Breed-related differences in altered BRCA1 expression,phenotype and subtype in malignant canine mammary tumors

BRCA1 is a high-penetrance breast cancer susceptibility gene and BRCA1-associated breast cancer has a high familial prevalence that is more common among certain populations of humans. A similar high prevalence also exists for canine mammary tumors (CMTs) and the objective of this study was to determine the breed-related differences in malignant CMTs. Comparative analyses of the expression of various prognostic factors for CMTs, including BRCA1, estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor-2 (HER-2) were conducted on 139 malignant CMT cases from five breeds with the highest prevalence of CMTs in Korea.Significant breed-related differences were observed in the expression of BRCA1 (P = 0.003), histological grade (P = 0.038), and extensive lymphatic invasion (P = 0.042). The Shih Tzu breed had the highest proportion of dogs with malignant CMT and strong overexpression of BRCA1. Cytoplasmic and membranous expression of BRCA1 was associated with the ER negative (P = 0.004), PR negative (P = 0.046), and triple negative (ER, PR, and HER-2 negative; P = 0.016) phenotype and the basal-like molecular subtype (P = 0.019) in Shih Tzu dogs. Since these features are similar to BRCA1-related human breast cancer, dogs with BRCA1-associated CMT, particularly Shih Tzu dogs, may serve as a suitable spontaneous model, although additional molecular studies are needed.     

Analytical validation of a portable human Accu-Chek glucometer in honeybee hemolymph

Glucose and trehalose are the main energy sources used by honeybees (Apis mellifera) for daily activities. However, there is no validated point-of-care method to reliably measure both sugars. We performed an analytical validation of a portable human glucometer (Accu-Chek; Roche) for glucose measurement in honeybee hemolymph compared to a reference method (GluCH, UniCel DxC 600; Beckman Coulter). We used 30 pooled hemolymph samples collected from the antennae of anesthetized honeybees and diluted 1:4 in 0.9% saline. We evaluated dilution linearity, spike recovery, and inter- and intra-assay imprecision. Glucose concentration was measured over time (2 h, 4 h, 8 h, 12 h, 1 d, 2 d, 3 d, 7 d, 21 d, 28 d) at various storage temperature (25°C, 4°C, −20°C, −80°C). The trehalose concentration was measured indirectly by trehalase hydrolyzation. Glucose concentrations measured by both instruments had a strong correlation (0.985, p < 0.0001) and a bias of −7.33 mmol/L (±1.96SD: 13.70 to −28.36), with linear agreement at <20 mmol/L (physiologic value: 100 mmol/L). The accuracy of the glucometer decreased at >20 mmol/L. Recovery of 115–130% of diluted spikes indicated good specificity. Inter- and intra-assay imprecision were 2.50% and 2.21%, respectively. Glucose concentrations fluctuated in stored samples dependent on time and temperature; however, glucose concentrations were constant with storage at −80°C for ≥28 d. The Accu-Chek glucometer is an adequate instrument to measure honeybee glucose concentration in hemolymph diluted with 0.9% NaCl, with good accuracy and precision at <20 mmol/L. Hemolymph storage at −80°C is suitable for long-term conservation of glucose.     

Effect of heat stress and feeding management on growth performance and physiological responses of finishing pigs

