Canine-specific tail-in, head-out sperm agglutination.

An interesting pattern of tail-in, head-out sperm agglutination was identified in a Brucella canis seronegative subfertile dog. Centrifuged seminal plasma from this dog could induce a similar pattern of agglutination in six other dogs, but not in ejaculates from a single stallion and two rams. The agglutination pattern was short-lived and appeared to depend on motility of spermatozoa, although intensity of agglutination may have been affected by concentration of agglutinating factor.

A novel rare CUBN variant and three additional genes identified in Europeans with and without diabetes: results from an exome-wide association study of albuminuria.

AIMS/HYPOTHESIS: Identifying rare coding variants associated with albuminuria may open new avenues for preventing chronic kidney disease and end-stage renal disease, which are highly prevalent in individuals with diabetes. Efforts to identify genetic susceptibility variants for albuminuria have so far been limited, with the majority of studies focusing on common variants. METHODS: We performed an exome-wide association study to identify coding variants in a two-stage (discovery and replication) approach. Data from 33,985 individuals of European ancestry (15,872 with and 18,113 without diabetes) and 2605 Greenlanders were included. RESULTS: We identified a rare (minor allele frequency [MAF]: 0.8%) missense (A1690V) variant in CUBN (rs141640975, ß = 0.27, p = 1.3 × 10-11) associated with albuminuria as a continuous measure in the combined European meta-analysis. The presence of each rare allele of the variant was associated with a 6.4% increase in albuminuria. The rare CUBN variant had an effect that was three times stronger in individuals with type 2 diabetes compared with those without (pinteraction = 7.0 × 10-4, ß with diabetes = 0.69, ß without diabetes = 0.20) in the discovery meta-analysis. Gene-aggregate tests based on rare and common variants identified three additional genes associated with albuminuria (HES1, CDC73 and GRM5) after multiple testing correction (pBonferroni < 2.7 × 10-6). CONCLUSIONS/INTERPRETATION: The current study identifies a rare coding variant in the CUBN locus and other potential genes associated with albuminuria in individuals with and without diabetes. These genes have been implicated in renal and cardiovascular dysfunction. The findings provide new insights into the genetic architecture of albuminuria and highlight target genes and pathways for the prevention of diabetes-related kidney disease.

Swept-frequency ultrasonic phase evaluation of adhesive bonding in tri-layer structures.

As modern aerospace and automotive designs continually strive for higher performance, and thus rely on advanced composite structures where adhesive bonding is a preferred method of joining, the need for a robust quantitative nondestructive bond strength measurement method has increased. As such, advanced nondestructive evaluation methods have been researched for increased sensitivity to weak interfacial bonding and ultimately to detect "kissing" bonds. In this work, a phase-based method for interrogating bonded joints and detecting weak adhesion is developed by using swept-frequency phase measurements of ultrasonic waves reflected from an adhesive joint and modeling adhesive interfaces as a distributed spring system. The method's sensitivity to bond strength is explored by ultrasonic phase evaluation of tri-layer joints with bond quality varied by controlling ultraviolet light exposure and extracting interfacial stiffness constants of the bonds. Mechanical tensile tests found each joint failed adhesively, allowing a linear correlation to be drawn between interfacial stiffness and tensile strength, consistent with previous theoretical research. The ultrasonic phase measurement method identifies intermediate bond strengths, rather than simply detecting good or bad bonds. This technique has the potential for the verification of bond quality in lightweight aerospace and automotive designs utilizing advanced composite structures with adhesive attachments.

A constant-frequency ultrasonic phase method for monitoring imperfect adherent/adhesive interfaces.

