Comparison of hematologic values and transforming growth factor-beta and insulin-like growth factor concentrations in platelet concentrates obtained by use of buffy coat and apheresis methods from equine blood

OBJECTIVE: To evaluate the buffy coat and apheresis methods for preparation of platelet concentrates from equine blood by comparing platelet and growth factor concentrations. ANIMALS: 15 mature mixed-breed geldings. PROCEDURE: Whole blood samples were collected and processed by use of a buffy coat or apheresis method to obtain platelet poor and platelet concentrated fractions. The PCV, WBC count, and platelet count were compared among whole blood samples, platelet poor fractions, concentrates obtained by use of the apheresis method (ie, apheresis platelet concentrates), and concentrates obtained by use of the buffy coat method (ie, buffy coat platelet concentrates). Concentrations of transforming growth factor-beta (ie,TGF-beta1 andTGF-beta2) and insulin-like growth factor were compared between buffy coat and apheresis platelet concentrates. RESULTS: Platelet concentrations were 8.9-fold and 5.2-fold greater in buffy coat and apheresis platelet concentrates, respectively, compared with whole blood. Platelet concentrations were 13.1-fold greater in filtered apheresis platelet concentrates, compared with whole blood. TGF-beta1 concentrations were 2.8-fold and 3.1-fold greater in buffy coat and apheresis platelet concentrates, respectively, and TGF-beta1 concentrations were 10.5-fold greater in filtered apheresis platelet concentrates, compared with whole blood. TGF-beta2 concentrations were 3.6-fold greater in apheresis platelet concentrates, compared with whole blood. Platelet concentrations correlated with growth factor concentrations across all blood and platelet fractions. White blood cell counts had a significant positive correlation with TGF-beta1 concentration in buffy coat platelet concentrates. CONCLUSIONS AND CLINICAL RELEVANCE: Platelets and TGF-beta1 can be concentrated reliably from equine blood by use of buffy coat or apheresis methods, without modification of the protocols used for humans.     

Short- and long-term effects of platelet-rich plasma upon healthy equine joints: Clinical and laboratory aspects

This study aimed to verify whether transient inflammatory reactions incited by the administration of intra-articular platelet-rich plasma (PRP) affected joint components through short- and long-term in vivo evaluation of inflammatory biomarkers and extracellular matrix degradation products in synovial fluid. The effects of PRP were analyzed in a short phase protocol (SPP) and in a prolonged phase protocol (PPP), using saline-injected joints as controls. In the SPP, higher white blood cell counts and prostaglandin E₂ and total protein concentrations were observed in the synovial fluid of PRP-treated joints (P < 0.05). There were no differences between the interleukin-1β, interleukin-1 receptor antagonist protein, tumor necrosis factor-α, chondroitin sulfate, or hyaluronic acid concentrations between PRP and saline injected joints. In the PPP, there were no differences in evaluated parameters between groups. PRP injection elicits a mild and self-limiting inflammatory response shortly after administration, without long-term deleterious effects on joint homeostasis.     

Vector‐borne disease and its relationship to hematologic abnormalities and microalbuminuria in retired racing and show‐bred greyhounds

BackgroundReference intervals for platelets and white blood cell (WBCs) counts are lower in greyhounds than other breeds. Proteinuria is common. Vector‐borne diseases (VBD) cause thrombocytopenia, leukopenia, and proteinuria. Racing greyhounds are commonly exposed to vectors that carry multiple organisms capable of chronically infecting clinically healthy dogs.Hypothesis/ObjectivesVector‐borne disease prevalence is higher in retired racing greyhounds than in show‐bred greyhounds. Occult infection contributes to breed‐related laboratory abnormalities.AnimalsThirty National Greyhound Association (NGA) retired racing and 28 American Kennel Club (AKC) show‐bred greyhounds.MethodsPeripheral blood was tested for Anaplasma, Babesia, Bartonella, Ehrlichia, hemotropic Mycoplasma, and Rickettsia species using PCR. Antibodies to Anaplasma, Babesia, Bartonella, Ehrlichia, and Rickettsia species and Borrelia burgdorferi were detected using immunofluorescence and ELISA assays. Complete blood counts, semiquantitative platelet estimates, and microalbuminuria concentration were determined.ResultsSeven of 30 NGA and 1/28 AKC greyhounds tested positive for ≥1 VBD (P = .05). More positive tests were documented in NGA (10/630) than in AKC dogs (1/588; P = .02). Exposure to Bartonella species (3/30), Babesia vogeli (2/30), Ehrlichia canis (1/30), and infection with Mycoplasma hemocanis (3/30) occurred in NGA dogs. Platelet counts or estimates were >170 000/μL. White blood cell counts <4000/μL (4/28 AKC; 5/30 NGA, P > .99; 1/8 VBD positive; 8/51 VBD negative, P = .99) and microalbuminuria (10/21 AKC; 5/26 NGA, P = .06; 1/8 VBD positive; 14/25 VBD negative, P = .41) were not associated with VBD.Conclusions and Clinical ImportanceThe prevalence of thrombocytopenia and B. vogeli exposure was lower than previously documented. Larger studies investigating the health impact of multiple VBD organisms are warranted.     

