G3= from 2 to 3 years (n= 43) and as per sex i.e. females (n= 79) and males (n= 51). All the overall mean values and RIs for all the studied attributes were within the reference ranges provided in earlier studies for sheep. Bilirubin, Alkaline Phosphatase (ALP) and creatinine were significantly (P=0.05) lower in females as compared to those in males. However, uric acid was significantly (P=0.05) higher for females 52.8±2.6mmols/L as compared to 44.5±2.9mmols/L for males. The remaining attributes (Alanine Transaminase, Aspartate Aminotransferase, Total Proteins, Albumin, Glucose, Low Density Lipoproteins, High Density Lipoproteins and Urea) did not differ -significantly (P=0.05) between male and female sheep under study. For age-related results, only ALP, glucose and HDL were significantly (P=0.05) different between the three studied age groups of Sipli sheep. The ALP and glucose were higher in G1 animals (up till 1 year) whereas HDL was lower in G2 animals (from 1 to 2 years) as compared to their counter-part groups 1 and 3. The data can be beneficial in diagnostic/prognostic purpose for veterinarians, academicians, researchers and all other stakeholders of sheep-rearing sector in Pakistan as well as for other indigenous sheep breeds being reared under similar climatic conditions throughout the world. [ABSTRACT FROM AUTHOR]

Even though several studies have reported how stress induces detrimental pathophysiological responses that are linked to increase in disease susceptibility, no study has evaluated the impact of overcrowding on the serum chemistry of Heterobranchus longifilis. This study aimed at evaluating the impact of overcrowding on serum chemistry in Heterobranchus longifilis. Briefly, 30 apparently healthy Heterobranchus longifilis weighing between 1 kg and 1.5 kg were randomly divided into 2 groups of 15 catfish, each designated as groups 1 and 2. Group 1 (control group) was not subjected to stress, while group 2 was subjected to 3 hours of overcrowding in a 120 L plastic water tank at 201 g/L of water. Blood samples were collected from the caudal vein into non-heparinized tubes before stress, immediately after stress and 24 hours after stress. Sera were collected, and serum chemistry and electrolytes were analyzed. At 24 hours after stress, alanine aminotransferase, alkaline phosphatase, aspartate aminotransferase, creatinine, uric acid, total bilirubin, conjugated bilirubin, total protein, albumin, total cholesterol, triglyceride, chloride, potassium, and sodium were significantly higher in group 2 compared to the control group. Based on the findings of this study, it was concluded that overcrowding of Heterobranchus longifilis for 3 hours has a significant impact on its health status. [ABSTRACT FROM AUTHOR]

This study was conducted to evaluate the effects of various combinations of Lactobacillus species (L. rhamnosus, L. paracasei, and L. plantarum) on closely associated variables of production of laying hens, nitrogenous compounds in manure, the serum concentration of specific chemicals, and liver uric acid (UA) concentrations at peak lay. White Leghorns W-36 (32-week-old) were randomly assigned to five treatments for 8 weeks. Treatments were T1, the Control, a commercial feed; T2, the Control + L. paracasei + L. plantarum; T3, the Control + L. paracasei + L. rhamnosus; T4, the Control + L. plantarum + L. rhamnosus and T5, the Control + L. paracasei + L. plantarum + L. rhamnosus. Each bacterial species was included at 3.33 × 1011cfu/kg feed for a total of 6.66 x 1011 cfu/kg feed for T2-T4 and a total of 1.0 × 1012 cfu/kg feed for T5. Major effects among combinations of probiotics on production were not noted. The interaction of Probiotics by Week (Probiotics*Time) affected feed intake (P = 0.0007) and feed conversion ratio (FCR, P = 0.0049) due to fluctuation by week. Significant effects of time were also recorded for a gradual increase in body weight (BW, P = 0.0007); lowest and greatest feed intake at weeks 2 and 7, respectively (P < 0.0001); an increase in egg production (P = 0.0007) and maximum FCR at week 7 (P < 0.0001). Ammonia (NH3) concentration, ammonium nitrogen (NH4-N), total Kjeldahl nitrogen (TKN), and total nitrogen remained unaffected at P < 0.05. Although there were fluctuations, a trend emerged for the reduction of TKN. Combinations of probiotics did not affect NH3, UA, total protein (TP), albumin (ALB), creatine kinase (CK), and UA in the liver. Temporal (Time as a fixed effect) effects were noted for all nitrogenous compounds present in manure. For ammonia, temporal effects were significant due to fluctuation over time. Week 0 had the lowest value followed by weeks 4 and 8. Week 6 had the greatest value. For ammonium nitrogen, week 8 had the lowest value followed by week 0 and 4 with the next highest value. Week 6 had the greatest value. For TKN, week 4 had the lowest value followed by weeks 6 and 8. Week 0 had the greatest value. For TN, weeks 4, 6, and 8 had similar and lowest values followed by week 0 having the greatest value. However, an overall reduction in NH4-N, TKN, and TN was noted. Fluctuations in NH3 (P = 0.0033) and CK (P = 0.0085) were noted for Time. There was also a trend (P = 0.0706) for the increase of UA in serum. Two or more species of probiotics with yeast should be investigated. If the combination is applicable for increasing production measurements and reducing nitrogenous and serum compounds, the most appropriate time to feed the probiotics from day 1 to the end of production should be investigated.

The study aimed to analyze the impact of calcium butyrate supplementation in calf starter on growth performance indices associated with early rumen development to decrease the volume of milk or milk replacer feeding and enhance early starter intake in Holstein calves. For this purpose, twelve Holstein calves were randomly assigned into three treatments (n = 4/treatment); a control without coated calcium butyrate, T1, and T2 treatments supplemented with coated calcium butyrate 3 g and 6 g per day/head, respectively. Body weight was measured at days 7, 14, 21, 28, 35, 42, 49, and 56 of the trial, and the average daily weight gain and feed conversion ratio were determined. Blood samples were collected at 14, 28, 42, and 56 days of trial for serological parameters. Gut morphometry was performed at the end of trial at slaughtering by collecting duodenal samples. Furthermore, the meat was also evaluated for its quality parameters including pH and tenderness after slaughtering. The results indicated that the feed intake, average daily weight gain, feed conversion ratio, and gut morphometric parameters involving villus height and crypts depth of calves were improved in coated calcium butyrate–supplemented groups. Furthermore, the supplementation of calf starter with coated calcium butyrate significantly enhanced serum concentrations of glucose and total protein. Besides, Beta hydroxy butyrate (BHBA) levels of blood were also found to be elevated in both treatment groups. However, it was revealed that coated calcium butyrate supplementation had no significant effect on meat quality parameters. In conclusion, the supplementation of calf starter with coated calcium butyrate could improve calf performance. [ABSTRACT FROM AUTHOR]

Blood samples are routinely collected from wild populations in remote locations with limited electricity, minimal diagnostic capabilities, and extreme environmental conditions. Under these conditions, serum samples may be stored for prolonged time under varied temperatures prior to processing, which could affect the ability to interpretation the results. This study’s objective was to evaluate the effects of delayed processing of serum samples and varied storage temperatures on biochemical values in African savanna elephants (Loxodonta africana). Blood samples were collected from six elephants managed by the North Carolina Zoo. For each elephant, seven red top tubes were collected. One serum sample for each elephant was analyzed on Day 0 (control group). The remaining samples were stored under different temperatures including room temperature (23 °C), refrigeration (2.2 °C), and incubation (32.2 °C), with samples from each temperature group being analyzed on Day 5 and Day 10. Many of analytes (10 out of 18) did not change significantly regardless of storage temperature or time. Refrigeration improved stability in an additional four analytes over prolonged storage. We conclude that if serum is properly separated shortly after collection, many serum biochemical analytes can be accurately measured even after suboptimal serum storage, but refrigeration and prompt evaluation are still required for some analytes.

