Serum Chemistry Analysis Research Articles

Evaluation of Relationship Between C-Reactive Protein, Leukocyte Count and Platelet Indices in Dogs with Leukocytosis

A complete blood count and acute-phase protein analyses are the laboratory tests that are used in veterinary medicine practice. Leukocytes, also known as white blood cells (WBCs), are the primary biological components of inflammatory and immunological responses. C-reactive protein (CRP) is a powerful predictor of inflammation and/or infection in veterinary medicine. Starting at that point, the investigation of the alterations and relationship between WBC count, platelet indices, and blood serum CRP levels in dogs with leukocytosis was the aim of this study. A total of 135 bloodwork records, from January 2018 to December 2022, of dogs were analyzed. The inflammatory panel of complete blood count data, including WBC, PLT, MPV, PDW, PCT, and MPV/PLT, and CRP levels in blood serum chemistry analyses were investigated. The inflammatory panel of complete blood count data was categorized according to CRP levels in blood serum chemistry analyses. There was no significant difference in platelet indices according to the blood serum levels of CRP in the dogs’ bloodwork records. The WBC count in dogs with an equal or above 10 mg/dL CRP level was significantly higher than the other levels of CRP. A moderately positive correlation was detected between CRP and WBC in the bloodwork of all dogs, undivided according to inflammatory status, and of dogs with an inflammatory status. Consequently, in veterinary clinical practice, C-reactive protein measures in dogs may be more applicable in cases where an important rise in WBC over the cut-off value is observed in the complete blood count, commonly used to diagnose inflammatory or infectious status.

Serum chemistry, thyroxine concentration, and blood cell morphology of wild, young Blackchin Guitarfish Glaucostegus cemiculus sampled on the eastern Mediterranean coastline.

The Blackchin Guitarfish Glaucostegus cemiculus is endemic to the Mediterranean Sea and is critically endangered, but relevant routine laboratory data are unavailable. Our objectives were to determine the packed cell volume (PCV), comprehensive serum chemistry analytes, and serum total thyroxine (sTT4) concentration; compare serum albumin and serum globulin concentrations as measured by two different methods; and describe the blood cell morphology of healthy, free-ranging Blackchin Guitarfish. Wild Blackchin Guitarfish were captured using a seine net. Blood samples for serum chemistry and hematological analyses were obtained and measured using routine laboratory methods. The fish were tagged and released. This study included 43 Blackchin Guitarfish (17 males and 26 females) that were younger than 6 months as estimated based on total length and body weight. The median PCV (n = 23) was 22% (minimum-maximum [min-max] = 15-25%). Median sTT4 (n = 10) measured by chemiluminescence immunoassay was 7.86 nmol/L (min-max = 7.52-9.57 nmol/L). The study included a comprehensive, 25-analyte serum chemistry analysis (e.g., serum iron and unbound and total iron-binding capacity) and a morphological description of all blood cells. Serum electrophoresis (SEP; n = 13) yielded a consistent serum albumin-migrating protein fraction and four globulin fractions. Serum electrophoretograms corroborating these results are presented. In Blackchin Guitarfish, the serum albumin-migrating fraction measured by SEP combined with serum total protein concentration yields a much higher albumin concentration compared to that measured by bromocresol green spectrophotometry. The true identity of this albumin-migrating fraction remains to be identified. The analytes' calculated 2.5-97.5% interpercentile intervals should be considered as reference intervals applying to Blackchin Guitarfish of similar age but should be applied cautiously to adult fish.

Local administration of irinotecan using an implantable drug delivery device stops high-grade glioma tumor recurrence in a glioblastoma tumor model

The treatment for Glioblastoma is limited due to the presence of the blood brain barrier, which restricts the entry of chemotherapeutic drugs into the brain. Local delivery into the tumor resection margin has the potential to improve efficacy of chemotherapy. We developed a safe and clinically translatable irinotecan implant for local delivery to increase its efficacy while minimizing systemic side effects. Irinotecan-loaded implants were manufactured using hot melt extrusion, gamma sterilized at 25 kGy, and characterized for their irinotecan content, release, and drug diffusion. Their therapeutic efficacy was evaluated in a patient-derived xenograft mouse resection model of glioblastoma. Their safety and translatability were evaluated using histological analysis of brain tissue and serum chemistry analysis. Implants containing 30% and 40% w/w irinotecan were manufactured without plasticizer. The 30% and 40% implants showed moderate local toxicity up to 2- and 6-day post-implantation. Histopathology of the implantation site showed signs of necrosis at days 45 and 14 for the 30% and 40% implants. Hematological analysis and clinical chemistry showed no signs of serious systemic toxicity for either implant. The 30% implants had an 80% survival at day 148, with no sign of tumor recurrence. Gamma sterilization and 12-month storage had no impact on the integrity of the 30% implants. This study demonstrates that the 30% implants are a promising novel treatment for glioblastoma that could be quickly translated into the clinic.Graphical

Prebiotic fiber blend supports growth and development and favorable digestive health in puppies.