The importance of manual white blood cell differential counts and platelet estimates in elephant hematology: blood film review is essential

Unique features of elephant hematology are known challenges in analytical methodology like two types of monocytes typical for members of the Order Afrotheria and platelet counts of the comparatively small elephant platelet. To investigate WBC differential and platelet data generated by an impedance-based hematology analyzer without availability of validated species-specific software for recognition of elephant WBCs and platelets, compared to manual blood film review. Blood samples preserved in ethylenediaminetetraacetic acid (EDTA) of 50 elephants (n = 35 Elephas maximus and n = 15 Loxodonta africana) were used. A Mann-Whitney test for independent samples was used to compare parameters between methods and agreement was tested using Bland-Altman bias plots. All hematological variables, including absolute numbers of heterophils, lymphocytes, monocytes, eosinophils, basophils, and platelets, were significantly different (p < 0.0001) between both methods of analysis, and there was no agreement using Bland-Altman bias plots. Manual review consistently produced higher heterophil and monocyte counts as well as platelet estimates, while the automated analyzer produced higher lymphocyte, eosinophil, and basophil counts. The hematology analyzer did not properly differentiate elephant lymphocytes and monocytes, and did not accurately count elephant platelets. These findings emphasize the importance of manual blood film review as part of elephant complete blood counts in both clinical and research settings and as a basis for the development of hematological reference intervals.     

Development of gates to measure the immature platelet fraction in C57BL/6J mice using the Sysmex XN-V series multispecies hematology analyzer

The immature platelet fraction (IPF) is a measure of newly released platelets, which has been used as a marker of platelet production in multiple human studies but is not widely available in multispecies analyzers. We developed gates to measure the IPF in diluted and undiluted murine blood samples on the Sysmex XN-1000V multispecies hematology analyzer. IPF gates were created using undiluted and diluted (1/10) blood samples obtained from adult and newborn (postnatal day 10, P10) C57BL/6J wild-type (WT) mice, and from 3 murine models of thrombocytopenia: c-MPL^−/− mice, which lack the thrombopoietin receptor (hyporegenerative); antibody-mediated thrombocytopenia; and acute inflammation-induced thrombocytopenia. P10 mice were chosen because, at their size, we could consistently obtain (by terminal phlebotomy) the blood volume needed to run an undiluted sample. The undiluted blood IPF gate successfully differentiated between mechanisms of thrombocytopenia in both adult and P10 mice. For diluted samples, 2 IPF gates were generated: a thrombocytopenic (T) gate, which performed well in samples with platelet counts (PCs) <800 × 10⁹/L in adult mice and <500 × 10⁹/L in newborn mice, and a non-thrombocytopenic (NT) gate, which performed well in samples with PCs above these thresholds. PCs and IPFs measured in diluted blood using these gates agreed well with those measured in undiluted blood and had good reproducibility. These diluted gates allow for the accurate measurement of PCs and IPFs in small (10 µL) blood volumes, which can be obtained easily from adult and newborn mice as small as P1 to assess platelet production serially.     