This study aimed to determine whether pig responses to heat stress (HS) were directly due to heat exposure (regardless of feeding level and pattern) or were indirectly due to the reduction of feed intake (FI) and to determine if increasing feeding frequency (splitting heat increments) can improve pig response to HS. A total of 48 pigs (66.1 ± 1.7 kg) were allocated to four groups in three replicates. After 7 d in thermoneutral (TN) conditions (22 °C; period 1 [P1; day −7 to −1]), pigs were placed in either TN or HS (32 °C) conditions for 20 d (period 2 [P2; day 0 to 19]). The diet was provided either ad libitum (AL; 2 distributions/d) or pair-fed (PF8; 8 distributions/d) using HS–AL pigs as the reference group. Thus, the four experimental groups were TN–AL, HS–AL, TN–PF8, and HS–PF8. The daily ration of PF8 pigs was distributed at every 90-min intervals from 0900 to 1930 hours. Data were analyzed using the PROC MIXED procedure with replicate (n = 3), experimental group (n = 4), and their interactions as fixed effects, and the REPEATED statement was used for repeated measures data. Pigs had a similar average daily feed intake (ADFI) during P1 (P > 0.05). In P2, HS–AL and PF8 pigs had lower ADFI (−19%), average daily gain (−25%), and final body weight (−6.1 kg) than TN–AL pigs (P < 0.01). TN–AL pigs had thicker backfat than TN–PF8 pigs (P < 0.05), while the HS pigs had intermediate results. HS pigs had a higher perirenal fat percentage based on the contrast analysis between PF8 pigs (P < 0.05). Thermoregulatory responses of pigs increased with HS exposure but did not differ between HS or between TN groups (P > 0.05). For TN pigs, variation in muscle temperature (T_muscle) depended on feeding and physical activity, while for HS pigs, T_muscle gradually increased throughout the day. The T_muscle of PF8 pigs increased with each additional meal but plateaued earlier for HS–PF8 than TN–PF8 pigs; an increase in T_muscle per meal was also lower in HS–PF8 than TN–PF8 (P < 0.05). Exposure to HS decreased plasma T3 and T4 (P < 0.05) and increased plasma creatinine (P < 0.05). Between the PF8 groups, HS pigs also had a transient increase in plasma insulin on day 8 (P < 0.05). The effect of HS on FI decreased the growth rate of pigs but there are heat-induced effects, such as altered physiological responses, which might explain the direct HS effects seen in other literature especially in terms of increased adiposity. The increased feed provision frequency in the present study did not improve the HS response of pigs.     

Effect of a single injection of gonadotropin-releasing hormone (GnRH) and human chorionic gonadotropin (hCG) on testicular blood flow measured by color doppler ultrasonography in male Shiba goats

Although color Doppler ultrasonography has been used to evaluate testicular blood flow in many species, very little has been done in goat. Eight male Shiba goats were exposed to a single intramuscular injection of either gonadotropin-releasing hormone (GnRH group; 1 µg/kg BW) or human chorionic gonadotropin (hCG group; 25 IU/kg BW). Plasma testosterone (T), estradiol (E2) and inhibin (INH) were measured just before (0 hr) and at different intervals post injection by radioimmunoassay. Testis volume (TV) and Doppler indices, such as resistive index (RI) and pulsatility index (PI) of the supratesticular artery, were measured by B-mode and color Doppler ultrasonography, respectively. The results indicated an increase in testicular blood flow in both groups, as RI and PI decreased significantly (P<0.05), but this increase was significant higher and earlier in hCG group (1 hr) than in the GnRH group (2 hr). A high correlation was found for RI and PI with both T (RI, r= −0.862; PI, r= −0.707) and INH in the GnRH group (RI, r=0.661; PI, r=0.701). However, a significant (P<0.05) correlation was found between E2 and both RI (r= −0.610) and PI (r= −0.763) in hCG group. In addition, TV significantly increased and was highly correlated with RI in both groups (GnRH, r= −0.718; hCG, r= −0.779). In conclusion, hCG and GnRH may improve testicular blood flow and TV in Shiba goats.     

Preanalytical variables affecting the measurement of serum paraoxonase-1 activity in horses

Paraoxonase-1 (PON-1) activity is a new inflammatory and oxidative marker. Technical effects and biological factors could affect the accuracy of PON-1 activity measurement. We investigated the effects of storage at different temperatures, repeated freeze–thaw cycles, interferences from hemolytic, lipemic, and icteric samples, and seasonal effects on PON-1 activity in horses. We evaluated 2 substrates with an automated spectrophotometer. Ten equine serum samples were stored under different conditions. Although storage at room (21°C) or refrigeration (4°C) temperature induced a statistically significant decrease (p < 0.05) in PON-1 activity, this is not diagnostically relevant. PON-1 activity in frozen samples (−20°C) was stable for short-term storage; diagnostically significant (p < 0.01) fluctuations were observed after 1 mo. Four repeated freeze–thaw cycles were assessed, and all cycles affected PON-1 activity (p < 0.01); however, this was diagnostically significant only after the 4th cycle. Hemolysis induced an overestimation of PON-1 activity; lipemia and hyperbilirubinemia did not change PON-1 activity. Thirty-four horses were sampled monthly for 1 y, and PON-1 activity was higher in autumn (p < 0.05) and winter (p < 0.05) than in spring and summer.     