A healthy gastrointestinal (GI) microbiome has been shown to be essential for proper nutrient absorption and metabolism, maintenance of intestinal epithelial integrity and osmolarity, gut immunomodulation, and overall health. One of the most effective ways to promote a healthy GI microbiome is through dietary interventions, such as the addition of prebiotics. Prebiotics are substrates that are selectively utilized by the host GI microbiome through fermentation to confer a health benefit. However, research on prebiotics in companion animals is limited, especially in growing animals. Thus, this study was conducted to assess the effects of a novel prebiotic fiber blend on key parameters related to intestinal health and growth in puppies. Twenty-two puppies at least 4 months of age but not older than 10 months were fed a commercially available dry food during a prefeed period, and then fed a similarly formulated test food with the addition of the prebiotic fiber blend for a minimum of 90 days. Serum and fecal samples were collected at the end of the prefeed period and throughout the test period. Puppies fed the test food grew as expected for puppies of this age. Complete blood count and serum chemistry analyses were clinically normal for all animals. Fecal score increased linearly, fecal moisture decreased linearly, and pH exhibited a cubic trend throughout the study duration. There was a linear increase in short-chain fatty acids throughout the study, which is associated with favorable digestive and overall health. The inflammatory cytokine interleukin-7 decreased linearly and interleukin-18 trended towards linear decrease. This study showed that puppies continued to grow and develop normally, and experienced serum and stool characteristics indicative of improved GI health when fed a growth food fortified with a novel prebiotic fiber blend. Furthermore, these results contribute to the overall understanding of the effects of prebiotics on the GI health of growing companion animals.

Abstract B179: 2-Deoxy-D-glucose (2-DG) combination with ketogenic diet induces sedation and hypothermia in mice that becomes lethal

Abstract Introduction. The term “ketogenic” was coined for the diet that specifically consists of a 4:1 ratio of fats to proteins. 2-Deoxy-d-glucose (2-DG) is a synthetic glucose analog that potently inhibits glycolysis. It has been hypothesized that 2-DG in combination with KD, should improve the anticancer effects of either therapeutic approach. The therapeutic benefit of such a combination was supported by studies of Voss et al. (Int J Mol Sci, 2018. 19(8): PMC6121440), showing that continued combination of KD and anti-glycolytic therapy is feasible, and KD increased tolerance to 2-DG. However, in contrast to this report, our preliminary studies revealed a dose-dependent reduction in the survival of mice of receiving inhibition of glycolysis (2-DG) and KD. The objective of this study was to assess the toxic limitation of the combination of glycolysis inhibition and ketogenic diet. Methods. CD-1 mice were maintained on either a standard diet (SD) or a ketogenic diet (KD). Some groups were treated with of 2-DG at 1.5 g/kg via the oral or intraperitoneal route or vehicle. For temperature monitoring, the animals were implanted subcutaneously with a temperature microchip UCT-2112. Body temperature was recorded manually for each mouse using a contactless, handheld URH-1HP receiver. Alternatively, for continuous monitoring of the body temperature, the UID Mouse Matrix was used. Mice exhibiting treatment-related toxicity symptoms, such as prolonged hypothermia or laborious breathing, and euthanized following blood collection for hematology and serum chemistry analysis. Key organs were also collected for histopathology analysis. Results. Previously, we reported that 2-DG reduces the survival of mice maintained on KD (median survival 9 days p=0.0022), while it is well tolerated by mice receiving regular chow (no deaths reported). Here, we report that the administration of 2-DG induces transient sedation and hypothermia. CBC analysis revealed that the number of platelets in the KD + 2-DG group was significantly decreased compared to the SD + 2-DG group and the KD + vehicle group (p<0.001). The reduction of platelets was associated with a two- to three-fold increase of LDH in the KD + 2-DG group. Histopathological analysis of the lung tissue revealed the presence of diffuse congestion in blood vessels and dilation of the alveolar capillaries consistent with interstitial pneumonia. Similarly, degenerative and congestive hepatic parenchyma was observed in animals treated with 2-DG and KD. Preliminary experiments involving supplementation of 2-DG-treated animals with D-glucose or D-mannose failed to provide a rescue effect. Conclusion. A highly reproducible, dose-dependent toxicity leading to rapid death of treated animals maintained on KD was observed following 2-DG administration, while the same doses appeared to be safe in mice fed a standard diet. The toxic effects induced by the combination treatment included acute thrombocytopenia and acute interstitial pneumonia; however, delineation of the exact mechanism of toxicity requires additional studies. Citation Format: Rafal Zielinski, Moises Martinez Castillo, Damian Grybowski, Krzysztof Grela, Roberto Cardenas-Zuniga, Izabela Fokt, Stanislaw Skora, Gerardo Ramirez, Waldemar Priebe. 2-Deoxy-D-glucose (2-DG) combination with ketogenic diet induces sedation and hypothermia in mice that becomes lethal [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr B179.