Thromboelastography platelet mapping in healthy dogs using 1 analyzer versus 2 analyzers

The objective of this study was to describe the results of thromboelastography platelet mapping (TEG-PM) carried out using 2 techniques in 20 healthy dogs. Maximum amplitudes (MA) generated by thrombin (MA_thrombin), fibrin (MA_fibrin), adenosine diphosphate (ADP) receptor activity (MA_ADP), and thromboxane A₂ (TxA₂) receptor activity (stimulated by arachidonic acid, MA_AA) were recorded. Thromboelastography platelet mapping was carried out according to the manufacturer’s guidelines (2-analyzer technique) and using a variation of this method employing only 1 analyzer (1-analyzer technique) on 2 separate blood samples obtained from each dog. Mean [± standard deviation (SD)] MA values for the 1-analyzer/2-analyzer techniques were: MA_thrombin = 51.9 mm (± 7.1)/52.5 mm (± 8.0); MA_fibrin = 20.7 mm (± 21.8)/23.0 mm (± 26.1); MA_ADP = 44.5 mm (± 15.6)/45.6 mm (± 17.0); and MA_AA = 45.7 mm (± 11.6)/45.0 mm (± 15.4). Mean (± SD) percentage aggregation due to ADP receptor activity was 70.4% (± 32.8)/67.6% (± 33.7). Mean percentage aggregation due to TxA₂ receptor activity was 77.3% (± 31.6)/78.1% (± 50.2). Results of TEG-PM were not significantly different for the 1-analyzer and 2-analyzer methods. High correlation was found between the 2 methods for MA_fibrin [concordance correlation coefficient (r) = 0.930]; moderate correlation was found for MA_thrombin (r = 0.70) and MA_ADP (r = 0.57); correlation between the 2 methods for MA_AA was lower (r = 0.32). Thromboelastography platelet mapping (TEG-PM) should be further investigated to determine if it is a suitable method for measuring platelet dysfunction in dogs with thrombopathy.     

Preslaughter diet management in sheep and goats: effects on physiological responses and microbial loads on skin and carcass

Sixteen crossbred buck goats (Kiko x Spanish; BW = 32.8 kg) and wether sheep (Dorset x Suffolk; BW = 39.9 kg) were used to determine the effect of preslaughter diet and feed deprivation time (FDT) on physiological responses and microbial loads on skin and carcasses. Experimental animals were fed either a concentrate (CD) or a hay diet (HD) for 4 d and then deprived of feed for either 12-h or 24-h before slaughter. Blood samples were collected for plasma cortisol and blood metabolite analyses. Longisimus muscle (LM) pH was measured. Skin and carcass swabs were obtained to assess microbial loads. Plasma creatine kinase activity (863.9 and 571.7 ± 95.21 IU) and non-esterified fatty acid concentrations (1,056.1 and 589.8 ± 105.01 mEq/L) were different (P < 0.05) between sheep and goats. Species and diet treatments had significant effects on the ultimate pH of LM. Pre-holding total coliform (TCC) and aerobic plate counts (APC) of skin were significantly different between species. Goats had lower (P < 0.05) TCC (2.1 vs. 3.0 log₁₀ CFU/cm²) and APC (8.2 vs. 8.5 log₁₀ CFU/cm²) counts in the skin compared to sheep. Preslaughter skin E. coli counts and TCC were different (P < 0.05) between species. Goats had lower (P < 0.05) counts of E. coli (2.2 vs. 2.9 log₁₀ CFU/cm²) and TCC (2.3 vs. 3.0 log₁₀ CFU/cm²) in the skin compared with those in sheep. Diet, species, and FDT had no effect (P > 0.05) on E. coli and TCC in carcass swab samples. The APC of carcass swab samples were only affected (P < 0.05) by the FDT. The results indicated that preslaughter dietary management had no significant changes on hormone and blood metabolite concentrations and sheep might be more prone for fecal contamination than goats in the holding pens at abattoir.     

Effects of low-dose G-CSF formulation on hematology in healthy horses after long-distance transportation

The present study evaluated the effects of single-dose filgrastim on hematology in 16 healthy horses after long-distance transportation. Horses were assigned to receive filgrastim (0.23 µg/kg, SC, once; G-CSF group; n=8) or saline (0.9% NaCl) solution (0.3 ml, SC, once; control group; n=8) ≤ 1 hr before transportation. Horses were transported 2,530 km using commercial vans over the course of approximately 44 hr. Clinical examinations and hematologic analyses were performed on all horses before and after transportation. Because the post-transportation white blood cell counts and bacillary neutrophil to segmented neutrophil ratio were significantly higher in the G-CSF group, filgrastim may have promoted the mobilization of neutrophils from marrow. Filgrastim deserves a further study for efficacy in preventing horse shipping fever.     