The leaves of Lawsonia inermis Linn are used in the treatment of many diseases such as diabetes, poliomyelitis, measles and gynecological disorders such as menorrhagia, vaginal discharge and leucorrhoea. This study was designed to investigate the safety evaluation of Lawsonia inermis Linn leaves (acutely and sub-chronically) on physiological, biochemical and histopathological changes seen in Wistar rat. Acutely, female rats were divided into four groups (n=3) and treated as thus A (untreated control); B (1000 mg/kg); C (2000 mg/kg) and D (5000 mg/kg). Sub-chronically, 25 male Wistar rats were grouped into five (n=5). Groups: A (control), B (100 mg/kg); C (200 mg/kg); D (400 mg/kg) and E (800 mg/kg). Lawsonia inermis Linn leaves have a wide safety margin (>5000mg/kg) and no mortality or visible toxic reaction was observed in acute phase. Lawsonia inermis extract did not inhibit physiological weight gain, except the highest dose that caused some weight loss. Haematological result showed that PVC, RBC, haemoglobin and platelets had no significant (P>0.05) effect unlike white blood cell and differentials (neutrophils, lymphocytes and monocytes) which decrease significantly (P

Mycotoxins are secondary metabolites produced by pathogenic fungi under suitable environmental conditions. Moringa oleifera and vitamin E are known antioxidants and immunostimulants. In present experimental study, M. oleifera and vitamin E were administered in birds feed to mitigate the toxico-pathological impacts of aflatoxin B1 in white Leghorn layer males. A total of 120 white Leghorn layer male birds were equally divided into six experimental groups. Group A was kept as negative control, group B was fed 400ppb of aflatoxin, while D and F groups were fed normal feed and supplemented with vitamin E (100ppm) and 1 %M. oleifera, respectively. The birds in C and E groups were given 400 ppb AFB1 in feed and supplemented with vitamin E and M. oleifera, respectively. During the 60 days of experimental trial, hematobiochemical and histopathological parameters were studied. The data obtained was analysed by analysis of variance test and group means were compared by DMR test using M STAT C statistical software. During and at the end of experimental trial, male layer birds were slaughtered, collected kidney and liver were fixed in 10% neutral buffered formalin to investigate gross and histopathological alternations. There were significant alterations in hematological and serum biochemical parameters of aflatoxin treated birds. Serum enzymes including ALT, AST and LDH values were significantly increased in group B in contrast to control group A. The histopathological changes in hepatic and renal parenchyma induced by AFB1 were prominent and no such changes were seen in A, D and F groups, while C and E group showed mild necrotic changes which was an indication of ameliorative effects of vitamin E and M. oleifera. It may be concluded that vitamin E and M. oleifera have potential to mitigate toxicity of aflatoxin B1. [ABSTRACT FROM AUTHOR]

Background: Evaluation of blood parameters in captive non‐human primates (NHPs) is crucial for monitoring their health and ensuring that their environment meets their physiological requirements. Methods: We performed hemogram, serum biochemistry, and parasitological exams in 20 howler monkeys and 21 capuchin monkeys. Results: In both species, over 50% of the individuals presented at least one parasite. There was a negative effect of age on red blood cell (RBC), white blood cell, platelets, total protein, globulin, and alkaline phosphatase, and a positive effect on the A:G ratio, gamma‐glutamyl transferase, and mean platelet volume (MPV). Capuchin monkeys presented the highest platelets and alanine aminotransferase (ALT) values and howler monkeys presented the highest MPV, aspartate aminotransferase, ALT, amylase, glucose, bilirubin, and triglycerides values. We observed an interaction between species and sex on RBC, Htc, mean corpuscular hemoglobin concentration, and cholesterol. Conclusions: Species differences found in blood parameters may reflect differences in physiological adaptations associated with ecological and morphological traits and are clinically relevant for evaluating animal health and the suitability of breeding programs. [ABSTRACT FROM AUTHOR]

Gladiolus psittacinus (GP) is an important medicinal plant in folk medicine where its corm is used for treatment of diabetes mellitus. Despite this, there is paucity of scientific information to justify its use as antidiabetic drug. Hence, this study was designed to explore antidiabetic, antihyperlipidemic and effects of aqueous extract of Gladiolus psittacinus (AGP) on hyperglycemia-associated oxidative stress in pancreas, kidney and liver of diabetic rats. Diabetes mellitus (DM) was induced in rats using streptozotocin 50 mg/kg (i.p.). Normal and diabetic rats were treated orally with AGP once a day for 14 days. Antidiabetic effects were evaluated on body weight, fasting blood glucose concentration (FBGC), lipid profiles and serum chemistry. Also, protective effects of AGP were also determined on markers of oxidative stress, antioxidant enzymes and histopathology of pancreas, kidney and liver of diabetic rats. Treatment with AGP emanated to significant decrease of FBGC (552.67–157.33 mg/dL), increase in body weight (100.01–133.76 g) and positive modulation of lipid parameters in diabetic rats. The alteration in the contents of markers of liver and kidney function were significantly modulated in the diabetic rats upon treatment. Also, oxidative damage and antioxidant depletions in pancreas, kidney and liver were significantly mitigated in treated diabetic rats. Structural aberrations in the histopathology slides of pancreas, kidney and liver were improved upon treatment. It can be concluded that AGP could be used in the treatment of diabetes mellitus and its related ailments, thereby justifying its usage in traditional medicine. [ABSTRACT FROM AUTHOR]

ABSTRACT We analyzed the influence of the season, the environment, and the sex, as well as the relation of body mass (BM) in the serum albumin (ALB), aspartate aminotransferase (AST), creatinine (C), creatine kinase (CK), phosphorus (P), total calcium (tCa), total protein (TP), urea (U), uric acid (UA), calcium:phosphorus ratio (Ca:P), and the globulin value (GV) of thirty individuals of Phrynops geoffroanus of the urban area of Cuiabá, Mato Grosso, Brazil. The modeling of biochemical parameters was performed using the Generalized Additive Models for Location, Scale and Shape (GAMLSS) to verify the influence of variables considered in this study on each of the biochemical parameters analyzed. The season influenced AST, CK, C, tCa, Ca:P and UA. The environment influenced tCa, Ca:P, U and UA. On the other hand, CK, tCa, P, Ca:P and U differed significantly between males and females. Regarding the BM, a relationship of this variable was observed with CK, C, tCa, P, U, UA and Ca:P. We concluded that the season, environment, sex, and body mass can influence the biochemical parameters of P. geoffroanus, and these factors should be routinely considered in the interpretation of laboratory results.