Factors affecting performance response of pigs exposed to different challenge models: a multivariate approach

Factors associated with the severity with which different challenge models (CMs) compromise growth performance in pigs were investigated using hierarchical clustering on principal components (HCPC) analysis. One hundred seventy-eight studies reporting growth performance variables (average daily gain [ADG], average daily feed intake [ADFI], gain:feed [GF], and final body weight [FBW]) of a Control (Ct) vs. a Challenged (Ch) group of pigs using different CMs (enteric [ENT], environmental [ENV], lipopolysaccharide [LPS], respiratory [RES], or sanitary condition [SAN] challenges) were included. Studies were grouped by similarity in performance in three clusters (C1, C2, and C3) by HCPC. The effects of CM, cluster, and sex (males [M], females [F], mixed [Mi]) were investigated. Linear (LRP) and quadratic (QRP) response plateau models were fitted to assess the interrelationships between the change in ADG (∆ADG) and ADFI (∆ADFI) and the duration of challenge. All variables increased from C1 through C3, except for GF, which decreased (P < 0.05). LPS was more detrimental to ADG than ENV, RES, and SAN models (P < 0.05). Furthermore, LPS also lowered GF more than all the other CMs (P < 0.05). The ∆ADG independent of ∆ADFI was significant in LPS and SAN (P < 0.05), showed a trend toward the significance in ENT and RES (P < 0.10), and was not significant in ENV (P > 0.10), while the ∆ADG dependent on ∆ADFI was significant in ENT, ENV, and LPS only (P < 0.05). The critical value of ∆ADFI influencing the ∆ADG was significant in pigs belonging to C1 (P < 0.05) but not C2 or C3 (P > 0.10). The ∆ADG independent of duration post-Ch (irreparable portion of growth) was significant in C1 and C2 pigs, whereas the ∆ADFI independent of duration post-Ch (irreparable portion of feed intake) was significant in C1 pigs only (P < 0.05). Moreover, the time for recovery of ADG and ADFI after Ch was significant in pigs belonging to C1 and C2 (P < 0.05). Control F showed reduced ADG compared with Ct-M, and Ch-F showed reduced ADFI compared with Ch-M (P < 0.05). Moreover, the irreparable portion of ΔADG was 4.8 higher in F (−187.7; P < 0.05) compared with M (−39.1; P < 0.05). There are significant differences in growth performance response to CM based on cluster and sex. Furthermore, bacterial lipopolysaccharide appears to be an appropriate noninfectious model for immune stimulation and growth impairment in pigs.     

Phylogenetic analysis and characterization of Korean bovine viral diarrhea viruses

Thirty-six bovine viral disease viruses (BVDVs) were identified in bovine feces (n = 16), brains (n = 2), and aborted fetuses (n = 18) in Korea. To reveal the genetic diversity and characteristics of these Korean strains, the sequences of their 5′-untranslated regions (5′-UTRs) were determined and then compared with published reference sequences. Neighbor-joining phylogenetic analysis revealed that most of the Korean viruses were of the BVDV subtypes 1a (n = 17) or 2a (n = 17). The remaining strains were of subtypes 1b (n = 1) and 1n (n = 1). This analysis indicates that the 1a and 2a BVDV subtypes are predominant and widespread in Korea. In addition, the prevalence of BVDV-2 was markedly higher in aborted fetuses than in other samples and was more often associated with reproductive problems and significant mortality in cattle.     