In utero heat stress alters the postnatal innate immune response of pigs

The effects of in utero heat stress (IUHS) range from decreased growth performance to altered behavior, but the long-term impact of IUHS on postnatal innate immune function in pigs is unknown. Therefore, the study objective was to determine the effects of early gestation IUHS on the immune, metabolic, and stress response of pigs subjected to an 8 hr lipopolysaccharide (LPS) challenge during postnatal life. Twenty-four pregnant gilts were exposed to thermoneutral (TN; n = 12; 17.5 ± 2.1 °C) or heat stress (HS; n = 12; cyclic 26 to 36 °C) conditions from days 6 to 59 of gestation, and then TN conditions (20.9 ± 2.3 °C) from day 60 of gestation to farrowing. At 12 wk of age, 16 IUHS and 16 in utero thermoneutral (IUTN) pigs were selected, balanced by sex and given an intravenous injection of LPS (2 µg/kg BW mixed with sterile saline [SAL] and injected at 2 µL/kg BW) or SAL (2 µL/kg BW). Body temperature was monitored every 30 min, and blood was obtained at 0, 1, 2, 3, 4, 6, and 8 hr following the LPS challenge. Blood samples were analyzed for glucose, insulin, non-esterified fatty acids (NEFA), cortisol, and cytokine concentrations. In addition, white blood cell counts were determined at 0 and 4 hr. Hour 0 data were used as covariates. Body temperature was increased (P < 0.01) in LPS (40.88 ± 0.08 °C) vs. SAL (39.83 ± 0.08 °C) pigs. Eosinophils tended to be decreased overall (P = 0.09; 43.9%) in IUHS vs. IUTN pigs. Glucose concentrations were reduced overall (P = 0.05; 5.9%) in IUHS vs. IUTN pigs. The NEFA concentrations tended to be greater (P = 0.07; 143.4%) in IUHS-LPS pigs compared with all other treatments, and IUTN-LPS pigs tended to have greater (127.4%) circulating NEFA concentrations compared with IUTN-SAL and IUHS-SAL pigs. Cortisol was increased (P = 0.04) in IUHS-LPS compared with IUTN-LPS pigs at 3 hr (21.5%) and 4 hr (64.3%). At 1 hr, tumor necrosis factor α was increased (P = 0.01; 115.1%) in IUHS-LPS compared with IUTN-LPS pigs. Overall, interleukin-1β (IL-1β) and interleukin-6 (IL-6) were greater (P < 0.04; 281.3% and 297.8%, respectively) in IUHS-LPS pigs compared with all other treatments, and IUTN-LPS pigs had increased IL-1β and IL-6 concentrations compared with IUTN-SAL and IUHS-SAL pigs. In summary, IUHS altered the postnatal cytokine, metabolic, and physiological stress response of pigs during postnatal life, which may have negative implications toward the innate immune response of IUHS pigs to pathogens.     

Diurnal variation in fecal concentrations of acid-detergent insoluble ash and alkaline-peroxide lignin from cattle fed bermudagrass hays of varying nutrient content

Background

The effect of time of fecal sampling on the accuracy of acid-detergent insoluble ash (ADIA) and alkaline-peroxide lignin (APL) for the prediction of fecal output (FO) in cattle was evaluated. Eight ruminally cannulated cows (594 ± 35.5 kg) were allocated randomly to 4 bermudagrass [Cynodon dactylon (L.) Pers.] hay diets markedly different in crude protein concentration (79–164 g/kg) with 2 replicates per diet for 3 periods. Cows were offered hay individually at 20 g DM/kg of body weight daily in equal feedings at 08:00 and 16:00 h for a 10-d adaptation period followed by 5-d of total fecal collection. Fecal grab samples also were taken each day during the fecal collection period at 06:00, 12:00, 18:00, and 24:00 h either directly from the rectum or from freshly voided feces. Samples were composited within cow and time across the 5 d total fecal collection period. Additionally, forage, ort, and fecal samples were analyzed for concentrations of APL and ADIA.