PSIX-2 Effects of Diets Supplemented with Bioactive Peptides on Nutrient Digestibility, Immune Cell Responsiveness, and Fecal Characteristics, Microbiota, and Metabolites of Adult Cats

Abstract Bioactive peptides (BP) are recognized for their ability to function as antioxidants and maintain lipid stability. They may have positive health effects, including antihypertensive, anti-inflammatory, antimicrobial, osteoprotective, gut health and immunomodulatory properties. Our objective was to determine the apparent total tract digestibility (ATTD) of bioactive peptide-containing kibble diets and assess how the fecal characteristics, metabolites, and microbiota were affected in adult cats. All procedures were approved by the University of Illinois IACUC. Twelve adult cats (4-5 yr; 4.84 kg avg. BW) were used in a replicated 4x4 Latin square design (n = 12/group) and fed to maintain BW. In each experimental period, cats were allotted to one of four extruded kibble experimental diets: control (4% dried chicken), chicken (4% chicken BP), marine1 (2% marine BP), and marine2 (4% marine BP). Health was confirmed by baseline serum chemistry and complete blood count analysis. Each experimental period lasted 28 days, with a 20-day adaptation phase, 5-day fecal collection [for nutrient digestibility, microbiota, metabolites, IgA, calprotectin, lactoferrin, pH, and dry matter (DM) analysis], 2-day blood collection (for serum chemistry, complete blood count, and immune cell functionality analysis), and 1 day for transport stress testing. For transport stress, cats were transported in a vehicle in individual carriers for 45 minutes. Upon return, their behavior was evaluated using the cat stress score (CSS). Saliva samples were collected within 15 minutes to measure cortisol concentrations and blood samples were collected after 3 hours to measure concentrations of oxidative stress markers [superoxide dismutase (SOD) and malondialdehyde (MDA)] and inflammatory cytokines. Fecal microbiota data were evaluated using QIIME2. All other data were analyzed using the Mixed Models procedure of SAS, with P < 0.05 being considered significant. No significant differences were observed in animal health outcomes, with all cats remaining healthy within reference ranges. There were no differences in fecal characteristics, metabolites, and microbiota diversity. Cats fed the marine2 diet had a 3.61% greater (P < 0.05) ATTD of DM and a 2.5% greater organic matter digestibility than those fed the control diet. The relative abundance of fecal Blautia was greater (P < 0.05) in cats fed the control diet than those fed the chicken diet, while relative abundance of fecal Slackia was greater (P < 0.05) in cats fed the control diet than those fed the chicken or marine1 diets. In comparison with cats fed the marine1 diet, the relative abundances of fecal Sellimonas and Bifidobacterium were less (P < 0.05) in cats fed the control diet. Neither transport stress nor diet impacted the oxidative stress markers, cytokines, cortisol, or CSS. Furthermore, the diet did not affect immune cell functionality. In conclusion, including BP, in particular marine2 at 4% inclusion to feline diets may increase nutrient digestibility and modify fecal microbiota.

Pathological, biochemical and hormonal changes associated with prolonged administration of methanol extract of <i>Garcinia kola</i> seed in adult male Wistar rats.

Garcinia kola (GK) seed is believed to possess aphrodisiac properties hence its traditional use to manage erectile dysfunctions in man. However, there has been conflicting reports of its long-term effects on the male reproductive organs. In this study, the effect of varying doses of methanol extract of GK on testicular function and structure in Wistar rats was evaluated. Forty adult male rats (350-360g) were randomly divided into 4 groups. Groups 1, 2 and 3 received oral doses of 600mg/kg, 400mg/kg and 100mg/kg of the extract daily, respectively for 70 days while group 4 (control group) received normal saline (0.9% NaCl). Blood samples were collected fortnightly from each rat for serum chemistry and testosterone analysis. At termination, the testes, epididymides and livers were harvested, weighed and fixed in 10% neutral buffered formalin and processed for histopathology. The results were presented as mean ± standard error of mean (SEM) and analyzed using one way analysis of variance (ANOVA) at P< 0.05. Dose-related pathological changes were observed in the testes and epididymides. Testicular weight decreased significantly (p<0.05) in 400mg/kg and 600mg/kg groups compared to the control group. There was testicular necrosis associated with distortion of the seminiferous tubular wall and reduced seminiferous tubular diameter. Seminiferous tubules had reduced germinal epithelial thickness along with an absence of viable spermatids and spermatozoa. Serum testosterone levels decreased progressively (p < 0.01) in groups 1 and 2 while those of groups 3 and 4 remain unchanged. The results established that ingestion of methanol extracts of GK seed at the dose of 400mg/kg and 600mg/kg produced significant gonadal pathology in adult male Wistar rats despite the absence of remarkable hepatic biochemical and structural changes.