Abstract Background Sampling from a peripheral intravenous catheter (PIVC) might be a more efficient and less traumatic collection of blood for serum biochemistry (SB) or CBC than direct venipuncture (DV). Agreement between results of samples obtained by these methods has not been evaluated in dogs. Objectives The primary objectives were to determine whether sampling from PIVC could be used in place of DV for dogs. We hypothesized DV and PIVC samples would have clinically equivalent SB and CBC results. Animals Sixty‐one client‐owned dogs were included in each study arm. Methods This was a partially randomized method‐comparison study. Paired DV and PIVC samples obtained within 1 to 2 minutes after, or approximately 24 hours after, placement of a PIVC in a cephalic vein were evaluated for agreement and bias using percentage difference plots (with a priori application of consensus total allowable error), Bland‐Altman analysis, Passing‐Bablok regression analysis, Wilcoxon signed rank test, and McNemar's test. Results There was statistically and clinically acceptable agreement and no bias between sampling methods for the majority of results. Analytes with the most frequent disagreement were aspartate aminotransferase, total bilirubin, potassium, bicarbonate, and leukocyte differential counts, as well as red blood cell count, hemoglobin, hematocrit, and packed cell volume in the hospitalized PIVC sampling group. Few observed differences would change clinical decision making. Conclusions and Clinical Importance PIVC sampling can provide generally acceptable SB and CBC results for most dogs, but clinicians should be aware of a few values for which disparate results might occasionally be obtained.

Blood-based diagnostics are critical for many medical decisions, but mostly rely on venepuncture, which can be inconvenient and painful. The Onflow Serum Gel (Loop Medical SA, Vaud, Lausanne, Switzerland) is a novel blood collection device that utilises needle-free technology to collect capillary blood. In this pilot study, 100 healthy participants were enrolled and provided two Onflow collected specimens and one venous blood specimen. Five chemistry analytes (AST, ALT, LDH, potassium, creatinine) and haemolysis were measured per specimen, and laboratory analyte results were compared. Onflow was found to be more acceptable than venepuncture with lower pain ratings, and 96.5% of participants would use the Onflow method again. All phlebotomists (100%) found Onflow intuitive and user-friendly, with ~1 mL of Onflow blood successfully collected from 99% of participants in <12 min (mean: 6 min, 40 s) and 91% collected on the first attempt. ALT and AST analytes showed no difference in performance, while creatinine generated a negative bias (−5.6 µmol/L), and increased variability was noted with potassium (3.6%CV) and LDH (6.7%CV), although none were clinically relevant. These differences may be due to 35% of Onflow collected specimens having "mild" haemolysis. Onflow is a promising alternative blood collection device that should now be evaluated in participants with expected abnormal chemistries and as an option for self-collection. [ABSTRACT FROM AUTHOR]

Blood samples are routinely collected from wild populations in remote locations with limited electricity, minimal diagnostic capabilities, and extreme environmental conditions. Under these conditions, serum samples may be stored for prolonged time under varied temperatures prior to processing, which could affect the ability to interpretation the results. This study's objective was to evaluate the effects of delayed processing of serum samples and varied storage temperatures on biochemical values in African savanna elephants (Loxodonta africana). Blood samples were collected from six elephants managed by the North Carolina Zoo. For each elephant, seven red top tubes were collected. One serum sample for each elephant was analyzed on Day 0 (control group). The remaining samples were stored under different temperatures including room temperature (23 °C), refrigeration (2.2 °C), and incubation (32.2 °C), with samples from each temperature group being analyzed on Day 5 and Day 10. Many of analytes (10 out of 18) did not change significantly regardless of storage temperature or time. Refrigeration improved stability in an additional four analytes over prolonged storage. We conclude that if serum is properly separated shortly after collection, many serum biochemical analytes can be accurately measured even after suboptimal serum storage, but refrigeration and prompt evaluation are still required for some analytes. [ABSTRACT FROM AUTHOR]

The objective was to provide an updated profile of the bovine acute-phase response to include recent advancements in technologies and expanded hematological, cytokine, and serum chemistry variables. Beef steers (n = 32; body weight [BW] = 251 ± 19.5 kg) were fitted with indwelling jugular catheters 1 d before lipopolysaccharide (LPS; 0.25 µg LPS/kg BW from Escherichia coli O111:B4) administration to facilitate serial blood collection. Rectal temperature was measured using indwelling probes, and ocular temperature was measured using infrared thermal imaging. Blood samples were collected for subsequent analysis of serum chemistry, hematology, and cytokine concentrations. Pearson correlation of rectal temperature and ocular infrared temperature was 0.61 (P < 0.01) and the Spearman correlation coefficient was 0.56 (P < 0.01). Interactions of hour × method were observed for ocular and rectal measurements of body temperature in response to endotoxin exposure. Maximum observed temperature was 39.6 °C at 2.5 h for both rectal and ocular measurements. Body temperature differed by method at hours 0.5, 2.5, 4.5, 7.5, 12.5, 36.5, and 47.5 (P < 0.01), but were not different otherwise. All variables of serum chemistry and complete blood count were influenced by LPS administration, except creatinine, serum glucose, and percent basophils (P ≤ 0.02). Alanine aminotransferase and alkaline phosphatase peaked at hour 2 relative to LPS administration, returned to baseline at hour 12 and continued to decrease below the baseline value at hour 48 (P < 0.01). Total protein concentration decreased 3% in response to LPS (P = 0.01). Total white blood cell count decreased 75% after LPS administration at hour 1 (P < 0.01). Lymphocyte count recovered to baseline at hour 6; sooner than neutrophil count at hour 36. Serum cortisol concentration increased 294% relative to baseline at hour 1 followed by a sustained decrease and return to normal concentration at hour 4 (P < 0.01). Additionally, circulating cytokine concentrations changed with time in response to the LPS challenge, excluding aFGF, bFGF, IGF-1, IL-2, IL-4, MCP-1, and ANG-1 (P ≤ 0.08). Maximum observed concentration of TNF-α at hour 1 was 117% greater than the pre-challenge value (P < 0.01). Data presented herein add to existing works to understand the endocrine and immune responses of beef steers administered exogenous LPS, and incorporate recent technologies, additional biomarkers, and an expanded cytokine profile that can be used as referential data in future research. [ABSTRACT FROM AUTHOR]

Shark assessments in the Mediterranean Sea are still scarce, and serum chemistry and haematological data have yet to be reported for wild dusky (Carcharhinus obscurus) or sandbar (Carcharhinus plumbeus) shark populations in the Mediterranean Sea. Herein, blood samples were obtained from adult dusky (n  = 23) and sandbar (n  = 14) sharks from an aggregation site near the Hadera power and desalination plants in Israel in the winters of 2016–20. Several serum chemistry analytes were characterized with relation to stress, body size and environmental conditions. Glucose concentrations were higher, while total cholesterol concentrations were lower in dusky sharks than in sandbar sharks, potentially due to distinct metabolic pathways utilized during the capture-related activity by both species. However, differences in sex and size are noted and should be considered. The blood cell morphology of both species was consistent with previous findings for sandbar sharks. Atypical monocytes were noted in one dusky shark. Preliminary and exploratory reference intervals for female dusky sharks were calculated for glucose, triglycerides, total cholesterol, total protein and creatine kinase. These data must be viewed with caution due to the potential influence of capture-related stress on analyte concentrations and activities and the fact that only females were employed in the calculations. Moreover, the sampling site is adjacent to coastal power and desalination plants, which may significantly affect shark physiology. Although limited, this novel database on dusky and sandbar shark serum chemistry and haematology aspects is essential as a first attempt to obtain data on these species in the eastern Mediterranean Sea and for future conservation and long-term biomonitoring efforts. [ABSTRACT FROM AUTHOR]

This case report investigates the occurrence of green discoloration in serum and citrate plasma samples collected from a male adult patient following a multivisceral organ transplant. In collected samples, it was necessary to investigate the influence of sample discoloration on the results of laboratory tests and to determine the appropriate approach to sample management. Hematology, coagulation and blood gas analysis showed no flags, but the biochemical lipemia index was susceptible to positive interference, necessitating dilution of the native sample. Despite the green discoloration, both native and diluted samples exhibited minimal interference on routine clinical chemistry analyses, demonstrating the reliability of the laboratory test results. This case report underscores the influence of preanalytical factors on the results of laboratory tests, the need for a thorough assessment of the sample adequacy for laboratory testing and the strict application of appropriate guidelines in the sample management in order to make an accurate diagnosis and ensure optimal patient care., Competing Interests: Potential conflict of interest None declared., (Croatian Society of Medical Biochemistry and Laboratory Medicine.)