A primary mechanism of adhesive bond failure is a degradation of the adherent/adhesive interfacial stiffness from unwanted contamination or exposure to those environmental factors, which reduce adhesion quality. Substantial research has been conducted on the assessment of adhesively bonded structures and the detection of "kissing" bonds. Advanced ultrasonic assessment methods to interrogate bonded joints and measure interfacial stiffness using a distributed spring interface model have been developed. Amplitude-based ultrasonic methods have traditionally been used in adhesive bond quality assessment, but recent advancements in ultrasonic phase measurements allow for high measurement resolution with low-uncertainty. In this work, an ultrasonic phase technique for the monitoring of adhesively-bonded interfaces is demonstrated. Constant frequency measurements are obtained from the ultrasonic phase of the reflection coefficient from the adhesive bond with a glass adherent, where the degree of cure is controlled by exposure to ultraviolet light. A peak in the phase of the reflection coefficient, as predicted by the interfacial spring model, is measured experimentally. It is shown that the peak phase predicts the interfacial stiffness when some frequency dependent threshold value is crossed. With knowledge of the acoustic impedances of both materials at the interface, the interfacial stiffness is determined by an inverse algorithm involving measurements of ultrasonic phase shifts of bonded joint reflections. By monitoring the interface of bonded structures and coatings, this method permits a nondestructive inspection of bond strength from structural construction through its service life.

Sexual Risk Taking Intentions Under the Influence of Relationship Motivation, Partner Familiarity, and Sexual Arousal.

Condom negotiation is typically a dyadic process, influenced by a host of factors that can impact sexual health decision-making. The current paper investigated the influence of sexual arousal, relationship motivation, and partner familiarity on sexual risk taking intentions in women and men. In Study 1, 331 participants were recruited online and responded to items assessing condom use intentions. Participants higher in relationship motivation were more likely to consider perceived partner desire for condom use when considering initiating condom negotiation. In Study 2, 169 undergraduate students participated in a sexual arousal manipulation and responded to scenarios depicting sexual encounters with more or less familiar hypothetical partners. Participants reported greater sexual risk taking intentions with more (vs. less) familiar hypothetical partners. Men (vs. women) showed greater risk taking intentions overall. Higher sexual arousal was associated with increased sexual risk-taking intentions, regardless of gender. With less familiar partners, participants higher in relationship motivation showed somewhat greater concern that insisting on condom use would interfere with the sexual encounter, though concern was greatest among men for both more and less familiar partner types. These findings offer some support for the notion that a stronger orientation toward forming long-term relationships can facilitate sexual risk taking behavior.

Research Note: It's not just stress-fecal contamination of plumage may affect feather corticosterone concentration.

The feather corticosterone concentration (fCORT) is increasingly used to assess long-term stress in birds as this indicator provides the potential to retrospectively evaluate the adrenocortical activity of a bird during the growth period of a feather over several weeks in one sample. However, there is still a lack of knowledge concerning external factors which can potentially influence fCORT in birds. The aim of the present study was to determine whether fCORT in laying hens is affected by previous fecal contamination of the plumage. Fully grown primaries 5 (P5s) of laying hens (n = 40) were used for the study. To test the effects of fecal contamination on fCORT, freshly defecated droppings from laying hens were collected and mixed manually. In order to simulate practical and at the same time standardized contamination, the upper surface of the previously determined middle third of the right P5 of each hen then was evenly coated with the paste prepared from fresh feces. The treated feathers were stored for 24 h protected from light at room temperature under a film cover to prevent evaporation. Thereafter, the applied layer of excreta was removed manually. Contralateral P5s of the same individuals were stored under identical conditions and served as controls. Both treatment and control feathers were washed in an aqueous soap solution, rinsed several times in pure water and air-dried subsequently. After pulverization and methanolic extraction, fCORT was analyzed by ELISA. The mean fCORT of treatment P5s (reference 'feather length': 12.88 ± 3.16 pg/mm; reference 'feather weight': 7.81 pg/mg ± 1.86 pg/mg) were significantly higher (P < 0.001) than those of control P5s (reference 'feather length': 9.76 ± 2.42 pg/mm; reference 'feather weight': 5.93 ± 1.44 pg/mg). Our results show that previous contamination with excreta can increase fCORT, which was detectable even after a washing procedure prior to analysis. In conclusion, fecal contamination of feathers is a significant influencing factor that has to be considered when applying fCORT measurements.

