Epidemiology: Culture of bovine bone marrow progenitor cells in vitro

Summary

In vitro methylcellulose cultures of bovine bone marrow progenitor cells were developed. An existing technique described for bovine species was compared to a method for human tissue and further adapted during subsequent experiments. Bovine bone marrow samples were collected at the slaughterhouse, and mononuclear cells were separated by gradient centrifugation (1.077 g/ml specific density and 400g). The use of 3% bovine leucocyte‐conditioned medium, produced by stimulation of blood lymphocytes with 4 pg/ml concanavalin A and harvested on day 4 of culture, gave better results than the use of supernatant of the human bladder carcinoma 5637, which is widely used in human bone marrow cultures. However, bovine leucocyte‐conditioned medium was not added to erythroid cultures because inhibitory effects were observed. Erythroid colonies were stimulated with erythropoietin, and hemin was added to enable microscopic identification. Reduced oxygen tension was necessary to induce growth of erythroid colonies. This was not necessary for myeloid cultures. In conclusion, the results of this study show that the growth of myeloid and erythroid colonies in methylcellulose‐based medium requires different culture conditions, which are different from the culture conditions for human cells.     

Over-processed meat and bone meal and phytase effects on broilers challenged with subclinical necrotic enteritis: Part 3. Bone mineralization and litter quality

This study was conducted to determine the effect of necrotic enteritis (NE), phytase level and meat and bone meal (MBM) processing on bone mineralization of broilers and litter quality. Ross 308 male broiler chicks (n = 768) were allotted to 48 pens with 16 birds each. There were 8 dietary treatments in a 2 × 2 × 2 factorial arrangement. Factors were NE challenge (no or yes), phytase level (500 or 5,000 FTU/kg), and MBM (as-received or over-processed). Half of the birds were challenged with field strains of Eimeria spp. at d 9 and 10⁸ CFU per mL of Clostridium perfringens strain EHE-NE18 on d 14 and 15. The middle toe, tibia and femur of 2 birds per pen were excised at d 16 and 29 for determination of ash, breaking strength (BS) and bone mineralization. At d 42, all were assessed for hock burns and litter was scored and assessed for dry matter (DM). At d 16, challenged birds had lower toe ash (P < 0.01), femur ash (P < 0.001), tibia ash (P < 0.001) and tibial BS (P < 0.001) than unchallenged birds. At d 16, challenged birds fed high phytase and over-processed MBM had higher toe Mn than those fed low phytase and as-received MBM. At d 29 unchallenged birds fed high phytase and as-received MBM had a higher toe Mn than those fed over-processed MBM. At d 16, a phytase × MBM interaction was detected for femur Zn concentration (P < 0.05), where a higher level of Zn was observed in the high phytase group fed over-processed MBM. At d 16, tibial Ca (P < 0.05) and P (P < 0.05) were lower in the challenged whereas the femur K (P < 0.001), Mn (P < 0.01) and Na (P < 0.001) were higher in the challenged at d 16. At d 42, challenged birds had higher litter DM (P = 0.058) and fewer hock burns than those unchallenged (P < 0.05). In conclusion, NE impaired bone traits but high phytase and over-processed MBM increased bone mineral contents. Cases of hock burns may be lower under NE incidences due to lower livability of birds reducing litter wetness.     

Impact of four fiber-rich supplements on nutrient digestibility,colostrum production,and farrowing performance in sows