Aptamers have shown potential for diagnosing clinical markers and targeted treatment of diseases. However, their limited stability and short half-life hinder their broader applications. Here, a real sample assisted capture-SELEX strategy is proposed to enhance the aptamer stability, using the selection of specific aptamer towards PD-L1 as an example. Through this developed selection strategy, the aptamer Apt-S1 with higher binding affinity and specificity towards PD-L1 was obtained as compared to the aptamer Apt-A2 which was screened by the traditional capture-SELEX strategy. Moreover, Apt-S1 exhibited a greater PD-L1 binding associated conformational change than Apt-A2, indicating its suitability as a biorecognition element. These findings highlight the potential of Apt-S1 in clinical applications requiring robust and specific targeting of PD-L1. Significantly, Apt-S1 exhibited a lower degradation rate in 10% diluted serum or pure human serum, under the physiological temperature and pH value, compared to Apt-A2. This observation suggested that Apt-S1 possesses higher stability and is more resistant to damage caused by the serum environmental factors, highlighting the superior stability of Apt-S1 over Apt-A2. Furthermore, defatted and deproteinized serum were used to investigate the potential reasons for the improved stability of Apt-S1. The results hinted that the pre-adaptation to nucleases present in serum during the selection process might have contributed to its higher stability. With its improved stability, higher affinity and specificity, Apt-S1 holds great potential for applications in PD-L1 assisted cancer diagnosis and treatment. Meanwhile, the results obtained in this work provide further evidence of the advantages of the real capture-SELEX strategy in improving aptamer stability compared to the traditional strategy., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Using Fourier Transform Infrared spectroscopy (FTIR), it is possible to show chemical composition of materials and / or profile chemical changes occurring in tissues, cells, and body fluids during onset and progression of diseases. For diagnostic application, the use of blood would be the most appropriate in biospectroscopy studies since, (i) it is easily accessible and, (ii) enables frequent analyses of biochemical changes occurring in pathological states. At present, different studies have investigated potential of serum, plasma and sputum being alternative biofluids for lung cancer detection using FTIR. However, until now, it has not been shown which biofluid; among serum and plasma, that can serve as the best material medium for detecting lung cancer with highest levels of accuracy. In this study, plasma and serum isolated from blood consenting participants without lung cancer symptoms (controls) and lung cancer patients. The samples were measured using FTIR and subsequently analyzed by machine learning (ML) algorithms in order to show which fluids (serum or plasma) would better enhance detection of lung cancer. Higher absorbances values of PO 2- , CH 2 , CH 3 and amides vibrations in FTIR spectra of both serum and plasma samples, collected from lung cancer patients were observed in comparison to individuals without lung cancer symptoms (controls). Principal component analysis (PCA) of FTIR spectra showed plasma and serum samples collected from lung cancer patients and individuals without lung cancer symptoms were better differentiated in fingerprinting region (from 800 to 1800 cm - 1 ) when compared to lipid region (2800-3000 cm - 1 ). Moreover, also sensitivity specificity and accuracy calculated by logistic regression (LR) and receive operating characteristic (ROC) showed higher values for fingerprint range (800-1800 cm - 1 ) in comparison with lipids (2800-3000 cm - 1 ) one for both, serum and plasma. However, using these methods differences between serum and plasma were not existed. From the all obtained results, it was visible, that both fluids could be used in detected lung cancer using FTIR. Moreover, it was also showed that fingerprint range gave a better distinction between the studied patient groups than the lipid range. This was noticeable for both serum and plasma., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Invasive aspergillosis (IA) is a life-threatening infection. Early and specific diagnosis is pivotal to ensure adequate therapy. Antigen testing from blood is a widespread and convenient diagnostic approach. Various tests for the detection of Aspergillus antigen as well as for the panfungal antigen β-1,3-D-glucan (BDG) are available, for which comprehensive comparisons are still lacking. Blood samples of 82 proven/probable (11/71) IA patients and 52 controls were tested using two enzyme-linked immunosorbent assays (ELISAs) (Bio-Rad and Euroimmun), one chemiluminescent immunoassay (CLIA) (Vircell), one BDG assay (Fujifilm Wako), and two point of care (PoC) assays (Immy sōna and OLM). PoC assays were evaluated visually and used automated read out systems. Of the 82 IA patients, 37 had received solid organ transplantation (SOT) and 25 hematopoietic stem cell transplant (HSCT). Sensitivities and specificities for the eight test systems ranged from 27% to 71% and from 64% to 100%. Estimating a 10% prevalence of IA, test performance would have resulted in positive and negative predictive values of 14%-100% and 91%-95%. Areas under the curve (AUCs) for all tests except GM were below 0.7. When the cut-off values for quantitative tests were normalized to a specificity close to 95%, sensitivities ranged from 14% to 40%. The use of automated read out systems for the PoC assays had a significant impact. Combining different tests did not result in better test strategies. Sensitivity of Aspergillus antigen testing from single serum samples is low. Due to specificity issues, the majority of tests is not suited for screening purposes. The different assays can meet different needs in different diagnostic settings., Competing Interests: J.W. received financial support (research grant) from Pfizer outside of this study, technical and financial support by Fujifilm Wako Chemicals Europe and Euroimmun Medizinische Labordiagnostika for past projects outside this study, and speaker fees from Pfizer, Wako, und Gilead. J.P. serves as President of the Austrian Society of Medical Mycology, received speaker fees from Gilead, Pfizer, Swedish Orphan BioVitrum, and Associates of Cape Cod, and holds stock in AbbVie Inc. and Novo Nordisk, all unrelated to the submitted work. M.H. received research funding from Gilead, Astellas, MSD, IMMY, Mundipharma, Scynexis, F2G, and Pfizer, all outside of the submitted work. J.H. received speaker fees from Pfizer, Gilead, Associates of Cape Cod, BD, and Biomerieux, research funding from Pfizer and Gilead, and technical support for projects outside this study from Associates of Cape Cod, Vircell, Virion\Serion, IMMY, and OLM. K.D. received technical and financial support by Fujifilm Wako Chemicals Europe and by Euroimmun Medizinische Labordiagnostika for past projects outside this study and technical support by Vircell, OLM, and IMMY for past projects outside this study. T.S., I.K., and S.S. have no conflicts of interest.