Research Note: A sip of stress. Effects of corticosterone supplementation in drinking water on feather corticosterone concentrations in layer pullets.

The measurement of feather corticosterone concentrations (fCORT) is a comparatively new method for the evaluation of stress in wild and captive birds and may be a useful indicator in animal welfare research. The aim of this study was to assess the suitability of fCORT as an indicator of stress, and for this purpose a prolonged stress period was experimentally simulated by oral intake of corticosterone via drinking water and corticosterone concentrations were analyzed in feathers grown during this period. Layer pullets of both a control group (n = 20) and a CORT group (n = 20) were offered drinking water ad libitum throughout the entire experimental phase. The drinking water of the CORT group was supplemented with corticosterone at a concentration of 20 mg/l from the 64th to the 114th day of life. The vaned parts of the primaries 5 (P5s) were clipped on d 114 and fCORT was analyzed by ELISA after extraction. Body weights increased from day 64 until d 114 in both groups, however, at the end of the experiment, mean body mass in the CORT group was significantly lower than in the control group (P < 0.001). Pullets of the CORT group also showed shorter and lighter P5s as well as a retarded molt of the primaries. The supplementation of drinking water with corticosterone increased the average fCORT in the P5s of the CORT pullets compared with the control group (median: 110.3 pg/mm [interquartile range (IQR): 47.2] vs. 10.0 pg/mm [IQR: 2.5], P < 0.001). The results show that experimentally increased systemic corticosterone concentrations over a period of seven weeks in layer pullets are reflected in corticosterone concentrations of feathers grown during that time. This indicates that the measurement of fCORT may be a useful and minimally invasive tool for the evaluation of long-term stress in chicken and provides the basis for further investigations on its use in animal welfare research.

Surgical uterine drainage and lavage as treatment for canine pyometra.

Pyometra is a common post-oestral syndrome in bitches. Classical treatment consists of either ovariohystorectomy or medical intervention. Surgical uterine drainage and lavage via direct trans-cervical catheterisation using a 5% povidone-iodine in saline solution was performed successfully in 8 bitches with pyometra. All bitches conceived and whelped without complications subsequent to this treatment. It is concluded that this method offers an effective alternative treatment for canine pyometra with shorter recovery times as well as good clinical recovery and pregnancy rates in bitches destined for further breeding.

Curtailing parturition observation and performing preparturient cesarean section in bitches.

Properly planned elective cesarean section (CS) in bitches is considered safe and justified for some breeds. Therefore, planning a scheduled (fixed date and time) preparturient CS in bitches belonging to a subpopulation where a CS is unavoidable, seems justified. The first day of cytological diestrus (D0) was used to predict the date of parturition. The aims of this study were to (1) compare the mortality of puppies delivered by preparturient CS to those delivered by parturient CS, (2) determine the extent to which a preparturient CS (performed at 08:00 on D57, while the cervix is closed) would shorten gestation, (3) compare the level of progesterone at the time of preparturient CS to that at the time of parturient CS, (4) compare the change in hematocrit before and after CS for parturient- and preparturient CSs. Out of 99 gestations for which D0 was known and a preparturient CS planned, the CS was performed at the scheduled time in 61%, before the scheduled time because cervical dilatation had started in 32% and before the scheduled time in 7% because the bitches had started showing signs of impending parturition, although their cervices were still closed. This study showed high neonatal survival ratios and good Apgar scores after preparturient CS with 99% of puppies delivered by preparturient CS born alive and 98.8% surviving to 2 h. Performing preparturient CSs shortened gestation by four to 52 h. Progesterone level was significantly higher at the time of preparturient CSs than at the time of parturient CSs. Further studies are required to determine whether preparturient CSs on D57 are routinely safe in all breeds, does not affect long term survival of the puppies and to timeously identify bitches in which parturition starts prior to the date and time for the scheduled elective preparturient CS.

Oxidatively generated modifications to nucleic acids in vivo: Measurement in urine and plasma.