Hematology and Serum biochemistry of Broiler Chickens Fed Red Sorghum (Sorghum bicolor (L.) Moench) Based Diets supplemented with Complex Enzyme (Kingzyme®) in Girei, Adamawa State, Nigeria

The study was designed to study the Hematology and Serum biochemistry of Broiler Chickens Fed Red Sorghum (Sorghum bicolor (L.) Moench) Based Diets supplemented with Complex Enzyme (Kingzyme®). Two hundred (200) chicks (day-old) of mixed sex was randomly allotted to five dietary treatments (T1, T2, T3, T4 and T5) supplemented with enzyme complex (Kingzyme®) at the levels of 0.0g, 0.2g, 0.5g, 0.8g and 1.1g kg per Kg of feed respectively in a Completely Randomized Design (CRD). Each treatment was replicated four (4) times to contain ten (10) birds each. Feed and water were provided ad libitum accompanied with standard management procedures for eight weeks. At end of the eight weeks’ experimental period, blood samples were collected for hematological and serum chemistry analysis. Data were analyzed using Statitix Analytical Software, Version 10; and the treatment means were compared using Turkey HSD.Except the RBC, MCV, MCH and Eosinophil which showed significant (P < 0.05) difference across the treatment groups, all other hematological parameters were not significantly (P > 0.05) affected. The TP, Alb and Glb were also not significantly (P > 0.05) affected. The Urea and Glucose were not significantly (P > 0.05) affected. The Creatinine and Cholesterol were significantly (P > 0.05) affected across the dietary treatment. Based on the result of the study, it was concluded that Red Sorghum-based diet supplemented with enzyme complex (Kingzyme®) do not have any significant effect on hematological and serum biochemistry of broiler chickens.

1847-PUB: Neutrophil Count Predicts Cardiovascular and Metabolic Health Risk Factors in a Population without Diabetes

Subclinical inflammation is implicated in cardio-metabolic conditions such as impaired glucose metabolism, diabetes, and hypertension. Neutrophils are first line of immune defense call and play a role in systemic inflammation. We therefore hypothesized that neutrophil quantity will be positively associated with cardio-metabolic risk factors among individuals without diabetes. Purpose: This study sought to determine the relevance of neutrophil count as a biomarker for cardio-metabolic health. Methods: Forty participants without diabetes (11 males, 29 females; age 28.4±10.7 years; BMI 28.2±6.0 kg/m2, fasting blood glucose 97.28±11.09 mg/dL) participated in the study. Following overnight fasting, participants’ blood pressure and resting heart rate were measured, and venous blood draw was performed. All participants completed a 3-hour oral glucose tolerance test with 75g glucose drink. Hematological and serum chemistry analyses were performed. Body composition was measured by dual X-ray absorptiometry. Homeostatic model assessment for insulin resistance (HOMA-IR) was computed from standard equations. Spearman’s correlation was performed to determine relationship between neutrophil count and markers of cardio-metabolic health. Results: Neutrophil count is positively correlated with resting heart (r=0.5, p=0.004), triglycerides (r=0.5, p=0.002), triglycerides-HDL ratio (r=0.5, p=0.002), VLDL (r=0.5, p=0.001), Globulin (r=0.3, p=0.03), BUN/creatinine ratio (r=0.5, p=0.0004), fasting insulin (r=0.7, p=0.004), fasting blood glucose (r=0.5, p=0.0008), 2-hour OGTT glucose (r=0.5, p=0.003), glucose area under the curve (r=0.5, p=0.0001), percent body fat (r=0.6, p=0.0003), HOMA-IR (r=0.6, p=0.008) and negatively correlated with albumin/globulin ratio (r=-0.4, p=0.01), HDL (r=-0.3, p=0.04). Conclusion: Neutrophil count is a biomarker for cardio-metabolic health risk factors in a population without diabetes. Disclosure J.Labadah: None. J.Apaflo: None. G.Narvaez: None. V.M.Rocha: None. A.Mossayebi: None. S.Bajpeyi: None. Funding The University of Texas at El Paso

Effects of a milk oligosaccharide biosimilar on fecal characteristics, microbiota, and bile acid, calprotectin, and immunoglobulin concentrations of healthy adult dogs treated with metronidazole.

In recent dog and cat experiments, a novel milk oligosaccharide biosimilar (GNU100) positively modulated fecal microbiota and metabolite profiles, suggesting benefits to gastrointestinal health. The objective of this study was to investigate the effects of GNU100 on the fecal characteristics, microbiota, and bile acid (BA) concentrations of healthy adult dogs treated with antibiotics. Twelve healthy adult female dogs (mean age: 3.74 ± 2.4 yr) were used in an 8-wk crossover design study (dogs underwent both treatments). All dogs were fed a control diet during a 2-wk baseline, then randomly allotted to 1 of 2 treatments (diet only or diet + 1% GNU100) for another 6 wk. From weeks 2 to 4, dogs were orally administered metronidazole (20 mg/kg BW) twice daily. Fecal scores were recorded daily and fresh fecal samples were collected at weeks 2, 4, 5, 6, and 8 for measurement of pH, dry matter, microbiota populations, and BA, immunoglobulin A, and calprotectin concentrations. On weeks 0, 4, and 8, blood samples were collected for serum chemistry and hematology analysis. All data were analyzed as repeated measures using the Mixed Models procedure of SAS version 9.4, with significance considered P < 0.05. Metronidazole increased (P < 0.0001) fecal scores (looser stools) and modified (P < 0.05) fecal microbiota and BA profiles. Using qPCR, metronidazole reduced fecal Blautia, Fusobacterium, Turicibacter, Clostridium hiranonis, and Faecalibacterium abundances, and increased fecal Streptococcus and Escherichia coli abundances. DNA sequencing analysis demonstrated that metronidazole reduced microbial alpha diversity and influenced the relative abundance of 20 bacterial genera and families. Metronidazole also increased primary BA and reduced secondary BA concentrations. Most antibiotic-induced changes returned to baseline by week 8. Fecal scores were more stable (P = 0.01) in GNU100-fed dogs than controls after antibiotic administration. GNU100 also influenced fecal microbiota and BA profiles, reducing (P < 0.05) the influence of metronidazole on microbial alpha diversity and returning some fecal microbiota and secondary BA to baseline levels at a quicker (P < 0.05) rate than controls. In conclusion, our results suggest that GNU100 supplementation provides benefits to dogs treated with antibiotics, providing more stable fecal scores, maintaining microbial diversity, and allowing for quicker recovery of microbiota and secondary BA profiles which play an essential role in gut health.