This study aimed to investigate the impact of dietary fiber (DF) sources on sow and litter performance, and apparent total tract digestibility (ATTD) of gross energy (GE) and nutrients. A total of 48 sows were stratified for body weight at mating and randomly assigned to one of four DF sources (mixed fiber [MF], palm kernel expellers [PKE], sugar beet pulp [SBP], or soy hulls [SH]) and fed the diet from mating until farrowing. Within DF treatments, sows were supplemented with one of two extra energy sources (glycerol or sugar dissolved in water), whereas a third group (control) received water from day 108 of gestation until farrowing. The number of total born, live-born, and stillborn pigs; birth time and birth weight of the pigs; farrowing duration; and farrowing assistance (FA) were recorded. Live-born pigs were weighed again at 12 and 24 h after birth to record weight gain, which was used to estimate intake and yield of colostrum. Blood samples were collected once daily from day −3 relative to farrowing until day 1 after farrowing in sows and once from selected pigs right after birth. Fecal samples were collected on day 114 of gestation and colostrum at 0, 12, 24, and 36 h after onset of farrowing. Intake of soluble and insoluble nonstarch polysaccharides (NSP) was greater for SBP (P < 0.001) and PKE (P < 0.001) supplemented sows, respectively, when compared with other groups. Farrowing duration and stillbirth rate were not affected by DF sources, but PKE and SH supplemented sows had greater FA than SBP and MF supplemented sows (P < 0.001). Extra energy supplement did not improve the farrowing performance. Concentration (P = 0.02) and output (P = 0.04) of dry matter in colostrum, and ATTD of GE (P < 0.001) and crude protein (CP; P < 0.001) were lower for PKE supplemented sows than in sows from the remaining groups. Intake of insoluble NSP correlated negatively with ATTD of GE (P < 0.001) and CP (P < 0.001). Concentrations of glucose (P < 0.001), lactate (P < 0.001), CO₂ (P < 0.001), and HCO₃ (P < 0.001) in sows blood were increased with time progress relative to farrowing. Newborn pigs from PKE supplemented sows had greater concentration of lactate (P = 0.02) and lower blood pH (P = 0.02) than the remaining treatments. In conclusion, PKE supplement reduced ATTD of GE and CP, and concentration and output of dry matter in colostrum but increased FA. Results of this experiment indicated that the use of PKE as a fiber source for late gestating sows should be avoided.     

Detecting endotoxin activity in bovine serum using an automated testing system

The aim of the present study was to compare the ability of the commercially available portable test system (PTS^TM) to detect endotoxin activity in bovine serum, with that of the traditional LAL-kinetic turbidimetric (KT) and chromogenic (KC) assays. Prior to testing, serum samples, which were obtained from endotoxin-challenged cattle, were diluted 1:20 in endotoxin-free water and heated to 80°C for 10 min. The performance of the PTS^TM was not significantly different from that of the traditional LAL-based assays. The results using PTS^TM correlated with those using KT (r²=0.963, P<0.001) or KC assays (r²=0.982, P<0.001). Based on these findings, the PTS^TM could be applied as a simplified system to assess endotoxin activity in bovine serum.     

Effect of a carbohydrase admixture in growing pigs fed wheat-based diets in thermoneutral and heat stress conditions