Objectives: In most countries the majority of patients are in outpatient care. In difference to hospitalized patients, their blood samples often take hours after collection to centrifugation. The study investigates the release of potassium and the development of pseudohyperkalemia in lithium heparin (Li-Hep) and serum blood collection tubes over time., Methods: From 201 donors 4 serum and 4 Li-Hep blood collection tubes were taken each. After 0.5, 4, 6 and 8h whole blood was centrifuged, and potassium levels were determined. To simulate the preanalytic conditions, the samples with a storage time >0.5h were shaken on a standard shaker for 1h and stored at 4-8°C for the remaining time., Results: Over time, significant more potassium was released before centrifugation from the Li-Hep plasma than from serum (1.21 vs 0.94 mmol/L). After 6h, the two groups were no longer highly statistically significantly different (potassium mean: 5.01 mmol/L in serum group, 4.92 mmol/L in Li-Hep group). In the Li-Hep group 164 donors developed a pseudohyperkalemia after 8h, compared to 76 in the serum group., Significance: The decision as to which material is best suited should not only be based on which value comes closest to the physiological situation immediately after blood collection. The subsequent preanalytic circumstances must also be considered. Serum tubes appear to be at least as suitable for potassium determination as Li-Hep tubes. In terms of patient blood management, serum provides the possibility of performing a wider range of analyses in the outpatient setting., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Reuter et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Lack of uniformity in infection models complicates preclinical development. The COMBINE protocol has standardized the murine neutropenic pneumonia model. Herein we provide benchmark efficacy data of humanized exposures of tigecycline and levofloxacin in plasma and epithelial lining fluid (ELF) against a collection of Klebsiella pneumoniae and Pseudomonas aeruginosa., Methods: Following the COMBINE protocol, plasma and ELF human-simulated regimens (HSRs) of tigecycline 100 mg followed by 50 mg q12h and levofloxacin 750 mg once daily were developed and confirmed in the murine neutropenic pneumonia model. Tigecycline HSRs were tested against seven K. pneumoniae isolates. Levofloxacin HSRs were assessed against 10 K. pneumoniae and 9 P. aeruginosa. The change in cfu/lung over 24 h for each treatment was calculated. Each isolate was tested in duplicate against both the plasma and ELF HSRs on separate experiment days., Results: Tigecycline 1.8 and 3 mg/kg q12h achieved humanized exposures of serum and ELF, respectively. Levofloxacin 120 and 90 mg/kg q8h led to fAUC exposures in plasma and ELF similar to in humans. Both tigecycline regimens were ineffective across the MIC range. Levofloxacin regimens achieved multilog kill against susceptible isolates, and no appreciable cfu/lung reductions in isolates with an MIC of ≥32 mg/L. Differences in cfu/lung were evident between the levofloxacin plasma and ELF HSRs against isolates with MICs of 4 and 8 mg/L., Conclusions: Administering HSRs of tigecycline and levofloxacin based on both serum/plasma and ELF in the COMBINE pneumonia model resulted in cfu/lung values reasonably aligned with MIC. These data serve as translational benchmarks for future investigations with novel compounds., (© The Author(s) 2024. Published by Oxford University Press on behalf of British Society for Antimicrobial Chemotherapy.)

Successful implementation of Safe and Sustainable by Design (SSbD) and grouping approaches requires simple, reliable, and cost-effective assays to facilitate hazard screening at early stages of product development. Especially for nanomaterials (NMs), which exist in many different forms, efficient hazard screening is of utmost importance. Oxidative potential (OP), which is the ability of a substance to induce reactive oxygen species (ROS), is an important indicator of the potential to induce oxidative damage and oxidative stress. A frequently used assay to measure OP of NMs is the ferric reducing ability of serum (FRAS) assay. Although the widely used cuvette-based FRAS protocol is considered a robust assay, its low throughput makes the screening of multiple materials challenging. Here, we adapt the original cuvette-based FRAS assay protocol, into a 96-well format and thereby improve its user-friendliness, simplicity, and screening capacity. The adapted protocol allows for the screening of multiple NMs per plate, and multiple plates per day, where the original protocol allows for the screening of one NM dose-range per day. When comparing the two protocols, the adapted protocol showed slightly decreased assay precision as compared to the original protocol. The results obtained with the adapted protocol were compared using eight reference NMs in an interlaboratory study and showed acceptably low intra- and interlaboratory variation. We conclude that the adapted FRAS assay protocol is suitable to be used for hazard screening to facilitate SSbD and grouping approaches.

ABSTRACT: A dorsal recumbency syndrome (DRS) has been recently described in market-age broiler chickens. Affected broilers fall onto their backs, and are unable to right themselves, and eventually die of cardiopulmonary insufficiency. These broilers are referred to as turtle chickens. A previous report and anecdotal evidence suggest that breast myopathies, such as woody breast (WB), may be associated with DRS due to impaired contractility of the pectoral muscles. In this study, we aimed to provide additional evidence to document DRS in broilers, and its possible association with breast myopathies.A total of 64 broilers (Ross 708), 33 DRS-affected and 31 controls, were culled between 42 and 48 d of age from 3 different commercial farms over 4 visits. All broilers underwent postmortem analysis; breast muscles were scored grossly and/or histologically to determine the presence and severity of myopathies, and sera were used to determine the level of aspartate aminotransferase (AST) and creatine kinase (CK).A gross diagnosis of WB was moderately associated with DRS broilers, and DRS broilers displayed a greater microscopic severity of lesions (P < 0.001) in the Pectoralis major, as typically observed with WB. Levels of AST and CK were greater (P < 0.001) in the sera of DRS-affected compared to control broilers, consistent with muscular damage. The frequency of cardiac changes, such as mild hydropericardium and right ventricular dilation, or severity of microscopic pulmonary lesions, such as edema, were not significantly different between the 2 groups. The odds of DRS increased with the histology score of the P. major (OR = 1.37, 95% CI 1.02–1.85).The data presented in this study support an association between DRS and muscular damage of the P. major, suggesting that WB may predispose broilers to DRS. DRS might be a cause of broiler death, and this syndrome could be responsible for significant financial loss to the farmers and to the whole poultry industry.

Simple Summary: A preparation of the Macleaya cordata extract (Sangrovit®) provides a unifying taste to feed for cattle. However, information is needed to see if the effect occurs when fed in milk replacer or starter feed to calves and residues of the extract end up in the tissues. Male and female calves (~5 d of age; 5/sex/group) were provided Sangrovit® in milk replacer and starter feed at control (0.0 Sangrovit®), 2.0 and 4.0 g (D1), 5.0 and 10.0 g (D2) and 10.0 and 20.0 g (D3) Sangrovit®/day, respectively, for 90 days. Body weight, feed intake and blood-derived endpoints were evaluated along with tissue residue levels of Sangrovit®. Compared to the control group, Sangrovit® tended to increase milk-replacer intake (4%). Average daily gain did not change, and some blood chemistry and hematology parameters changed with Sangrovit® intake, but were within the ranges of healthy calves. Residues of Sangrovit® were located in the edible tissues of the calves. Sangrovit® can be added to milk replacer and calf starter feeds with no adverse effects on feed intake, health, or growth of the calves. This study examined the effects of Sangrovit®, a Macleaya cordata plant extract (MCE) preparation on feed intake, growth, blood chemistry, and tissue-residue levels of calves. Twenty male and 20 female calves (~5 d of age) were assigned to one of four daily Sangrovit® doses: 0.0 and 0.0 (CTL), 2.0 and 4.0 (D1), 5.0 and 10.0 (D2), and 10.0 and 20.0 (D3) g/calf in pre-weaning (5 to 49 d of age) and post-weaning (50 to 95 d of age) periods, respectively. Sangrovit® doses were fed in milk replacer pre-weaning and top-dressed on calf starter post-weaning. Milk replacer and calf starter intake was recorded daily. Body weight, hematology, and serum chemistry were measured at 5, 49, and 95 d of age. Calves were slaughtered at 95 d of age for MCE tissue residue analysis. Compared to CTL, D1 increased milk-replacer intake (4.90 to 5.09 L/day), but decreased calf starter intake pre- (0.65 to 0.53 kg/d) and post-weaning (3.42 to 3.20 kg/d). No Sangrovit® dose affected average daily gain. The hematology and blood chemistry of all treatment groups fell within the ranges of healthy calves. Results showed no adverse effects of MCE on health and growth performance of calves when fed up to 10.0 g/calf/day pre-weaning and up to 20.0 g/calf/day post-weaning. [ABSTRACT FROM AUTHOR]