Dietary Ground Flaxseed Increases Serum Alpha-Linolenic Acid Concentrations in Adult Cats.

Simple SummaryThe use of flaxseed in complete pet foods as a source of omega-3 fatty acids, and in particular alpha-linolenic acid, has been studied extensively in dogs. However, to date, no scientific published literature exists on the impacts of feeding whole ground flaxseed as an ingredient in cat food on nutrient digestibility, fatty acid bioavailability, and overall health in cats. In addition, dietary flaxseed inclusion has been challenged as providing little nutritional value for cats by not changing serum fatty acid levels and lowering nutrient digestibility. Therefore, the objective of this study was to directly evaluate the effects of adding ground flaxseed to dry cat food on fatty acid bioavailability, stool consistency, nutrient digestibility, and overall health in adult cats. These results clearly exhibit that a flaxseed-supplemented diet resulted in no changes in stool quality or nutrient digestibility, and elicited an increase in serum alpha-linolenic acid concentrations in healthy cats. Overall, these results clearly demonstrate that ground flaxseed can be used to modulate serum fatty acid concentrations and serve as a bioavailable source of alpha-linolenic acid in commercial cat food.We evaluated effects of dietary ground flaxseed on fecal and serum alpha-linolenic acid (ALA) concentrations, nutrient digestibility, and stool quality in female and male adult cats (n = 20 (8 males, 12 females); 3.95 ± 1.49 years of age (mean ± SD); 3.88 ± 0.82 kg BW). We hypothesized that adding ground flaxseed would increase serum ALA compared with feeding no flax, without changing nutrient digestibility. Cats were fed as-is 2.6% added-flaxseed (flax, n = 10) or no-flax (control, n = 10) diets (2.66 vs. 0.78% ALA of total fatty acids; crude protein 35%, fat 20%, fiber 3% as-fed) twice daily to maintain body weight for 28 days. Fecal collections were conducted on days 23–27 for total-tract nutrient digestibility, stool quality (scale 1–5; 1 = watery diarrhea, 5 = hard, dry, crumbly) and long-chain fatty acid (LCFA) analyses. Blood was collected on days 0, 14, and 28 for serum LCFA and chemistry analysis. Digestibility and fecal data were analyzed by ANOVA (SAS v9.4, Cary, NC, USA) and a repeated measures ANOVA for serum ALA. Flax-fed cats, compared with control-fed, had greater (p < 0.05) serum ALA after 14 days (4.00 vs. 0.71 µg/mL) and 28 days (7.83 and 3.67 µg/mL). No differences were observed in stool quality, and dry matter, protein, fat, and ALA digestibility. However, metabolizable energy was greater in the flax vs. control diet (4.18 vs. 3.91 kcal/g; p < 0.05). Overall, these data demonstrate that ground flaxseed added to cat diets increases serum ALA within 14 days, with no detriments to nutrient digestibility. We conclude that flaxseed can be used as a bioavailable source of ALA in cat diets.

Toxic Effects of High-dose Meloxicam and Carprofen on Female CD1 Mice.

The nonsteroidal anti-inflammatory drugs meloxicam and carprofen are commonly used as analgesics in mice. The current recommended doses of meloxicam at 0.2-1.0 mg/kg once daily and carprofen at 5-10 mg/kg twice daily may not be adequate to provide analgesia in mice. Several studies have suggested that doses up to 20 mg/kg of meloxicam and carprofen are needed to provide analgesic efficacy. This study investigated the clinical safety of these higher doses of meloxicam and carprofen by evaluating their potential for renal and gastrointestinal toxicity. Female CD-1 mice were given 20 mg/kg of either meloxicam, carprofen, or an equivalent volume of saline subcutaneously once daily for 3 or 7 d. On day 4, mice treated for 3 d were euthanized, and on days 8 and 15, mice treated for 7 d were euthanized. Blood was collected by cardiocentesis for serum chemistry analysis. Feces was collected from the colon for fecal occult blood testing, and tissues were collected for histopathology. No clinically significant changes in serum chemistry profiles were found in the drug-treated mice at any time point as compared with the saline controls. Fecal occult blood and histologic evidence of gastritis was associated with meloxicam administration in mice evaluated at days 4 and 8. By day 15, there was no association with meloxicam treatment and the presence of fecal occult blood or gastritis. There was no association between fecal occult blood and gastritis in the carprofen or saline-treated mice regardless of the treatment durations. These findings suggest that 20 mg/kg of meloxicam in mice causes gastric toxicity when given for 3 or 7 d and should be used cautiously; however, carprofen at 20 mg/kg appears to have minimal toxic effects with regard to the parameters measured.

