A comparative study of the effects of glycerol and hydroxyethyl starch in canine red blood cell cryopreservation

Hydroxyethyl starch (HES) is a nonpenetrating extracellular cryoprotectant. In contrast to glycerol, it does not require labor-intensive removal from thawed red blood cells (RBCs) prior to transfusion. In this study, we compared glycerol and HES, and assessed HES as a substitute for glycerol in cryopreserved canine RBCs. The RBCs were preserved for 2 months in liquid nitrogen using a 20% (w/v) glycerol solution, and variable concentrations of HES solution. We evaluated the two cryoprotectants by the percentage of post-thaw hemolysis from the total free hemoglobin, saline stability, osmotic fragility, and by observing the erythrocyte morphology using a scanning electron microscope after thawing. The optimal concentration of HES was 12.5% (w/v) for the cryopreservation of canine RBCs. The thaw hemolysis, saline stability, and osmotic fragility index were 25.6 +/- 4.7%, 87.8 +/- 6.9%, and 0.445 +/- 0.024% NaCl respectively. These parameters resemble the results of RBCs frozen in a 20% (w/v) glycerol solution, which are 24.7 +/- 5.2%, 99.2 +/- 0.1%, and 0.485 +/- 0.023% NaCl respectively. From a morphological point of view, 12.5% (w/v) HES showed the best cryoprotection of RBCs compared to the other concentrations of HES. These results suggest that HES could be a possible substitute for glycerol for the cryopreservation of canine RBCs.     

Effects of warming basal ends of Carolina poplar (Populus × canadensis Moench.) softwood cuttings at controlled low-air-temperature on their root growth and leaf damage after planting

We investigated the effects of warming the basal ends of Carolina poplar (Populus × canadensis Moench.) softwood cuttings at controlled low-air-temperature on their root growth and leaf damage after planting. The warming treatment was applied to the cuttings by soaking 10 mm beyond the cut end in warmed water maintained at 30 °C in a cold chamber maintained at an air temperature of 10 °C and a photosynthetic photon flux density (PPFD) of 10 μmol m^?2 s^?1 (near the light compensation point at 10 °C) until rooting was observed. The warmed cuttings were then grown in a growth chamber at an air temperature of 30 °C, relative humidity 85–90 %, and a PPFD of 100 μmol m^?2 s^?1. Control cuttings were grown in the growth chamber throughout the experiment. Rooting occurred simultaneously for both warmed and control cuttings, irrespective of air temperature. Root development was greater and leaf damage, evaluated on the basis of extent of necrosis, was less for warmed cuttings than for control cuttings. The reduction of leaf damage for warmed cuttings probably resulted from reduced post-planting water stress and leaf senescence, because of improved root development as a result of the pre-planting warming treatment. This technique could improve the propagation of cuttings of woody plants, because it would ensure that the cuttings are ready to develop roots with minimum loss of carbohydrates, irrespective of weather conditions.     

Compliance rate of livestock vehicles with welfare standards, and behavior of feeder cattle at loading ramp

The objective of the present study was to assess cattle welfare during transportation. Vehicle inspection and observation of cattle behavior during loading operations were conducted at two major livestock markets (T and M) in Japan. Market T provided young feeder cattle (Wagyu and cross‐bred aged 6.8–9.0 months) mainly for regions farther than 1500 km. Market M provided young cattle (Wagyu aged 6.6–11.2 month) and calves (crossbred and Holstein aged 0.7–1.5 month) for nearby regions within 500 km. Market T had loading platforms 1.0 m high, whereas market M had partial allocation of platforms and forced most transporters to load cattle from the ground. Vehicles were inspected according to the welfare standards for beef cattle of the RSPCA. Number of vehicles inspected was 36 and 31 in markets T and M, respectively. Cattle hesitations (kneeling down, slipping, balking, backing down, turning around, jumping and eliminating) were observed at the loading ramp. Vehicles inspected at the markets complied with most requirements of the welfare standards, but non‐compliance was found in two requirements: in market M, 71.0% of vehicles had the loading ramp at a >20% incline, whereas 17.1% of vehicles did in market T (P < 0.001). Slope of the loading ramp was steeper in market M than in market T (33.9 ± 17.3% vs. 14.9 ± 8.9%, P < 0.001). Market M had higher proportion of vehicles that did not comply with the requirement ‘Both loading ramps and tail boards must be appropriately designed and covered with litter, to prevent animals from falling off or slipping’ compared with market T (83.9% vs. 17.1%, P < 0.001). Higher frequencies were observed in two kinds of hesitating behavior in market M than in market T (both P < 0.01): mean frequencies (times/head) of slipping and balking were 1.3 ± 1.2 and 2.0 ± 1.8 in market M, and 0.2 ± 0.2 and 0.7 ± 0.5 in market T, respectively. Steeper loading ramp was correlated with higher frequencies of kneeling down (r = 0.53), slipping (r = 0.59), balking (r = 0.45) and backing down (r = 0.42) (all P < 0.05).     