The efficacy of exogenous carbohydrases in pig diets has been suggested to depend on enzyme activity and dietary fiber composition, but recent evidence suggests other factors such as ambient temperature might be important as well. Therefore, we investigated the effect of heat stress (HS) on the efficacy of a multienzyme carbohydrase blend in growing pigs. Ninety-six (barrows: gilts; 1:1) growing pigs with initial body weight (BW) of 20.15 ± 0.18 kg were randomly assigned to six treatments, with eight replicates of two pigs per pen in a 3 × 2 factorial arrangement: three levels of carbohydrase (0, 1X, or 2X) at two environmental temperatures (20 °C or cyclical 28 °C nighttime and 35 °C day time). The 1X dose (50 g/tonne) provided 1,250 viscosimetry unit (visco-units) endo-β-1,4-xylanase, 4,600 units α-l-arabinofuranosidase and 860 visco-units endo-1,3(4)-β-glucanase per kilogram of feed. Pigs were fed ad libitum for 28 d and 1 pig per pen was sacrificed on day 28. There was no enzyme × temperature interaction on any response criteria; thus, only main effects are reported. Enzyme treatment quadratically increased (P < 0.05) BW on day 28, average daily gain (ADG) (P < 0.05), and average daily feed intake (ADFI) (P < 0.05) with the 1X level being highest. HS reduced the BW at day 14 (P < 0.01) and day 28 (P < 0.01), ADG (P < 0.01), and ADFI (P<0.001). There was a trend of increased feed efficiency (G:F) (P < 0.1) in the HS pigs. HS increased apparent jejunal digestibility of energy (P < 0.05) and apparent ileal digestibility of calcium (P < 0.01). At day 1, HS reduced serum glucose (P < 0.001) but increased nonesterified fatty acid (P < 0.01). In the jejunum, there was a trend of increased villi height by carbohydrases (P < 0.1), whereas HS reduced villi height (P < 0.05). HS increased the jejunal mRNA abundance of IL1β in the jejunum (P < 0.001). There was a trend for a reduction in ileal MUC2 (P < 0.1) and occludin (P < 0.1) by HS, and a trend for increased PEPT1 (P < 0.1). There was no effect of HS on alpha diversity and beta diversity of the fecal microbiome, but there was an increase in the abundance of pathogenic bacteria in the HS group. In conclusion, HS did not alter the efficacy of carbohydrases. This suggests that carbohydrases and HS modulate pig performance independently.     

Culture of bovine bone marrow progenitor cells in vitro.

In vitro methylcellulose cultures of bovine bone marrow progenitor cells were developed. An existing technique described for bovine species was compared to a method for human tissue and further adapted during subsequent experiments. Bovine bone marrow samples were collected at the slaughterhouse, and mononuclear cells were separated by gradient centrifugation (1.077 g/ml specific density and 400 g). The use of 3% bovine leucocyte-conditioned medium, produced by stimulation of blood lymphocytes with 4 microg/ml concanavalin A and harvested on day 4 of culture, gave better results than the use of supernatant of the human bladder carcinoma 5637, which is widely used in human bone marrow cultures. However, bovine leucocyte-conditioned medium was not added to erythroid cultures because inhibitory effects were observed. Erythroid colonies were stimulated with erythropoietin, and hemin was added to enable microscopic identification. Reduced oxygen tension was necessary to induce growth of erythroid colonies. This was not necessary for myeloid cultures. In conclusion, the results of this study show that the growth of myeloid and erythroid colonies in methylcellulose-based medium requires different culture conditions, which are different from the culture conditions for human cells.     

Effectiveness of combined acupuncture and manual therapy relative to no treatment for canine musculoskeletal pain

Despite the rise in popularity of both acupuncture and manual therapy in veterinary medicine, and the increasing number of Canadian veterinarians practising these techniques, there is little research demonstrating their effectiveness. In this repeated measures, therapeutic trial, 47 client-owned dogs with naturally occurring lameness were assessed for clinical response to treatment. Owners were blinded to the treatment schedule and completed questionnaires to assess their dogs’ comfort and mobility. Comparison between pre- and post-treatment results demonstrated that combined acupuncture and manual therapy provides immediate short-term improvement in comfort and mobility, as demonstrated by owner observed changes in play behavior (P = 0.015), walking (P < 0.001), trotting (P = 0.002), jumping (P < 0.001), descending stairs (P = 0.003), rising from a lying position (P < 0.001), and reduced stiffness after rest (P < 0.001) or following exercise (P < 0.001). Mood and attitude also improved, but did not attain statistical significance.     