BACKGROUND: The oxidized guanine nucleosides, 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) and 8-oxo-7,8-dihydroguanosine (8-oxoGuo), derived from DNA and RNA, respectively, were used to investigate the importance of oxidative stress to nucleic acids in vivo. High urinary excretion of 8-oxodG is associated with cancer development, whereas high urinary excretion of 8-oxoGuo is associated with mortality in type 2 diabetes. Like creatinine, these small water-soluble molecules are not reabsorbed in the kidney. Therefore, 8-oxo nucleoside/creatinine reciprocal concentration ratios are identical in plasma and urine. The total amount of 8-oxo guanine nucleosides excreted by the kidneys is the product of plasma concentration and glomerular filtration rate. METHODS: With relevant equations and an estimated glomerular filtration rate, the 24-h urinary excretion of 8-oxodG and 8-oxoGuo was calculated in 2679 subjects with type 2 diabetes, displaying good correlation with the measured urinary 8-oxo nucleoside/creatinine ratio: DNA oxidation r = 0.86 and RNA oxidation r = 0.84 (p < 0.05 for both). RESULTS: Survival analyses based on the quartiles of the 8-oxodG/creatinine ratio and the quartiles of calculated 24-h urinary excretion rate of the 2679 subjects gave similar hazard ratio estimates for death due to all causes. This finding was similar for the 8-oxoGuo hazard ratio estimates. CONCLUSIONS: This study shows that oxidatively generated modifications to DNA and RNA in vivo can be measured using 1) a spot urine sample, normalized to urinary creatinine, 2) 24-h urine, or 3) a single plasma sample based on concentrations of 8-oxo nucleoside and creatinine and glomerular filtration rate.

The precision of predicting the time of onset of parturition in the bitch using the level of progesterone in plasma during the preparturient period.

Precise prediction of the time of onset of parturition in the bitch is of clinical importance. Many parturition management cases in clinical practice are presented in the last two weeks of pregnancy without a reliable estimate of the parturition date. The aim of this study was to assess the value of a single progesterone level in the blood plasma from a preparturient bitch to predict the time of onset of parturition. The temporal relationship between the decrease in the plasma progesterone levels and the time of cervical dilatation (TCD)-which correlates to the onset of stage 1 of parturition-was evaluated in 25 bitches in the preparturient period. Among bitches destined to reach TCD within 12 h there is a 2% probability of having a plasma progesterone level of 15.8 nmoL/L or above and a 6% probability of having a level of 8.7 nmoL/L or above. Conversely, if the level is below 8.7 nmoL/L there is a 99% probability of reaching TCD within 48 h and if the level is below 3.18 nmoL/L there is a 100% probability of reaching TCD within 24 h. These results allow the veterinary obstetrician to make prompt decisions in the management of parturition.

Is the biparietal diameter of fetuses in late gestation too variable to predict readiness for cesarean section in dogs?

Correct assessment of readiness for cesarean section is essential for timing elective cesarean section during late pregnancy in the bitch. In humans, biparietal diameter is sufficiently precise and accurate and used in a clinical setting daily. The objectives of this study were to determine whether fetal biparietal diameter in late gestation in the dog could be used to predict readiness for cesarean section by having reached a minimum cut-off value and to correlate the biparietal diameter to birth weight. The biparietal diameter of 208 puppies in 34 litters from 31 English bulldog bitches and 660 puppies in 78 litters from 70 Boerboel bitches were measured immediately after delivery by cesarean section, performed at full term, using digital calipers. At the same time the birth weight of the same 208 English bulldog puppies and 494 of the same Boerboel puppies in 59 litters from 54 bitches was measured by means of an electronic scale. With a cesarean section, all the puppies in a litter are delivered simultaneously and readiness for cesarean section must be determined for a litter. The minimum, median and maximum biparietal diameter varied from 21.1 to 47.8, 32.9 to 50.0 and 34.2-58.2 mm, respectively, among English bulldog litters and from 18.4 to 48.7, 35.5 to 49.7 and 39.8-54.3 mm among Boerboel litters. This large variation suggests that biparietal diameter is too variable within and among litters to be useful as a means of determining readiness for cesarean section.