Investigation of single‐nucleotide polymorphisms in the NR3C1a glucocorticoid receptor gene in Cocker Spaniels with primary immune thrombocytopenia

BackgroundIn dogs, 6 single‐nucleotide polymorphisms (SNPs) have been described in the glucocorticoid receptor gene NR3C1a, 2 of which were nonsynonymous SNPs in exons 2 and 8. The clinical importance of these SNPs is unknown.ObjectivesTo investigate whether SNPs in NR3C1a are associated with clinical outcome in Cocker Spaniels with primary immune thrombocytopenia (pITP).AnimalsTwenty‐four Cocker Spaniels with pITP presented to a referral center. Dogs were classified as slow (n = 11) or fast responders (n = 12) based on time required after initiating glucocorticoid treatment to achieve a platelet count >70 000/μL.MethodsDeoxyribonucleic acid was extracted from stored blood samples before amplification by PCR and sequencing of exons 2 and 8 of NR3C1a. Associations between genotype and clinical response variables were investigated.ResultsNeither previously identified nonsynonymous SNPs were identified. The synonymous SNP NR3C1a:c.798C>T in exon 2 was found at an increased prevalence compared to a previous report. No difference was found in prevalence of any genotype at NR3C1a:c.798C>T between fast and slow responders (P = .70).Conclusions and Clinical ImportanceNone of the previously reported nonsynonymous SNPs in exons 2 and 8 of the NR3C1a gene were detected in our cohort of Cocker Spaniels with pITP. The synonymous SNP NR3C1a:c.798C>T in exon 2 was reported at a higher frequency than previously, but was not associated with outcome measures that estimated responsiveness to glucocorticoids.     

Effect of storage time and temperature on the total protein concentration and electrophoretic fractions in equine serum

Serum protein electrophoresis (SPE) is a technique that could be considered one of the most useful diagnostic aids available to the clinician. The effect of storage time and temperature on the total proteins and electrophoretic fractions (albumin, α₁-, α₂-, β₁-, β₂-, and γ-globulins) was assessed in 24 healthy horses. All samples, collected by jugular vein puncture, were centrifuged and divided into 4 aliquots. The 1st aliquot was analyzed within 3 h from collection (time 0), the 2nd was refrigerated at +4°C for 24 h, the 3rd was refrigerated at +4°C for 48 h, and the last was frozen at −20°C for 48 h. One-way repeated-measures analysis of variance (ANOVA) showed a significant effect (P < 0.05) of the different storage conditions on the concentrations of all the parameters studied and significant variations in the percentages of albumin, α₁-globulins, α₂-globulins, and γ-globulins. Compared with time 0 the total protein concentration increased significantly after 48 h at −20°C, the albumin percentage decreased after 48 h at −20°C, the α₁-globulin percentage increased after 24 h at +4°C, the α₂-globulin percentage increased after 48 h at +4°C and at −20°C, and the γ-globulin percentage increased after 48 h at −20°C. The results should help veterinary practitioners handle and store equine serum samples appropriately. Further investigations at different storage times and temperatures could be useful.     

Antibiotic resistance,antimicrobial residues,and bacterial community diversity in pasture-raised poultry,swine, and beef cattle manures

Animal manure can be a source of antibiotic-resistant genes (ARGs) and pharmaceutical residues; however, few studies have evaluated the presence of ARG in pasture-raised animal production systems. The objective of this study was to examine changes in microbiome diversity and the presence of antibiotic residues (ABRs) on three farms that contained a diverse range of animal species: pasture-raised poultry (broiler and layer), swine, and beef cattle. Total bacterial communities were determined using 16S rRNA microbiome analysis, while specific ARGs (sulfonamide [Sul; Sul1] and tetracycline [Tet; TetA]) were enumerated by qPCR (real-time PCR). Results indicated that the ARG abundances (Sul1 [P < 0.05] and TetA [P < 0.001]) were higher in layer hen manures (16.5 × 10⁻⁴ and 1.4 × 10⁻⁴ µg kg⁻¹, respectively) followed by broiler chickens (2.9 × 10⁻⁴ and 1.7 × 10⁻⁴ µg kg⁻¹, respectively), swine (0.22 × 10⁻⁴ and 0.20 × 10⁻⁴ µg kg⁻¹, respectively) and beef cattle (0.19 × 10⁻⁴ and 0.02 × 10⁻⁴ µg kg⁻¹, respectively). Average fecal TetA ABR tended to be greater (P = 0.09) for broiler chickens (11.4 µg kg⁻¹) than for other animal species (1.8 to 0.06 µg kg⁻¹), while chlortetracycline, lincomycin, and oxytetracycline ABRs were similar among animal species. Furthermore, fecal microbial richness and abundances differed significantly (P < 0.01) both among farms and specific species of animal. This study indicated that the microbial diversity, ABR, ARG concentrations, and types in feces varied from farm-to-farm and from animal species-to-animal species. Future studies are necessary to perform detailed investigations of the horizontal transfer mechanism of antibiotic-resistant microorganisms (ARMs) and ARG.     