Background: Sampling from a peripheral intravenous catheter (PIVC) might be a more efficient and less traumatic collection of blood for serum biochemistry (SB) or CBC than direct venipuncture (DV). Agreement between results of samples obtained by these methods has not been evaluated in dogs. Objectives: The primary objectives were to determine whether sampling from PIVC could be used in place of DV for dogs. We hypothesized DV and PIVC samples would have clinically equivalent SB and CBC results. Animals: Sixty‐one client‐owned dogs were included in each study arm. Methods: This was a partially randomized method‐comparison study. Paired DV and PIVC samples obtained within 1 to 2 minutes after, or approximately 24 hours after, placement of a PIVC in a cephalic vein were evaluated for agreement and bias using percentage difference plots (with a priori application of consensus total allowable error), Bland‐Altman analysis, Passing‐Bablok regression analysis, Wilcoxon signed rank test, and McNemar's test. Results: There was statistically and clinically acceptable agreement and no bias between sampling methods for the majority of results. Analytes with the most frequent disagreement were aspartate aminotransferase, total bilirubin, potassium, bicarbonate, and leukocyte differential counts, as well as red blood cell count, hemoglobin, hematocrit, and packed cell volume in the hospitalized PIVC sampling group. Few observed differences would change clinical decision making. Conclusions and Clinical Importance: PIVC sampling can provide generally acceptable SB and CBC results for most dogs, but clinicians should be aware of a few values for which disparate results might occasionally be obtained. [ABSTRACT FROM AUTHOR]

The effects of yeast cell wall (YCW) supplementation on growth performance, cell-mediated immune response, blood metabolites, and health scores in early-weaned male buffalo (Bubalus bubalis) calves were evaluated. Forty Nili Ravi buffalo calves were randomly divided into four groups and supplemented with four dietary treatments, namely control (animals without prebiotics), YCW-2 (yeast cell wall fed at 2 g/calf/day), YCW- 4 (yeast cell wall fed at 4 g/calf/day), and cMOS-4 (commercial mannan-oligosaccharide fed at 4 g/calf/day). Milk intake, dry matter intake (DMI), and health scores were recorded daily, whereas body weight (body weight) and structural developments (hip height, wither height, heart girth, and body length) were recorded weekly. Feed efficiency and average daily gain (ADG) were calculated at the end of the experiment. Blood samples were collected fortnightly to determine glucose, non-esterified fatty acids, beta-hydroxybutyric acid (βHBA), blood urea nitrogen (BUN), catalase, malondialdehyde, creatinine, hepatic enzymes, cholesterol, triglycerides, and total serum protein profile. The cell-mediated immune response was determined by the application of dinitrochlorobenzene directly to the skin. The results of the study revealed that supplementation of YCW and cMOS increased DMI, body weight, feed efficiency, ADG, and structural growth in buffalo calves, whereas faecal scores were significantly improved in supplemented groups compared to the control. Glucose, BUN, and βHBA improved in supplemented animals more than in the control group, indicating bioactivity that contributed to the improvement of gut health. Supplementation with YCW improved the growth performance, physiological responses, and gut health of early-weaned male buffalo calves. [ABSTRACT FROM AUTHOR]

The coati (Nasua nasua, Linnaeus 1766) is a generalist species, feeding on often-discarded human food in dumpsters around ecological tourism sites. We investigated the body weight and some blood chemistry variables related to the diet of wild coatis from three parks: Parque Municipal das Mangabeiras (PM), Parque Nacional do Caparaó (PNC) e Estação Ecológica Água Limpa (EEAL). We tested the plasma of 53 coatis for high-density lipoprotein (HDL), alanine transaminase (ALT), aspartate transaminase (AST), gamma-glutamyl transferase (GGT), cholesterol (Chol), triglycerides (Trig), and alkaline phosphatase (ALP). Male and female adults did not significantly differ on the weight (P > 0.05) and blood chemistry indexes (P > 0.05). The adult coatis of the PM were heavier than the adult coatis of the other two parks. There were significant differences in HDL (P < 0.04), AST (P < 0.001), ALT (P < 0.001), and GGT (P < 0.001) between adults of the three parks. Only ALT and ALP were significantly different (P < 0.05) among the young coatis. The results suggested the coatis of the three parks have different health status. The consumption of discarded human food seems to affect body weight of the PM coatis. The coatis from PNC and EEAL had blood chemistry profiles suggestive of liver disorder. We recommend carrying on environmental education programs to visitors and additional clinical investigations on coatis from these parks. [ABSTRACT FROM AUTHOR]

Background: Solanum erianthum leaves extract has been used to treat sexually-transmitted diseases, malaria, and leprosy. This study assessed the toxicity and safety of S. erianthum extract in rats. Methods: Treatment with 250, 500 or 750 mg/kg of the aqueous, ethanolic and methanolic extracts in the rats had different effects on the biochemical activities of the liver, heart and kidneys, and based on the hematological and histopathological changes observed after short-term (30 days) and long-term (60 days) exposure. Results: The serum biochemical parameters examined were AST, ALT, and ALP concentrations in the albino rats treated with the extracts of S. Erianthum at various concentrations. The extract showed significantly different effects in the treated versus untreated rats (P

This study evaluated the effect of environmental enrichment on the performance, behaviour and welfare aspects; blood biochemistry; carcass and meat quality traits of broiler chickens. A total of 450 straight run broiler chicks (Ross-308) were divided into 5 treatment groups having 6 replicates of 15 birds each under a completely randomised design. Treatments were environmental enrichment (EE) tools and consisted of C = control group; R = red ball for EE; G = green ball for EE; B = blue ball for EE and M = mirror for EE. These environmental enrichment tools were provided throughout the experimental period (0 to 35 days). Mean feed intake per bird was higher in all treatment groups except the blue balls group; weight gain and feed conversion ratio were better in the green and blue ball groups. Broiler chickens reared under different environmental enrichment were more active and they exhibited maintenance behaviour (preening, dust bathing and wing stretching, or scratching) more frequently. Regarding welfare traits, lower incidence of toe damage, footpad dermatitis and hock burn was observed in birds having different environmental enrichment tools as compared to the control group. The birds reared with red balls as environmental enrichment showed the lowest values for glucose, cholesterol, total protein, albumin and globulin amongst all the treatment groups. Birds reared with green balls had the highest body weight at slaughter, dressed weight, carcass yield and liver weight. Breast meat of environmentally enriched treated groups was lighter and had lower ultimate pH. It was concluded that the addition of environmental enrichment tools (visual, structural and plastic) motivates the birds for physical activities and improves the performance of broiler chickens. [ABSTRACT FROM AUTHOR]

The Pacific mackerel Scomber japonicus aquaculture is receiving growing attention, and further development of this industry is expected to protect wild stocks. Information on S. japonicus hematology is very limited. Therefore, the present study aimed to provide the hematological and serum chemistry profiles associated with sex and some seasonal variations in Pacific mackerel from Todos Santos Bay, Baja California, Mexico. Fish were sampled in April (spring) and August (summer). Results showed that the condition factor and all blood parameters analyzed in male and female mackerel were not affected by sex (P > 0.05). The seasonality effect on the mackerel was observed in some blood parameters such as mean corpuscular hemoglobin, total protein, globulin, and triglycerides that exhibited lower values in April compared to August (P < 0.05). While white blood cell count was higher in August (P < 0.05) compared with April. The fish sampled during the two sampling times were possibly a heterogeneous group of ages (1 to 3 years) with lengths ranging from 20 to 30 cm. Consequently, there is an urgent need to follow doing reference interval databases to assess Pacific mackerel health status under different conditions. [ABSTRACT FROM AUTHOR]