Results

Fecal concentrations of ADIA and APL were not affected by sampling time (P ≥ 0.22), even though diet affected (P < 0.01) fecal ADIA and APL concentrations. There were no diet × sampling time interactions (P ≥ 0.60). Estimates of FO and dry matter digestibility (DMD) from ADIA and APL were not affected (P ≥ 0.16) by sampling time or the diet × sampling time interaction (P ≥ 0.74). Estimates of FO and DMD from markers from different sampling times or all different combinations of sampling time were not different (P ≥ 0.72) from those of total collection among internal markers.

Conclusion

Little variation in concentrations of ADIA and APL in daily fecal excretion across time increases flexibility in fecal grab sampling schedules for predicting FO and DMD.     

Comparison of methods for determining platelet numbers and volume in Cavalier King Charles spaniels

OBJECTIVE: The purpose of this study was to compare platelet concentration in cavalier King Charles spaniels (CKCS) measured by different methods commonly used in veterinary hospitals and commercial laboratories. METHODS: Blood samples obtained from 41 CKCS [corrected] were analysed by impedance cell counter, laser cell counter and microscopic estimation. Quantitative buffy coat analysis was performed only on 17 samples, selected from CKCS [corrected] that had low platelet counts detected by cell counters. Platelet counts, platelet estimations and platelet parameters using these different methods were compared. RESULTS: The median platelet number was lower when estimated using impedance cell counter (1363x10(9)/I) with respect to laser cell counter (1723x10(9)/I), microscopic estimation (238x10(9)/I) [corrected] or quantitative buffy coat analyser (292x10(9)/I) [corrected] (P<0.01). Although impedance cell counter, laser cell counter and microscopic estimation were positively correlated, there was no acceptable agreement among methods. CKCS [corrected] with macrothrombocytes in blood smears had significantly lower counts on impedance cell counter, laser cell counter and microscopic estimation. The percentages of CKCS [corrected] with platelet count < 100x10(9)/I [corrected] were 34.1 per cent (impedance cell counter), 26.8 per cent (laser cell counter), 22.0 per cent (microscopic estimation) (not statistically different) and 5.8 per cent (quantitative buffy coat analyser) (P<0.05). CLINICAL SIGNIFICANCE: CKCS [corrected] with macrothrombocytosis have low platelet counts on impedance cell counters, laser cell counters and microscopic estimation. CKCS [corrected] with low platelet counts may have a normal platelet crit detected by a quantitative buffy coat analyser and thus a normal circulating platelet mass.     

Comparison of performance of F1 Romanov crossbred ewes with wool and hair breeds during fall lambing and body weight and longevity through six production years

The objective of this study was to evaluate wool (Dorset and Rambouillet) and hair (Dorper, Katahdin, and White Dorper) breeds for their ability to complement Romanov germplasm in an annual fall lambing system by estimating direct maternal grandsire and sire breed effects on economically important lamb and ewe traits. After 3 yr of evaluation under spring lambing, ewes of the five F1 types were transitioned to spring mating, exposed to composite terminal sires, and evaluated under a barn lambing system at 4, 5, and 6 yr of age. A total of 527 first generation crossbred (F1) ewes produced 1,151 litters and 2,248 lambs from 1,378 May exposures. After accounting for differences in dam age, birth type, and sex, lamb survival to weaning was unaffected by maternal grandsire breed (P = 0.30). However, lambs born to 50% Dorset (16.8 ± 0.21 kg) or 50% White Dorper ewes (16.8 ± 0.28 kg) were heavier at weaning than those born to 50% Katahdin dams (13.8 ± 0.32 kg; P < 0.001). Additionally, lambs born to 50% Dorset ewes were heavier than those born to 50% Rambouillet (16.0 ± 0.22 kg) and 50% Dorper ewes (15.7 ± 0.33; P ≤ 0.03), but no other pairwise maternal grandsire breed differences were observed (P ≥ 0.06). Ewe body weight (n = 3,629) was recorded prior to each of six possible mating seasons and, across ages, was greatest for Dorset- and Rambouillet-sired ewes (56.7 ± 0.44 and 56.5 ± 0.45 kg, respectively), intermediate for Dorper- and White Dorper-sired ewes (54.7 ± 0.78 and 54.1 ± 0.64 kg, respectively), and least for Katahdin-sired ewes (51.5 ± 0.45 kg). Fertility after spring mating (0.80 ± 0.03 to 0.87 ± 0.02), litter size at birth (1.46 ± 0.09 to 1.71 ± 0.07), and litter size at weaning (1.25 ± 0.06 to 1.46 ± 0.06) were not impacted by sire breed (P ≥ 0.16). Ewe longevity, assessed as the probability of being present after 6 production years, was also not affected by sire breed (0.39 ± 0.03 to 0.47 ± 0.03; P = 0.44). Rambouillet-sired ewes weaned more total weight of lamb (21.5 ± 0.94 kg) than Katahdin-sired ewes (17.8 ± 0.94 kg; P = 0.05), but no other sire breed differences were detected (P ≥ 0.07). Results demonstrated that incorporating the Romanov into a crossbreeding system is a practical means of improving out-of-season ewe productivity.     