Safety and Clinical Response Following a Repeat Intraarticular Injection of Tin-117m (117mSn) Colloid in Dogs with Elbow Osteoarthritis.

ObjectiveTo determine if a repeat intraarticular (IA) injection of a tin-117m colloid radiosynoviorthesis (RSO) agent can be safely given in the same joint 12 months after an initial injection for treatment of canine elbow osteoarthritis (OA), and to evaluate the pain reduction effect of the repeat injection.Methods and MaterialsNine client owned dogs with grade 1 or 2 elbow OA were given an IA injection of tin-117m colloid in both elbows, one of which had been treated ≤12 months earlier with the same RSO device. Treatment safety was evaluated by joint fluid analysis at baseline (BL) and at 180 days after treatment, and by urinalysis, CBC, and serum chemistry analysis of diagnostic samples obtained at BL and 180 days. Radiographs, computed tomography, and MRI scans were obtained at BL and 180 days to determine if disease progression differed in elbows given one versus two injections. Clinical response to treatment was assessed subjectively by dog owner responses to the Canine Brief Pain Inventory (CBPI) survey at BL, 90 and 180 days, and objectively by investigator-conducted force plate (FP) analysis of dogs at BL, 90, and 180 days.ResultsAll post-treatment urinalysis, CBC and clinical chemistry results were within normal ranges. Joint fluid analysis showed a significant (P=0.0411) reduction in the percentage of monocytes at 180 days, consistent with the tin-117m colloid mode of action of apoptosis of pro-inflammatory macrophages at the injection site. There was no significant difference in OA progression in elbows given one or two injections. The treatment success rate was 55.5% (5/9) on day 90 as determined either by CBPI responses or FP analysis, and 66.6% (6/9) on day 180 as determined by FP analysis.ConclusionThe tin-117m colloid can be safely given as a repeat injection 12 months after an initial injection, and can potentially provide a durable therapeutic response in dogs with elbow OA.

A double-blinded placebo-controlled evaluation of adipose-derived mesenchymal stem cells in treatment of canine atopic dermatitis.

Mesenchymal stem cells (MSCs) have emerged as a new therapy for various immune-mediated inflammatory diseases. In this study we perform the first double-blinded, placebo-controlled evaluation of the efficacy of adipose-derived allogenic canine MSCs for the treatment of canine atopic dermatitis (cAD). Enrolled canine patients were randomly divided into placebo (PBS saline), low-dose (5 × 105 cells/kg), and high-dose (5 × 106 cells/kg) treatment groups. Each patient received three subcutaneous MSCs treatments or PBS saline at four-week intervals with injections at five sites. Patients were monitored by physical exams, pruritus visual analog scales (PVAS) signed by the primary caretaker, canine atopic dermatitis extent and severity index-4 (CADESI-4) scores by two veterinarians, and complete blood count and serum chemistry analysis along with laboratory analysis for potential biomarkers. Patients were kept off any immune-modulating drugs during the study period, and oral antibiotics and topicals were used for managing pruritus and secondary infections. The PVAS scores and the serum miR-483 levels were significantly lower in the high dose group compared to the placebo group at day90 post first-treatment. The CADESI-4 scores of the high dose group also showed downward trends. No severe adverse effects were observed in any patient in this study. The high dose MSC treatment is efficacious in alleviating the clinical signs of cAD until 30days after the last subcutaneous administration of MSCs, and miRNA-483 may be a reliable prognostic biomarker for cAD. The MSCs efficacy and potential biomarkers should be further explored by a larger scale clinical trial.

Serum biochemistry and haematology in wild and captive bearded seals (Erignathus barbatus) from Svalbard, Norway

BackgroundHealth assessment of seals in captivity include haematology and serum biochemistry measurements. Because such parameters differ between species, it is crucial to have species-specific reference values for the interpretation of clinical samples. Furthermore, differences in nutrition and environment, life cycles as well as seasonal/annual cycles and varying physiological conditions can potentially affect serum chemistry and haematology parameters. Blood samples from four captive adult bearded seals (initially caught as pups in Svalbard, Norway, now held at Polaria, an Arctic experience centre in Tromsø, Norway) collected over a 16-month period were analysed for haematology (n = 22) and serum chemistry (n = 25) parameters. Serum chemistry analyses were also conducted on blood samples from 74 wild bearded seals (1995–2007) collected from Svalbard, Norway.ResultsWe found higher activity of creatine kinase (CK) and higher concentrations of cortisol in the wild animals when compared to the captive seals, probably reflecting the physical restraint and concomitant stress induced during sampling. For the captive bearded seals, we did not find marked differences in haematology or serum chemistry parameters throughout the different seasons of sampling.ConclusionsThis study presents haematology and serum chemistry reference values for captive and wild bearded seals. Comparing physiological parameters for captive seals with wild seals indicated that having wild-caught bearded seals under the conditions offered at Polaria for several years did not markedly affect physiological parameters of the animals, and that training may have helped to alleviate stress associated with blood sampling and veterinary inspection.