The effects of creep feed composition and form and nursery diet complexity on small intestinal morphology and jejunal mucosa-specific enzyme activities after weaning in pigs

Fifty-six litters from first-parity sows standardized to 12 piglets were used to determine the effects of creep feed composition and form and the provision of low- or high-complexity nursery diets on the evolution of small intestinal histomorphology and jejunal mucosa-specific enzyme activities postweaning. At 5 d of age, litters (initial bodyweight [BW] 2.31 ± 0.61 kg) were assigned to one of four creep feeding regimens (n = 14): 1) commercial creep feed (COM), 2) liquid milk replacer (LMR), 3) pelleted milk replacer (PMR), or 4) no creep feed (NO). At weaning (21 d of age), six pigs per litter were provided a HIGH- (contained highly digestible animal proteins) or LOW- (contained corn and soybean meal as main protein sources) complexity nursery diet (n = 7). At 21, 28, and 59 d of age, two pigs per pen (one castrated male and one female) were euthanized, and ileal and jejunal segments for histomorphological measurements and jejunal mucosal scrapings were collected to determine specific mucosa enzyme activities. At weaning, pigs provided COM had a greater ileal absorptive capacity (M) than LMR or NO, which were not different (14.1 vs. 10.4 and 10.5 ± 0.9 μm²; P < 0.05); PMR was intermediate. On days 28 and 59, M was not different among pigs regardless of creep feed treatments. Pigs fed LOW had reduced jejunal villus height (VH; P < 0.001) and M (P < 0.001) on day 28 vs. day 21. The VH and M were not different for pigs fed HIGH or LOW by the end of the nursery period. For all dietary treatments except COM-HIGH and COM-LOW, jejunal mucosal maltase-specific activity was not different between days 21 and 28 of age but greater on day 59 (P < 0.05). For pigs that received COM-HIGH, maltase-specific activity was not different between days 21 and 28 but greater on day 59 than day 28 (P < 0.05). For pigs that received COM-LOW, maltase-specific activity was not different between days 21, 28, and 59. Regardless of creep or nursery treatment, sucrase-specific activity was the greatest on day 59, followed by days 21 and 28 (P < 0.001), and lactase-specific activity was greater on day 21 than on days 28 and 59 (P < 0.001), which were not different. Therefore, pigs that provided LOW diet had greater villus atrophy and reduced M during the first week after weaning vs. pigs that provided HIGH, regardless of creep feeding regimen, but were able to recover by the end of the nursery period.     

Wingless-type mouse mammary tumor virus integration site regulation of bovine theca cells

Ovarian paracrine mediation by components of the wingless-type mouse mammary tumor virus integration site ligands (WNT1 to 11) and their receptors, frizzled family members (FZD1 to 10), has been proposed. Secreted truncated forms of FZD proteins (e.g., secreted frizzled-related protein 4 [SFRP4]) block the action of WNT ligands. Dickkopf-1 (DKK1) is another WNT antagonist, and R-spondin-1 (RSPO1) is one of a group of four secreted proteins that enhance WNT/β-catenin signaling. Our hypothesis was that granulosa cells signal theca cells (TCs) via SFRP4, DKK1, RSPO1, and WNT secretion to regulate TC differentiation and proliferation. Therefore, in vitro experiments were conducted to study the effects of WNT family member 3A (WNT3A), WNT5A, RSPO1, DKK1, insulin-like growth factor 1 (IGF1), bone morphogenetic protein 7 (BMP7), Indian hedgehog (IHH), and fibroblast growth factor 9 (FGF9) on bovine TC proliferation and steroidogenesis. TCs of large (8 to 20 mm) and small (3 to 6 mm) follicles were collected from bovine ovaries; TC monolayers were established in vitro and treated with various doses of recombinant human WNT3A, WNT5A, RSPO1, DKK1, IGF1, FGF9, BMP7, IHH, and/or ovine luteinizing hormone (LH) in serum-free medium for 48 h. In experiment 1, using LH-treated TC, IGF1, IHH, and WNT3A increased (P < 0.05) cell numbers and androstenedione production, whereas WNT3A and BMP7 inhibited (P < 0.05) progesterone production. In experiment 2, FGF9 blocked (P < 0.05) the WNT3A-induced increase in androstenedione production in LH plus IGF1-treated TC. In experiment 3, RSPO1 further increased (P < 0.05) LH plus IGF1-induced progesterone and androstenedione production. In experiment 4, SFRP4 and DKK1 alone had no significant effect on TC proliferation or progesterone production of large-follicle TC but both blocked the inhibitory effect of WNT5A on androstenedione production. In contrast, DKK1 alone inhibited (P < 0.05) small-follicle TC androstenedione production whereas SFRP4 was without effect. We conclude that the ovarian TC WNT system is functional in cattle, with WNT3A increasing proliferation and androstenedione production of TC.     