Background: The integration of paper-based analytical devices with lab-on-a-chip technology has opened up new possibilities for miniaturized, rapid, low cost and portable diagnostic testing in resource limited settings. This work introduces an integrated lab-on-a-chip platform coupling paper-based analytical devices (LOC-PADs) for dual usage: electrochemically controlled solid-phase microextraction (EC-SPME) and determination of naproxen (NAP) by potentiometric ion-selective electrode (ISE) in biological samples., Results: Conductive papers were successfully fabricated by a chemical procedure. Thereupon, a conducting molecularly imprinted polymer (CMIP) film based on PPy and an anionic model drug (NAP) as a template were applied to the conductive paper substrate via electrochemical methods. A microfluidic chip device was employed to assess the analytical performance of the fabricated paper-based CMIP. The designed chip is divided into two parts with a microfluidic channel between them. The first chamber is an extraction zone for EC-SPME of NAP by using the CMIP paper as a selective sorbent. The second chamber is the detection zone and consists of a paper-based NAP ion selective sensor for potentiometric detection of drug. The analytical process was investigated and optimized for each chamber. Remarkable selectivity towards NAP was achieved in the concentration range from 4.0 × 10 -7 to 1.0 × 10 -2  mol L -1 with determination coefficient (R 2  = 0.99) and the limit of detection (LOD) was 2.0 × 10 -7  mol L -1 . Anionic Nernstian compliance was gained -58.5 ± 0.5 mV decade -1 , and response time in the detection step was 7s for ISE. Satisfactory spiking recovery values within the acceptable range of 90-110 % with RSDs ≤7 % were achieved for the analysis of real samples with fully portable LOC device., Significance: The LOC-PADs were favorably used for selective extraction and determination of NAP in serum and saliva samples, respectively. EC-SPME caused sample clean-up, reduced or eliminated different interferences and enhanced selective response of ISE as detection zone of device. Integration of EC-SPME and ISE in a single chip enabled easy and fast determination of trace amounts of NAP in limited real sample volumes, and fully portable advantages., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This study investigated the effects of Agrimoniae Herba-Coptidis Rhizoma(XHC-HL)-medicated serum on the proliferation, migration, invasion, and apoptosis of human colorectal cancer HT29 and HCT116 cells via the autophagy mediated by lysosome-associated membrane protein type 2A(LAMP2A). Bioinformatics analysis was conducted to explore the role of LAMP2A in the development and progression of colorectal cancer. Western blot(WB) was used to detect the expression of LAMP2A protein in colorectal cancer cell lines. Lentiviral transfection was utilized to construct LAMP2A knockdown in HT29 and overexpression in HCT116 colorectal cancer cell models. Real-time fluorescence quantitative polymerase chain reaction(real-time qPCR) was performed to assess transfection efficiency. HT29 and HCT116 cells were treated with different concentrations of XHC-HL-medicated serum. The cell counting kit-8(CCK-8) assay was used to detect cell proliferation and determine the optimal concentration and duration of medicated serum intervention. HT29 cells were divided into a normal control(NC) group, an XHC-HL(medicated serum treatment) group, and an XHC-HL+shLAMP2A(medicated serum treatment+LAMP2A knockdown) group. HCT116 cells were divided into a NC group, an XHC-HL group, and an XHC-HL+LAMP2A(medicated serum treatment+LAMP2A overexpression) group. CCK-8 was used to measure cell viability. Colony formation assay was employed to assess cell proliferation ability. Scratch and Transwell migration assays were conducted to evaluate cell migration ability, and Transwell invasion assay was used to detect cell invasion ability. Flow cytometry was adopted to determine apoptosis rates. WB and real-time qPCR were employed to detect the effect of XHC-HL on the protein and mRNA expression of LAMP2A, heat shock cognate protein 70(HSC70), heat shock protein 90(HSP90), and glyceraldehyde-3-phosphate dehydrogenase(GAPDH) in colorectal cancer cells. Differential expression analysis revealed that LAMP2A expression was significantly higher in colorectal cancer patients compared to that in normal controls. Survival analysis indicated that the key molecule of chaperone-mediated autophagy(CMA), LAMP2A, was closely associated with colorectal cancer progression. Gene set enrichment analysis showed that patients with high LAMP2A expression significantly upregulated tumor progression-related signaling pathways such as angiogenesis and immune suppression. Immune infiltration analysis found that patients with high LAMP2A expression had fewer CD8 T cell infiltrations in their tumor microenvironment. XHC-HL-medicated serum inhibited the viability of HT29 and HCT116 cells, with the optimal intervention concentration and duration being 20% and 48 hours, respectively. Compared to the NC group, XHC-HL inhibited the proliferation, migration, and invasion of HT29 and HCT116 cells, and induced apoptosis. The medicated serum treatment with LAMP2A knockdown further inhibited colorectal cancer cell proliferation, invasion, and migration, and promoted apoptosis, whereas overexpression of LAMP2A reversed the inhibitory effects of the medicated serum on proliferation, migration, and invasion, and reduced apoptosis rates. XHC-HL-medicated serum inhibited CMA by upregulating the protein and mRNA expression of LAMP2A, HSC70, and HSP90 and downregulating substrate protein GAPDH expression via the autophagy mediated by LAMP2A. In conclusion, XHC-HL-medicated serum inhibits the proliferation, migration, and invasion of colorectal cancer cells and induces apoptosis by downregulating the expression of the key CMA molecule LAMP2A and inhibiting CMA activity.