A digital, constant-frequency pulsed phase-locked-loop instrument for real-time, absolute ultrasonic phase measurements.

A digitally controlled instrument for conducting single-frequency and swept-frequency ultrasonic phase measurements has been developed based on a constant-frequency pulsed phase-locked-loop (CFPPLL) design. This instrument uses a pair of direct digital synthesizers to generate an ultrasonically transceived tone-burst and an internal reference wave for phase comparison. Real-time, constant-frequency phase tracking in an interrogated specimen is possible with a resolution of 0.000 38 rad (0.022°), and swept-frequency phase measurements can be obtained. Using phase measurements, an absolute thickness in borosilicate glass is presented to show the instrument's efficacy, and these results are compared to conventional ultrasonic pulse-echo time-of-flight (ToF) measurements. The newly developed instrument predicted the thickness with a mean error of -0.04 µm and a standard deviation of error of 1.35 µm. Additionally, the CFPPLL instrument shows a lower measured phase error in the absence of changing temperature and couplant thickness than high-resolution cross-correlation ToF measurements at a similar signal-to-noise ratio. By showing higher accuracy and precision than conventional pulse-echo ToF measurements and lower phase errors than cross-correlation ToF measurements, the new digitally controlled CFPPLL instrument provides high-resolution absolute ultrasonic velocity or path-length measurements in solids or liquids, as well as tracking of material property changes with high sensitivity. The ability to obtain absolute phase measurements allows for many new applications than possible with previous ultrasonic pulsed phase-locked loop instruments. In addition to improved resolution, swept-frequency phase measurements add useful capability in measuring properties of layered structures, such as bonded joints, or materials which exhibit non-linear frequency-dependent behavior, such as dispersive media.

Current approaches to avoid the culling of day-old male chicks in the layer industry, with special reference to spectroscopic methods.

The negative correlation between fattening and laying performance prevents breeding improvement in both laying performance and meat yield. Therefore, specialized chicken lines have been bred in order to achieve either an efficient production of high-quality eggs or high growth rates. As a result, day-old male chicks are culled in the layer hatchery, which poses animal welfare and ethical problems. Breeding companies, scientific groups, and hatcheries are attempting to resolve this issue, with a common aim to find feasible alternatives for the routine killing of male layer chicks. Some approaches aim to influence the sex ratio, while others target at the economically feasible use of the male layer offspring, such as the fattening of "laying hen brothers" or crossbreedings of layers and broilers to create "dual-purpose chickens." Another approach is the sex determination prior to hatch. One of the prerequisites of in ovo sex determination is a practicable method that can be used in industry. The analysis needs to be rapid, cost-efficient, and highly precise; in addition, negative impacts on hatching rate, animal health, and/or performance parameters should be limited. Furthermore, sex determination should be performed before the sensory nervous system's response of the chick embryo to certain or potentially harmful stimuli is developed, which according to current knowledge is before the d 7 of incubation.

Selective release of excreted DNA sequences from phytohemagglutinin-stimulated human peripheral blood lymphocytes. Effects of trypsin and divalent cations.