Effects of cooled floor pads combined with chilled drinking water on behavior and performance of lactating sows under heat stress

This study was conducted to evaluate whether cooled floor pads combined with chilled drinking water could alleviate negative impacts of heat stress on lactating sows. Thirty sows (Landrace × Yorkshire, Parity = 1 to 6) were housed in individual farrowing stalls in two rooms with temperatures being controlled at 29.4°C (0700–1900 hours) and 23.9°C (1900–0700 hours). Sows in one room (Cool), but not in the other room (Control) were provided cooled floor pads (21–22°C) and chilled drinking water (13–15°C). Behavior of sows (15 sows/treatment) was video recorded during farrowing, and days 1, 3, 7, 14, and 21 after farrowing. Videos were viewed continuously to register the birth time of each piglet, from which total farrowing duration and birth intervals were calculated. The number of drinking bouts and the duration of each drinking bout were registered for each sow through viewing videos continuously for 2 h (1530–1730 hours) each video-recording day. Postures (lying laterally, lying ventrally, sitting, and standing) were recorded by scanning video recordings at 5-min intervals for 24 h each video-recording day, and time budget for each posture was calculated. Rectal temperature and respiration rate were measured for all sows the day before and after farrowing, and then once weekly. Sow and litter performance was recorded. Data were analyzed using the Glimmix procedure of SAS. The cooling treatment did not affect sow behavior or litter performance. Sows in the Cool room had lower rectal temperature (P = 0.03) and lower respiration rate (P < 0.001), consumed more feed (P = 0.03), tended to have reduced weight loss (P = 0.07), and backfat loss (P = 0.07) during lactation than sows in the Control room. As lactation progressed, sows increased drinking frequency (P < 0.001) and time spent lying ventrally (P < 0.0001), standing (P < 0.001), and sitting (P < 0.0001), and decreased time spent lying laterally (P < 0.0001) in both Cool and Control rooms. While cooled floor pads combined with chilled drinking water did not affect sow behavior, they did alleviate heat stress partially, as indicated by decreased rectal temperature, respiration rate, weight, and backfat loss, and increased feed intake in lactating sows.     

Comparison of the bovine blood gas parameters produced with three types of portable blood gas analyzers

BackgroundA definite diagnosis should be made in the bovine practice field, however, it was difficult to perform laboratory analysis immediately. Currently, three types of portable blood gas analyzers are available in Korea.ObjectivesThis study aimed to evaluate the correlations among these three analyzers.MethodsSeventy-two plasma samples from Holstein-Friesian cows were used for blood gas analysis, and three instruments (EDAN i15 Vet, VETSCAN i-STAT, and EPOC) were operated simultaneously. Moreover, plasma calcium levels were compared between these portable analyzers and blood chemistry device, which is usually used in a laboratory environment. Pearson analysis was performed to confirm the correlation of each parameter produced with the three instruments and blood chemistry analyzer.ResultsAs results, high correlation was observed in parameters of pH, pO₂, potassium ion, ionized calcium, and glucose (p < 0.001, r > 0.7). In addition, pCO₂ showed a moderate correlation among the three analyzers (p < 0.001, r > 0.5), and there was no correlation among all instruments for sodium ions. There was also a high correlation between ionized calcium from the three portable devices and total calcium from the biochemistry analyzer (p < 0.001, r > 0.9).ConclusionsIn conclusion, there was a high correlation between results from the three different blood gas analyzers used in the bovine clinical field in Korea. Thus, a consistent diagnosis can be made even with different equipment if the operator is aware of the strengths and weaknesses of each piece of equipment and operates it properly.     