Comparison of behavior, physical condition and performance of laying hens in four molting methods

The objective of the present study was to evaluate the welfare level of hens in four molting methods by comparing their behavior, physical condition and performance. In total, 96 White Leghorn layers (72 weeks of age) were used. The four molting methods consisted of 10 days of feeding and the first 3 days of water withdrawal WFW); 10 day feed withdrawal (WF) followed by ad libitum access to a layer diet from day 11, 28 days of restricted feeding (RF) and 28 days without withdrawing their feed (NW). The RF and NW hens were fed a low energy diet. The WFW and WF hens showed two clear phases of behavior during the 10‐day fasting period. Explorative and stereotyped behavior increased for WFW and WF hens just after their fast. Standing‐resting then increased for WFW and WF hens while lying‐resting decreased. In contrast, RF and NW hens did not show clear behavioral changes during the 28‐day molt period. The weight loss of the NW hens was more gradual compared with the other three methods. No significant differences were seen in any productive trait among the four methods during the postmolt period. In conclusion, the welfare level of RF and NW, especially NW, was higher compared with WFW and WF.     

Somatic cell and innate immune responses in mammary glands of lactating cows to intramammary infusion of Bifidobacterium breve at pre-drying off period

Intramammary infusion of Bifidobacterium breve (B. breve)-induced somatic cell (SC) counts, chemiluminescent response (CL), lactoferrin (LF) concentrations and mastitis-causing pathogens from quarters with subclinical mastitis were measured to evaluate innate immune response of mammary glands in dairy cows at 3 to 4 weeks before drying off. SC counts in 7 quarters of 7 control cows and 5 quarters of 6 cows with mastitis increased markedly on day 1 and SC values in control cows were significantly (P<0.05) increased and returned to pre-infusion levels on day 5 after B. breve-infusion. CL values in both groups increased markedly on day 1 and then decreased after B. breve-infusion; however, CL values in cows with mastitis did not return to normal levels on day 5 and at postpartum. The CL values were highly correlated with their SC counts in milk from both groups. LF concentrations increased toward day 3 after B. breve-infusion and were higher in cows with mastitis. B. breve-infusion eliminated 16.6% (1/6) of pathogens from 6 quarters with chronic subclinical mastitis. B. breve-induced SC responses in quarters from 3 cows with mastitis showed characteristic patterns of recovery, persistent and new infections. B. breve-induced SC counts in quarters from the cows in the pre-drying off were lower (25.7–70.6%) than those of the cows in mid-lactation. The intrinsic innate immune response in cows on pre-drying off may be decreased and appears to be insufficient to eliminate pathogens from mammary gland in the pre-drying off.     

Cytosolic glutamine synthetase is present in the phloem sap of rice (Oryza sativa L.)

The presence of glutamine synthetase (GS) in the rice sieve tube was examined. Proteins in the rice phloem sap from leaf sheaths were separated by sodium dodecylsulfate–polyacrylamide gel electrophoresis, transferred to polyvinylidine difluoride membranes and immunoblotted with anti-GS1 antibody. A cross-reacting band, thought to be GS1, was detected in the phloem sap. Moreover, the phloem sap contained a significant amount of GS transferase activity. Previous studies have shown that the concentrations of substrates and cofactors in the rice phloem sap are sufficient for cytosolic GS reaction. These data suggest that physiologically active GS1 is present in rice phloem sap, which might convert glutamate to glutamine in vivo .     

Survey of the Occurrence of Equine Deep Digital Flexor Tendon Contraction (Clubfoot) in the Main Thoroughbred Breeding Area in Japan

At 114 ranches in the Hidaka District of Hokkaido, a total of 1,118 Thoroughbred foals that were born from January to June 2003 were examined at different times from May to September of the same year to ascertain the occurrence of clubfoot. Clubfoot was seen in a total of 179 foals (16%) at 89 ranches (78%). Also, 124 of the 179 foals (69.3%) had clubfoot of grade II or higher, and it was found that grade I was likely to be overlooked or rapidly advance. In the present study, the occurrence of clubfoot was higher than expected in the investigated region. Hence, it is necessary to establish accurate diagnostic techniques and clubfoot guidelines to minimize the impact.