Recent advances in mass spectrometry have indicated that the water-soluble antioxidant vitamin C differentially modulates the abundance of various proteins in the hepatic tissue of female and male mice. In this study, we performed LC-MS/MS to identify and quantify proteins that correlate with serum vitamin C concentrations in the whole brain, heart, liver, and spleen tissues in mice deficient for the enzyme L-Gulonolactone oxidase required for vitamin C synthesis in mammals. This work shows for the first time that various biological processes affected by a vitamin C deficiency are not only sex specific dependent but also tissue specific dependent even though many proteins have been identified and quantified in more than three organs. For example, the abundance of several complex III subunits of the mitochondrial electron transport chain correlated positively with the levels of serum vitamin C only in the liver and not in the other tissues examined in this study even though such proteins were identified in all the organs analyzed. Western blot analyses on the Uqcrc1 and Uqcrfs1 complex III subunits validated the mass spectrometry results. Interestingly, the ferritin subunits represented the few quantified protein complexes that correlated positively with serum vitamin C in all the organs examined. Concomitantly, serum ferritin light chain 1 was inversely correlated with vitamin C levels in the serum. Thus, our study provides an initial comprehensive atlas of proteins significantly correlating with vitamin C in four organs in mice that will be a useful resource to the scientific community., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Aumailley, Lebel. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Objective: The 0bjective is to compare treatment effects between undiluted autologous platelet-rich plasma (APRP) and autologous serum (AS) in patients with moderate-to-severe dry eye disease (DED)., Methods and Analysis: A single-centre, randomised, double-masked, non-inferiority clinical trial was conducted. 96 adult DED patients with an Ocular Surface Disease Index (OSDI) Score of ≥23 and/or Oxford staining grade of ≥2 were randomised to receive either 100% APRP (n=48) or 100% AS (n=48) for 4 weeks. Primary outcomes included OSDI Score and ocular surface staining measured by Oxford grading scale at 4 weeks. Secondary outcomes included fluorescein tear break-up time, Schirmer's test, meibum quality and expressibility, and adverse events. The 95% CI for the mean difference in OSDI scores between groups was estimated to assess non-inferiority of the OSDI score at a prespecified margin of 4.18 points., Results: At week 4, there was no significant difference in decreased OSDI scores between groups, with the mean difference (100% APRP-100% AS) of 1.41 (95% CI -1.26, 4.08, p=0.299). The upper limit was less than the prespecified margin, indicating non-inferiority of 100% APRP vs 100% AS. The probabilities of achieving an Oxford grade 0-1 after treatment were not significantly different between groups, with an OR of 0.61 (95% CI 0.25, 1.52, p=0.288). No significant differences in secondary outcomes were observed between groups., Conclusion: In the short-term, 100% APRP was not inferior to 100% AS in reducing dry eye symptoms and ocular surface staining in moderate-to-severe DED., Trial Registration Number: NCT04683796., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2024. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Background: Sepsis is a recognized global health challenge that places a considerable disease burden on countries. Although there has been some progress in the study of sepsis, the mortality rate of sepsis remains high. The relationship between serum osmolality and the prognosis of patients with sepsis is unclear., Method: Patients with sepsis who met the criteria in the Medical Information Mart for Intensive Care IV database were included in the study. Hazard ratios (HRs) and 95% confidence intervals (CIs) were determined using multivariable Cox regression. The relationship between serum osmolality and the 28-day mortality risk in patients with sepsis was investigated using curve fitting, and inflection points were calculated., Results: A total of 13,219 patients with sepsis were enrolled in the study; the mean age was 65.1 years, 56.9 % were male, and the 28-day mortality rate was 18.8 %. After adjusting for covariates, the risk of 28-day mortality was elevated by 99% (HR 1.99, 95%CI 1.74-2.28) in the highest quintile of serum osmolality (Q5 >303.21) and by 59% (HR 1.59, 95%CI 1.39-1.83) in the lowest quintile (Q1 ≤285.80), as compared to the reference quintile (Q3 291.38-296.29). The results of the curve fitting showed a U-shaped relationship between serum osmolality and the risk of 28-day mortality, with an inflection point of 286.9 mmol/L., Conclusion: There is a U-shaped relationship between serum osmolality and the 28-day mortality risk in patients with sepsis. Higher or lower serum osmolality is associated with an increased risk of mortality in patients with sepsis. Patients with sepsis have a lower risk of mortality when their osmolality is 285.80-296.29 mmol/L., (© 2024. The Author(s).)

Introduction: Serum is the International Federation of Clinical Chemistry (IFCC)-recommended matrix for the measurement of lactate dehydrogenase (LD); however, many laboratories opt for lithium heparin plasma to achieve quicker turnaround times and minimize tube usage. When introducing the new Sigma-Strong IFCC-recommended LDH2 assay from Abbott Laboratories on lithium-heparin collected samples, we observed a rise in the patient median LD activity as well as several samples exhibiting falsely elevated values., Materials and Methods: 120 + serum and plasma samples from consenting patients were collected and evaluated for complete blood count and lactate dehydrogenase using two different assays. Aggregated patient results before and after introduction of the LDH2 assay were compared., Results: Mean LD was 14% higher in plasma than in serum when using the LDH2 assay but only 5% higher when using the previous LDH legacy assay from Abbott Laboratories. Similarly, platelets and leukocytes were 10-30 times higher in plasma than in serum. Aggregated lactate dehydrogenase patient results demonstrated a dramatic increase in patient median following introduction of the LDH2 assay. Various experiments were tried to reduce cellular interference, but the only viable solution we found, apart from reverting to the LDH legacy assay, was to utilize serum tubes., Conclusion: We conclude that lithium-heparin plasma leads to falsely elevated lactate dehydrogenase activity when using the LDH2 assay. These errors can be prevented by using serum collected in gel separator tubes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.)

The concept of enzyme stability is typically used to refer to an enzyme's thermostability - its ability to retain structure and activity as temperature increases. For a therapeutic enzyme, other measures of stability may also be critical, particularly its ability to retain function in human serum at 37 °C, which we refer to as serum stability. Here, we describe an in vitro assay to assess the serum stability of the wildtype Homo sapiens adenosine deaminase I (HsADA1) enzyme using an absorbance-based microplate procedure. Specifically, this manuscript describes the preparation of buffers and reagents, a method arranging for the coincubation of HsADA1 in serum, a method to analyze the test samples using a microplate reader, and an accompanying analysis to determine the fraction of activity that an HsADA1 enzyme retains in serum as a function of time. We further discuss considerations to adapt this protocol to other enzymes, using an example of a Homo sapiens kynureninase enzyme, to help aid the protocol's adaptation to other enzymes where serum stability is of interest.

Fluorinated liquid-crystal monomers (FLCMs) are a potential emerging class of persistent, bioaccumulative, and toxic compounds. Humans inevitably ingest FLCMs via food and the environment. However, there are limited studies on internal exposure biomonitoring of FLCMs. Herein, we evaluated the estimated daily intakes (EDIs) of FLCMs in the general population based on serum residue levels. For the first time, 38 FLCMs were detected in 314 serum samples from the general population in Beijing, with a median value of 132.48 ng/g of lipid weight (lw). BDPrB is a predominant FLCM in serum. The median EDI of ∑ 38 FLCMs in the general residents was 37.96 pg/kg bw/day. The residual levels of most FLCMs were higher in urban than in suburban areas ( p < 0.05). The concentrations of EFPEB, EDPrB, EDFPBB, and PDTFMTFT in serum showed positive associations with blood glucose (GLU) ( r = 0.126-0.275, p < 0.05). Logistic regression analysis showed that FLCMs were significantly positively correlated with dyslipidemia, with an odds ratio of 2.19; BDPrB was significantly positively correlated with hyperglycemia (OR: 2.48). Overall, the present study suggests the occurrence of FLCMs in the nonoccupational population, and the exposure of certain FLCMs may cause abnormal blood glucose and lipid levels.

Label-free Surface Enhanced Raman Spectroscopy (SERS) is a rapid technique that has been extensively applied in clinical diagnosis and biomedicine for the analysis of biofluids. The purpose of this approach relies on the ability to detect specific "metabolic fingerprints" of complex biological samples, but the full potential of this technique in diagnostics is yet to be exploited, mainly because of the lack of common analytical protocols for sample preparation and analysis. Variation of experimental parameters, such as substrate type, laser wavelength and sample processing can greatly influence spectral patterns, making results from different research groups difficult to compare. This study aims at making a step toward a standardization of the protocols in the analysis of human serum samples with Ag nanoparticles, by directly comparing the SERS spectra obtained from five different methods in which parameters like laser power, nanoparticle concentration, incubation/deproteinization steps and type of substrate used vary. Two protocols are the most used in the literature, and the other three are "in-house" protocols proposed by our group; all of them are employed to analyze the same human serum sample. The experimental results show that all protocols yield spectra that share the same overall spectral pattern, conveying the same biochemical information, but they significantly differ in terms of overall spectral intensity, repeatability, and preparation steps of the sample. A Principal Component Analysis (PCA) was performed revealing that protocol 3 and protocol 1 have the least variability in the dataset, while protocol 2 and 4 are the least repeatable., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)