Impact of meloxicam on respiratory virus titers and health outcomes when administered concurrently with a modified live respiratory vaccine in abruptly weaned beef steers

Abruptly weaned crossbred steer calves (N = 271) were used in a randomized, blinded 2-arm clinical trial to assess the impact of a long-acting non-steroidal anti-inflammatory drug on bovine herpesvirus type 1, bovine respiratory syncytial virus, parainfluenza virus type 3, and coronavirus titers and health outcomes when administered concurrently with a modified live respiratory vaccine upon arrival at a feedlot. Treatment groups included a control (saline; n = 135) and an experimental group (injectable meloxicam; n = 136). Viral antibody titers and body weight were measured on arrival, day 7, and day 21, along with a final weight on day 45. Body weight and antibody titers for all viruses increased over time (P < 0.001); however, there were no differences by treatment group or a significant group × time interaction when evaluated using repeated measures analysis of variance. Interestingly, the use of meloxicam was associated with increased treatment risk (P < 0.05). In conclusion, the administration of meloxicam may adversely affect health; however, a decreased vaccine response is likely not a contributing factor.     

Effects of extended postmortem aging and intramuscular location on protein degradation,muscle fiber morphometrics,and tenderness of beef longissimus lumborum and semitendinosus steaks

The objective of this study was to determine effects of extended aging and intramuscular location on Warner-Bratzler shear force (WBSF), muscle fiber cross-sectional area (CSA), and protein degradation of semitendinosus (ST) and longissimus lumborum (LL) steaks. Left ST and LL were removed from 40 carcasses at 6 d postmortem. The ST was fabricated into five locations (LOC), with LOC 1 being most proximal and LOC 5 being most distal. The posterior LL was fabricated into 3 LOC, with LOC 1 being most anterior. Vacuum sealed ST steaks were aged 7, 14, 28, 56, or 112 d postmortem, while LL steaks were aged 7, 28, or 112 d postmortem at 2 ± 1 °C. A steak from each LOC was assigned to WBSF or laboratory analyses. There were no Day of Aging (DOA) × LOC interactions for all dependent variables (P > 0.06). There were DOA effects for ST and LL WBSF values and degraded 38-kDa desmin (DES; P < 0.01). Day-7 ST-steak WBSF value was greater than all other days (P < 0.01) and day-14 steaks had greater WBSF value than remaining days (P < 0.05). Day-28 ST-steak WBSF values were greater than day 56 and 112 (P < 0.01), which did not differ (P = 0.53). In the LL, day-7 steaks had greater WBSF values than the other two timepoints (P < 0.01) and day-28 steaks had greater (P < 0.01) WBSF values than day-112 steaks. Degraded ST 38-kDa DES content was less on day 7 and 14 compared to all other days (P < 0.03), but did not differ (P = 0.79) from each other. Days 28 and 56 38-kDa DES content was less than day 112 (P < 0.01), but did not differ (P = 0.34) from each other. Degraded LL 38-kDa DES content was less on day 7 than day 28 and 112 (P < 0.02), which did not differ (P = 0.67). There were LOC effects for only ST WBSF and muscle fiber CSA (P < 0.05). Semitendinosus steak LOC 1 and 2 had greater WBSF values than all other locations (P < 0.01), but did not differ (P = 0.32) from each other. Semitendinosus steak LOC 3 and 5 had greater WBSF values than LOC 4 (P < 0.01), but did not differ (P = 0.85) from each other. The CSA of all ST fiber types were largest in LOC 1 compared to all other fiber types (P < 0.01). The CSA of all LOC 2 and 3 fiber types was greater than LOC 4 and 5 (P < 0.01), but were not different from each other (P > 0.81), and LOC 4 had greater CSA than LOC 5 (P < 0.01). Steak aging WBSF value improvements seemed proteolysis catalyzed, while the ST intramuscular tenderness gradient was more likely due to muscle fiber CSA.     