Assessing single-nucleotide polymorphism selection methods for the development of a low-density panel optimized for imputation in South African Drakensberger beef cattle

A major obstacle in applying genomic selection (GS) to uniquely adapted local breeds in less-developed countries has been the cost of genotyping at high densities of single-nucleotide polymorphisms (SNP). Cost reduction can be achieved by imputing genotypes from lower to higher densities. Locally adapted breeds tend to be admixed and exhibit a high degree of genomic heterogeneity thus necessitating the optimization of SNP selection for downstream imputation. The aim of this study was to quantify the achievable imputation accuracy for a sample of 1,135 South African (SA) Drakensberger cattle using several custom-derived lower-density panels varying in both SNP density and how the SNP were selected. From a pool of 120,608 genotyped SNP, subsets of SNP were chosen (1) at random, (2) with even genomic dispersion, (3) by maximizing the mean minor allele frequency (MAF), (4) using a combined score of MAF and linkage disequilibrium (LD), (5) using a partitioning-around-medoids (PAM) algorithm, and finally (6) using a hierarchical LD-based clustering algorithm. Imputation accuracy to higher density improved as SNP density increased; animal-wise imputation accuracy defined as the within-animal correlation between the imputed and actual alleles ranged from 0.625 to 0.990 when 2,500 randomly selected SNP were chosen vs. a range of 0.918 to 0.999 when 50,000 randomly selected SNP were used. At a panel density of 10,000 SNP, the mean (standard deviation) animal-wise allele concordance rate was 0.976 (0.018) vs. 0.982 (0.014) when the worst (i.e., random) as opposed to the best (i.e., combination of MAF and LD) SNP selection strategy was employed. A difference of 0.071 units was observed between the mean correlation-based accuracy of imputed SNP categorized as low (0.01 < MAF ≤ 0.1) vs. high MAF (0.4 < MAF ≤ 0.5). Greater mean imputation accuracy was achieved for SNP located on autosomal extremes when these regions were populated with more SNP. The presented results suggested that genotype imputation can be a practical cost-saving strategy for indigenous breeds such as the SA Drakensberger. Based on the results, a genotyping panel consisting of ~10,000 SNP selected based on a combination of MAF and LD would suffice in achieving a <3% imputation error rate for a breed characterized by genomic admixture on the condition that these SNP are selected based on breed-specific selection criteria.     

The Effect of Single Nucleotide Polymorphisms in the Tumor Necrosis Factor-α Gene on Reproductive Performance and Immune Function in Dairy Cattle