CHRONIC PANCREATIC DISEASE IN THE LESSER KUDU (TRAGELAPHUS IMBERBIS): A REPORT OF 16 CASES IN THE UNITED STATES.

A review of archival cases at Northwest ZooPath from 1995-2018 identified 16/96 (17%; eight females, eight males) lesser kudus (Tragelaphus imberbis) with chronic pancreatic disease (CPD) from three institutions, all of which originated from an initial founder stock of 12 animals. Ages at time of death or euthanasia ranged from 2 to 132 mo (average age = 69 mo). All cases had varying degrees of pancreatic acinar cell depletion, ductular hyperplasia, and fibrosis. Serum fructosamine, glucose, insulin, and insulin to glucose ratio collected close to time of death from 13 affected animals were not significantly different from controls (n = 19). Of these analytes, receiver operating characteristic analysis identified fructosamine as the best-performing analyte with an area under the curve 0.671 (95% confidence interval = 0.480-0.828; sensitivity = 58.3%, specificity = 84.2%; cutoff point >352 µmol/L) in the diagnosis of CPD. With a 15% prevalence, there was a positive predictive value of 41% and a negative predictive value of 92%, indicating that the probability of false positives is high, but of false negatives is low. An etiologic agent was not identified by histology (n = 16), transmission electron microscopy (n = 1), or enterovirus PCR (n = 2). Serum zinc and hepatic heavy metal analyses were judged to be within normal limits. Chronic pancreatic disease is considered an important cause of morbidity and mortality in the lesser kudu; serum chemistry analysis warrants further investigation in its use for diagnosis. The etiopathogenesis is not understood, but the absence of obvious causes, the occurrence also in very young animals, and the inbred lineage of lesser kudus in the United States suggest a genetic basis for this disease.

Haematological and serological response of broiler chickens to varying levels of direct fed microbes as feed additive

Several efforts are being made to find the possibilities of utilizing Direct fed microbes (DFM) that has a potential to replace antibiotics as a result of its ability to prevent internal colonization of enteropathogenic enzymes, stimulate intestinal immunity of broiler chickens and reduce stress in animals. A studywas conducted using 120, one day-old Ross 308 broiler chicks to evaluate blood and serum chemistry response to direct fed microbes. The broiler chicks were obtained from a reputable hatchery and randomly assigned to four dietary treatments, each with three replicates of ten birds per treatment. The treatments were T1, T2, T3 and T4, with levels of inclusion of direct fed microbes (DFM) at 0, 2.5, 5% and 7.5%, respectively. The birds were assigned to these treatment diets, feed and water was given ad libitum throughout the duration of the experiment which lasted for 56 days. The experimental design was Completely Randomized Design (CRD). At the end of the experiment blood samples were collected from the chickens and taken to the laboratory for haematological and serum chemistry analysis. The haematological characteristics of the broiler chickens showed that there was no significant (P&gt;0.05) in haemoglobin (Hb), Packed Cell Volume (PCV), Red Blood Cell (RBC), White Blood Cell (WBC), Mean Corpuscular Volume (MCV), Mean Corpuscular Haemoglobin (MCH) and Mean Cell Haemoglobin Concentration (MCHC). The WBC portrayed decreasing trend as the level of DFM increased, also the Hb, MCV and MCH portrayed an increasing trend as the DFM increased in the treatment diet up to the 5% inclusion level. The serum chemistry result shows that there were significant differences (P&lt;0.05) in total serum protein and globulin whereas no significant differences (P&gt;0.05) were observed in serum albumin, glucose, urea and creatinine. It could be concluded that fed diet 2 containing 2.5% DFM performed better as they compete favourably with the control. Broiler chickenss fed diet 2 also had a balanced physiological system

Comprehensive pulmonary toxicity assessment of cetylpyridinium chloride using A549 cells and Sprague-Dawley rats.

Cetylpyridinium chloride (CPC), a quaternary ammonium compound and cationic surfactant, is used in personal hygiene products such as toothpaste, mouthwash, and nasal spray. Although public exposure to CPC is frequent, its pulmonary toxicity has yet to be fully characterized. Due to high risks of CPC inhalation, we aimed to comprehensively elucidate the in vitro and in vivo toxicity of CPC. The results demonstrated that CPC is highly cytotoxic against the A549 cells with a half-maximal inhibitory concentration (IC50 ) of 5.79 μg/ml. Following CPC exposure, via intratracheal instillation (ITI), leakage of lactate dehydrogenase, a biomarker of cell injury, was significantly increased in all exposure groups. Further, repeated exposure of rats to CPC for 28 days caused a decrease in body weight of the high-exposure group and the relative weights of the lungs and kidneys of the high recovery group, but no changes were evident in the histological and serum chemical analyses. The bronchoalveolar lavage fluid (BALF) analysis showed a significant increase in proinflammatory cytokines interleukin (IL)-6, IL-1β, and tumor necrosis factor (TNF)-α levels. ITI of CPC induced focal inflammation of the pulmonary parenchyma in rats' lungs. Our study demonstrated that TNF-α was the most commonly secreted proinflammatory cytokine during CPC exposure in both in vitro and in vivo models. Polymorphonuclear leukocytes in the BALF, which are indicators of pulmonary inflammation, significantly increased in a concentration-dependent manner in all in vivo studies including the ITI, acute, and subacute inhalation assays, demonstrating that PMNs are the most sensitive parameters of pulmonary toxicity.