We studied the synthesis of excreted DNA sequences and their release from phytohemagglutinin-stimulated human peripheral blood lymphocytes under conditions permitting optimal cell growth. Cells were labeled by constant exposure to low specific activity [3H]thymidine. Excreted DNA sequences were synthesized during the period of logarithmic cell growth and moved slowly from the high molecular weight chromosomal DNA fraction into the low molecular weight cell DNA fraction (Hirt supernate) from which they could be specifically released by treating the cells briefly with small amounts of various proteases; 1 microgram/ml trypsin for 5 min was optimal. On day 5 of culture, 13.3 +/- 6.9% of the total cellular acid-precipitable [3H]thymidine was released by this treatment. Trypsin-induced release was partially and reversibly inhibited by incubating the cells for 16 h with 5 mM dibutyryl-cyclic AMP. Cells incubated in the absence of divalent cations spontaneously released this Hirt supernatant DNA; after maximal release had occurred under these circumstances, additional trypsin treatment caused no further release of DNA. Trypsin-induced DNA release could be completely and reversibly inhibited by incubating the cells in the presence of 10 mM calcium. Trypsin-released DNA was isolated and analyzed by reassociation kinetics. A major component, representing 54% of the DNA, reassociated with a C0t1/2 of 68 mol.s/liter (the value at which DNA association is 50% complete). The reassociation of this DNA was studied in the presence of an excess of DNA isolated from stimulated lymphocytes on day 3 in culture, and in the presence of an excess of resting lymphocyte DNA. The high molecular weight fraction of day-3 cell DNA contained three times more copies of the trypsin-released DNA major component as compared to resting lymphocyte DNA. Hirt supernatant DNA isolated from day-5 stimulated lymphocytes reassociated in an intermediate component representing 34% of the DNA with a Cot1/2 of mol.s/liter; after cells were treated with trypsin, this component could no longer be identified in the Hirt supernatant fraction, presumably because it had been released into the incubation medium. These data describe a quantitatively reproducible system with which synthesis and release of excreted DNA sequences can be studied.

The effect of homologous prostatic fluid on motility and morphology of dog epididymal spermatozoa extended and frozen in Biladyl with Equex STM paste or Andromed.

Although dog prostatic fluid decreases the longevity of ejaculated dog spermatozoa, it also increases their rate of motility and their fertility after vaginal insemination, as well as the fertility of epididymal spermatozoa after uterine insemination. These findings indicate a need to further characterize the effects of prostatic fluid on dog spermatozoa. This study was done to determine the effects (P<0.05) of homologous prostatic fluid added prior to cooling, after thawing, or at both times to epididymal spermatozoa from 21 dogs. The effects of two extenders were also determined. The one extender was Biladyl(*) with Equex STM paste(**) (BilEq) and the other Andromed(*) (Minitüb, Tiefenbach, Germany (*); Nova Chemical Sales, Scituate, MA, USA (**)). The response variables were percentage progressively motile spermatozoa (Prog) and morphology after thawing. Prog was measured at various times until 8h after extension (unfrozen spermatozoa) or until 2h after thawing. Prog after thawing was higher with BilEq than Andromed, when no prostatic fluid was added prior to cooling, and when prostatic fluid was added after thawing. BilEq resulted in a higher mean percentage of spermatozoa with bent principle pieces than Andromed and the addition of prostatic fluid prior to cooling resulted in lower mean percentages of cytoplasmic droplets and bent principle pieces than when none was added. The optimal combination was BilEq with prostatic fluid added prior to cooling (in order to inhibit the development of bent principle pieces) and after thawing (to achieve higher motility until 1h after thawing). This study shows that BilEq is more suitable for the freezing of epididymal spermatozoa than Andromed and that prostatic fluid improves the freezability and post-thaw longevity of epididymal spermatozoa frozen in BilEq.

The precision of peri-oestrous predictors of the date of onset of parturition in the bitch.

Precise prediction of the date of onset of parturition in the bitch is clinically important. The study compared the precision with which four peri-oestrous predictors predict the date of onset of parturition. The predictors evaluated in 24 bitches were: the date of the first or only day of the LH surge, the date on which the concentration of progesterone in the blood plasma first exceeded 6 nmol/L, the date on which the concentration of progesterone in the blood plasma first exceeded 16 nmol/L and the date of onset of cytological dioestrus. Among the 24 bitches, the date of onset of cytological dioestrus predicted the date of onset of parturition with greater precision than the other three predictors. Following the evaluation of another 218 intervals between the onset of cytological dioestrus and the date of onset of parturition, it was shown that the onset of cytological dioestrus predicted the date of onset parturition with a precision of ±1 d, ± 2 d and ±3 d in 88%, 99% and 100% of the 242 pregnancies. This study concludes that the first day of cytological dioestrus is a useful predictor of the date of onset of parturition.