The present study aimed to assess the effect of polymorphisms in the tumor necrosis factor α (TNF-α) promoter (A/A, A/G and G/G) and exons (T/T, T/C and C/C) on immune function and reproductive performance in dairy cows. The occurrence of the first postpartum ovulation within 3 weeks in the cows with the TNF-α promoter A/G and G/G genotypes was higher than in the A/A group. Among the different TNF-α exon genotypes, the occurrence of early first postpartum ovulation was higher in the T/C and C/C genotype groups than in the T/T group. Single nucleotide polymorphisms (SNPs) in the TNF-α gene did not affect the rate of artificial insemination (AI) or duration from parturition to next conception (days open). The apoptosis rate of polymorphonuclear leukocytes (PMNs) did not differ among the TNF-α promoter genotypes, but the PMN transmigration rate was significantly higher for the A/A and A/G genotypes than for the G/G genotype. Interleukin 8 (IL-8) mRNA expression in PMNs and peripheral blood mononuclear cells (PBMCs) before culture was significantly higher for the A/A genotype compared with the G/G genotype. There were no significant differences between the genotypes in the mRNA expression of TNF-α, IL-6, IL-1β, and toll-like receptor 4 (TLR4) in PMNs and PBMCs before and 4 h after culture. IL-8 and IL-1β production by PBMCs cultured for 4 h was significantly higher for the animals with the A/A genotype than for those with the G/G genotype. On the other hand, no significant difference was observed in IL-8 and IL-1β production by PMNs among different TNF-α genotypes. Taken together, these results suggest that SNP in the TNF-α gene affects immune function and reproductive performance in dairy cows.     

Effects of a novel consensus bacterial 6-phytase variant on the apparent ileal digestibility of amino acids,total tract phosphorus retention,and tibia ash in young broilers

The effect of a novel consensus bacterial 6-phytase variant (PhyG) on apparent ileal digestibility (AID) of amino acids (AA) and phosphorus (P) utilization in young broilers when added to diets with high phytate-P (PP) content without added inorganic phosphate (Pi) and deficient in digestible (dig) AA and metabolizable energy (ME) was investigated. A total of 256 Ross 308 male broilers were assigned to 4 treatments (8 birds/cage, 8 cages/treatment) in a completely randomized design. Treatments comprised a positive control (PC, 2,975 kcal/kg ME, 3.7 g/kg dig P, 2.83 g/kg PP, 8.4 g/kg Ca, 10.6 g/kg dig lysine), a negative control (NC) without added Pi (ME −68 kcal/kg, crude protein −10 g/kg, dig AA −0.1 to −0.4 g/kg, Ca −2.0 g/kg, dig P −2.2 g/kg, Na −0.4 g/kg vs. PC), and NC plus 500 or 1,000 FTU/kg of PhyG. Test diets were corn/soy/rapeseed-meal/rice-bran-based and fed from 5 to 15 d of age. Ileal digesta and tibias were collected on day 15. Excreta was collected during days 12 to 15 to determine P retention. The NC (vs. PC) reduced (P < 0.05) P retention (−10.4% units), tibia ash (−14.3% units), weight gain (−109 g), feed intake (−82 g) and increased FCR (from 1.199 to 1.504), confirming that the NC was extremely deficient in nutrients and energy. Phytase addition to the NC linearly (P < 0.001) improved performance, but did not fully recover it to the level of the PC due to the severe nutrients/energy reduction in NC. Phytase linearly increased P retention (P < 0.001), tibia ash (P < 0.001), AID of dry matter (P < 0.05), nitrogen (P < 0.01), gross energy (P < 0.05), and all 17 individual AA (P < 0.01). At 1,000 FTU/kg, phytase increased (P < 0.05) P retention vs. PC and NC (+14.5 and +24.9% units, respectively) and increased tibia ash vs. NC (+13.8% units), equivalent to PC. The NC decreased AID of Cys, Gly, Thr, and Met vs. PC (P < 0.05). At 1,000 FTU/kg, phytase increased AID of all 17 AA vs. NC (P < 0.01), equivalent to PC. At 1,000 FTU/kg, AID AA responses (above NC) ranged from +4.5% (Met) to +15.0% (Cys), being maximal for essential Thr (+10.4%) and Val (+8.2%) and non-essential Cys (+15.0%) and Gly (+10.4%). The results highlight the efficacy of PhyG at a dose level of 500 to 1,000 FTU/kg in young broilers for improving the ileal digestibility of nitrogen, AA, and energy alongside P retention and tibia ash. The performance data emphasize the need to consider digestible nutrient intake as a response variable in exogenous enzyme studies.     