Osteogenic potential of mesenchymal cells derived from canine umbilical cord matrix co-cultured with platelet-rich plasma and demineralized bone matrix

Canine mesenchymal cells (MSCs) derived from Wharton''s jelly were co-cultured, then supplemented or not supplemented with platelet rich plasma (PRP) and demineralized bone matrix (DBM) to verify osteogenic differentiation. Osteoblastic differentiation followed by mineralized bone matrix production was found to be significantly higher (p < 0.05) when MSCs were associated with PRP/DBM in culture after 14-21-days of induction. Osteopontin and osteocalcin gene expression were significantly superior (p < 0.05) under the same culture conditions after 21 days of observation. In conclusion, addition of PRP to DBM co-cultured with MSCs successfully induced osteogenesis in vitro.     

Blood and milk cellular immune responses of mastitic non-periparturient cows inoculated with Staphylococcus aureus

Lymphocyte function and phenotype of peripheral blood and mammary gland cells were evaluated in non-periparturient cows before and at 1, 4 to 8 and 9 to 14 d after inoculation with Staphylococcus aureus, as expressed by percentage of CD3+, CD2+, and CD45R+ cells, antigen density of these markers per lymphocyte, and mitogen-induced blastogenesis. Milk bacterial counts and somatic cell counts (SCC) were also assessed. Mitogen-induced blastogenic responses were strong in blood and weak in mammary gland cells in all observations and positively correlated with the percent of CD45R+ cells. Significantly greater percentages of milk CD3+ lymphocytes and increased CD3, CD2, and CD45R antigen density per cell were observed after challenge. The blood CD3 and CD2 antigen density per lymphocyte and the milk CD2+ lymphocyte percent were negatively correlated with SCC (P ≤ 0.01). No mastitis (SCC ≤ 500 000 cells/mL) was observed in cows showing blood lymphocyte CD2 and CD3 antigen density indices ≥ 2.5 and 6, respectively. Forty-one percent of SCC values were predicted by the combined blood CD2 and milk CD3 antigen density (P ≤ 0.01). These findings support the hypotheses that mitogen-induced lymphocyte blastogenesis is not a valid test to assess mammary gland immunocompetence and that CD2 expression may facilitate immune responses by decreasing the number of T cell receptors required to achieve full activation.     

Thrombocytes,Leucocytes and Packed Red Cell Volume in Piglets During the First two Weeks of Life

Number of platelets, leucocytes and packed red cell volume (PGV) have been determined in normal piglets during the first 2 weeks of life.A mean platelet count of 337 ± 79 at birth decreased during the first 2 days to 241 ± 48, followed by increasing numbers the next week to a maximum of 578 ±128 at 10 days after birth. Then a slight decrease to 492 ±115 on the 15th day was observed.The leucocyte counts were highest the first 30 hrs. of life, the mean count being 16.9. During this period, however, there was as wide a range as from 3.6 to 46.3. For the rest of the period the mean varied from 8.0 to 11.8, the counts ranging from 4.5 to 20.0.The mean PGV, which was highest at birth, decreased from a mean of 40.8 ± 6.1 to 32.9 ± 5.0 during the first 2 days. At 7 days of age the mean was 31.5 ± 3.8 but increased during the following week to 39.9 ± 3.6 on the 15th day.     