Research Note: Comparison of chicken blood chemistry and electrolyte parameters between the portable i-STAT1 clinical analyzer and VetScan VS2 serum biochemistry panel using Hy-Line commercial white-egg laying hens

The i-STAT1 clinical analyzer has become an increasingly popular tool in clinical production animal medicine as it can provide pen-side results in a cost effective and timely manner when compared to standard benchtop serum biochemistry blood gas and chemistry analyses. This study compares the results of the portable Abbott i-STAT1 analyzer and the Abaxis VetScan VS2 for glucose (Glu, mg/dL), ionized Ca (mmol/L), Na (mmol/L), and K (mmol/L) values. Three genetically distinct commercial varieties (CV) of Hy-Line white-egg laying hens are used in this study (Hy-Line W-36, Hy-Line W-80, and Hy-Line W-80+). Thirty blood samples (n = 10 per CV) were obtained in the production house from the brachial vein and concurrently analyzed by the i-STAT1 portable device. Serum from 22 of these same samples was analyzed via VetScan VS2, a benchtop serum clinical biochemistry analyzer, using VetScan Avian/Reptilian Profile Plus reagent rotors. A paired T-test was used to test for statistical differences in means between the 2 instruments for each of the parameters. Parameters with significant mean differences were then subject to correlation and regression analysis to further evaluate relationships between the results from the 2 methods. Significant differences between means were found for Glu, Na, and K levels. Ca levels were found to be not directly comparable by the 2 analysis instruments. This comparison elucidates the importance of clinical analyzer validations when applying different strategies of diagnostic medicine in poultry.

Longitudinal In Vivo Imaging of Atherosclerotic Disease Development in The apoE Deficient Zucker Rat

One obstacle hindering the development of therapies to treat diabetic vasculopathies, is access to an affordable and appropriate small animal model that recapitulates human pathology. There are currently no well‐established small animal models to study diabetes and atherosclerosis combined. Longitudinal in vivo imaging of inflammation in atherosclerosis is a promising tool to evaluate progression of cardiovascular complications. However, vascular size of current mouse models of atherosclerosis does not allow for detailed molecular imaging. Radiolabeled DANBIRT has been used as an in vivo molecular imaging tool on environmental exposure, aging, and atherosclerosis. Our group has modified the lipid metabolism in Lean Zucker (LZ: Lepr+/?) rats to promote dyslipidemia and development of atherosclerosis by knocking out the apolipoprotein E (apoE) gene using CRISPR/Cas9. We hypothesize that apoE knockout, LZ rats (apoE−/−, Lepr+/?) will develop atherosclerosis with dietary exposure to high fat, high cholesterol (HFHC) diet. [111In]‐DANBIRT will be able to localize inflammation in vivo and allow longitudinal monitoring of vascular atherosclerotic disease using SPECT/CT imaging.MethodsapoE−/−;LZ adult rats were imaged for 1hr post injection of ~900 μCi of [111In]‐DANBIRT for SPECT/CT imaging using a NanoSPECT/CT®. Rats were placed on either HFHC or standard rodent chow for 10 weeks starting at 12 weeks of age (baseline). apoE+/+;LZ under normal and HFHC diet were used as controls. Regions of Interests (ROI) are identified and determined for muscle (background tissue), carotid arteries, aortic arch, and descending aorta. Activity is decay corrected and compared between and among experimental groups. Serum chemistry and histologic analysis of en face aortas and aortic root were performed. Data is represented as mean ± SEM. Two‐way ANOVA with Tukey’s correction was performed.ResultsAll apoE−/− rats developed severe hypercholesterolemia particularly under a HFHC diet. After 10 weeks of HFHC diet apoE−/−, LZ rats have higher [111In]‐DANBIRT uptake in the descending aorta uniquely compared to baseline (p&lt;0.05) and to normal diet fed rats (ns). Histologic analysis showed apoE−/−, LZ have increased significant atherosclerosis on the aortas and the aortic sinus, which correlates to our imaging results. apoE+/+, LZ rats did not develop atherosclerosis disease as expected.ConclusionTo our knowledge, we have the first polygenic rat model able to develop atherosclerosis translationally under HFHC. [111In]‐DANBIRT longitudinally evidenced atherosclerotic disease in the abdominal aorta in vivo correlated to ex vivo histologic analysis.Support or Funding InformationSupported by the NORC PPF (P30 DK056350) to EMB and the CRTG (Burroughs Wellcome Fund) to RIM.