Influence of a novel consensus bacterial 6-phytase variant on mineral digestibility and bone ash in young growing pigs fed diets with different concentrations of phytate-bound phosphorus

A 20-d experiment was conducted to test the hypothesis that phytase increases nutrient digestibility, bone ash, and growth performance of pigs fed diets containing 0.23%, 0.29%, or 0.35% phytate-bound P. Within each level of phytate, five diets were formulated to contain 0, 500, 1,000, 2,000, or 4,000 phytase units (FTU)/kg of a novel phytase (PhyG). Three reference diets were formulated by adding a commercial Buttiauxella phytase (PhyB) at 1,000 FTU/kg to diets containing 0.23%, 0.29%, or 0.35% phytate-bound P. A randomized complete block design with 144 individually housed pigs (12.70 ± 4.01 kg), 18 diets, and 8 replicate pigs per diet was used. Pigs were adapted to diets for 15 d followed by 4 d of fecal collection. Femurs were collected on the last day of the experiment. Results indicated that diets containing 0.35% phytate-bound P had reduced (P < 0.01) digestibility of Ca, P, Mg, and K compared with diets containing less phytate-bound P. Due to increased concentration of total P in diets with high phytate, apparent total tract digestible P and bone ash were increased by PhyG to a greater extent in diets with 0.29% or 0.35% phytate-bound P than in diets with 0.23% phytate-bound P (interaction, P < 0.05). At 1,000 FTU/kg, PhyG increased P digestibility and bone P more (P < 0.05) than PhyB. The PhyG increased (P < 0.01) pig growth performance, and pigs fed diets containing 0.35% or 0.29% phytate-bound P performed better (P < 0.01) than pigs fed the 0.23% phytate-bound P diets. In conclusion, the novel phytase (i.e., PhyG) is effective in increasing bone ash, mineral digestibility, and growth performance of pigs regardless of dietary phytate level.     

Colony Formation in Culture by Bovine Granulopoietic Progenitor Cells

Calf bone marrow cells cultured in a semi-solid medium of 0.8% methyl cellulose produced colonies of granulocytic cells and macrophages by seven days. A prerequisite for colony growth was the presence of serum obtained from a calf three hours after intravenous injection of endotoxin. Three morphological types of colonies were seen but cell types within these types of colonies did not differ. Cultured cells were identified by morphological and cytochemical characteristics.

Optimum growth occurred when serum from endotoxin stimulated calves and fetal calf serum were present in a volumetric ratio of 7:3. Inhibition of colony growth occurred when endotoxin-stimulated serum was present at greater than optimum concentration. Normal calf serum, fetal calf serum, mouse L-cell conditioned medium and bovine urine did not stimulate significant colony growth when 8.0 x 10⁴ marrow cells were cultured.

There was a linear relationhip between the number of marrow cells in the cultures and the number of colonies produced. Colony forming efficiency ranged from 13 to 59 colonies per 10⁵ cells plated.

The behaviour of calf colony forming units in suspension culture was similar to that reported for mouse colony forming units.

     

Melatonin alters bovine uterine artery hemodynamics,vaginal temperatures,and fetal morphometrics during late gestational nutrient restriction in a season-dependent manner