Effect of storage of wet brewer’s grains with incremental levels of salt on apparent total tract nutrient digestibility and purine derivative excretion in dairy heifers

Objectives of this study were to evaluate apparent total tract nutrient digestibility and purine derivative (PD) excretion in dairy heifers limit-fed diets containing wet brewer’s grains (WBG) treated with salt. A 12-wk replicated 4 × 4 Latin square was conducted using 8 Holstein heifers of 224.5 ± 19.4 d of age, and body weight (BW) of 219.2 ± 28.1 kg (mean ± SD). Fresh WBG were treated with 0%, 0.8%, 1.6%, and 2.4% salt and stored for 4 d before being fed. Salt was added either to the WBG or separately to equalize the amount of salt in the diet. The diet contained 9% grass silage, 47% corn silage, 19% corn meal, 17.6% WBG and salt, 2% soybean meal, and 3% mineral mix. Diets were formulated to be limit-fed at 2.15% of BW, provide 14% crude protein (CP) and 2.27 Mcal metabolizable energy (ME)/kg of dry matter (DM). Heifers were adapted to diets for 14 d followed by a 7-d collection period. Dry matter intake (DMI) was recorded daily during the collection week while BW was recorded once a week. Urine and fecal samples were collected during the last 4 d of the collection period. Acid insoluble ash was used as an internal marker to determine apparent nutrient digestibility. Weight loss of WBG during storage was determined from days 1 to 11 and initial and final yeast and mold counts were determined. Final yeast counts were similar among treatments while final mold counts tended to be lesser (P = 0.07) for the 0.8% and 1.6% salt treatments. Urinary volume was similar among treatments while allantoin (P = 0.14), and uric acid (P < 0.01) and total PD excretion tended to increase (P = 0.13) quadratically. DMI was varied by treatment (linear, quadratic, and cubic effects P < 0.01). Heifers fed the 0.8% treatment had the least DMI. Nonfiber carbohydrate (NFC) digestibility linearly decreased (P < 0.04) as salt increased. Digestibilities of DM, and organic matter (OM), tended to decrease (P < 0.10) with increasing levels of salt added to WBG. Fat digestibility was quadratic with the greatest value for the 1.6% treatment. Treating WBG with salt reduced its deterioration based on lesser mold counts for the 0.8% and 1.6% treatments. These treatments had resulted in greater fat digestibility and tended to have increased PD excretion suggesting improved microbial protein synthesis.     

Acute lead poisoning in western Canadian cattle — A 16-year retrospective study of diagnostic case records

This study describes the epidemiology of acute lead poisoning in western Canadian cattle over the 16-year period of 1998 to 2013 and reports background bovine tissue lead concentrations. Case records from Prairie Diagnostic Services, Western College of Veterinary Medicine, identified 525 cases of acute lead toxicity over the investigational period. Poisonings were influenced by year (P < 0.0001) and month (P < 0.0001). Submissions were highest in 2009 (15.6%), 2001 (11.2%), and 2006 (9.9%). Most cases were observed during May, June, and July (62.3%). Cattle 6 months of age and younger were frequently poisoned (53.5%; P < 0.0001). Beef breeds were predominantly poisoned. Mean toxic lead concentrations (mg/kg wet weight) in the blood, liver, and kidney were 1.30 ± 1.70 (n = 301), 33.5 ± 80.5 (n = 172), and 56.3 ± 39.7 (n = 61). Mean normal lead concentrations in the blood, liver, and kidney were 0.036 ± 0.003 mg/kg (n= 1081), 0.16 ± 0.63 mg/kg (n = 382), and 0.41 ± 0.62 mg/kg (n = 64).     

Effects of dietary iron level on growth performance,hematological status,and intestinal function in growing-finishing pigs

This study investigated the different addition levels of iron (Fe) in growing-finishing pigs and the effect of different Fe levels on growth performance, hematological status, intestinal barrier function, and intestinal digestion. A total of 1,200 barrows and gilts ([Large White × Landrace] × Duroc) with average initial body weight (BW; 27.74 ± 0.28 kg) were housed in 40 pens of 30 pigs per pen (gilts and barrows in half), blocked by BW and gender, and fed five experimental diets (eight replicate pens per diet). The five experimental diets were control diet (basal diet with no FeSO₄ supplementation), and the basal diet being supplemented with 150, 300, 450, or 600 mg/kg Fe as FeSO₄ diets. The trial lasted for 100 d and was divided into the growing phase (27 to 60 kg of BW) for the first 50 d and the finishing phase (61 to 100 kg of BW) for the last 50 d. The basal diet was formulated with an Fe-free trace mineral premix and contained 203.36 mg/kg total dietary Fe in the growing phase and 216.71 mg/kg in the finishing phase based on ingredient contributions. And at the end of the experiment, eight pigs (four barrows and four gilts) were randomly selected from each treatment (selected one pig per pen) for digesta, blood, and intestinal samples collection. The results showed that the average daily feed intake (P = 0.025), average daily gain (P = 0.020), and BW (P = 0.019) increased linearly in the finishing phase of pigs fed with the diets containing Fe. On the other hand, supplementation with different Fe levels in the diet significantly increased serum iron and transferrin saturation concentrations (P < 0.05), goblet cell numbers of duodenal villous (P < 0.001), and MUC4 mRNA expression (P < 0.05). The apparent ileal digestibility (AID) of amino acids (AA) for pigs in the 450 and 600 mg/kg Fe groups was greater (P < 0.05) than for pigs in the control group. In conclusion, dietary supplementation with 450 to 600 mg/kg Fe improved the growth performance of pigs by changing hematological status and by enhancing intestinal goblet cell differentiation and AID of AA.     