Effect of long-term feeding of graded levels of deoxynivalenol on performance,nutrient utilization,and organ health of grower-finisher pigs (35 to 120 kg)

The objective of this study was to evaluate the effect of long-term feeding of graded levels of deoxynivalenol (DON) on performance, nutrient utilization, and organ health of grower-finisher pigs. A total of 240 mixed-sex grower-finisher pigs (35.9 ± 1.1 kg initial body weight, BW) were randomly assigned to 1 of 4 dietary treatments (6 pigs/pen; 10 pens/treatment) for 77 d. Diets consisted of a control diet without DON (CONT) and diets containing 1, 3, or 5 ppm DON (DON1, DON3, or DON5). Nitrogen-balance was determined in 1 pig/pen during weeks 6 and 12 of the study. Growth performance measures were taken weekly for average daily feed intake (ADFI), average daily gain (ADG), and gain:feed (GF) until day 77. Blood samples were collected on days 0, 14, 42, 56, and 84 from 1 pig/pen for analysis of indicators of liver and kidney function. On day 7, ADG and ADFI for pigs fed DON3 and DON5 diets were lower (P < 0.05) compared with DON1- and CONT-fed pigs. Overall, ADG and ADFI (days 0 to 77) were lower in DON3- and DON5-fed pigs compared with CONT and DON1 pigs (P < 0.05), with no difference in GF (P > 0.05). Final BW was reduced in DON3- and DON5-fed pigs (P < 0.05) compared with CONT and DON1, which were not different (P > 0.05). No significant (P > 0.05) treatment effects were observed on carcass characteristics. In the grower-phase, protein deposition (PD) was reduced in DON3 and DON5 pigs compared with CONT and DON1 pigs (P < 0.05). In the finisher phase, PD was not affected by dietary treatment (P > 0.05). There was no effect of dietary treatment on the majority of selected serum chemistry (P > 0.05). In summary, pigs exposed to diets containing > 1 ppm DON had reduced growth performance with little or no effect on nitrogen utilization, organ health, or carcass characteristics, suggesting that the negative effects of DON may be largely due to depressed feed intake.     

Significant association of serum autoantibodies to TYMS,HAPLN1 and IGFBP5 with early stage canine malignant mammary tumours

Canine mammary tumours (CMTs) are the most prevalent neoplasms in female dogs. Despite the high incidence of such tumours, a lack of easily accessible biomarkers still impedes early diagnosis of malignant CMTs. Herein we identify thymidylate synthetase (TYMS), hyaluronan and proteoglycan link protein 1 (HAPLN1) and insulin‐like growth factor‐binding protein 5 (IGFBP5) as CMT antigens eliciting corresponding autoantibodies in CMT cases. We establish enzyme‐linked immunosorbent assays (ELISAs) to detect autoantibodies to TYMS (TYMS‐AAb), HAPLN1 (HAPLN1‐AAb) and IGFBP5 (IGFBP5‐AAb) in sera from 81 dogs with malignant CMTs (41 in Stage I), 24 with benign CMTs and 35 healthy controls. Levels of all the three autoantibodies are elevated in the malignant group compared with the healthy or the benign group; notably, the elevated autoantibody levels significantly correlate with the stage‐I CMTs. For discriminating malignant CMTs from healthy control, the area under curve (AUC) of TYMS‐AAb is 0.694 with specificity of 82.9% and sensitivity of 50.6%. The AUC of utilising HAPLN1‐AAb for distinguishing the stage‐I CMTs from healthy controls is 0.711 with specificity of 77.1% and sensitivity of 58.5%. In differentiating malignant CMTs from the benign, the AUC of IGFBP5‐AAb reaches 0.696 with specificity of 70.8% and sensitivity of 67.9%, and a combination of IGFBP5‐AAb and TYMS‐AAb increases the AUC to 0.72. Finally, the AUC of combined HAPLN1‐AAb and IGFBP5‐AAb in discriminating the stage‐I CMTs from the benign achieves 0.731. Collectively, this study highlights a significant association of the three serum autoantibodies with early stage malignant CMTs.     

