Effects of niacin and betaine on bovine mammary and uterine cells exposed to thermal shock in vitro.

The objective of this study was to investigate the direct effects of feed supplements niacin and betaine on the heat shock responses of in vitro cultured cells derived from bovine mammary and uterine tissues. First, we determined the mRNA expression profiles of the niacin receptor (GPR109A) in bovine tissues (liver, skin, uterus, udder, and ovary) and in cells derived from bovine mammary epithelium (mammary alveolar cells, MAC-T; bovine mammary epithelial cells, BMEC) and endometrium (bovine endometrial cells, BEND). We found that GPR109A was distributed in all examined tissues and cells, and the highest expression was in cells from skin and udder. Second, we evaluated the effects of niacin treatment on the mRNA abundance of heat shock proteins 70 and 27 (HSP70 and HSP27) in MAC-T, BMEC, and BEND under thermoneutral conditions and heat stress, and whether these effects were associated with alterations in the mRNA expression of prostaglandin E2 synthesis-related genes, including cyclooxygenase 1 and 2 (COX-1 and COX-2) and microsomal prostaglandin E synthase 1 and 2 (mPGES-1 and mPGES-2). Quantitative PCR data indicated that niacin suppressed HSP70 mRNA expression in BMEC and both HSP70 and HSP27 in BEND under thermoneutral conditions. Only COX-2 expression was downregulated by niacin in BMEC; other prostaglandin E2 synthesis-related genes stayed unaltered in BMEC and BEND. The mRNA abundance of HSP70, COX-1, COX-2, and mPGES-1 were elevated in niacin-treated MAC-T. During heat stress, niacin increased mRNA levels of HSP70 and HSP27 in MAC-T and HSP27 in BEND, but decreased HSP70 in BMEC. Although mPGES-2 was stimulated by niacin in BEND, the mRNA expression of prostaglandin E2 synthesis-related genes were consistent with neither HSP70 nor HSP27 expression patterns in niacin-treated BMEC and MAC-T. These data suggest that the effects of niacin on heat shock protein expression and prostaglandin E2 synthesis were not well coupled in these cells. Finally, we tested the effects of betaine treatment on viability and apoptosis in BMEC. Compared with control cultures, viability was higher in betaine-treated cells at 8 h under thermoneutral conditions and at 16 h in heat stress, and apoptotic rates were lower at 8 h. Our data support a dual role for niacin in regulating heat shock protein expression in normal and heat-shocked cells derived from mammary and uterine tissues, and positive effects of betaine in regulating mammary cell viability during heat stress.

Evaluation of dietary betaine in lactating Holstein cows subjected to heat stress.

Betaine (BET), a natural, organic osmolyte, improves cellular efficiency by acting as a chaperone, refolding denatured proteins. To test if dietary BET reduced the effect of heat stress (HS) in lactating dairy cows, multiparous, lactating Holstein cows (n=24) were blocked by days in milk (101.4±8.6 d) and randomly assigned to 1 of 3 daily intakes of dietary BET: the control (CON) group received no BET, mid intake (MID) received 57mg of BET/kg of body weight, and high dose (HI) received 114mg of BET/kg of body weight. Cows were fed twice daily and BET was top-dressed at each feeding. Cows were milked 2 times/d and milk samples were taken daily for analysis. Milk components, yield, feed intake, and water intake records were taken daily. Rectal temperature and respiration rate were taken 3 times/d at 0600, 1400, and 1800h. Cows were housed in environmentally controlled rooms and were allowed acclimation for 7d at thermoneutral (TN) conditions with a mean temperature-humidity index of 56.6. Cows were then exposed to 7d of TN followed by 7d of HS represented by a temperature-humidity index of 71.5 for 14d. This was followed by a recovery period of 3d at TN. Dietary BET increased milk yield during the TN period. No differences were found between BET and CON in total milk production or milk composition during HS. The increase in water intake during HS was not as great for cows fed BET compared with controls. The cows on CON diets had higher p.m. respiration rate than both MID and HI BET during HS, but lower rectal temperature compared with BET. No difference was found in serum glucose during TN, but cows given HI had elevated glucose levels during HS compared with CON. No differences were found in serum insulin levels between CON and BET but an intake by environment interaction was present with insulin increasing in HI-treated lactating dairy cows during HS. The heat shock response [heat shock protein (HSP) 27 and HSP70] was upregulated in bovine mammary epithelial cells in vitro. Blood leukocyte HSP27 was downregulated at the HI dose under TN conditions and HSP70 was upregulated at the HI dose and this effect was increased by HS. No effect was seen with the MID dose with HSP27 or HSP70. The lack of effect of BET at MID may be associated with uptake across the gut. We conclude that BET increased milk production under TN conditions and was associated with reduced feed and water intake and slightly increased body temperatures during HS of cows fed BET. The effect of BET on milk production was lost during HS with HI BET, whereas serum glucose levels increased during HS.

A dose-response evaluation of rumen-protected niacin in thermoneutral or heat-stressed lactating Holstein cows.