Evaluation of adverse reactions in dogs following intravenous mesenchymal stem cell transplantation

Background

Recent studies have assessed the therapeutic potential and drawbacks of mesenchymal stem cells (MSCs). The adverse reactions of intravenous transplantation of bone marrow (BM)-derived MSCs were examined at varying doses and frequencies of administration.Nine healthy beagle dogs were purchased from a commercial laboratory. The dogs were distributed equally (n = 3 per group) and randomly into three groups. All dogs received allogeneic BM-derived MSCs: 2 × 10⁶ once (group A), 2 × 10⁷ once (group B), and 2 × 10⁶ for three consecutive days (group C). Various laboratory examinations, multi-detector computed tomography features and histopathology were evaluated to clarify the clinical and diagnostic features of adverse reactions of MSCs administration, prior to receiving MSCs (pre procedure) and on days 1, 3, and 7 post transplantation.

Results

Only one dog had clinical signs during and after MSCs transplantation. Dogs receiving 2 × 10⁶ MSCs showed increased numbers of lymphocytes but the total white blood cell counts were not elevated (P < 0.01). Multi-detector computed tomography (MDCT) revealed pulmonary parenchymal changes in one dog and histopathologic examination revealed pulmonary parenchymal edema and hemorrhage in four dogs. The presence of pulmonary thromboembolism was not detected in either examination.

Conclusions

We considered the presence of pulmonary edema and hemorrhage as possible adverse reactions after intravenous MSCs transplantation; however these results should be cautiously interpreted.     

Immune cell kinetics in the ovine abomasal mucosa following hyperimmunization and challenge with Haemonchus contortus

Sheep were sensitized by repeated infection with Haemonchus contortus L3, followed by a 12 week rest period, and an abomasal cannula was surgically implanted in all sheep. Seven of the sensitized sheep were subsequently challenged with 50 000 H. contortus L3 while 4 control sheep were challenged with saline. Biopsy samples were taken using a fibreoptic endoscope on days 0, 1, 2, 3, 5, 7 and 28 after challenge and leukocyte subpopulations quantified by (immuno)histology. Differential blood cell counts were performed on the same days. At the end of the trial, sheep showed significantly reduced worm burdens compared to unsensitized control sheep, confirming their resistance status. Both blood and tissue eosinophils, as well as tissue γδ TCR⁺ cells were rapidly elevated by day 1 post L3 challenge (pc), peaking at day 3 pc. There was a slight increase in tissue CD4 T cells at day 2 pc, peaking at day 3 pc while no significant changes in CD8 T cells were observed. B cells (CD45R⁺) increased later into challenged tissues with a peak at 5 days pc. All tissue lymphocyte subpopulations as well as tissue and blood eosinophils were reduced by day 7 pc before increasing again at day 28 pc, suggesting separate responses to larval and adult antigens. In contrast, globule leukocytes and mucosal mast cells only showed one peak at day 5 pc and 28 pc, respectively. Unexpectedly, globule leukocytes correlated significantly with tissue eosinophils but not mucosal mast cells. The results are consistent with an early eosinophil-mediated killing of L3, possibly recruited by IL-5 produced by γδ T cells. In contrast to post-mortem studies, abomasal cannulation allowed sequential analysis of both early and late time points in the same animal, providing a more complete picture of cellular interactions at both peripheral and local sites, and their correlation with the different stages of parasite development.