The objectives were to examine melatonin-mediated changes in temporal uterine blood flow (UBF), vaginal temperatures (VTs), and fetal morphometrics in 54 commercial Brangus heifers (Fall, n = 29; Summer, n = 25) during compromised pregnancy. At day 160 of gestation, heifers were assigned to one of the four treatments consisting of adequately fed (ADQ-CON; 100% National Research Council [NRC]; n = 13), global nutrient restricted (RES-CON; 60% NRC; n =13), and ADQ or RES supplemented with 20 mg/d of melatonin (ADQ-MEL, n = 13; RES-MEL, n = 15). In the morning (0500 hours; AM) and afternoon (1300 hours; PM) of day 220 of gestation, UBF was determined via Doppler ultrasonography, while temperature data loggers attached to progesterone-free controlled internal drug releases were used to record VTs. At day 240 of gestation, heifers underwent cesarean sections for fetal removal and morphometrics determination. The UBF and VT data were analyzed using repeated measures of analysis of variance (ANOVA), while the morphometrics was analyzed using the MIXED procedure of SAS. Seasons were analyzed separately. In Fall, a nutrition by treatment interaction was observed, where the RES-CON heifers exhibited reduced total UBF compared with ADQ-CON (5.67 ± 0.68 vs. 7.97 ± 0.54 L/min; P = 0.039). In Summer, MEL heifers exhibited increased total UBF compared with the CON counterparts (8.16 ± 0.73 vs. 6.00 ± 0.70 L/min; P = 0.048). Moreover, there was a nutrition by treatment by time interaction in VT for Fall and Summer heifers (P ≤ 0.005). In Fall, all groups had decreased VT in the morning compared with the afternoon (P < 0.05). Whereas, in Summer, VT increased for ADQ-CON and RES-CON (P < 0.0001) from morning to afternoon, the ADQ-MEL and RES-MEL remained constant throughout the day (P = 0.648). Furthermore, the RES-MEL-PM exhibited decreased VT compared with ADQ-CON-PM (38.91 ± 0.09 vs. 39.26 ± 0.09 °C; P = 0.018). Lastly, in Fall, a main effect of nutrition was observed on fetal weights, where the RES dams had fetuses with decreased body weight when compared with ADQ (24.08 ± 0.62 vs. 26.57 ± 0.64 kg; P = 0.0087). In Summer, a nutrition by treatment interaction was observed on fetal weights where the RES-CON dams had fetuses with reduced weight when compared with ADQ-CON and RES-MEL (P < 0.05). In summary, nutrient restriction decreased UBF and melatonin supplementation increased UBF depending on the season. Additionally, melatonin appeared to decrease VT and rescue fetal weights when supplemented in the Summer.     

Relationship between the Lactic Acid Content and Sour Taste of Broiler Broth and the Broth of Choshu-Kurokashiwa—a Japanese Jidori Chicken

The present study aimed to determine whether lactic acid content is associated with the intensity of the sour taste of Choshu-Kurokashiwa broth. Chicken broth was prepared using breast (pectoralis major) and leg (thigh + drumstick) meat of male and female Choshu-Kurokashiwa and broilers. These broths were assessed by a screened and trained panel and analyzed for sour taste substances (lactic, citric, pyruvic, malic, succinic, acetic, phosphoric, aspartic, and glutamic acids) and pH. The sensory sour taste was significantly higher in the Choshu-Kurokashiwa breast broth than in the broiler breast broth (P>0.001), and no significant difference was observed in the leg broths among the breeds (P<0.05). Choshu-Kurokashiwa breast broth had a significantly higher lactic acid content than broiler breast broth (P<0.001). The leg meat broth of male Choshu-Kurokashiwa had a significantly lower lactic acid content than that of female Choshu-Kurokashiwa and broiler leg meat (P<0.01). The sensory sour taste score was significantly and positively correlated with lactic acid content in the breast broth (P<0.001), but not in the leg broth (P<0.1). No other organic acids were detected. Phosphoric acid and glutamic acid contents were higher in broiler broth than in Choshu-Kurokashiwa broth for both breast and leg meat (P<0.001). In the breast broth, the aspartic acid content was not significantly different (P>0.1), and in the leg broth, it was higher in broiler and female Choshu-Kurokashiwa broth than in male Choshu-Kurokashiwa broth (P<0.001). The present study suggests that high lactic acid content contributes to the sour taste of the Choshu-Kurokashiwa breast broth and demonstrated that the lactic acid content is an essential indicator for determining the sour taste of Choshu-Kurokashiwa breast meat.