Inhibition of prolactin promotes secondary skin follicle activation in cashmere goats

The aim of this study was to investigate the involvement of prolactin (PRL) on development of secondary skin follicles in cashmere goats. Goats were randomly assigned to either a bromocriptine treatment or control group. Samples of cashmere fiber, blood, and skin were collected from all goats after 1 mo. The results indicated that the length, growth rate, and diameter of fibers were not influenced (P > 0.05) by the inhibition of PRL resulting from the treatment with bromocriptine. There was a tendency for increases in total follicle number, primary and secondary follicle numbers, and in the ratio of secondary to primary follicles following treatment with bromocriptine, but these differences were not significant (P > 0.05). The percentage of active secondary follicles in anagen was increased (P < 0.05) in the bromocriptine-treated goats, but there was no effect of treatment on the percentage of active primary follicles. Bromocriptine decreased (P < 0.05) circulating concentrations of PRL and Insulin-like growth factor 1 (IGF1) and increased (P < 0.05) those of melatonin (MT), but there was no effect of this treatment on the serum concentrations of cortisol, growth hormone, tetraiodothyronine, and triiodothyronine. In bromocriptine-treated goats, mRNA expressions of PRL and MT membrane receptor 1a (MTNR1a) were decreased (P < 0.05) and mRNA expression of MT nuclear receptor (RORα) was increased (P < 0.05), but there was no effect of the treatment on expression of long PRL receptor, short PRL receptor, MT membrane receptor 1b and IGF1. It is concluded that inhibition of PRL promotes secondary hair follicle development in the anagen phase, possibly by downregulating MTNR1a and up-regulating RORα gene expression in the skin.     

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.     

A longer adaptation period to a functional amino acid-supplemented diet improves growth performance and immune status of Salmonella Typhimurium-challenged pigs

We recently showed that dietary supplementation with key functional amino acids (FAA) improves growth performance and immune status of Salmonella Typhimurium (ST)-challenged pigs. It is not known if ST-challenged pigs will benefit from a longer adaptation period to FAA. The objective of this study was to evaluate the effects of different adaptation periods to diets containing FAA above requirements for growth on performance and immune response of weaned pigs subsequently challenged with ST. A total of 32 mixed-sex weanling pigs (11.6 ± 0.3 kg) were randomly assigned to 1 of 4 dietary treatments, being a basal amino acid (AA) profile fed throughout the experimental period (FAA−) or a functional AA profile (FAA+; Thr, Met, and Trp at 120% of requirements) fed only in the postinoculation (FAA+0), for 1 wk pre- and postinoculation (FAA+1), or throughout the experimental period (FAA+2). After a 14-d adaptation period, pigs were inoculated with ST (2.15 × 10⁹ CFU/mL). Growth performance, body temperature, fecal score, acute-phase proteins, oxidant/antioxidant balance, score for ST shedding in feces and intestinal colonization, and fecal and digesta myeloperoxidase (MPO) were measured pre- and postinoculation. Postinoculation body temperature and fecal score, serum haptoglobin, plasma superoxide dismutase (SOD), malondialdehyde (MDA), and fecal MPO were increased while serum albumin and plasma reduced glutathione (GSH):oxidized glutathione (GSSG) were reduced compared to pre-inoculation (P < 0.05). Average daily gain and G:F were greater in FAA+2 pigs compared to FAA− pigs (P < 0.05). Serum albumin was higher in FAA+2 and FAA+1 compared to FAA+0 and FAA− pigs (P < 0.05) while FAA+2 pigs had lower haptoglobin compared to FAA− (P < 0.05). Plasma SOD was increased and GSH:GSSG was decreased in FAA− pigs compared to the other treatments (P < 0.05). Score for ST shedding in feces was progressively lower from d 1 to 6 regardless of treatment (P < 0.05) and was lower in FAA+2 pigs compared to FAA− and FAA+0 (P < 0.05). Counts of ST in colon digesta were higher in FAA− and FAA+0 pigs compared to FAA+2 (P < 0.05). Fecal and colonic digesta MPO were lower in FAA+2 and FAA+1 pigs compared to FAA− (P < 0.05). These results demonstrate a positive effect of a longer adaptation period to FAA-supplemented diets on performance and immune status of weaned pigs challenged with Salmonella.