Estimated impact of multiple conceptuses per follicle on fecundity in the bitch.

Multiple conceptuses from one follicle due to multi-ovular follicles or monozygotic twins inflate the ratio between the numbers of conceptuses and corpora lutea and possibly the effect ascribed to experimental treatment. The aim of this study was to estimate the impact of multiple conceptuses per follicle on fecundity in the bitch. The numbers of conceptuses and corpora lutea of 105 bitches originated from 7 published experimental studies and of 132 from ovariohysterectomies done in a private practice and a welfare organisation. Ninety five bitches, producing 597 conceptuses, had at least as many conceptuses as corpora lutea; 83 had as many, 10 had one more and 2 had 2 more. Bootstrap resampling on these 95 bitches let us to conclude that 12.6% of bitches are expected to produce more oocytes that are each capable of yielding a conceptus than the number of follicles that ovulate and that 2.35% of conceptuses are expected to be in excess of the number of follicles that ovulate. Applying the results of the current study to 2 earlier studies showed that multiple conceptuses from one follicle may, at maximum, have increased the effect ascribed to prostatic fluid by 0.022 and 0.024 conceptuses per corpus luteum, which is minor in relation to the magnitude of the ascribed effects of 0.34 and 0.22 conceptuses per corpus luteum. The number of conceptuses relative to the number of corpora lutea provides a valid and precise measurement of fertility in the bitch.

Puppy survival and vigor associated with the use of low dose medetomidine premedication, propofol induction and maintenance of anesthesia using sevoflurane gas-inhalation for cesarean section in the bitch.

The safety of an anesthetic protocol consisting of medetomidine hydrochloride (7 µg/kg iv) as premedicant, propofol, (1-2 mg/kg iv) as induction agent and sevoflurane, at 2% in oxygen for maintenance of anesthesia was studied in 292 cesarean sections (CSs) and 2232 puppies delivered. Medetomidine effects were reversed using atipamezole hydrochloride at 50 µg/puppy sc immediately following delivery and in the bitch iv immediately following surgery. The protocol's safety for puppies was expressed using survival immediately, 2 h and 7 d after delivery, and Apgar scores (measurement starting 15 min after delivery of the last puppy). The maternal survival rate was established immediately, 2 h and 7 d after cesarean section (CS). The CSs included 148 on Boerboel, 84 on English bulldog and 60 on other purebred bitches, which resulted in 1378, 541 and 313 puppies, respectively. Boerboel, English bulldog and other purebred bitches yielded 97.39%, 96.67% and 91.69% live puppies at delivery, 95.43%, 88.35% and 89.78% alive by 2 h and 89.19%, 79.11% and 84.03% alive by 7 d. Sixteen (1.16%), 32 (5.59%) and 4 (1.28%) malformed Boerboel, English bulldog and other purebred puppies were euthanized. Thirty five, 18 and 26, Boerboel, English bulldog and other purebred puppies were stillborn respectively, of which 12, 9 and 15, respectively had been discovered dead upon ultrasound examination immediately before CS. After correction for fetuses found dead on ultrasound examination and malformed euthanized puppies, 98.21%, 95.60% and 94.30% of Boerboel, English bulldog and other purebred puppies survived until 2 h and 91.78%, 87.17% and 88.26% until 7 d. Two-hour survival rates are negatively correlated to the proportion of puppies in a litter with scores of 8 or below (r = 0.14, P = 0.01, n = 292 litters) and tends to be positively correlated to the lowest Apgar score in a litter (r = 0.11, P = 0.05, n = 292 litters). This study shows that medetomidine hydrochloride in the protocol used is a safe premedicant in bitches prior to cesarean section and is associated with good puppy vigor as well as 2 h and 7 d puppy survival rates. The use of medetomidine as premedicant permitted use of less than half the dose of propofol usually required as induction agent.