Twenty-four multiparous high-producing dairy cows (40.0±1.4kg/d) were used in a factorial design to evaluate effects of 2 environments [thermoneutral (TN) and heat stress (HS)] and a dose range of dietary rumen-protected niacin (RPN; 0, 4, 8, or 12g/d) on body temperature, sweating rate, feed intake, water intake, production parameters, and blood niacin concentrations. Temperature-humidity index values during TN never exceeded 68 (stress threshold), whereas temperature-humidity index values during HS were above 68 for 24h/d. The HS environment increased hair coat and skin, rectal, and vaginal temperatures; respiration rate; skin and hair coat evaporative heat loss; and water intake and decreased DMI (3.5kg/d), milk yield (4.1kg/d), 4% fat-corrected milk (2.7kg/d), and milk protein yield (181.7g/d). Sweating rate increased during HS (12.7g/m(2) per h) compared with TN, but this increase was only 10% of that reported in summer-acclimated cattle. Niacin supplementation did not affect sweating rate, dry-matter intake, or milk yield in either environment. Rumen-protected niacin increased plasma and milk niacin concentrations in a linear manner. Heat stress reduced niacin concentration in whole blood (7.86 vs. 6.89µg/mL) but not in milk. Reduced blood niacin concentration was partially corrected by dietary RPN. An interaction existed between dietary RPN and environment; dietary RPN linearly increased water intake in both environments, but the increase was greater during HS conditions. Increasing dietary RPN did not influence skin temperatures. During TN, supplementing 12g/d of RPN increased hair coat (unshaved skin; 30.3 vs. 31.3°C at 1600h) but not shaved skin (32.8 vs. 32.9°C at 1600h) temperature when compared with 0g/d at all time points, whereas the maximum temperature (18°C) of the room was lower than skin temperature. These data suggest that dietary RPN increased water intake during both TN and HS and hair coat temperature during TN; however, core body temperature was unaffected. Thus, encapsulated niacin did not improve thermotolerance of winter-acclimated lactating dairy cows exposed to moderate thermal stress in Arizona.

Effects of adjustable and stationary fans with misters on core body temperature and lying behavior of lactating dairy cows in a semiarid climate.

Cows readily seek shade to reduce solar heat load during periods of high ambient temperature. Typically, auxiliary cooling systems are oriented to maximize cooling for shaded cows. However, when a shade structure is oriented north-south, stationary fan and mister cooling systems are unable to track shade as the sun's angle shifts throughout the day, and thus can become ineffective. The FlipFan Dairy Cooling System (Schaefer Ventilation Equipment, Sauk Rapids, MN) employs fans and misters that follow shade and compensate for wind speed by rotating on a horizontal axis. Multiparous, lactating Holstein cows (n=144) on a commercial dairy in Arizona were cooled by a fixed system comprised of stationary fans and misters acting as control or the adjustable FlipFan operated for 16.5 h/d (0830 to 0100 h). Core body temperatures (CBT) of 64 cows (4 pens/treatment; 8 cows/pen; 6d) and lying behavior of 144 cows (4 pens/treatment; 18 cows/pen; 5d) were collected by intravaginal and leg data loggers, respectively. Cows were balanced by milk production, blocked by days in milk, and randomly assigned to pen within block. Pen was the experimental unit. In a second experiment, isothermal maps were developed using a fixed system of thermal data loggers arranged in the shaded areas of the pens at different times of day and were analyzed for differences in the temperature-humidity index (THI) achieved by each cooling treatment. Ambient conditions consisted of a mean temperature of 33.0°C, mean relative humidity of 40.3%, and mean THI of 80.2. Mean 24-h CBT for FlipFan was lower than control (38.9 vs. 39.1±0.04°C). A treatment × time interaction was observed in which CBT of FlipFan was 0.4°C lower than control from 0600 to 0800h and 1500 to 1600h. Cows cooled by FlipFan spent more time lying down compared with those cooled by control (9.5 vs. 8.6 h/d). Cows under FlipFan had more frequent lying bouts than did those under control (12.8 vs. 10.7 bouts/d). Lower CBT and decreased standing time are consistent with the findings of other studies when ambient heat load was reduced. In the second experiment, the FlipFan system achieved a lower THI in the morning and evening (5.9 and 1.7%, respectively), and the THI also tended to be 0.8% lower in the afternoon compared with that of control. Results indicate that FlipFan is more effective than a stationary fan and mister system at decreasing CBT, increasing lying time and bouts, and providing a more desirable microenvironment for cows throughout the day in a semiarid environment.

Ofloxacin containing combined drug regimens in the treatment of multibacillary leprosy.

The results of ofloxacin containing combined drug regimens in the treatment of 60 multibacillary leprosy cases from January 1989 to June 1995 are reported. The objective of the trial is to compare the antileprotic property of ofloxacin and rifampicin in multibacillary leprosy patients and to study the killing rate of M. leprae by ofloxacin and rifampicin before mass treatment can be recommended. The complications and side-effects of ofloxacin and rifampicin were of a mild nature and both drugs were well tolerated. Moderate to marked clinical improvement was noticed in a short period with ofloxacin containing regimens in multibacillary leprosy patients. No persisters were detected in any of the 33 specimens (of mouse footpads) that had been obtained after treatment for 6 months. Ofloxacin if added to the currently used WHO recommended MB-MDT regimen may shorten the duration of treatment. Ofloxacin, therefore, may be considered as a suitable alternative in suspected/proven rifampicin resistant cases and where rifampicin is contraindicated. The results were evaluated on the basis of the clinical conditions, mycobactericidal effectiveness, signs of drug toxicity and side effects.

Combined chemotherapy trials with regimens containing ofloxacin and rifampicin for multibacillary leprosy patients.

The results of combined chemotherapy trials with regimens containing ofloxacin and rifampicin for the treatment of 60 multibacillary leprosy cases from January 1989 to September 1995 was reported. Clinical improvement was achieved by all regimens from the end of the first month. Most patients continued to improve for 3 years. Bacterial indices were gradually reduced during the treatment. Patients on regimens containing rifampicin were clear of M. leprae at the end of the 5th year. The complications and side-effects of ofloxacin and rifampicin were trivial and both drugs were well tolerated. Ofloxacin added to the current WHO recommended M.B.-MDT regimen may shorten the duration of treatment. Ofloxacin may be an alternative in suspected/proven rifampicin resistant cases or rifampicin contraindicated.